KR101897340B1 - Composition for diagnosing, preventing or improving aging of skin using HAPLN1 - Google Patents
Composition for diagnosing, preventing or improving aging of skin using HAPLN1 Download PDFInfo
- Publication number
- KR101897340B1 KR101897340B1 KR1020160115670A KR20160115670A KR101897340B1 KR 101897340 B1 KR101897340 B1 KR 101897340B1 KR 1020160115670 A KR1020160115670 A KR 1020160115670A KR 20160115670 A KR20160115670 A KR 20160115670A KR 101897340 B1 KR101897340 B1 KR 101897340B1
- Authority
- KR
- South Korea
- Prior art keywords
- hapln1
- skin
- protein
- aging
- skin aging
- Prior art date
Links
- 101001079904 Homo sapiens Hyaluronan and proteoglycan link protein 1 Proteins 0.000 title claims abstract 9
- 102100028084 Hyaluronan and proteoglycan link protein 1 Human genes 0.000 title claims abstract 9
- 239000000203 mixture Substances 0.000 title abstract description 27
- 230000032683 aging Effects 0.000 title abstract description 19
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 13
- 230000037303 wrinkles Effects 0.000 claims abstract description 12
- 229920002674 hyaluronan Polymers 0.000 claims description 36
- 229960003160 hyaluronic acid Drugs 0.000 claims description 36
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 34
- 238000000034 method Methods 0.000 claims description 13
- 239000004480 active ingredient Substances 0.000 claims description 5
- 230000037394 skin elasticity Effects 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 59
- 230000009759 skin aging Effects 0.000 abstract description 38
- 230000036541 health Effects 0.000 abstract description 18
- 235000013376 functional food Nutrition 0.000 abstract description 16
- 239000002537 cosmetic Substances 0.000 abstract description 13
- 239000000556 agonist Substances 0.000 abstract description 11
- 239000003795 chemical substances by application Substances 0.000 abstract description 8
- 101150014382 Hapln1 gene Proteins 0.000 abstract description 6
- 230000001737 promoting effect Effects 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 230000006872 improvement Effects 0.000 abstract description 2
- 108700026220 vif Genes Proteins 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 description 44
- 210000003491 skin Anatomy 0.000 description 34
- 241000699670 Mus sp. Species 0.000 description 26
- 241000699666 Mus <mouse, genus> Species 0.000 description 24
- 210000001519 tissue Anatomy 0.000 description 16
- 239000012472 biological sample Substances 0.000 description 11
- 101710128038 Hyaluronan synthase Proteins 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- 230000001965 increasing effect Effects 0.000 description 9
- 108020004999 messenger RNA Proteins 0.000 description 9
- 108091005735 TGF-beta receptors Proteins 0.000 description 8
- 102000016715 Transforming Growth Factor beta Receptors Human genes 0.000 description 8
- 238000009826 distribution Methods 0.000 description 8
- 230000002500 effect on skin Effects 0.000 description 8
- 239000000843 powder Substances 0.000 description 8
- 108010035532 Collagen Proteins 0.000 description 7
- 102000008186 Collagen Human genes 0.000 description 7
- 229920001436 collagen Polymers 0.000 description 7
- -1 for example Substances 0.000 description 7
- 238000009472 formulation Methods 0.000 description 7
- 238000002965 ELISA Methods 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 6
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 6
- 239000000839 emulsion Substances 0.000 description 6
- 239000002953 phosphate buffered saline Substances 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 5
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 5
- 108010050808 Procollagen Proteins 0.000 description 5
- 238000011529 RT qPCR Methods 0.000 description 5
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 description 5
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 description 5
- 230000003213 activating effect Effects 0.000 description 5
- 239000000090 biomarker Substances 0.000 description 5
- 230000037319 collagen production Effects 0.000 description 5
- 239000006071 cream Substances 0.000 description 5
- 210000002744 extracellular matrix Anatomy 0.000 description 5
- 239000000546 pharmaceutical excipient Substances 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 238000001262 western blot Methods 0.000 description 5
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 4
- 238000008157 ELISA kit Methods 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 241000283074 Equus asinus Species 0.000 description 4
- 101100451392 Homo sapiens HAPLN1 gene Proteins 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 4
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 4
- 238000012790 confirmation Methods 0.000 description 4
- 239000003085 diluting agent Substances 0.000 description 4
- 238000003125 immunofluorescent labeling Methods 0.000 description 4
- 108700041430 link Proteins 0.000 description 4
- 239000006210 lotion Substances 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- 230000037311 normal skin Effects 0.000 description 4
- 210000001626 skin fibroblast Anatomy 0.000 description 4
- 239000007921 spray Substances 0.000 description 4
- 239000000454 talc Substances 0.000 description 4
- 229910052623 talc Inorganic materials 0.000 description 4
- 235000012222 talc Nutrition 0.000 description 4
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 4
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 3
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 3
- 101100451393 Mus musculus Hapln1 gene Proteins 0.000 description 3
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 108010067787 Proteoglycans Proteins 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 210000003489 abdominal muscle Anatomy 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004204 blood vessel Anatomy 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 235000010980 cellulose Nutrition 0.000 description 3
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 239000008101 lactose Substances 0.000 description 3
- 239000000314 lubricant Substances 0.000 description 3
- 239000012188 paraffin wax Substances 0.000 description 3
- 239000006072 paste Substances 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000012110 Alexa Fluor 594 Substances 0.000 description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 2
- 108020005544 Antisense RNA Proteins 0.000 description 2
- 108091023037 Aptamer Proteins 0.000 description 2
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- LCGLNKUTAGEVQW-UHFFFAOYSA-N Dimethyl ether Chemical compound COC LCGLNKUTAGEVQW-UHFFFAOYSA-N 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000320 Hyaluronan Synthases Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 244000299461 Theobroma cacao Species 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000000440 bentonite Substances 0.000 description 2
- 229910000278 bentonite Inorganic materials 0.000 description 2
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 2
- SESFRYSPDFLNCH-UHFFFAOYSA-N benzyl benzoate Chemical compound C=1C=CC=CC=1C(=O)OCC1=CC=CC=C1 SESFRYSPDFLNCH-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000010805 cDNA synthesis kit Methods 0.000 description 2
- 239000000378 calcium silicate Substances 0.000 description 2
- 229910052918 calcium silicate Inorganic materials 0.000 description 2
- 235000012241 calcium silicate Nutrition 0.000 description 2
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 239000003184 complementary RNA Substances 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000003599 food sweetener Nutrition 0.000 description 2
- 210000003194 forelimb Anatomy 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 230000002163 immunogen Effects 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 238000001114 immunoprecipitation Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000003141 lower extremity Anatomy 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000000377 silicon dioxide Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 235000010356 sorbitol Nutrition 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000003765 sweetening agent Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 239000008158 vegetable oil Substances 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000001993 wax Substances 0.000 description 2
- JNYAEWCLZODPBN-JGWLITMVSA-N (2r,3r,4s)-2-[(1r)-1,2-dihydroxyethyl]oxolane-3,4-diol Chemical compound OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O JNYAEWCLZODPBN-JGWLITMVSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- WNWHHMBRJJOGFJ-UHFFFAOYSA-N 16-methylheptadecan-1-ol Chemical class CC(C)CCCCCCCCCCCCCCCO WNWHHMBRJJOGFJ-UHFFFAOYSA-N 0.000 description 1
- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 206010015548 Euthanasia Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 101500025614 Homo sapiens Transforming growth factor beta-1 Proteins 0.000 description 1
- 102100040203 Hyaluronan synthase 1 Human genes 0.000 description 1
- 102100029425 Hyaluronan synthase 3 Human genes 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 101001078441 Mus musculus Hyaluronan and proteoglycan link protein 1 Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 229940123573 Protein synthesis inhibitor Drugs 0.000 description 1
- 235000005764 Theobroma cacao ssp. cacao Nutrition 0.000 description 1
- 235000005767 Theobroma cacao ssp. sphaerocarpum Nutrition 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 239000006096 absorbing agent Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000013334 alcoholic beverage Nutrition 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000010775 animal oil Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 229960002903 benzyl benzoate Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000005827 chlorofluoro hydrocarbons Chemical class 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 1
- 229940093471 ethyl oleate Drugs 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 150000002337 glycosamines Chemical class 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 238000013095 identification testing Methods 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 229940029329 intrinsic factor Drugs 0.000 description 1
- VMPHSYLJUKZBJJ-UHFFFAOYSA-N lauric acid triglyceride Natural products CCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCC)COC(=O)CCCCCCCCCCC VMPHSYLJUKZBJJ-UHFFFAOYSA-N 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- CQDGTJPVBWZJAZ-UHFFFAOYSA-N monoethyl carbonate Chemical compound CCOC(O)=O CQDGTJPVBWZJAZ-UHFFFAOYSA-N 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 239000012457 nonaqueous media Substances 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 235000013550 pizza Nutrition 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000001294 propane Substances 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 239000000007 protein synthesis inhibitor Substances 0.000 description 1
- 238000000575 proteomic method Methods 0.000 description 1
- 239000000700 radioactive tracer Substances 0.000 description 1
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000004003 subcutaneous fat Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000516 sunscreening agent Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000002511 suppository base Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/728—Hyaluronic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/50—Polysaccharides, gums
- A23V2250/51—Polysaccharide
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/20—Dermatological disorders
Abstract
본 발명은 노화된 개체의 HAPLN1 단백질에 대한 것으로, 상기 HAPLN1 단백질은 노화 시 발현양이 감소하며, 투여 시 주름을 포함한 피부 노화를 개선 및 회복시키는 효과가 우수하므로, 이 발현 차이를 이용하여 피부 노화를 진단할 수 있는 피부 노화 측정용 바이오마커 조성물, 키트, HAPLN1 단백질 또는 HAPLN1 유전자의 발현 수준을 검출하는 단계를 포함하는 피부 노화 개선제 스크리닝 방법을 제공할 수 있다. 더불어 HAPLN1 단백질, 이를 코딩하는 유전자 및 HAPLN1 단백질 또는 유전자의 발현 촉진 또는 기능을 활성화하는 효능제로 이루어진 군에서 선택된 어느 하나 이상을 포함하는 피부 노화 예방 또는 개선용 약학 조성물, 화장료 조성물 또는 건강기능식품 및 주름 개선용 화장료 조성물 또는 건강기능식품을 제공할 수 있다.The present invention relates to a HAPLN1 protein of an aged individual, wherein the HAPLN1 protein has a reduced expression amount upon aging and has an excellent effect of improving and restoring skin aging including wrinkles upon administration. Therefore, A kit for detecting skin aging, a kit, a HAPLN1 protein or a HAPLN1 gene, which can diagnose a skin aging-improving agent. A pharmaceutical composition for preventing or improving skin aging comprising a HAPLN1 protein, a gene encoding the HAPLN1 protein, a gene encoding the HAPLN1 protein, and a HAPLN1 protein or an agonist for promoting the expression or function of the gene, A cosmetic composition for improvement or a health functional food.
Description
본 발명은 히알루론산과 프로테오글리칸 연결 단백질 1(Hyaluronan and proteoglycan link protein 1, HAPLN1)을 이용한 피부 노화 측정용 바이오마커 조성물과 피부 노화 예방 또는 개선용 약학 조성물, 화장료 조성물 또는 건강기능식품에 대한 것이다.The present invention relates to a biomarker composition for measuring skin aging using hyaluronic acid and proteoglycan-linked protein 1 (HAPLN1), a pharmaceutical composition for preventing or improving skin aging, a cosmetic composition or a health functional food.
피부노화에 따른 가장 뚜렷한 현상 중의 하나는 진피층 세포외기질(extracellular matrix)의 감소이며 이로 인해 주름이 발생한다. 진피층 세포외기질은 콜라겐(collagen), 엘라스틴(elastin), 당단백질(glycoprotein) 등 다양한 단백질이 주요 성분으로 포함되어있다. 이들 단백질 중 90% 이상은 콜라겐이며, 그 중에서도 약 90% 정도가 타입 1 콜라겐(type 1 collagen)이다. 노화로 인해 진피층 세포외기질에서 타입 1 콜라겐의 양이 약 70% 감소하는 현상이 확인되었다.One of the most prominent phenomena of skin aging is the reduction of the extracellular matrix of the dermis and wrinkles. The extracellular matrix of the dermal layer contains various proteins such as collagen, elastin, and glycoprotein. More than 90% of these proteins are collagen, and about 90% of them are
히알루론산(Hyaluronic acid)은 아미노당과 우론산으로 이루어진 다당류의 일종으로 생체의 모든 조직에 분포하며, 체내 총량의 50% 정도가 피부에 분포되어 있다. 히알루론산은 최대 1000배 질량의 물과 결합할 수 있으며, 진피조직 세포외기질을 채워주는 역할을 한다. 또한, 히알루론산은 여러 가지 당단백질과 프로테오글리칸(proteoglycan) 간의 응집(aggregate)을 형성하여 여러 성장 인자(growth factor)들을 저장하거나 보존하는 역할을 한다. 임상연구 결과에 따르면, 히알루론산은 진피 섬유아세포(dermal fibroblast)의 증식 및 콜라겐 생성, 성장 인자의 분비를 증가시키는데, 나이가 증가하면서 피부 중 40% 이상 감소하는 것으로 알려졌다.Hyaluronic acid (Hyaluronic acid) is a kind of polysaccharide composed of amino sugar and uronic acid. It is distributed in all tissues of living body, and about 50% of the total amount in the body is distributed in the skin. Hyaluronic acid can bind up to 1000 times the mass of water, filling the extra-cellular extracellular matrix. In addition, hyaluronic acid acts as an aggregate between various glycoproteins and proteoglycan to store or preserve various growth factors. Clinical studies have shown that hyaluronic acid increases dermal fibroblast proliferation and collagen production and secretes growth factors, which are known to decrease by more than 40% in the skin with increasing age.
피부 노화는 크게 내인성 피부노화(intrinsic aging)와 외인성 피부노화(extrinsic aging)의 2가지로 나뉜다. 외인성 피부 노화는 자외선, 흡연 등 주로 외부 환경적인 요인에 의해 야기되며, 내인성 피부 노화는 나이를 먹음에 따라 동반되는 피부 노화이다. 현재 외인성 피부 노화를 방지하기 위해 자외선 차단 기능을 가진 향장품 등 여러 방법이 존재하지만 내인성 피부 노화를 방지하기 위한 방법은 아직 없는 실정이다. 따라서, 내인성 피부 노화를 예방하거나 노화된 피부를 개선시키기 위한 노력이 필요하다. Skin aging is divided into two types: intrinsic aging and extrinsic aging. Exogenous skin aging is mainly caused by external environmental factors such as ultraviolet rays and smoking, and aging of the skin is aging accompanied by aging. Currently, there are many ways to prevent aging of the extrinsic skin, such as a facial with an ultraviolet shielding function, but there is no way to prevent the aging of the endogenous skin. Therefore, efforts should be made to prevent aging of the endogenous skin or to improve aged skin.
본 발명의 목적은 피부 노화 측정용 바이오마커 조성물을 제공하는 데에 있다.It is an object of the present invention to provide a biomarker composition for measuring skin aging.
본 발명의 다른 목적은 피부 노화 측정용 키트를 제공하는 데에 있다.Another object of the present invention is to provide a kit for measuring skin aging.
본 발명의 또 다른 목적은 피부 노화 개선제의 스크리닝 방법을 제공하는 데 에 있다.Another object of the present invention is to provide a screening method of a skin aging-improving agent.
본 발명의 또 다른 목적은 피부 노화 예방 또는 개선용 약학 조성물을 제공하는 데에 있다.It is still another object of the present invention to provide a pharmaceutical composition for preventing or ameliorating skin aging.
본 발명의 또 다른 목적은 피부 노화 예방 또는 개선용 화장료 조성물, 또는 주름 개선용 화장료 조성물을 제공하는 데에 있다.Another object of the present invention is to provide a cosmetic composition for preventing or improving skin aging, or a cosmetic composition for improving wrinkles.
본 발명의 또 다른 목적은 피부 노화 예방 또는 개선용 건강기능식품, 또는 주름 개선용 건강기능식품을 제공하는 데에 있다.Another object of the present invention is to provide a health functional food for preventing or improving skin aging, or a health functional food for improving wrinkles.
상기 목적을 달성하기 위하여, 본 발명은 생물학적 시료로부터 HAPLN1 단백질 또는 HAPLN1 유전자를 검출하는 제제를 포함하는 피부 노화 측정용 바이오마커 조성물을 제공한다.In order to achieve the above object, the present invention provides a biomarker composition for measuring skin aging comprising a preparation for detecting a HAPLN1 protein or a HAPLN1 gene from a biological sample.
또한, 본 발명은 생물학적 시료로부터 HAPLN1 단백질 또는 HAPLN1 유전자를 검출하는 제제를 포함하는 피부 노화 측정용 키트를 제공한다.In addition, the present invention provides a kit for measuring skin aging comprising an agent for detecting HAPLN1 protein or HAPLN1 gene from a biological sample.
또한, 본 발명은 생물학적 시료에 임의의 화합물을 처리하는 단계; 상기 생물학적 시료로부터 HAPLN1 단백질 또는 유전자의 발현 수준을 검출하는 단계; 및 상기 발현 수준을 임의의 화합물을 처리하지 않은 정상대조군에서의 동일 단백질 또는 유전자의 발현 수준과 비교하는 단계를 포함하는 피부 노화 개선제의 스크리닝 방법을 제공한다.The present invention also relates to a method of treating a biological sample, comprising: treating a biological sample with an arbitrary compound; Detecting an expression level of the HAPLN1 protein or gene from the biological sample; And comparing the expression level with an expression level of the same protein or gene in a normal control group in which no compound has been treated.
또한, 본 발명은 HAPLN1 단백질, 이를 코딩하는 유전자 및 HAPLN1 단백질 또는 유전자의 발현 촉진 또는 기능을 활성화하는 효능제로 이루어진 군에서 선택된 어느 하나 이상을 유효성분으로 포함하는 것을 특징으로 하는 피부 노화 예방 또는 개선용 약학 조성물을 제공한다.The present invention also provides a method for preventing or ameliorating skin aging, which comprises the HAPLN1 protein, a gene encoding the HAPLN1 protein, or a HAPLN1 protein or an effective agent for activating the expression or function of the gene, To provide a pharmaceutical composition.
또한, 본 발명은 HAPLN1 단백질 또는 HAPLN1 단백질의 발현 촉진 또는 기능을 활성화하는 효능제 중 어느 하나를 유효성분으로 포함하는 것을 특징으로 하는 피부 노화 예방 또는 개선용 화장료 조성물, 또는 주름 개선용 화장료 조성물을 제공한다.Also, the present invention provides a cosmetic composition for preventing or improving skin aging, or a cosmetic composition for improving wrinkles, which comprises an effective ingredient for promoting the expression of HAPLN1 protein or HAPLN1 protein or an agonist for activating the function do.
또한, 본 발명은 HAPLN1 단백질 또는 HAPLN1 단백질의 발현 촉진 또는 기능을 활성화하는 효능제 중 어느 하나를 유효성분으로 포함하는 것을 특징으로 하는 피부 노화 예방 또는 개선용 건강기능식품, 또는 주름 개선용 건강기능식품을 제공한다.The present invention also relates to a health functional food for preventing or ameliorating skin aging or a health functional food for improving wrinkles characterized by containing any one of an agonist for promoting the expression of HAPLN1 protein or HAPLN1 protein or an agonist for activating the function, .
상기 약학 조성물, 화장료 조성물 또는 건강기능식품은 히알루론산을 더 포함할 수 있다.The pharmaceutical composition, the cosmetic composition or the health functional food may further comprise hyaluronic acid.
본 발명에 따르면, HAPLN1 단백질은 노화 개체에서 발현양이 감소하며, 투여 시 주름을 포함한 피부 노화를 개선 및 회복시키는 효과가 우수하므로, 본 발명은 HAPLN1 단백질 또는 HAPLN1 유전자를 검출하는 제제를 포함하는 피부 노화를 진단할 수 있는 피부 노화 측정용 바이오마커 조성물, 키트, HAPLN1 단백질 또는 유전자의 발현 수준을 검출하는 단계를 포함하는 피부 노화 예방 또는 개선제 스크리닝 방법, 피부 노화 예방 또는 개선용 약학 조성물, 화장료 조성물 또는 건강기능식품 및 주름 개선용 화장료 조성물 또는 건강기능식품으로 유용하게 제공될 수 있다.According to the present invention, the HAPLN1 protein has a reduced expression amount in an aging individual, and has an excellent effect of improving and restoring skin aging including wrinkles upon administration. Therefore, the present invention relates to a skin comprising a preparation for detecting HAPLN1 protein or HAPLN1 gene A method for screening a preventive or remedy for skin aging comprising the step of detecting the expression level of a biomarker composition, a kit, a HAPLN1 protein or a gene for skin aging measurement capable of diagnosing aging, a pharmaceutical composition for preventing or improving skin aging, A health functional food, a wrinkle improving cosmetic composition or a health functional food.
도 1은 노화 마우스(Old, O)와 어린 마우스(Young, Y)간의 비동시성 개체결합(Heterochronic parabiosis) 쌍에서 콜라겐의 생성을 확인한 결과이고,
도 2는 노화 마우스(Old, O)와 어린 마우스(Young, Y)간의 노화 마우스(Old, O)와 젊은 마우스(Young, Y)간의 비동시성 개체결합(Heterochronic parabiosis) 쌍에서 히알루론산(Hyaluronic acid, HA)의 생성을 확인한 결과이고,
도 3은 피부노화와 관련된 내인성 인자를 확인한 결과이고,
도 4는 노화 마우스와 어린 마우스 피부의 프로콜라겐의 mRNA 양 및 분포를 비교한 결과이고,
도 5는 노화 마우스와 어린 마우스 피부의 히알루론산 농도 및 분포를 비교한 결과이고,
도 6은 노화 마우스와 어린 마우스 피부의 HAPLN1 농도 및 분포를 비교한 결과이고,
도 7은 노화 마우스에서 HAPLN1 투여로 인한 콜라겐 증가를 확인한 결과이고,
도 8은 노화 마우스에서 HAPLN1 투여로 인한 히알루론산 증가를 확인한 결과이고,
도 9는 사람 정상 피부 섬유아세포(NHDF)에서 HAPLN1 처리로 인한 TGF-β 유도 콜라겐 증가를 확인한 결과이고,
도 10은 사람 정상 피부 섬유아세포에서 HAPLN1 처리로 인한 콜라겐 합성 신호 전달 경로 중 TGF-β 수용체 Ⅱ의 단백질 발현양의 증가를 확인한 결과이고,
도 11은 사람 정상 피부 섬유아세포에서 HAPLN1 처리로 인한 TGF-β 수용체 Ⅱ의 안정성 증가를 확인한 결과이고,
도 12는 사람 정상 피부 섬유아세포에서 HAPLN1 처리로 인한 TGF-β 유도 히알루론산 합성 효소(HAS2, hyaluronic acid synthase 2) 단백질 발현양의 증가를 확인한 결과이다.FIG. 1 shows the results of confirming the generation of collagen in a pair of Heterochronic parabiosis between an aged mouse (Old, O) and a young mouse (Young, Y)
FIG. 2 is a graph showing the effect of hyaluronic acid (Hyaluronic acid) on the pair of Heterochronic parabiosis between an aged mouse (Old, O) and a young mouse (Young, Y) , And HA)
FIG. 3 is a result of confirming endogenous factors related to skin aging,
FIG. 4 shows the results of comparing mRNA levels and distribution of procollagen in aged and young mouse skin,
FIG. 5 shows the results of comparing the concentration and distribution of hyaluronic acid in aged and young mouse skin,
FIG. 6 shows the results of comparing HAPLN1 concentration and distribution of aged and young mouse skin,
FIG. 7 shows the results of confirming collagen increase due to administration of HAPLN1 in aged mice,
8 is a result of confirming the increase of hyaluronic acid due to administration of HAPLN1 in the aged mouse,
FIG. 9 shows the results of confirming TGF-β induced collagen increase due to HAPLN1 treatment in human normal skin fibroblast (NHDF)
FIG. 10 shows the results of confirming an increase in the expression level of TGF-β receptor II in the collagen synthesis signal transduction pathway due to HAPLN1 treatment in human normal skin fibroblasts,
11 is a result of confirming the increase in the stability of TGF-beta receptor II due to HAPLN1 treatment in human normal skin fibroblasts,
FIG. 12 shows the results of confirming an increase in the amount of TGF-β induced hyaluronic acid synthase (HAS2) protein expression in human normal skin fibroblasts due to HAPLN1 treatment.
본 발명의 발명자들은 노화 마우스와 어린 마우스의 단백질 발현을 비교한 결과, 노화 마우스에서 HAPLN1의 발현이 감소된다는 것을 발견하여 본 발명을 완성하였다. 상기 HAPLN1는 히알루론산을 프로테오글리칸에 연결하여 히알루론산을 안정화하는 단백질로서, HAPLN1의 피부 노화 개선 효과는 현재 보고된 바가 없다. The inventors of the present invention have found that the expression of HAPLN1 is reduced in aged mice as a result of comparing protein expression between aged and young mice, thus completing the present invention. HAPLN1 is a protein that stabilizes hyaluronic acid by connecting hyaluronic acid to a proteoglycan. HAPLN1 has not been reported to improve skin aging at present.
본 발명은 생물학적 시료로부터 HAPLN1 단백질 또는 HAPLN1 유전자를 검출하는 제제를 포함하는 피부 노화 측정용 바이오마커 조성물을 제공한다.The present invention provides a biomarker composition for measuring skin aging comprising a preparation for detecting a HAPLN1 protein or a HAPLN1 gene from a biological sample.
상기 HAPLN1 단백질은(mouse HAPLN1 accession: NP_038528.3, Gene ID:12950; Human HAPLN1 accession: AAH57808.1, Gene ID: 1404)이나, 이에 제한되는 것은 아니다.The HAPLN1 protein (mouse HAPLN1 accession: NP_038528.3, Gene ID: 12950, Human HAPLN1 accession: AAH57808.1, Gene ID: 1404), but is not limited thereto.
상기 제제는 항체, 안티센스 RNA(antisense RNA), 앱타머(Aptamer) 및 화합물로 이루어진 군에서 선택된 어느 하나이나, 이에 제한되는 것은 아니다.The agent may be any one selected from the group consisting of an antibody, an antisense RNA, an aptamer, and a compound, but is not limited thereto.
상기 피부 노화는 피부 진피층의 세포외 기질 감소로 인해 발생한 주름일 수 있다.The skin aging may be wrinkles caused by a decrease in the extracellular matrix of the dermal layer of the skin.
더불어 본 발명은 생물학적 시료로부터 HAPLN1 단백질 또는 HAPLN1 유전자를 검출하는 제제를 포함하는 피부 노화 측정용 키트를 제공한다.In addition, the present invention provides a kit for measuring skin aging comprising an agent for detecting a HAPLN1 protein or a HAPLN1 gene from a biological sample.
상기 제제는 항체, 안티센스 RNA(antisense RNA), 앱타머(Aptamer) 및 화합물로 이루어진 군에서 선택된 어느 하나이나, 이에 제한되는 것은 아니다.The agent may be any one selected from the group consisting of an antibody, an antisense RNA, an aptamer, and a compound, but is not limited thereto.
상기 항체는 다클론성 항체(polyclonal antibody) 또는 단일클론 항체(monoclonal antibody)이다.The antibody is a polyclonal antibody or a monoclonal antibody.
상기 다클론성 항체는 당업자에 알려진 방법에 따라 면역원으로 상기 유전자에 의해 발현된 단백질 또는 그 단편을 외부 숙주에 주사함으로써 제조할 수 있다. 외부 숙주는 마우스, 랫트, 양 또는 토끼와 같은 포유동물을 포함한다. 면역원은 근내, 복강내 또는 피하 주사방법으로 주사되며, 일반적으로 항원성을 증가시키기 위한 보조제(adjuvant)와 함께 투여된다. 외부숙주로부터 정기적으로 혈청을 채취하여 향상된 역가 및 항원에 대한 특이성을 보이는 혈청을 수거하거나 이로부터 항체를 분리정제한다.The polyclonal antibody can be prepared by injecting an external host with a protein or fragment thereof expressed by the gene as an immunogen according to methods known to those skilled in the art. External hosts include mammals such as mice, rats, sheep or rabbits. The immunogen is injected intraperitoneally, intraperitoneally or subcutaneously, and is generally administered with an adjuvant to increase the antigenicity. Serum is collected periodically from an external host to collect the sera showing improved titer and specificity for the antigen or isolate and purify the antibody therefrom.
상기 단일클론 항체는 당업자에 알려진 융합에 의한 불멸화된 세포주 생성기술에 의해 제조될 수 있다. 예를 들어, 상기 유전자에 의해 발현된 단백질을 마우스에 면역화시키거나 펩타이드를 합성하여 소혈청 알부민과 결합시켜 마우스에 면역화시킨다. 마우스에서 분리된 항원-생산 B 림프구를 인간 또는 마우스의 골수종(myeloma)과 융합하여 불멸화된 하이브리도마(hybridoma)를 생성하며, 상기 하이브리도마 세포를 가지고 간접적인 효소 결합 면역흡착 분석법(enzyme-linked immunoabsorbent assays, ELISA)을 사용하여 모노클노날 항체의 생성 여부를 확인하고 양성 클론을 택하여 배양한 후 항체를 분리정제하거나 랫트의 복강에 주입한 후 복수를 채취함으로써, 단일클론 항체를 제조할 수 있다.Such monoclonal antibodies can be produced by immortalized cell line generation techniques by fusion as known to those of skill in the art. For example, a protein expressed by the gene is immunized with a mouse or a peptide is synthesized and bound to bovine serum albumin and immunized with a mouse. An antigen-producing B lymphocyte isolated from a mouse is fused with a myeloma of a human or a mouse to generate an immortalized hybridoma, and an enzyme-linked immunosorbent assay (ELISA) linked immunoabsorbent assays (ELISA) to identify monoclonal antibodies. After positive clones were selected, the antibodies were isolated and purified, or injected into the abdominal cavity of rats, and the ascites was collected to obtain monoclonal antibodies have.
상기 단일클론 항체는 일반적으로 알칼라인 포스파타아제(alkaline phosphatase, AP) 또는 호올스래디쉬 퍼옥시다제(horseradish peroxidase, HRP) 등의 효소가 결합된 2차 항체 및 이들의 기질을 사용하여 발색반응 시킴으로써 정량분석할 수도 있고, 또는 직접 상기 단백질에 대한 단일클론 항체에 AP 또는 HRP 효소 등이 결합된 것을 사용하여 정량분석할 수 있다.The monoclonal antibody can be quantitatively assayed by using a secondary antibody to which an enzyme such as alkaline phosphatase (AP) or horseradish peroxidase (HRP) is conjugated, or a substrate thereof, Or may be quantitatively analyzed using a monoclonal antibody against the protein directly coupled with an AP or HRP enzyme or the like.
상기 단백질과 항체의 반응은 웨스턴 블랏(western blot), 면역침강법(Immunoprecipitation, IP), 효소 결합 면역흡착 분석법(enzyme-linked immunoabsorbent assays, ELISA) 및 면역염색법(Immunohistochemistry, IHC)등의 단백질 확인 실험을 통해 확인할 수 있으나, 이에 제한되는 것은 아니다.The reaction of the protein with the antibody can be carried out using a protein identification test such as western blot, immunoprecipitation (IP), enzyme-linked immunoabsorbent assays (ELISA) and immunohistochemistry (IHC) However, the present invention is not limited thereto.
상기 항체를 포함하는 키트는 전형적으로 동결건조 형태의 항체와 버퍼, 안정화제, 불활성 단백질 등을 포함할 수 있으며, 상기 항체는 방사종(radionuclides), 형광원(fluorescors), 효소(enzymes) 등에 의해 표지화될 수 있다.The kit comprising the antibody may typically comprise a lyophilized form of the antibody and a buffer, a stabilizer, an inert protein, and the like, wherein the antibody is radionuclides, fluorescors, enzymes, etc. Lt; / RTI >
또한, 본 발명은 생물학적 시료에 임의의 화합물을 처리하는 단계; 상기 생물학적 시료로부터 HAPLN1 단백질 또는 유전자의 발현 수준을 검출하는 단계; 및 상기 발현 수준을 임의의 화합물을 처리하지 않은 정상대조군에서의 동일 단백질 또는 유전자의 발현 수준과 비교하는 단계를 포함하는 피부 노화 개선제의 스크리닝 방법을 제공한다.The present invention also relates to a method of treating a biological sample, comprising: treating a biological sample with an arbitrary compound; Detecting an expression level of the HAPLN1 protein or gene from the biological sample; And comparing the expression level with an expression level of the same protein or gene in a normal control group in which no compound has been treated.
상기 생물학적 시료는 조직, 세포, 전혈, 혈청 및 혈장으로 이루어진 군에서 선택된 어느 하나 이상일 수 있다.The biological sample may be any one or more selected from the group consisting of tissue, cells, whole blood, serum, and plasma.
더불어 본 발명은 HAPLN1 단백질, 이를 코딩하는 유전자 및 HAPLN1 단백질 또는 유전자의 발현 촉진 또는 기능을 활성화하는 효능제로 이루어진 군에서 선택된 어느 하나 이상을 유효성분으로 포함하는 것을 특징으로 하는 피부 노화 예방 또는 개선용 약학 조성물을 제공한다.In addition, the present invention relates to a pharmaceutical composition for preventing or improving skin aging characterized by comprising at least one selected from the group consisting of HAPLN1 protein, a gene coding therefor, and HAPLN1 protein or an agonist for promoting the expression or function of gene Lt; / RTI >
상기 약학 조성물은 히알루론산(hyaluronic acid, HA)을 더 포함할 수 있다.The pharmaceutical composition may further comprise hyaluronic acid (HA).
상기 약학 조성물은 통상적인 방법에 따라 겔제, 유제, 주사제, 산제, 과립제, 에어로솔제, 페이스트제, 경피흡수제 및 패치제로 이루어진 군에서 선택된 하나 이상의 제형으로 제공될 수 있으나, 이에 제한되는 것은 아니다.The pharmaceutical composition may be provided in one or more formulations selected from the group consisting of gels, emulsions, injections, powders, granules, aerosols, pastes, percutaneous absorbers and patches according to a conventional method, but is not limited thereto.
본 발명의 다른 구체예에서, 상기 약학 조성물은 약학 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제, 붕해제, 감미제, 피복제, 팽창제, 윤활제, 활택제, 향미제, 항산화제, 완충액, 정균제, 희석제, 분산제, 계면활성제 및 결합제로 이루어진 군에서 선택되는 하나 이상의 첨가제를 추가로 포함할 수 있다.In another embodiment of the present invention, the pharmaceutical compositions may be formulated with suitable carriers, excipients, disintegrants, sweeteners, coatings, swelling agents, lubricants, lubricants, flavors, antioxidants, buffers, , A diluent, a dispersant, a surfactant, and a binder.
구체적으로 담체, 부형제 및 희석제는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 사용할 수 있으며, 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 조성물에 적어도 하나 이상의 부형제, 예를 들면, 전분, 칼슘카보네이트, 수크로스 또는 락토오스, 젤라틴 등을 섞어 조제할 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스티레이트, 탈크 같은 윤활제들도 사용할 수 있다. 경구를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 있으며 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제 등이 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기재로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.Specific examples of carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Solid formulations for oral administration may be in the form of tablets, pills, powders, granules, capsules These solid preparations can be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., into the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin which are commonly used simple diluents. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the suppository base, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.
상기 HAPLN1 단백질 및 효능제의 바람직한 투여량은 대상체의 상태 및 체중, 노화의 종류 및 정도, 약물 형태, 투여경로 및 기간에 따라 달라질 수 있으며 당업자에 의해 적절하게 선택될 수 있다. The desired dose of the HAPLN1 protein and agonist may vary depending on the condition and body weight of the subject, the kind and degree of aging, the drug form, the administration route and the period, and may be appropriately selected by those skilled in the art.
본 발명에 있어서, 상기 '대상체'는 인간을 포함하는 포유동물일 수 있으나, 이들 예에 한정되는 것은 아니다.In the present invention, the 'subject' may be a mammal including a human, but is not limited thereto.
또한, 본 발명은 HAPLN1 단백질 또는 HAPLN1 단백질의 발현 촉진 또는 기능을 활성화하는 효능제 중 어느 하나를 유효성분으로 포함하는 것을 특징으로 하는 피부 노화 예방 또는 개선용 화장료 조성물, 또는 주름 개선용 화장료 조성물을 제공한다.Also, the present invention provides a cosmetic composition for preventing or improving skin aging, or a cosmetic composition for improving wrinkles, which comprises an effective ingredient for promoting the expression of HAPLN1 protein or HAPLN1 protein or an agonist for activating the function do.
상기 화장료 조성물은 히알루론산을 더 포함할 수 있다.The cosmetic composition may further comprise hyaluronic acid.
상기 화장료 조성물은 유효성분인 HAPLN1 단백질 및 효능제외에 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함할 수 있다.The cosmetic composition may contain conventional auxiliary agents such as stabilizers, solubilizers, vitamins, pigments and flavoring agents, and carriers, except for the HAPLN1 protein and efficacy of the active ingredient.
상기 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 썬 크림, 유연 화장수, 수렴 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.The cosmetic composition may be prepared in any form conventionally produced in the art and may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, oil, powder foundation, emulsion foundation, Wax foundation, spray, and the like. However, the present invention is not limited thereto. More specifically, it can be manufactured in the form of a sun cream, a flexible lotion, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a pack, a spray or a powder.
상기 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation is a paste, cream or gel, an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component .
상기 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, chlorofluorohydrocarbons, propane / Or propellants such as dimethyl ether.
상기 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해 화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르가 있다. When the formulation is a solution or an emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, - butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.
상기 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, a microcrystalline cellulose , Aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.
또한, 본 발명은 HAPLN1 단백질 또는 HAPLN1 단백질의 발현 촉진 또는 기능을 활성화하는 효능제 중 어느 하나를 유효성분으로 포함하는 것을 특징으로 하는 피부 노화 예방 또는 개선용 건강기능식품, 또는 주름 개선용 건강기능식품을 제공한다.The present invention also relates to a health functional food for preventing or ameliorating skin aging or a health functional food for improving wrinkles characterized by containing any one of an agonist for promoting the expression of HAPLN1 protein or HAPLN1 protein or an agonist for activating the function, .
상기 건강기능식품은 히알루론산을 더 포함할 수 있다.The health functional food may further include hyaluronic acid.
상기 건강기능식품은 분말, 과립, 정제, 캡슐, 시럽 또는 음료의 형태로 제공될 수 있으며, 상기 건강기능식품은 유효성분인 본 발명에 따른 HAPLN1 단백질 및 효능제 이외에 다른 식품 또는 식품 첨가물과 함께 사용되고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합양은 그의 사용 목적 예를 들어 예방, 건강 또는 치료적 처치에 따라 적합하게 결정될 수 있다. The health functional food may be provided in the form of powder, granules, tablets, capsules, syrups or beverages. The health functional food may be used together with food or food additives other than the active ingredient HAPLN1 protein and agonist according to the present invention , And can be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.
상기 건강기능식품에 함유된 HAPLN1 단백질 및 효능제의 유효용량은 상기 약학조성물의 유효용량에 준해서 사용할 수 있으나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 범위 이하일 수 있으며, 유효성분은 안전성 면에서 아무런 문제가 없기 때문에 상기 범위 이상의 양으로도 사용될 수 있음은 확실하다.The effective dose of the HAPLN1 protein and the agonist contained in the health functional food may be used in accordance with the effective dose of the pharmaceutical composition. However, in the case of long-term ingestion for the purpose of health and hygiene, Range, and it is clear that the active ingredient can be used in an amount of more than the above range because there is no problem in terms of safety.
상기 건강기능식품의 종류에는 특별한 제한이 없고, 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등을 들 수 있다.There is no particular limitation on the kind of the above health functional food and examples thereof include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, , Drinks, alcoholic beverages and vitamin complexes.
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.
<실시예 1> 피부노화 내인성 인자의 탐색Example 1: Detection of skin aging endogenous factors
비동시성 개체결합(Heterochronic parabiosis)은 같은 종의 노화 마우스와 젊은 마우스의 혈관을 연결하여 혈액을 공유하는 실험법으로, 비동시성 개체결합 실험모델을 이용하여, 신경, 심장 및 근육의 노화를 일으키는 생체 내부 인자들을 탐색, 규명하는데 이용된다.Heterochronic parabiosis is an experiment in which blood vessels are connected by connecting blood vessels of aged mice and young mice of the same species. By using an asynchronous individual binding experiment model, the in vivo interior of the nerves, heart and muscles It is used to search for and identify factors.
따라서, 생체 내부에서 피부노화와 관련된 인자의 존재 여부를 확인하기 위해, 한국기초과학지원연구원으로부터 얻은 20개월령 노화 C57BL/6J 마우스와 중앙실험동물로부터 얻은 6주령 어린 C57BL/6J 마우스를 마취한 후 각 마우스의 피부 및 복강근육을 절개하여 개체결합(parabiosis)하였다. 어린 마우스의 경우 우측 앞발에서 뒷다리까지의 피부를 약 5cm 정도 절개 후 우측 복강근육을 1cm 정도 절개하였고, 노화 마우스의 경우 좌측 앞발에서 뒷다리까지의 피부를 5cm 정도를 절개 후 좌측 복강근육을 1cm 정도 절개하였다. 절개한 복강근육 및 피부를 서로 연결 및 봉합하여 개체결합하였고 3주 동안 유지 후 안락사한 후, 각 실험군 마우스의 혈장 및 피부조직을 채취하였다. 안와채혈로 혈액을 전량 취해 에틸렌디아민사아세트산(ethylenediaminetetraacetic acid, EDTA)-코팅 채혈관에서 30분 정도 배양한 후 10,000 xg의 속도로 원심분리하여 혈장을 분리하였다. 분리한 혈장은 -70℃에서 보관하였다. 피부 조직은 마우스 제모 후 목 경부의 피부(cervical skin)를 1cm × 1cm 사이즈로 채취하였다. Therefore, in order to confirm the existence of factors related to skin aging in the living body, 20-month old aged C57BL / 6J mice obtained from Korean Basic Science Research Institute and 6-week old young C57BL / 6J mice obtained from central experimental animals were anesthetized The skin and abdominal muscles of the mice were incised and parabiosed. In the case of the young mouse, the skin from the right forelimb to the hind limb was incised about 5 cm, and the right abdominal muscle was incised about 1 cm. In the case of the aged mouse, the skin from the left forelimb to the hind limb was incised about 5 cm. Respectively. The incised abdominal muscles and skin were ligated and sutured together and individualized. After 3 weeks of maintenance and euthanasia, the plasma and skin tissues of each experimental group were collected. All blood was taken with orbital blood and incubated in ethylenediaminetetraacetic acid (EDTA) -coated blood vessels for 30 minutes and centrifuged at 10,000 xg to separate plasma. The separated plasma was stored at -70 ° C. The skin tissue was removed from the mice and the cervical skin of the neck was collected in a size of 1 cm x 1 cm.
면역형광법(Immunofluorescence)을 위해, 채취한 피부조직을 10% 포르말린(formalin)에 고정하여 파라핀 섹션(paraffin section)을 제작하였다. 제작한 슬라이드를 파라핀 제거(deparaffinization) 및 재수화(rehydration)하였고 고트 항-마우스 프로-콜라겐 타입 1 알파 2 IgG(goat anti-mouse pro-collagen type Ⅰ α2(COL1A2) polyclonal IgG, Santa cruz biotechnology), 동키 항-고트 IgG(H+L) 알렉사 플루오르 594(donkey anti-goat IgG (H+L) Alexa Fluor 594, Life technologies)항체를 이용하여 면역형광염색을 하였다.For immunofluorescence, the collected skin tissue was fixed in 10% formalin to prepare a paraffin section. The slides were deparaffinized and rehydrated and the goat anti-mouse
qRT-PCR을 하기 위해, 분리한 피부조직을 0.25% 트립신/EDTA(trypsin/EDTA)에서 4℃, 밤새 배양한 후 진피층을 분리하였고, 트리졸 시약(trizol reagent)을 이용해서 총 mRNA를 추출하였다. 추출한 mRNA를 cDNA 합성 키트(Life technologies)를 이용하여 cDNA로 합성 후 proCOL1A1, proCOL1A2, proCOL3A1 및 GAPDH를 qRT-PCR법으로 정량하였다. For qRT-PCR, the dermal tissue was separated from the dermal layer after incubation at 4 ° C overnight in 0.25% trypsin / EDTA (trypsin / EDTA), and total mRNA was extracted using a trizol reagent . The extracted mRNA was synthesized with cDNA using cDNA synthesis kit (Life technologies), and proCOL1A1, proCOL1A2, proCOL3A1 and GAPDH were quantified by qRT-PCR method.
그 결과, 도 1 및 도 2와 같이, 노화 마우스(Old, O)와 젊은 마우스(Young, Y)간의 비동시성 개체결합 쌍 중의 노화 개체(YO-O)의 피부조직에서 콜라겐(collagen)(도 1)과 히알루론산(도 2)의 생성이 노화-노화 간의 비동시성 개체결합 쌍 중의 노화 개체(OO-O)보다 증가함을 확인하였다.As a result, as shown in Fig. 1 and Fig. 2, in the skin tissue of an aging individual (YO-O) in an asynchronous individual pair between an aged mouse (Old, O) and a young mouse 1) and hyaluronic acid (FIG. 2) were found to be higher than the aging individuals (OO-O) in the asynchronous individual pair between aging-aging.
따라서, 생체 내부에 피부노화와 관련된 내인성 인자가 존재할 수 있음을 확인하였다.Therefore, it has been confirmed that an intrinsic factor related to skin aging may be present inside the living body.
더불어 각 실험 개체의 마우스로부터 혈장을 분리하고 프로테옴 분석(SomaScan proteomics, 미국 SomaLogic사에 의뢰)을 통해 내인성 인자를 규명하였다. 그 결과 도 3과 같이, 노화와 관련성이 있는 단백질이 HAPLN1임을 확인하였다.Plasma was isolated from the mice of each experimental subject and endogenous factors were identified through proteomic analysis (SomaScan proteomics, US SomaLogic). As a result, as shown in Fig. 3, it was confirmed that the protein associated with aging was HAPLN1.
<실시예 2> 피부노화 상태에서 HAPLN1의 양적 감소 확인Example 2: Quantitative reduction of HAPLN1 in skin aging state
1. 피부노화 시 콜라겐 생성 감소1. Decreased collagen production during skin aging
C57BL/6J 수컷 20개월령 노화 마우스와 6주령 어린 마우스의 목 경부 부위의 피부조직을 상기 실시예 1과 동일한 방법으로 1cm × 1cm 사이즈로 채취하고 이의 총 mRNA를 추출하였다. 추출한 mRNA를 cDNA 합성 키트를 이용하여 cDNA로 합성한 후 프로콜라겐 타입 (proCOL1A1 및 proCOLIA2) 및 타입 Ⅲ(proCOL3A1) mRNA의 양을 qRT-PCR 방법으로 정량하였다.C57BL / 6J Male 20-month old aged mice and 6-week-old young mice were sampled in a size of 1 cm x 1 cm by the same method as in Example 1, and their total mRNAs were extracted. The extracted mRNA was synthesized by cDNA synthesis kit and quantified with qRT-PCR method for the amount of procollagen type (proCOL1A1 and proCOLIA2) and type III (proCOL3A1) mRNA.
그 결과, 도 4 중 A와 같이, 노화 마우스에서 proCOL1A1, proCOLIA2 및 proCOL3A1의 mRNA 수준이 어린 마우스 대비 각 1/4.3, 1/4.2, 1/5.6 수준으로 감소하였다. As a result, as shown in FIG. 4A, the levels of proCOL1A1, proCOLIA2 and proCOL3A1 mRNA in senescent mice were reduced to 1 / 4.3, 1 / 4.2, and 1 / 5.6, respectively,
또한, 상기 실시예 1과 동일한 방법으로 노화 마우스와 어린 마우스의 목 경부 부위의 피부 조직을 파라핀 섹션으로 제조하고 면역형광법을 수행하였다. 이를 공초점 현미경으로 관찰한 결과, 도 4 중 B와 같이 노화 마우스 피부에서 프로콜라겐 타입 Ⅰ의 분포 감소를 확인하였다. In addition, the skin tissue of the neck region of the aged mouse and the young mouse was prepared as a paraffin section in the same manner as in Example 1, and immunofluorescence was performed. As a result of observation with a confocal microscope, a decrease in the distribution of procollagen type I was observed in aged mouse skin as shown in Fig. 4B.
2. 피부노화 시 히알루론산 양 감소2. Reduction of hyaluronic acid in skin aging
C57BL/6J 수컷 20개월령 노화 마우스와 6주령 어린 마우스의 목 경부 부위의 피부 조직을 상기 실시예 1과 동일한 방법으로 1cm × 1cm 사이즈로 채취하여 효소면역측정법(enzyme-linked immunosorbent assay, ELISA)을 수행하였다. 먼저, 블레이드(blade)를 이용해 분리한 피부조직에서 피하지방을 제거 후 1X 프로테아제 억제제(protease inhibitor)를 참가한 인산완충식염수(Phosphate buffer saline, PBS)용액에서 균질화(homogenization)하였다. 히알루론산 ELISA 키트(hyaluronic acid ELISA kit, CUSABIO)를 이용하여 제조사의 지침에 따라 히알루론산을 정량하였다.C57BL / 6J Male 20-month-old aged mice and 6-week-old young mice were sampled at a size of 1 cm × 1 cm by the same method as in Example 1 and subjected to an enzyme-linked immunosorbent assay (ELISA) Respectively. First, subcutaneous fat was removed from the skin tissue using a blade, and homogenization was performed in phosphate buffered saline (PBS) solution supplemented with 1 × protease inhibitor. Hyaluronic acid was quantitated using a hyaluronic acid ELISA kit (CUSABIO) according to the manufacturer's instructions.
그 결과, 도 5 중 A와 같이, 노화 마우스에서 히알루론산의 농도가 어린 마우스 대비 1/4.2 수준으로 감소하였다. As a result, as shown in FIG. 5A, the concentration of hyaluronic acid in aged mice was reduced to 1 / 4.2 level compared with that of young mice.
또한, 노화 마우스와 어린 마우스의 피부 조직 절편을 상기 실시예 1과 동일한 방법으로 면역형광염색하였다. 이때, 항-히알루론산 항체인 쉽 항-히알루론산 폴리클로날 항체(sheep anti-hyaluronic acid polyclonal antibody, abcam) 및 동키 항-쉽 IgG H&L 알렉사 플루오르 488(donkey anti-sheep IgG H&L Alexa Fluor 488, abcam)를 이용하였다. 이를 공초점 현미경으로 관찰한 결과 도 5 중 B와 같이, 노화 마우스 피부에서 히알루론산의 분포 감소를 확인하였다. In addition, skin tissue sections of aged and young mice were subjected to immunofluorescent staining in the same manner as in Example 1 above. At this time, sheep anti-hyaluronic acid polyclonal antibody (abcam), an anti-hyaluronic acid antibody, and Donkey anti-sheep IgG H & L Alexa Fluor 488, abcam ) Were used. As a result of observation with a confocal microscope, a decrease in the distribution of hyaluronic acid was observed in the skin of aged mice as shown in B of FIG.
3. 피부 노화 시 HAPLN1의 발현 양 감소3. Decreased expression of HAPLN1 in skin aging
상기 실시예 2 중 2와 같은 방법으로 C57BL/6J 수컷 20개월령 노화 마우스와 6주령 어린 마우스의 목 경부 피부조직을 취해 PBS 용액에서 균질화한 후, 마우스 히알루론산 및 HAPLN1 ELISA 키트(mouse hyaluronan and proteoglycan link protein 1(HAPLN1) ELISA kit, CUSABIO)를 이용하여 HAPLN1의 농도를 측정하였다. The cervical skin tissues of C57BL / 6J male 20-month old aged and 6-week-old young mice were homogenized in PBS solution in the same manner as in Example 2, and then homogenized in mouse hyaluronic acid and HAPLN1 ELISA kit (mouse hyaluronan and proteoglycan link protein 1 (HAPLN1) ELISA kit, CUSABIO) was used to measure the concentration of HAPLN1.
측정 결과, 도 6 중 A와 같이 노화 마우스에서 HAPLN1의 농도가 어린 마우스 대비 1/4.2 수준으로 감소하였다. As a result, the concentration of HAPLN1 in aged mice was reduced to 1 / 4.2 level compared with that of young mice as shown in A of FIG.
또한, 노화 마우스와 어린 마우스의 피부 조직 절편을 상기 실시예 1과 동일한 방법으로 면역형광염색하였다. 이때, 고트 항-HAPLN1 폴리클로날 IgG(goat anti-HAPLN1 polyclonal IgG, santa cruz biotechnology) 및 동키 항-고트 IgG (H+L) 알렉사 플루오르 488(donkey anti-goat IgG (H+L) Alexa Fluor 594, Life technologies)를 이용하였다. In addition, skin tissue sections of aged and young mice were subjected to immunofluorescent staining in the same manner as in Example 1 above. At this time, the goat anti-HAPLN1 polyclonal IgG, santa cruz biotechnology and Donkey anti-goat IgG (H + L) Alexa Fluor 594 (H + L) , Life technologies) were used.
이를 공초점 현미경으로 관찰한 결과, 도 6 중 B와 같이 노화 마우스 피부에서 HAPLN1의 분포가 감소된 것을 확인하였다. As a result of observation with a confocal microscope, it was confirmed that the distribution of HAPLN1 in aged mouse skin was decreased as shown in FIG. 6B.
<실시예 3> HAPLN1 투여로 인한 피부노화 개선 효과 확인<Example 3> Confirmation of skin aging improvement effect by administration of HAPLN1
재조합 마우스 HAPLN1(Recombinant mouse HAPLN1, Cusabio biotech co.에서 구매)을 PBS에 녹여 C57BL/6J 수컷 20개월령 노화 마우스에 0.1 mg/kg의 용량으로 1일 1회 14일 동안 복강 투여하였다. 노화 마우스 및 C57BL/6J 수컷(6주령)에 같은 양의 PBS를 투여하여 대조군으로 비교하였다.Recombinant mouse HAPLN1 (purchased from Recombinant mouse HAPLN1, Cusabio biotech co.) Was dissolved in PBS and administered intraperitoneally to the C57BL /
1. 노화 마우스에서 HAPLN1 투여로 인한 콜라겐 증가 확인1. Confirmation of collagen increase due to HAPLN1 administration in aging mice
투여 종료 후, 상기 실시예 1과 동일한 방법으로 각 실험군에서 피부 조직을 취해서 진피층을 분리하여 RNA를 추출 후 프로콜라겐 타입 (proCOL1A1 및 proCOLIA2) 및 타입 Ⅲ(proCOL3A1)를 qRT-PCR 방법으로 정량하였다.After completion of the administration, skin tissues were taken from each experimental group in the same manner as in Example 1, and the dermal layer was separated to extract RNA. The procolase types (proCOL1A1 and proCOLIA2) and type III (proCOL3A1) were quantified by qRT-PCR method.
그 결과, 도 7 중 A와 같이, HAPLN1 투여군에서 proCOL1A1, proCOLIA2 및 proCOL3A1의 mRNA 수준이 비투여군 대비 각 2.6배, 2.5배, 2.8배 증가하여 어린 마우스와 비슷한 수준으로 회복하였음을 확인하였다. 또한, 피부조직 절편을 상기 실시예 2 중 1과 동일한 방법으로 항-프로콜라겐 타입 Ⅰ 항체으로 염색하고 공초점 현미경으로 관찰한 결과 도 7 중 B와 같이, HAPLN1 투여군에서 프로콜라겐 타입 Ⅰ 분포 수준이 비투여군에 비해 어린 마우스와 비슷한 수준으로 증가하였음을 확인하였다.As a result, it was confirmed that the levels of proCOL1A1, proCOLIA2 and proCOL3A1 mRNA were 2.6, 2.5, and 2.8 times higher than those of the non-treated group in the HAPLN1-treated group, respectively, as shown in FIG. In addition, skin tissue sections were stained with anti-procollagen type I antibody in the same manner as in Example 2, and observed with a confocal microscope. As a result, as shown in Fig. 7B, the distribution level of procollagen type I in the HAPLN1- And increased to a level similar to that of the untreated mice.
2. HAPLN1 투여로 인한 노화 마우스의 히알루론산 증가 확인2. Confirmation of hyaluronic acid increase in aging mice due to HAPLN1 administration
투여 종료 후, 상기 실시예 2 중 2와 같은 방법으로 각 실험군의 목 경부 피부조직을 취해 PBS 용액에서 균질화한 후, 히알루론산 ELISA 키트(hyaluronic acid ELISA kit, CUSABIO)를 이용하여 제조사의 지침에 따라 히알루론산을 정량하였다.After completion of the administration, the neck skin tissue of each experimental group was taken and homogenized in a PBS solution in the same manner as in Example 2, and then, using a hyaluronic acid ELISA kit (CUSABIO) according to the manufacturer's instructions Hyaluronic acid was quantified.
그 결과, 도 8 중 A와 같이, HAPLN1 투여군에서 히알루론산 수준이 비투여군 대비 1.5배 증가하여 어린 마우스와 비슷한 수준으로 회복하였음을 확인하였다.As a result, as shown in A of FIG. 8, it was confirmed that the hyaluronic acid level in the HAPLN1-treated group was 1.5-fold higher than that in the non-treated group and recovered to a level similar to that of the young mouse.
또한, 피부조직 절편을 상기 실시예 2 중 2와 동일한 방법으로 히알루론산을 면역형광염색하고 공초점 현미경으로 관찰한 결과 도 8 중 B와 같이, HAPLN1 투여군에서 히알루론산 분포 수준이 비투여군에 비해 어린 마우스와 비슷한 수준으로 증가하였음을 확인하였다.In addition, the skin tissue sections were subjected to immunofluorescent staining of hyaluronic acid in the same manner as in Example 2-2 above, and observed with a confocal microscope. As a result, as shown in B of FIG. 8, the hyaluronic acid distribution level in the HAPLN1- And it was confirmed to be similar to that of mouse.
<< 실시예Example 4> 4> HAPLN1HAPLN1 처리로 인한 Due to processing TGFTGF -- β1β1 수용체 Ⅱ, Receptor II, TGFTGF -- β1β1 유도 콜라겐 생성 및 히알루론산 합성 효소(HAS2) 증가 확인 Induction collagen production and hyaluronic acid synthase (HAS2) increase confirmation
1. NHDF 세포에서 HAPLN1 처리로 인한 TGF-β1 유도 콜라겐 생성 증가 확인1. Identification of increased TGF-β1 induced collagen production by HAPLN1 treatment in NHDF cells
NHDF(normal human dermal fibroblasts, 정상 사람 피부 섬유아세포) 세포주에서 HAPLN1이 TGF-β1 유도 proCOL1α2 생성에 영향을 주는지를 확인하기 위해 웨스턴 블랏법을 이용하여 측정하였다. TGF-β는 콜라겐 생성을 유도하는 인자로써(비특허문헌2), NHDF 세포주에 각 농도별(0, 25, 50, 100 ng/ml) 재조합 인간 HAPLN1(rhHAPLN1)을 처리하고 30분 후, 1 ng/ml 재조합 인간 TGF-β1을 24시간 동안 처리하였다. To determine whether HAPLN1 affects TGF-β1-induced proCOL1α2 production in normal human dermal fibroblasts (NHDF) cell lines, Western blotting was used. TGF-β was treated with recombinant human HAPLN1 (rhHAPLN1) at various concentrations (0, 25, 50, 100 ng / ml) in NHDF cell line as a factor inducing collagen production (Non-Patent Document 2) ng / ml recombinant human TGF-? 1 was treated for 24 hours.
그 결과, 도 9와 같이, TGF-β1 유도 proCOL1α2 생성이 HAPLN1 처리에 따라 증가하는 것을 확인하였다.As a result, as shown in FIG. 9, it was confirmed that the production of TGF-β1-induced proCOL1α2 was increased by treatment with HAPLN1.
2. NHDF 세포에서 HAPLN1 처리로 인한 TGF-β 수용체 Ⅱ 증가 확인2. Identification of TGF-β Receptor II Increase by HAPLN1 Treatment in NHDF Cells
HAPLN1이 TGF-β의 신호전달에 관여하는지를 확인하기 위해, NHDF 세포주에 각 농도별(0, 25, 50, 100 ng/ml) 재조합 인간 HAPLN1(rhHAPLN1)을 처리하고 24시간 후, TGF-β 수용체의 단백질 발현양을 웨스턴 블랏법을 이용하여 측정하였다. In order to confirm whether HAPLN1 is involved in TGF-β signaling, recombinant human HAPLN1 (rhHAPLN1) was treated with NHDF cell line (0, 25, 50, 100 ng / ml) Was measured by Western blotting.
그 결과, 도 10 중 A와 같이, 두 가지 수용체 중에 TGF-β 수용체 Ⅱ의 단백질 발현양이 HAPLN1 처리에 따라 농도 의존적으로 증가하는 것을 확인하였다. 같은 실험 조건에서 TGF-β 수용체의 mRNA 발현양을 qRT-PCR을 통하여 확인한 결과, 도 10 중 B 및 C와 같이 HAPLN1 처리에 따른 변화가 없는 것을 확인하였다. As a result, as shown in FIG. 10A, it was confirmed that the amount of TGF-β receptor II expressed in the two receptors increased in a concentration-dependent manner by the HAPLN1 treatment. In the same experimental conditions, the amount of mRNA expression of TGF-β receptor was confirmed by qRT-PCR, and it was confirmed that there was no change according to treatment with HAPLN1 as shown in B and C in FIG.
따라서, HAPLN1이 전사적 조절이 아닌 안정성 조절을 통해 TGF-β 수용체 Ⅱ의 양을 증가시켰는지를 확인하기 위해, 단백질 합성 억제제인 사이클로헥사마이드(cycloheximide)를 10 μM의 농도로 1시간 동안 선처리하고, 100 ng/ml rhHAPLN1을 6시간 또는 24시간 동안 처리한 후, TGF-β 수용체 Ⅱ의 단백질 발현 정도를 웨스턴 블랏법을 이용하여 확인하였다. Therefore, in order to confirm whether HAPLN1 increased the amount of TGF-β receptor II by controlling the stability rather than the transcriptional regulation, cycloheximide, a protein synthesis inhibitor, was pretreated for 1 hour at a concentration of 10 μM, ng / ml rhHAPLN1 was treated for 6 hours or 24 hours, and the degree of protein expression of TGF-beta receptor II was confirmed by western blotting.
그 결과, 도 11과 같이, 사이클로헥사마이드 존재 하에서도 rhHAPLN1 처리가 TGF-β 수용체 Ⅱ의 양을 증가시키는 것을 확인하였다. 따라서, HAPLN1은 TGF-β 수용체 Ⅱ의 안정성을 증가시킨다고 볼 수 있다. As a result, it was confirmed that treatment with rhHAPLN1 also increased the amount of TGF-? Receptor II in the presence of cyclohexamide, as shown in Fig. Thus, HAPLN1 is thought to increase the stability of TGF-beta receptor II.
3. 3. NHDFNHDF 세포에서 In a cell HAPLN1HAPLN1 처리로 인한 Due to processing TGFTGF -- β1β1 유도 히알루론산 Induced hyaluronic acid 생성효Generation 소(HAS2) 발현 증가 확인Increased expression of small (HAS2) expression
진피 섬유아세포에서는 히알루론산 생성효소 3가지(HAS1, HAS2 및 HAS3) 중 주로 HAS2가 존재하고 있다(비특허문헌3). NHDF 세포주에서 HAPLN1이 TGF-β1 유도 HAS2 발현에 영향을 주는지를 확인하기 위해, NHDF 세포주에 각 농도 별로(0, 25, 50, 100 ng/mL) 재조합 인간 HAPLN1(rhHAPLN1)을 처리하고 30분 후, 1 ng/mL의 재조합 인간 TGF-β1을 24시간 동안 처리한 뒤, 웨스턴 블랏법을 이용하여 HAS2의 단백질 발현양을 확인하였다. In dermal fibroblasts, HAS2 is mainly present among three kinds of hyaluronic acid producing enzymes (HAS1, HAS2 and HAS3) (Non-Patent Document 3). To determine whether HAPLN1 affects TGF-β1 induced HAS2 expression in NHDF cell lines, NHDF cell lines were treated with recombinant human HAPLN1 (rhHAPLN1) at each concentration (0, 25, 50, 100 ng / mL) , And 1 ng / mL of recombinant human TGF-
그 결과, 도 12와 같이, TGF-β1 유도 HAS2 발현이 HAPLN1 처리에 따라 증가하는 것을 확인하였다.As a result, as shown in Fig. 12, it was confirmed that TGF-β1-induced HAS2 expression increased with HAPLN1 treatment.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시예일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that such detail is solved by the person skilled in the art without departing from the scope of the invention. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
Claims (14)
상기 약학 조성물은 히알루론산을 더 포함하는 것을 특징으로 하는 피부 탄력 또는 주름 개선용 약학 조성물.8. The method of claim 7,
Wherein the pharmaceutical composition further comprises hyaluronic acid.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR20150127585 | 2015-09-09 | ||
| KR1020150127585 | 2015-09-09 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20170031060A KR20170031060A (en) | 2017-03-20 |
| KR101897340B1 true KR101897340B1 (en) | 2018-09-13 |
Family
ID=58502793
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020160115670A KR101897340B1 (en) | 2015-09-09 | 2016-09-08 | Composition for diagnosing, preventing or improving aging of skin using HAPLN1 |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR101897340B1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3887525A4 (en) * | 2020-02-03 | 2022-06-22 | Haplnscience Inc. | Composition for preventing or treating pulmonary diseases comprising hyaluronan and proteoglycan link protein 1 |
| WO2022146036A1 (en) | 2020-12-30 | 2022-07-07 | Haplnscience inc. | Medium composition for culturing animal cells for producing recombinant extracellular matrix protein and method of using the same |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2017403257B2 (en) | 2017-03-06 | 2021-09-30 | Haplnscience inc. | Composition for skin aging measurement, prevention, or alleviation, using HAPLN1 |
| KR20190024727A (en) | 2017-08-29 | 2019-03-08 | 중앙대학교 산학협력단 | Composition for cartilage regeneration comprising HAPLN1 |
| WO2019045451A1 (en) * | 2017-08-29 | 2019-03-07 | 중앙대학교 산학협력단 | Cartilage regeneration composition containing hapln1 as active ingredient |
| JP7182816B2 (en) * | 2019-04-30 | 2022-12-05 | ハプルサイエンス・インコーポレイテッド | Composition for prevention or treatment of hair loss containing HAPLN1 |
| JP7222001B2 (en) * | 2021-01-22 | 2023-02-14 | ハプルサイエンス・インコーポレイテッド | Composition for measuring, preventing or improving skin aging using HAPLN1 |
| CN117881412A (en) * | 2021-08-03 | 2024-04-12 | 中央大学校产学协力团 | Composition for preventing or treating fibrotic diseases comprising HAPLN1 |
| KR20240017998A (en) * | 2022-08-01 | 2024-02-13 | 주식회사 하플사이언스 | An anti-oxidant composition including HAPLN1 and a method for anti-oxidation of cells using the same |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070161022A1 (en) * | 2005-11-30 | 2007-07-12 | Kim Stuart K | Signatures for human aging |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120051180A (en) | 2010-11-12 | 2012-05-22 | 연세대학교 산학협력단 | Composition for preventing or improving skin aging and wrinkling comprising fermentation products prepared from alcoholic beverages byproducts |
-
2016
- 2016-09-08 KR KR1020160115670A patent/KR101897340B1/en active IP Right Grant
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070161022A1 (en) * | 2005-11-30 | 2007-07-12 | Kim Stuart K | Signatures for human aging |
Non-Patent Citations (2)
| Title |
|---|
| Arch Dermatol Res., (2015.03.), Vol. 307, pp 351-364.* |
| PLOS ONE, (2012), Vol. 7, Issue 11, e50393, pp 1-10. |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3887525A4 (en) * | 2020-02-03 | 2022-06-22 | Haplnscience Inc. | Composition for preventing or treating pulmonary diseases comprising hyaluronan and proteoglycan link protein 1 |
| WO2022146036A1 (en) | 2020-12-30 | 2022-07-07 | Haplnscience inc. | Medium composition for culturing animal cells for producing recombinant extracellular matrix protein and method of using the same |
| EP4070869A1 (en) | 2020-12-30 | 2022-10-12 | Haplnscience Inc. | Method of analyzing a monomer of a recombinant extracellular matrix protein by size exclusion chromatography |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20170031060A (en) | 2017-03-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101897340B1 (en) | Composition for diagnosing, preventing or improving aging of skin using HAPLN1 | |
| US9097725B2 (en) | Compositions and methods for modulating the immune system | |
| Schmidt et al. | Disulfide-bonded multimers of proteoglycan 4 (PRG4) are present in normal synovial fluids | |
| Suzuki et al. | Aberrant glycosylation of lumican in aortic valve stenosis revealed by proteomic analysis | |
| US11202749B2 (en) | Composition for skin aging measurement, prevention, or alleviation, using HAPLN1 | |
| JP6667437B2 (en) | Identification of novel polypeptide hormones for maintaining optimal weight and blood glucose | |
| Neame et al. | Pleiotrophin is an abundant protein in dissociative extracts of bovine fetal epiphyseal cartilage and nasal cartilage from newborns | |
| JP7222001B2 (en) | Composition for measuring, preventing or improving skin aging using HAPLN1 | |
| US11028128B2 (en) | Inhibition of the aggregation of transthyretin by specific binding of peptides to aggregation-driving segments | |
| US10729634B2 (en) | Sex hormone-binding globulin for use as a medicament | |
| Bruch et al. | Rat olfactory neurons express a 200 kDa neurofilament | |
| EP1867341A1 (en) | Therapeutic agent for non-alcoholic fatty liver disease, and screening method for drug candidate compound for treatment or prevention of non-alcoholic fatty liver disease | |
| Nyindodo-Ogari et al. | Giardia mitosomal protein import machinery differentially recognizes mitochondrial targeting signals | |
| KR102137531B1 (en) | Pharmaceutical composition for preventing or treating age related sarcopenia comprising inhibitors of PLD2 protein as an active ingredient | |
| US20100215583A1 (en) | Cell nucleus-entering compositions | |
| KR20230172930A (en) | Composition for preventing or treating of obesity comprising peptides derived from Neuronal growth regulator 1(NEGR1) | |
| KR20230132377A (en) | Composition for preventing or treating obesity, diabetes or NASH comprising SHLP2 | |
| CA3220075A1 (en) | Peptides derived from ruminococcus torques | |
| CN117750967A (en) | Peptides derived from ruminococcus sprain | |
| Bian et al. | Change of cardiac function in experimental autoimmune myocarditis is correlated with the expression of annexin VI | |
| Eyre et al. | Molecular Consequences of Defective SERPINH1/HSP47 in the Dachshund Natural Model of Osteogenesis Imperfecta Uschi Lindert1, Mary Ann Weis2, Jyoti Rai2, Frank Seeliger3, Ingrid Hausser4, Tosso Leeb5, David | |
| KR20160040553A (en) | Composition for treating and preventing multiple sclerosis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E902 | Notification of reason for refusal | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant |