JP7343864B2 - 線維芽細胞又は軟骨細胞におけるエラスチン産生促進剤 - Google Patents
線維芽細胞又は軟骨細胞におけるエラスチン産生促進剤 Download PDFInfo
- Publication number
- JP7343864B2 JP7343864B2 JP2021196566A JP2021196566A JP7343864B2 JP 7343864 B2 JP7343864 B2 JP 7343864B2 JP 2021196566 A JP2021196566 A JP 2021196566A JP 2021196566 A JP2021196566 A JP 2021196566A JP 7343864 B2 JP7343864 B2 JP 7343864B2
- Authority
- JP
- Japan
- Prior art keywords
- elastin
- gly
- pro
- val
- peptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 229920002549 elastin Polymers 0.000 title claims description 56
- 102000016942 Elastin Human genes 0.000 title claims description 54
- 108010014258 Elastin Proteins 0.000 title claims description 54
- 238000004519 manufacturing process Methods 0.000 title claims description 31
- 210000001612 chondrocyte Anatomy 0.000 title claims description 24
- 210000002950 fibroblast Anatomy 0.000 title description 20
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 43
- 230000001737 promoting effect Effects 0.000 claims description 24
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 23
- GIAZPLMMQOERPN-YUMQZZPRSA-N Val-Pro Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(O)=O GIAZPLMMQOERPN-YUMQZZPRSA-N 0.000 claims description 14
- 150000003839 salts Chemical class 0.000 claims description 14
- 239000003795 chemical substances by application Substances 0.000 claims description 10
- 108010003885 valyl-prolyl-glycyl-glycine Proteins 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 18
- 230000000694 effects Effects 0.000 description 15
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 12
- 210000001626 skin fibroblast Anatomy 0.000 description 11
- 230000004663 cell proliferation Effects 0.000 description 10
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 8
- 235000013305 food Nutrition 0.000 description 8
- 210000004072 lung Anatomy 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000000034 method Methods 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 238000001061 Dunnett's test Methods 0.000 description 5
- -1 amine salt Chemical class 0.000 description 5
- 239000012228 culture supernatant Substances 0.000 description 5
- 210000000629 knee joint Anatomy 0.000 description 5
- 210000003041 ligament Anatomy 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- OZFAFGSSMRRTDW-UHFFFAOYSA-N (2,4-dichlorophenyl) benzenesulfonate Chemical compound ClC1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=CC=C1 OZFAFGSSMRRTDW-UHFFFAOYSA-N 0.000 description 4
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 4
- 239000004480 active ingredient Substances 0.000 description 4
- 210000004177 elastic tissue Anatomy 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 241000283086 Equidae Species 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- 241000282887 Suidae Species 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 210000002808 connective tissue Anatomy 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 239000008194 pharmaceutical composition Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 210000003491 skin Anatomy 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 230000037303 wrinkles Effects 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- 241000269851 Sarda sarda Species 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 239000012298 atmosphere Substances 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 150000007942 carboxylates Chemical class 0.000 description 2
- 210000000845 cartilage Anatomy 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 230000001617 migratory effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000017423 tissue regeneration Effects 0.000 description 2
- 210000003556 vascular endothelial cell Anatomy 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- 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 description 1
- UUUHXMGGBIUAPW-UHFFFAOYSA-N 1-[1-[2-[[5-amino-2-[[1-[5-(diaminomethylideneamino)-2-[[1-[3-(1h-indol-3-yl)-2-[(5-oxopyrrolidine-2-carbonyl)amino]propanoyl]pyrrolidine-2-carbonyl]amino]pentanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-methylpentanoyl]pyrrolidine-2-carbon Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(C(C)CC)NC(=O)C(CCC(N)=O)NC(=O)C1CCCN1C(=O)C(CCCN=C(N)N)NC(=O)C1CCCN1C(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C1CCC(=O)N1 UUUHXMGGBIUAPW-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 102100026189 Beta-galactosidase Human genes 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 102100028065 Fibulin-5 Human genes 0.000 description 1
- 101710170766 Fibulin-5 Proteins 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 description 1
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 1
- HAEGAELAYWSUNC-WPRPVWTQSA-N Pro-Gly-Val Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O HAEGAELAYWSUNC-WPRPVWTQSA-N 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 1
- SJRUJQFQVLMZFW-WPRPVWTQSA-N Val-Pro-Gly Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SJRUJQFQVLMZFW-WPRPVWTQSA-N 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 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 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 108010064033 elastin-binding proteins Proteins 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 230000008717 functional decline Effects 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-M hydrogensulfate Chemical compound OS([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-M 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 238000010232 migration assay Methods 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000007065 protein hydrolysis Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000007665 sagging Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 230000037394 skin elasticity Effects 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/07—Tetrapeptides
-
- 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
- C07K5/06052—Val-amino acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/32—Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/33—Fibroblasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/05—Dipeptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- 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
- C07K5/10—Tetrapeptides
- C07K5/1002—Tetrapeptides with the first amino acid being neutral
- C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
- C07K5/101—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0655—Chondrocytes; Cartilage
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0656—Adult fibroblasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/998—Proteins not provided for elsewhere
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Rheumatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Virology (AREA)
- Developmental Biology & Embryology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Fodder In General (AREA)
- Cosmetics (AREA)
Description
96ウェルプレートに、培養フラスコ中で培養した皮膚線維芽細胞を3500cells/cm2になるように播種した(100μL/ウェル、培地:10%FBS DMEM)。37℃、5%CO2条件下で24時間培養した後、ウェル中の培養液を除去し、0.5%FBSを含むDMEMを添加して同条件下で更に24時間培養した。ウェル中の培養液を除去し、DPBSにより洗浄後、DPBSを除去した。0.5%FBSを含むDMEMにて、所定の各濃度となるように、ペプチドVal-Pro-Gly-Gly(VPGG)及びペプチドVal-Pro-Gly-Ala(VPGA)の溶液を調製し、100μLずつウェルに添加した。37℃、5%CO2の条件化で4日間培養を行い、培養上清を回収した。回収した培養上清は、実施例2においてエラスチン産生量の測定に使用した。0.5%FBSを含むDMEMを加え、cell counting kit-8(10μL/ウェル)により、細胞数を測定し、ペプチドを添加しない場合に対する細胞増殖率を算出した。
正常ヒト皮膚線維芽細胞におけるエラスチンの産生量の定量は、ELISA法を用いて行った。検量線作成のための標準物質として、可溶化エラスチン(牛項靱帯由来)を用いて、2000、1000、500、250、125、62.5、31.25、0ng/mLの濃度となるように、0.5%のFBSを含むDMEMで希釈した。このようにして調製した試料を、40μLずつ、Corning 96Well EIA/RIA Half Area Flat Bottom Plate (High binding)へ加えて、4℃で1時間30分インキュベートし、プレートに固定化した。併せて、実施例1において回収した各培養上清も、40μLずつ添加して、同様の手順により固定化した。ウェル内の液を捨て、PBSTで洗浄後、0.1%ゼラチンを含むブロッキングバッファー(150μL)で1時間ブロッキングした。PBSTで洗浄後、500倍希釈した一次抗体(ウサギ抗エラスチン抗体:Anti-tropo elastin (Rabbit-Poly)
PR385}を40μL加えて1時間インキュベートした。PBSTで洗浄後、1000倍希釈した二次抗体(HRPコンジュゲートヤギ抗ウサギ抗体:Peroxidase-Labeled Affinity Purified Antibody To Rabbit IgG (H+L))を40μL加えて30分間インキュベートした。ウェル内の液を捨て、PBSTで洗浄後、基質溶液(TMB Peroxidase substrate: Peroxidase Substrate Solution B = 1:1 solution)を40μL加え、5分間室温にてインキュベートした。1N H2SO4を40μL加え、酵素反応を停止した後、450nmの吸光度(A450)を測定し、検量線を元にエラスチンの産生量を算出し、これを実施例1における細胞数の測定結果で除することにより、細胞1個あたりのエラスチンの産生量を求めた。
96ウェルプレートに、培養フラスコ中で培養した肺線維芽細胞を2500cells/cm2になるように播種した(100μL/ウェル、培地:10%FBS DMEM)。37℃、5%CO2条件下で24時間培養した後、ウェル中の培養液を除去し、0.5%FBSを含むDMEMを添加して同条件下で更に24時間培養した。ウェル中の培養液を除去し、DPBSにより洗浄後、DPBSを除去した。0.5%FBSを含むDMEMにて、所定の各濃度となるようにVal-Pro-Gly-Gly及びVal-Pro-Gly-Alaの溶液を調製し、100μLずつウェルに添加した。37℃、5%CO2の条件化で3日間培養を行い、培養上清を回収した。回収した培養上清は、参考例4においてエラスチン産生量の測定に使用した。0.5%FBSを含むDMEMを加え、cell counting kit-8(10μL/ウェル)により細胞増殖率を算出した。
組織の修復、再生及び創傷治癒の過程にも関わる線維芽細胞の遊走活性にペプチドVal-Pro-Gly-Glyが及ぼす効果について検討した。CytoSelect(登録商標)24-well Cell Migration Assayキット(Cell Bionics社製)24ウェルプレートのウェルの底に、DMEM培地(対照)、ペプチドVal-Pro-Gly-Gly又は陽性対照としてのペプチドVal-Pro-Gly(伊藤浩行編著、「エラスチン-構造・機能・病理-」、日本エラスチン研究会(非売品)、p.124-137、2008年参照)を溶解したDMEM培地を、それぞれ500μLずつ添加した。次に、ヒト皮膚線維芽細胞(P4~5)をDMEM 培地で懸濁した細胞懸濁液(1×106 cells/mL)を調製し、メンブレンインサート(ポアサイズ8μm)内部に300μL添加した。インサートをウェルに挿入後、細胞培養インキュベーターにて37℃で、20時間インキュベートした(予備検討では6時間も設定)。反応後、インサートから培地を吸引し綿棒(2本両端/ウェル)を使って、メンブレンを透過しなかった非遊走細胞を除去した。細胞染色液を400μL添加して室温で10分反応後、インサートを蒸留水で洗浄し乾燥させ、光学顕微鏡で染色を確認した。次に空のウェルにインサートを移し、抽出液200μLを添加し、室温で10分撹捧しながら反応させた。反応液100μLを96ウェルプレートに移し、プレートリーダーで560nmの吸光度を測定した。
正常ヒト膝関節軟骨細胞(NHAc-kn:1.0×105 cells/mL)を24穴プレートに播種し(900μL/ウェル、培地:CBM+0.5%FBS)、5%CO2雰囲気で、加湿下、37℃で48時間インキュベートした。培地を交換後、ペプチドVPGG(参考例5)及びペプチドVP(実施例1)を添加し(100μL/ウェル、終濃度:0~5000nM)、5%CO2雰囲気で、加湿下、37℃で48時間インキュベートした。得られた培養物から細胞上清を回収し、PVDF膜にドットブロットした。5%スキムミルクを含むPBSTでブロッキング後、PBSTで洗浄した。一次抗体として、ウサギ抗エラスチン抗体と反応させ、PBSTで洗浄後、二次抗体としてHRPコンジュゲートヤギ抗ウサギ抗体と反応させ、PBSTで洗浄した。検出試薬ImmunoStar Zeta(和光純薬)と反応させ、得られたシグナル強度を元にエラスチンの産生量を求めた。
Claims (2)
- Val-Proで表されるアミノ酸配列からなるペプチド又はその塩を含む軟骨細胞に おけるエラスチン産生促進剤。
- 前記軟骨細胞が、ヒト軟骨細胞であることを特徴とする請求項1に記載の軟骨細胞におけるエラスチン産生促進剤。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2021196566A JP7343864B2 (ja) | 2017-10-13 | 2021-12-03 | 線維芽細胞又は軟骨細胞におけるエラスチン産生促進剤 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2017199547A JP7074995B2 (ja) | 2017-10-13 | 2017-10-13 | 線維芽細胞増殖促進剤及び線維芽細胞におけるエラスチン産生促進剤 |
JP2021196566A JP7343864B2 (ja) | 2017-10-13 | 2021-12-03 | 線維芽細胞又は軟骨細胞におけるエラスチン産生促進剤 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2017199547A Division JP7074995B2 (ja) | 2017-10-13 | 2017-10-13 | 線維芽細胞増殖促進剤及び線維芽細胞におけるエラスチン産生促進剤 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2022025155A JP2022025155A (ja) | 2022-02-09 |
JP7343864B2 true JP7343864B2 (ja) | 2023-09-13 |
Family
ID=66096837
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2017199547A Active JP7074995B2 (ja) | 2017-10-13 | 2017-10-13 | 線維芽細胞増殖促進剤及び線維芽細胞におけるエラスチン産生促進剤 |
JP2021196566A Active JP7343864B2 (ja) | 2017-10-13 | 2021-12-03 | 線維芽細胞又は軟骨細胞におけるエラスチン産生促進剤 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2017199547A Active JP7074995B2 (ja) | 2017-10-13 | 2017-10-13 | 線維芽細胞増殖促進剤及び線維芽細胞におけるエラスチン産生促進剤 |
Country Status (2)
Country | Link |
---|---|
US (1) | US20190111103A1 (ja) |
JP (2) | JP7074995B2 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113209271A (zh) * | 2021-05-11 | 2021-08-06 | 北京易扬三泰科贸有限公司 | 一种可以促进成纤维细胞增殖的组合物及其制备方法 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003104997A (ja) | 2001-09-28 | 2003-04-09 | Japan Tobacco Inc | 旨味を有する新規ペプチド、及びそれを旨味成分とする調味料 |
JP2007182414A (ja) | 2006-01-06 | 2007-07-19 | Kunio Suetsuna | 新規なかつお節ペプチド、l−バリル−l−プロリン及び血圧降下剤 |
JP2010202578A (ja) | 2009-03-03 | 2010-09-16 | Hayashikane Sangyo Kk | 皮膚改善剤、血管機能改善剤、並びにこれらを含む医薬組成物、食品、飼料及び化粧品 |
WO2013005362A1 (ja) | 2011-07-07 | 2013-01-10 | 高砂香料工業株式会社 | 風味改善ペプチド |
JP2015051953A (ja) | 2013-09-09 | 2015-03-19 | 国立大学法人山口大学 | 血管弛緩作用を有するペプチド及び血管弛緩剤 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4474851A (en) * | 1981-10-02 | 1984-10-02 | The University Of Alabama In Birmingham | Elastomeric composite material comprising a polypeptide |
US7494788B2 (en) * | 2005-07-11 | 2009-02-24 | Molecular Kinetics, Inc. | Entropic bristle domain sequences and their use in recombinant protein production |
JP2007151453A (ja) * | 2005-12-02 | 2007-06-21 | Hayashikane Sangyo Kk | エラスチン高含有可溶性ペプチドおよびその製造方法 |
JP5276813B2 (ja) * | 2006-08-25 | 2013-08-28 | 日本ハム株式会社 | エラスチン分解ペプチド並びにエラスチン及びその酵素分解ペプチドの製造方法 |
WO2008030968A2 (en) * | 2006-09-06 | 2008-03-13 | Phase Bioscience, Inc. | Fusion peptide therapeutic compositions |
JP2010155820A (ja) * | 2008-10-10 | 2010-07-15 | Hayashikane Sangyo Kk | 皮膚改善剤、血管改善剤、並びにこれらを含む医薬組成物、食品、飼料及び化粧品 |
KR101317420B1 (ko) * | 2010-03-11 | 2013-10-10 | 한국과학기술원 | 고분자량의 재조합 실크 또는 실크 유사 단백질 및 이를 이용하여 제조된 마이크로 또는 나노 크기의 거미줄 또는 거미줄 유사 섬유 |
-
2017
- 2017-10-13 JP JP2017199547A patent/JP7074995B2/ja active Active
-
2018
- 2018-10-09 US US16/155,015 patent/US20190111103A1/en not_active Abandoned
-
2021
- 2021-12-03 JP JP2021196566A patent/JP7343864B2/ja active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003104997A (ja) | 2001-09-28 | 2003-04-09 | Japan Tobacco Inc | 旨味を有する新規ペプチド、及びそれを旨味成分とする調味料 |
JP2007182414A (ja) | 2006-01-06 | 2007-07-19 | Kunio Suetsuna | 新規なかつお節ペプチド、l−バリル−l−プロリン及び血圧降下剤 |
JP2010202578A (ja) | 2009-03-03 | 2010-09-16 | Hayashikane Sangyo Kk | 皮膚改善剤、血管機能改善剤、並びにこれらを含む医薬組成物、食品、飼料及び化粧品 |
WO2013005362A1 (ja) | 2011-07-07 | 2013-01-10 | 高砂香料工業株式会社 | 風味改善ペプチド |
JP2015051953A (ja) | 2013-09-09 | 2015-03-19 | 国立大学法人山口大学 | 血管弛緩作用を有するペプチド及び血管弛緩剤 |
Non-Patent Citations (4)
Title |
---|
Agr. Biol. Chem.,1973年,Vol. 37, No. 10,pp. 2427-2428 |
Biochimica et Biophysica Acta,2005年,Vol. 1721,pp. 89-97 |
FISHERIES SCIENCE,2002年,Vol. 68,pp. 921-928 |
Int. J. Pept. Res. Ther.,2010年,Vol. 16,pp. 111-121 |
Also Published As
Publication number | Publication date |
---|---|
JP2019073465A (ja) | 2019-05-16 |
JP7074995B2 (ja) | 2022-05-25 |
JP2022025155A (ja) | 2022-02-09 |
US20190111103A1 (en) | 2019-04-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP4947475B2 (ja) | 鳥軟骨の水解物並びにその調製方法及び使用 | |
US9458197B2 (en) | Elastin digest compositions and methods utilizing same | |
CN108472336A (zh) | C型利尿钠肽变体在治疗骨骼发育不良中的用途 | |
CN104619352A (zh) | 含有hc-ha/ptx3复合物的组合物及其使用方法 | |
KR20090120451A (ko) | 천연(텔로펩티드) 태반 콜라겐 조성물 | |
CN108697769A (zh) | 用于治疗肥胖症的胰高血糖素和glp-1共激动剂 | |
US20170260511A1 (en) | Method for preparing a three-dimensionally cultured skin comprising dermis and epidermis, and the cultured skin made therefrom | |
KR20190101990A (ko) | 펩티드 wkdeagkplvk를 포함하는 조성물 | |
JP2010155820A (ja) | 皮膚改善剤、血管改善剤、並びにこれらを含む医薬組成物、食品、飼料及び化粧品 | |
US20200147142A1 (en) | Compositions and methods for enhancing the therapeutic potential of stem cells | |
JP7343864B2 (ja) | 線維芽細胞又は軟骨細胞におけるエラスチン産生促進剤 | |
CN109069546A (zh) | 可食用燕窝提取物和提取方法 | |
JP2010202578A (ja) | 皮膚改善剤、血管機能改善剤、並びにこれらを含む医薬組成物、食品、飼料及び化粧品 | |
CN106999535A (zh) | 用于治疗疾病和病症的NFκB活性的抑制剂 | |
CN102247589A (zh) | 海洋胶原肽在制备延缓皮肤衰老的药物、食品中的用途 | |
US10813954B2 (en) | Pharmaceutical composition for treating diabetes, comprising pancreatic islet cells and elastin-like artificial extracellular matrix | |
CN103509102B (zh) | 野生型人生长激素突变体 | |
JP2019206495A (ja) | ウマプラセンタエキスを有効成分とする剤 | |
US11186612B2 (en) | Substance P analog having progenitor cell or stem cell recruiting activity and method for progenitor cell or stem cell recruiting using the same | |
CN104511008A (zh) | 一种脑蛋白水解物制备方法 | |
JP2005281186A (ja) | 創傷治癒剤、腱または靭帯の損傷等に対する改善、治癒促進または予防剤、機能性食品ならびに医薬品 | |
CN105734102B (zh) | 胶原蛋白肽的制备方法 | |
CN115721638B (zh) | 月桂酰精氨酸乙酯盐酸盐在制备促伤口愈合药物中的应用 | |
CN109758449A (zh) | 杨梅素在制备用于治疗肺纤维化疾病的药物中的应用 | |
CN110179778B (zh) | 固绿在制备α-突触核蛋白聚集抑制剂中的用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20211204 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20211204 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20211209 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20221220 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230215 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230216 |
|
RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20230216 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230418 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20230725 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20230823 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7343864 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |