JP6411403B2 - Protein saccharification reaction inhibitor - Google Patents
Protein saccharification reaction inhibitor Download PDFInfo
- Publication number
- JP6411403B2 JP6411403B2 JP2016105996A JP2016105996A JP6411403B2 JP 6411403 B2 JP6411403 B2 JP 6411403B2 JP 2016105996 A JP2016105996 A JP 2016105996A JP 2016105996 A JP2016105996 A JP 2016105996A JP 6411403 B2 JP6411403 B2 JP 6411403B2
- Authority
- JP
- Japan
- Prior art keywords
- ages
- saccharification reaction
- reaction
- extract
- oph
- 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
- 102000004169 proteins and genes Human genes 0.000 title description 27
- 108090000623 proteins and genes Proteins 0.000 title description 27
- 239000002683 reaction inhibitor Substances 0.000 title description 11
- 102100032488 Acylamino-acid-releasing enzyme Human genes 0.000 claims description 31
- 230000000694 effects Effects 0.000 claims description 30
- 101710168439 Acylamino-acid-releasing enzyme Proteins 0.000 claims description 27
- 230000009471 action Effects 0.000 claims description 17
- 238000004132 cross linking Methods 0.000 claims description 14
- 239000003623 enhancer Substances 0.000 claims description 12
- 239000004480 active ingredient Substances 0.000 claims description 11
- 239000002537 cosmetic Substances 0.000 claims description 9
- 239000003814 drug Substances 0.000 claims description 7
- 235000013305 food Nutrition 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 239000003431 cross linking reagent Substances 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 6
- 235000013373 food additive Nutrition 0.000 claims description 6
- 239000002778 food additive Substances 0.000 claims description 6
- 230000000593 degrading effect Effects 0.000 claims description 5
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 4
- 229940127557 pharmaceutical product Drugs 0.000 claims description 4
- 108091005996 glycated proteins Proteins 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 description 52
- 238000005259 measurement Methods 0.000 description 23
- AIJULSRZWUXGPQ-UHFFFAOYSA-N Methylglyoxal Chemical compound CC(=O)C=O AIJULSRZWUXGPQ-UHFFFAOYSA-N 0.000 description 18
- LEQAOMBKQFMDFZ-UHFFFAOYSA-N glyoxal Chemical compound O=CC=O LEQAOMBKQFMDFZ-UHFFFAOYSA-N 0.000 description 16
- 239000000243 solution Substances 0.000 description 14
- 230000002401 inhibitory effect Effects 0.000 description 13
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 12
- 238000004519 manufacturing process Methods 0.000 description 12
- 239000000843 powder Substances 0.000 description 11
- 238000000034 method Methods 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- ZGCHLOWZNKRZSN-NTSWFWBYSA-N 3-deoxyglucosone Chemical compound OC[C@@H](O)[C@@H](O)CC(=O)C=O ZGCHLOWZNKRZSN-NTSWFWBYSA-N 0.000 description 8
- UHPMJDGOAZMIID-UHFFFAOYSA-N 3-deoxyglucosone Natural products OCC1OC(O)C(=O)CC1O UHPMJDGOAZMIID-UHFFFAOYSA-N 0.000 description 8
- 229940015043 glyoxal Drugs 0.000 description 8
- 239000012488 sample solution Substances 0.000 description 7
- 239000005711 Benzoic acid Substances 0.000 description 6
- 235000010233 benzoic acid Nutrition 0.000 description 6
- 210000000988 bone and bone Anatomy 0.000 description 6
- AYEKKSTZQYEZPU-RYUDHWBXSA-N pentosidine Chemical compound OC(=O)[C@@H](N)CCCCN1C=CC=C2N=C(NCCC[C@H](N)C(O)=O)N=C12 AYEKKSTZQYEZPU-RYUDHWBXSA-N 0.000 description 6
- 210000003491 skin Anatomy 0.000 description 6
- 238000001035 drying Methods 0.000 description 5
- 230000002708 enhancing effect Effects 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- WMBWREPUVVBILR-WIYYLYMNSA-N (-)-Epigallocatechin-3-o-gallate Chemical compound O([C@@H]1CC2=C(O)C=C(C=C2O[C@@H]1C=1C=C(O)C(O)=C(O)C=1)O)C(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-WIYYLYMNSA-N 0.000 description 4
- 108010061216 Acylaminoacyl-peptidase Proteins 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- WMBWREPUVVBILR-UHFFFAOYSA-N GCG Natural products C=1C(O)=C(O)C(O)=CC=1C1OC2=CC(O)=CC(O)=C2CC1OC(=O)C1=CC(O)=C(O)C(O)=C1 WMBWREPUVVBILR-UHFFFAOYSA-N 0.000 description 4
- 102000008100 Human Serum Albumin Human genes 0.000 description 4
- 108091006905 Human Serum Albumin Proteins 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 229940030275 epigallocatechin gallate Drugs 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- BVQVLAIMHVDZEL-UHFFFAOYSA-N 1-phenyl-1,2-propanedione Chemical compound CC(=O)C(=O)C1=CC=CC=C1 BVQVLAIMHVDZEL-UHFFFAOYSA-N 0.000 description 3
- 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 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 230000036252 glycation Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000004007 reversed phase HPLC Methods 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- HAMNKKUPIHEESI-UHFFFAOYSA-N aminoguanidine Chemical compound NNC(N)=N HAMNKKUPIHEESI-UHFFFAOYSA-N 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- BJRNKVDFDLYUGJ-RMPHRYRLSA-N hydroquinone O-beta-D-glucopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-RMPHRYRLSA-N 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N squalane Chemical compound CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- ABYZSYDGJGVCHS-ZETCQYMHSA-N (2s)-2-acetamido-n-(4-nitrophenyl)propanamide Chemical compound CC(=O)N[C@@H](C)C(=O)NC1=CC=C([N+]([O-])=O)C=C1 ABYZSYDGJGVCHS-ZETCQYMHSA-N 0.000 description 1
- 229940043375 1,5-pentanediol Drugs 0.000 description 1
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 1
- FLPJVCMIKUWSDR-UHFFFAOYSA-N 2-(4-formylphenoxy)acetamide Chemical compound NC(=O)COC1=CC=C(C=O)C=C1 FLPJVCMIKUWSDR-UHFFFAOYSA-N 0.000 description 1
- QCDWFXQBSFUVSP-UHFFFAOYSA-N 2-phenoxyethanol Chemical compound OCCOC1=CC=CC=C1 QCDWFXQBSFUVSP-UHFFFAOYSA-N 0.000 description 1
- QGZCUOLOTMJILH-UHFFFAOYSA-N 2h-tetrazol-2-ium;bromide Chemical compound [Br-].C1=N[NH+]=NN1 QGZCUOLOTMJILH-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 102000013455 Amyloid beta-Peptides Human genes 0.000 description 1
- 108010090849 Amyloid beta-Peptides Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- 241001424413 Lucia Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 235000019774 Rice Bran oil Nutrition 0.000 description 1
- 108010022999 Serine Proteases Proteins 0.000 description 1
- 102000012479 Serine Proteases Human genes 0.000 description 1
- 229920002125 Sokalan® Polymers 0.000 description 1
- 244000273928 Zingiber officinale Species 0.000 description 1
- 235000006886 Zingiber officinale Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000000996 additive effect Effects 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
- 229960000271 arbutin Drugs 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- QQFYLZXBFWWJHR-UHFFFAOYSA-M benzyl(triethyl)phosphanium;bromide Chemical compound [Br-].CC[P+](CC)(CC)CC1=CC=CC=C1 QQFYLZXBFWWJHR-UHFFFAOYSA-M 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000001058 brown pigment Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229960001631 carbomer Drugs 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 229940074979 cetyl palmitate Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000007806 chemical reaction intermediate Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 201000009101 diabetic angiopathy Diseases 0.000 description 1
- VJZWIFWPGRIJSN-XRHABHTOSA-N dilinoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O.CCCCC\C=C/C\C=C/CCCCCCCC(O)=O VJZWIFWPGRIJSN-XRHABHTOSA-N 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 229940008099 dimethicone Drugs 0.000 description 1
- 239000004205 dimethyl polysiloxane Substances 0.000 description 1
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000008397 ginger Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 229960005150 glycerol Drugs 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 229940075529 glyceryl stearate Drugs 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- PXDJXZJSCPSGGI-UHFFFAOYSA-N hexadecanoic acid hexadecyl ester Natural products CCCCCCCCCCCCCCCCOC(=O)CCCCCCCCCCCCCCC PXDJXZJSCPSGGI-UHFFFAOYSA-N 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- JXTPJDDICSTXJX-UHFFFAOYSA-N n-Triacontane Natural products CCCCCCCCCCCCCCCCCCCCCCCCCCCCCC JXTPJDDICSTXJX-UHFFFAOYSA-N 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000037311 normal skin Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- BJRNKVDFDLYUGJ-UHFFFAOYSA-N p-hydroxyphenyl beta-D-alloside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-UHFFFAOYSA-N 0.000 description 1
- WCVRQHFDJLLWFE-UHFFFAOYSA-N pentane-1,2-diol Chemical compound CCCC(O)CO WCVRQHFDJLLWFE-UHFFFAOYSA-N 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229960005323 phenoxyethanol Drugs 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 239000001818 polyoxyethylene sorbitan monostearate Substances 0.000 description 1
- 235000010989 polyoxyethylene sorbitan monostearate Nutrition 0.000 description 1
- 229940113124 polysorbate 60 Drugs 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 239000008165 rice bran oil Substances 0.000 description 1
- 238000011076 safety test Methods 0.000 description 1
- 229940032094 squalane Drugs 0.000 description 1
- 210000000434 stratum corneum Anatomy 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001256 tonic effect Effects 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
Images
Landscapes
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Description
本発明は、黒ガリンガル(Kaempheria parviflora)抽出物を含有する蛋白質糖化反応阻害剤などに関する。 The present invention relates to a protein saccharification reaction inhibitor containing black galingal (Kaempheria parviflora) extract and the like.
蛋白質糖化反応(以下、糖化反応と省略)は、L.C.Maillardがアミノ酸と還元糖を加熱すると褐色の色素が生成することを発見したことからメイラード反応として知られるようになった。近年、この糖化反応が老化現象、認知症、癌、高血圧、動脈硬化症などにも関与していることが明らかになっている。例えば、糖化反応により蛋白質は褐変化するが、これにより、肌などにくすみが生じることになる。また、糖化反応により皮膚や骨のコラーゲンが硬化することにより、皮膚や骨の弾力及びしなやかさが損なわれてしまう。そこで、生体に様々な影響を及ぼす糖化反応を阻害するための研究が種々行われている。 Protein saccharification reaction (hereinafter abbreviated as saccharification reaction) C. It became known as Maillard reaction because Maillard discovered that heating amino acids and reducing sugars produced brown pigments. In recent years, it has been revealed that this saccharification reaction is also involved in aging, dementia, cancer, hypertension, arteriosclerosis and the like. For example, the protein turns brown due to the saccharification reaction, which causes dullness on the skin and the like. Further, the skin and bone collagen is hardened by the saccharification reaction, so that the elasticity and flexibility of the skin and bone are impaired. Thus, various studies have been conducted to inhibit saccharification reactions that have various effects on living bodies.
例えば、特定の植物の抽出物が蛋白質糖化反応を抑制することについて有効であることが報告されている(特許文献1)。 For example, it has been reported that an extract of a specific plant is effective for suppressing a protein saccharification reaction (Patent Document 1).
上記事情を鑑み、蛋白質の糖化反応を阻害し得る天然素材を見出し、その抽出物を有効成分とする蛋白質糖化反応素材剤などを提供することを課題とする。 In view of the above circumstances, an object of the present invention is to find a natural material capable of inhibiting a glycation reaction of a protein and provide a protein saccharification reaction material agent or the like containing the extract as an active ingredient.
上記課題を解決するための手段として、以下の発明などを提供する。すなわち、黒ガリンガル(Kaempheria parviflora)抽出物を有効成分として含有する蛋白質糖化反応素材剤を提供する。また、上記の蛋白質糖化反応素材剤を含有する食品、食品添加物、医薬品、医薬部外品及び化粧品を提供する。 As means for solving the above problems, the following inventions and the like are provided. That is, the protein saccharification reaction raw material agent which contains a black galingal (Kaempheria parviflora) extract as an active ingredient is provided. The present invention also provides foods, food additives, pharmaceuticals, quasi drugs and cosmetics containing the protein saccharification reaction material.
また下記の発明を提供する。 The following invention is also provided.
項1.黒ガリンガル(Kaempheria parviflora)抽出物を有効成分として含有するAGEs架橋切断剤。
項2.医薬品又は医薬部外品である項1に記載のAGEs架橋切断剤。
項3.食品又は食品添加物である項1に記載のAGEs架橋切断剤。
項4.化粧品である項1に記載のAGEs架橋切断剤。Item 4.
項5.黒ガリンガル(Kaempheria parviflora)抽出物を有効成分として含有するOPH活性増強剤。
項6.糖化蛋白質分解作用又はAGEs分解作用を有する項5に記載のOPH活性増強剤。
項7.医薬品又は医薬部外品である項5又は6に記載のOPH活性増強剤。Item 7. Item 7. The OPH activity enhancer according to
項8.食品又は食品添加物である項5又は6に記載のOPH活性増強剤。
項9.化粧品である項5又は6に記載のOPH活性増強剤。
本発明により、生体に好ましくない影響を及ぼす蛋白質の糖化反応を阻害する蛋白質糖化反応阻害剤などを提供することができる。 According to the present invention, it is possible to provide a protein saccharification reaction inhibitor that inhibits a saccharification reaction of a protein that has an undesirable effect on a living body.
以下、本発明の実施の形態について説明する。なお、本発明は、これらの実施形態に何ら限定されるべきものではなく、その要旨を逸脱しない範囲において、種々なる態様で実施し得る。
<実施形態1>
Embodiments of the present invention will be described below. In addition, this invention should not be limited to these embodiments at all, and can be implemented in various modes without departing from the gist thereof.
<
本実施形態の蛋白質糖化反応阻害剤は、黒ガリンガル(Kaempheria parviflora)抽出物を有効成分として含有するものである。 The protein saccharification reaction inhibitor of this embodiment contains a black galingal (Kaempheria parviflora) extract as an active ingredient.
「黒ガリンガル(Kaempheria parviflora)」は、ショウガ科ケンペリア属の植物である。また、「クラチャイ・ダム((Krachai dam)」や「黒ショウガ(Black Ginger)」などとも称される。タイやラオスなど東南アジアでは古来より塊茎を薬用としてきた。また、近年では、乾燥及び粉末化し滋養強壮などの健康用途に日本でも使用されている。 “Kaempheria parviflora” is a plant belonging to the genus Kempelia in the family Ginger. Also known as “Krachai dam”, “Black Ginger”, etc. In Southeast Asia such as Thailand and Laos, tubers have been used for medicinal purposes since ancient times. It is also used in Japan for health uses such as nutrition and tonic.
黒ガリンガル抽出物は、主に塊茎を原料とし精製水、エタノール、メタノール、プロピルアルコール、イソプロピルアルコール、ブタノール、プロピレングリコールなどの溶媒を用いて抽出する。抽出は、塊茎を破砕したものを溶媒に浸漬してもよいし、破砕し粉体としてから浸漬してもよい。また、塊茎だけでなく葉や茎を用いてもよい。また、液体として抽出物を得た後にさらに乾燥等を施し粉体や粒体として得てもよい。 Black galling extract is mainly extracted from tubers using a solvent such as purified water, ethanol, methanol, propyl alcohol, isopropyl alcohol, butanol, and propylene glycol. Extraction may be carried out by immersing the crushed tuber in a solvent, or by pulverizing it into a powder. Further, not only tubers but also leaves and stems may be used. Moreover, after obtaining the extract as a liquid, it may be further dried to obtain powder or granules.
黒ガリンガルの抽出物を有効成分とする本実施形態の蛋白質糖化反応阻害剤は、さらに既知の方法を用いることにより、当該蛋白質糖化反応阻害剤を含有する食品、食品添加物、医薬品、医薬部外品、化粧品などとして提供することが可能である。 The protein saccharification reaction inhibitor of the present embodiment containing black gallingual extract as an active ingredient can be further used in a food, food additive, pharmaceutical, quasi-drug containing the protein saccharification reaction inhibitor by using a known method. Products, cosmetics, etc.
例えば、医薬品とする場合には、本実施形態の蛋白質糖化反応阻害剤を粉体や粒体としカプセルに充填したり、あるいは、賦形剤、結合剤、崩壊剤などを添加して打錠機等を用いて錠剤とすることができる。また、食品とする場合には、黒ガリンガルを適宜乾燥や破砕等を施してから湯で煮出すことで提供できる。また、医薬品のようにカプセルや錠剤のような形態で提供してもよいし、他の飲料、調味料、菓子等の各種の食品に蛋白質糖化反応阻害剤を添加した態様で提供することもできる。 For example, in the case of a pharmaceutical product, the protein saccharification reaction inhibitor of the present embodiment is powdered or granulated and filled into a capsule, or an excipient, a binder, a disintegrant and the like are added to the tablet press. Etc. can be used as tablets. Moreover, when it is set as a foodstuff, it can provide by simmering in black hot water after giving a black galling gal suitably drying or crushing. Moreover, it may be provided in the form of a capsule or a tablet like a pharmaceutical, or can be provided in a form in which a protein saccharification reaction inhibitor is added to various foods such as other beverages, seasonings, and confectionery. .
また、美容液、クリーム、ローションなどの化粧品とすることもできる。例えば、美容液とする場合には、本実施形態の蛋白質糖化反応阻害剤の他、水、コメヌカ油、ペンチレングリコール、グリセリン、スクワラン、パルミチン酸セチル、ダイマージリノール酸などを主成分とし、ヒアルロン酸Na、水添ナタネ油アルコール、カルボマー、キサンタンガム、水酸化K、ジメチコン、ポリソルベート−60、ステアリン酸グリセリル、水添ヒマシ油、フェノキシエタノール、尿素、アルギニン、アルブチン、クエン酸などを添加剤とする。そして、各成分を水溶性原料・油溶性原料に分けて溶解してから、それらを加熱して混合・乳化する。これを冷却しながらエキスなどの添加物を配合し、さらに低温になったところで精油や香料などの揮発性の高いものを添加する。その後、所定の安全性の検査(菌、pH、温度安定性、粘度等)を行い、瓶などに充填して製品として提供することができる。上述した種々の応用は、実施形態2のAGEsが関与する架橋切断剤、実施形態3の酸化蛋白質分解酵素活性の増強剤についても同様に適用できる。
<試験>
Moreover, it can also be set as cosmetics, such as a cosmetic liquid, cream, and lotion. For example, in the case of a cosmetic liquid, in addition to the protein saccharification reaction inhibitor of the present embodiment, water, rice bran oil, pentylene glycol, glycerin, squalane, cetyl palmitate, dimer dilinoleic acid and the like as main components, hyaluron Acid Na, hydrogenated rapeseed oil alcohol, carbomer, xanthan gum, hydroxylated K, dimethicone, polysorbate-60, glyceryl stearate, hydrogenated castor oil, phenoxyethanol, urea, arginine, arbutin, citric acid and the like are used as additives. Then, after each component is dissolved in a water-soluble raw material and an oil-soluble raw material, they are heated and mixed and emulsified. While cooling this, an additive such as an extract is blended, and a highly volatile substance such as essential oil or fragrance is added when the temperature becomes lower. Thereafter, a predetermined safety test (bacteria, pH, temperature stability, viscosity, etc.) is performed, and the product can be provided as a product after filling into a bottle or the like. The various applications described above can be similarly applied to the cross-linking cleaving agent in which the AGEs of
<Test>
≪1≫黒ガリンガル抽出液の調製
真空ポンプを用いずに40℃未満の真空状態にて乾燥(低温真空乾燥)させ粉末化して得た黒ガリンガル乾燥粉末2gと、高温状態で乾燥(高温乾燥)させ粉末化して得た黒ガリンガル乾燥粉末2gのそれぞれを、80mLの熱水で1時間抽出し、常温まで冷却後、濾紙を用いて濾過後、試料溶液とした。以降の測定には本抽出液を使用した。各試料溶液の固形分濃度は、各試料溶液5mLをアルミトレイに入れ、110℃に設定したインキュベーター内で4時間乾燥させた後の残重量を測定して算出した。低温真空乾燥による固形分濃度(mg/ mL)は7.87±0.18であり、高温乾燥による固形分濃度は7.63±0.15であり、両サンプルともほぼ同様であった。
≪1≫ Preparation of black gallingual extract 2 g of black gallingual dry powder obtained by drying (low temperature vacuum drying) in a vacuum state below 40 ° C without using a vacuum pump, and drying in a high temperature state (high temperature drying) Each 2 g of dry black gallingal powder obtained by pulverization was extracted with 80 mL of hot water for 1 hour, cooled to room temperature, filtered using filter paper, and used as a sample solution. This extract was used for subsequent measurements. The solid content concentration of each sample solution was calculated by measuring the remaining weight after 5 mL of each sample solution was placed in an aluminum tray and dried in an incubator set at 110 ° C. for 4 hours. The solid concentration (mg / mL) by low-temperature vacuum drying was 7.87 ± 0.18, and the solid concentration by high-temperature drying was 7.63 ± 0.15, which was almost the same for both samples.
≪2≫測定概要
AGEs(糖化最終生成物)は糖化反応における最終生成物の総称であり、その特徴の一つとして蛍光性を有する。蛍光性を有するAGEsにはペントシジン、クロスリン、ピロピリジンなどがある。
≪2≫ Measurement overview
AGEs (final product of saccharification) are a general term for final products in saccharification reactions, and have fluorescence as one of their characteristics. Examples of fluorescent AGEs include pentosidine, croslin, and pyropyridine.
糖化反応抑制作用はヒト血清アルブミン(HSA)-グルコース糖化反応系に試験サンプルを添加し、試験サンプルによる蛍光性AGEs、3-deoxyglucosone(3DG)、グリオキサール(GO)、メチルグリオキサール(MGO)、ペントシジン、CMLの生成阻害率を測定した。蛍光性AGEsは反応液には370nmの励起波長を照射したときの蛍光を440nmで測定した。3DG、GO、MGO、ペントシジンの測定にはHPLC法を使用した。CMLの測定にはELISA法を使用した。 Saccharification reaction inhibitory action is achieved by adding test samples to human serum albumin (HSA) -glucose saccharification reaction system, and fluorescent AGEs, 3-deoxyglucosone (3DG), glyoxal (GO), methylglyoxal (MGO), pentosidine, The inhibition rate of CML production was measured. Fluorescence AGEs were measured at 440 nm when the reaction solution was irradiated with an excitation wavelength of 370 nm. The HPLC method was used for the measurement of 3DG, GO, MGO, and pentosidine. The ELISA method was used for CML measurement.
糖化反応抑制作用のポジティブコントロールとしては糖化反応阻害剤の一種であるアミノグアニジンまたはエピガロカテキンガレート(EGCg)を使用した。 As a positive control of the saccharification reaction inhibitory effect, aminoguanidine or epigallocatechin gallate (EGCg), which is a kind of saccharification reaction inhibitor, was used.
≪3≫方法
in vitro 糖化反応
0.1 mol/L NaH2PO4-Na2HPO4リン酸緩衝液(pH7.4)、8 mg/mLヒト血清アルブミン(HSA)、0.2moL/Lグルコース糖化反応液中に、調製した各濃度のサンプルを1/10濃度になるように添加し、60℃で40時間インキュベートした。コントロールとしてはサンプルの代わりに蒸留水を添加したものを用いた。
≪3≫ Method
In vitro saccharification reaction
0.1 mol / L NaH2PO4-Na2HPO4 phosphate buffer solution (pH 7.4), 8 mg / mL human serum albumin (HSA), 0.2 mol / L glucose
蛍光性AGEs測定
糖化反応終了後、反応液中に生成した蛍光性AGEsをマイクロプレートリーダーで測定した(励起波長370nm/蛍光波長440nm)。
Measurement of fluorescent AGEs After completion of the saccharification reaction, fluorescent AGEs generated in the reaction solution were measured with a microplate reader (excitation wavelength 370 nm / fluorescence wavelength 440 nm).
3DG、GO、MGO測定
糖化反応終了後、反応液中に生成した3DG、GO、MGOを2.3-diaminonaphthalen(DAN)プレラベル化逆相HPLC法により定量した。
Measurement of 3DG, GO, MGO After completion of the saccharification reaction, 3DG, GO, MGO produced in the reaction solution was quantified by 2.3-diaminonaphthalen (DAN) prelabeled reverse phase HPLC method.
ペントシジン測定HPLC法の場合はScheijenaら(2009) の方法を参考に、反応液を塩酸加水分解後、逆相HPLCで測定した。(参考文献1「Scheijena et al, Journal of Chromatography B, 2009 ; 877 : 610-614」)
In the case of the HPLC method for measuring pentosidine, the reaction solution was hydrolyzed with hydrochloric acid and measured by reverse-phase HPLC with reference to the method of Scheijena et al. (2009). (
CML測定
反応液中に生成したCMLはELISA法(CircuLex CML/Nε-(carboxymethyl)lysine), サイクレックス)で測定した。
CML measurement CML generated in the reaction solution was measured by ELISA (CircuLex CML / Nε- (carboxymethyl) lysine), Cyclex).
AGEs生成阻害作用の算出
AGEsの生成阻害率(%)は、in vitro糖化反応系においてサンプルを添加した反応液(A)、グルコース水溶液の代わりに蒸留水を添加したもの(B)、サンプルを添加しない溶液のみを添加してインキュベーションしたもの(C)、ブランクとしてグルコースの代わりに蒸留水を添加したもの(D)として下記の式に従って算出した。
AGEs生成阻害率(%) = {1-(A -B)/(C -D)} ×100
抗AGEs活性はIC50(50%生成阻害濃度)を算出し、有効数字3桁で表示した。
IC50は値が小さいほど活性が強いことを示している。
Calculation of AGEs production inhibitory effect
The AGEs production inhibition rate (%) was determined by adding only the reaction solution (A) to which the sample was added in the in vitro saccharification reaction system, the addition of distilled water instead of the aqueous glucose solution (B), and the solution to which no sample was added. Incubation (C) and blank (D) added with distilled water instead of glucose were calculated according to the following formula.
AGE generation inhibition rate (%) = {1- (A -B) / (C -D)} × 100
The anti-AGE activity was calculated as IC50 (50% production inhibitory concentration) and displayed as three significant figures.
IC50 indicates that the smaller the value, the stronger the activity.
≪4≫結果
図1及び図2は、測定結果を表として示すものである。また、図3及び図4は、測定結果をグラフで示すものである。MGO及びペントシジンについては拡大したグラフも併せて示す。図示するように、低温真空乾燥粉末と高温乾燥粉末の各熱水抽出液にはAGEs生成抑制作用がみられた。その作用は蛍光性AGEs、3DG、GO、ペントシジンにおいて低温真空乾燥粉末が強かった。MGOの生成阻害作用は両サンプルともにアミノグアニジンと同等以上に強かった。CMLの生成阻害作用は高温乾燥粉末が約2倍強かった。
<< 4 >> Results FIG. 1 and FIG. 2 show the measurement results as a table. Moreover, FIG.3 and FIG.4 shows a measurement result with a graph. An enlarged graph is also shown for MGO and pentosidine. As shown in the figure, the AGEs formation inhibitory action was observed in each hot water extract of the low temperature vacuum dry powder and the high temperature dry powder. The action was strong in low-temperature vacuum-dried powders in fluorescent AGEs, 3DG, GO, and pentosidine. The inhibition of MGO production was as strong as or better than aminoguanidine in both samples. The high temperature dry powder was about twice as strong as CML formation inhibitory action.
図5は、糖化反応によるAGEsの生成過程の概略と、この生成過程において黒ガリンガルのAGEs生成阻害能の推定作用ポイントを示している。「×」を付した過程が推定作用ポイントである。また、○で囲んだ物質が本実施形態で生成阻害作用を評価した物質である。上述した測定結果から黒ガリンガルは生体内糖化反応の多経路を抑制する可能性があった。
<効果>
FIG. 5 shows an outline of the AGEs production process by saccharification reaction and the estimated action points of black gallingual AGEs production inhibition ability in this production process. The process marked with “x” is the estimated action point. Further, a substance surrounded by a circle is a substance whose production inhibitory action is evaluated in this embodiment. From the above-described measurement results, black galling gal may suppress multipaths in the glycation reaction in vivo.
<Effect>
本実施形態により、黒ガリンガル抽出物を有効成分とする蛋白質糖化反応阻害剤などを提供することができる。
<実施形態2>
According to the present embodiment, it is possible to provide a protein saccharification reaction inhibitor containing black gallingual extract as an active ingredient.
<
黒ガリンガル抽出物にAGEsが関与する架橋構造を分解(切断)する作用があるかについて検証する。実施形態1で示したように蛋白質は糖化反応により様々な経路でAGEsへ至る。
It is verified whether black gallingual extract has an action of decomposing (cutting) a crosslinked structure involving AGEs. As shown in
ここで、AGEsの生成に至る中間体である糖化反応中間体として例示したグリオキサール(GO)、メチルグリオキサール(MG)、3−デオキシグルコソン(3DG)は、いずれも分子内に2つのカルボニル基(C=0)を有するα−ジカルボニル化合物である。 Here, glyoxal (GO), methylglyoxal (MG), and 3-deoxyglucosone (3DG) exemplified as saccharification reaction intermediates, which are intermediates leading to the generation of AGEs, all have two carbonyl groups ( An α-dicarbonyl compound having C = 0).
このα−ジカルボニル化合物は反応性に富んでおり蛋白質間に架橋を形成し、例えば生体内のコラーゲン分子間で架橋を形成する。正常な状態の皮膚や骨は、酵素の作用を介して遺伝的に規定された部位に秩序立って形成される架橋(生理的架橋)により皮膚や骨の適正な柔軟性や強度を維持しているが、α−ジカルボニル化合物による無秩序で余分に形成される架橋は皮膚の硬化や骨の脆弱化をもたらす。 This α-dicarbonyl compound is highly reactive and forms a cross-link between proteins, for example, a cross-link between collagen molecules in vivo. Normal skin and bone maintain proper flexibility and strength of the skin and bone by the cross-links (physiological cross-links) that are orderedly formed at sites genetically defined through the action of enzymes. However, disorderly and excessively formed cross-linking by the α-dicarbonyl compound leads to skin hardening and bone weakening.
このようなα−ジカルボニル化合物を分解することは、AGEsの生成及び蓄積を抑制するとともに、糖化反応により形成された無秩序で余分な架橋を分解し、皮膚の硬化や骨の脆弱化などを回復させるために有効であると考えられる。 Degrading such an α-dicarbonyl compound suppresses the formation and accumulation of AGEs and also breaks up the disorderly and excessive cross-links formed by the saccharification reaction to restore skin hardening and bone weakness. It is considered effective for
このような架橋を分解する化合物としてN-フェナシルチアゾリウムブロミド(N-phenacylthiazolium bromide: PTB)が報告されている。PTBはαジケトン構造のC-C結合を切断分解することで血管内のAGEsの蓄積を抑制し、糖尿病性血管合併症の治療に寄与する可能性が示唆されている。このため本作用は糖化ストレスの治療的なアプローチとして注目されている。
<試験>
Such N crosslinking as a decomposing compound - phenacylthiazolium Lucia tetrazolium bromide (N-phenacylthiazolium bromide: PTB) have been reported. It has been suggested that PTB may contribute to the treatment of diabetic vascular complications by inhibiting the accumulation of AGEs in the blood vessel by cleaving the C - C bond of the α-diketone structure. For this reason, this action attracts attention as a therapeutic approach for glycation stress.
<Test>
≪1≫試験概要
以下の試験ではαジケトン構造を有する1-フェニル-1,2-プロパンジオン(1-phenyl-1,2-propanedione: PPD)をモデル基質とした反応系を使用して、AGEs架橋切断作用を評価する。ポジティブコントロールとしてはPTBを使用した。
≪1≫ Test Outline In the following test, AGEs were prepared using a reaction system using 1-phenyl-1,2-propanedione (PPD) having an α-diketone structure as a model substrate. Evaluate the cross-link cutting action. PTB was used as a positive control.
≪2≫方法
試験サンプルには各熱水抽出液(サンプル溶液)を使用した。架橋切断作用のポジティブコントロールとしてはPTBを使用した。
<< 2 >> Method Each hot water extract (sample solution) was used as a test sample. PTB was used as a positive control for the cross-linking cleavage action.
AGEs架橋切断作用の測定にはサンプル溶液または10mmol/L PTB、10 mmol/L PPD、0.2mol/Lリン酸緩衝液(pH7.4)を5:1:4の割合で混合し、37℃で8時間反応させた(n=3)。反応終了後、塩酸を加えて反応停止させた。
反応液は20℃、3,000×gで10分間遠心分離し、上清中の安息香酸量を逆相HPLCで分析した。反応液中の安息香酸量は、別途測定したサンプル中の安息香酸量を差し引いて求めた。1molのPPDは1molの安息香酸を生成することから、以下の式で架橋切断率を算出した。架橋切断の相対値はPTBの架橋切断率を100としたときの値を求めた。
架橋切断率(%)= {(A -B)/C} ×100
A:反応液中の安息香酸量
B:サンプル中の安息香酸量
C:反応に供したPPD量(基質量)
To measure the AGEs cross-linking activity, mix sample solution or 10mmol / L PTB, 10mmol / L PPD, 0.2mol / L phosphate buffer (pH7.4) at a ratio of 5: 1: 4 at 37 ℃. The reaction was performed for 8 hours (n = 3). After completion of the reaction, hydrochloric acid was added to stop the reaction.
The reaction solution was centrifuged at 20 ° C. and 3,000 × g for 10 minutes, and the amount of benzoic acid in the supernatant was analyzed by reverse phase HPLC. The amount of benzoic acid in the reaction solution was determined by subtracting the amount of benzoic acid in the sample measured separately. Since 1 mol of PPD produces 1 mol of benzoic acid, the crosslinking cleavage rate was calculated by the following formula. The relative value of cross-linking breakage was obtained when the cross-linking breakage rate of PTB was taken as 100.
Crosslink cutting rate (%) = {(A -B) / C} × 100
A: Amount of benzoic acid in the reaction solution
B: Benzoic acid content in the sample
C: Amount of PPD used for reaction (base mass)
≪3≫結果
図6は、測定結果を表として示すものである。また、図7は、測定結果をグラフとして示すものである。低温真空乾燥粉末と高温乾燥粉末の各熱水抽出液にはAGEs架橋切断作用がみられた。その作用は低温真空乾燥粉末が強かった。
<効果>
<< 3 >> Results FIG. 6 shows the measurement results as a table. Moreover, FIG. 7 shows a measurement result as a graph. AGEs cross-linking action was observed in each hot water extract of low temperature vacuum dry powder and high temperature dry powder. The action was strong in the low temperature vacuum dried powder.
<Effect>
本実施形態により、黒ガリンガル抽出物を有効成分とするAGEs架橋切断剤などを提供することができる。
<実施形態3>
According to the present embodiment, it is possible to provide an AGEs cross-linking cleaving agent containing black gallingual extract as an active ingredient.
<
黒ガリンガル抽出物に酸化蛋白質分解酵素の活性を増強する作用があるかについて検証する。酸化蛋白質分解酵素(oxidized protein hydrolase: OPH)は、蛋白質のN末端アシル化アミノ酸を遊離するセリンプロテアーゼの一種で、アシルアミノ酸遊離酵素(acylamino-acid releasing enzyme: AARE) 、アシル化ペプチド分解酵素(acylpeptide hydrolase: APH)などとも言われている。OPHはブタ肝臓、ラット脳、ヒト血液、角層などの生体組織に広く存在している。OPHは酸化蛋白質や糖化蛋白質を優先的に分解するとともにプロテアソームと協働して老化した蛋白質を分解すること、アルツハイマー病の原因であるアミロイドβを減少させることが報告されている。またOPHがAGEsを分解することも確認されている。
<試験>
We examine whether black gallingual extract has the effect of enhancing the activity of oxidized protease. Oxidized protein hydrolase (OPH) is a kind of serine protease that releases N-terminal acylated amino acids of proteins.Acylamino-acid releasing enzyme (AARE) hydrolase: APH). OPH is widely present in living tissues such as pig liver, rat brain, human blood, and stratum corneum. OPH has been reported to preferentially degrade oxidized proteins and glycated proteins, to degrade aging proteins in cooperation with the proteasome, and to reduce amyloid β, which causes Alzheimer's disease. It has also been confirmed that OPH degrades AGEs.
<Test>
≪1≫測定概要
本測定ではOPHとその反応基質であるN-acetyl-L-alanine p-nitro-anilide(AAPA)との反応系に試料溶液を添加し、OPHの酵素反応への影響を評価した。なお、サンプルについては実施形態1の試験と同様に調製した。
≪1≫ Outline of measurement In this measurement, sample solution was added to the reaction system of OPH and its reaction substrate, N-acetyl-L-alanine p-nitro-anilide (AAPA), and the effect of OPH on enzyme reaction was evaluated. did. The sample was prepared in the same manner as in the test of
≪2≫方法
OPHとしてacylamino-acid releasing enzyme (AARE)、OPHの反応基質としてN-acetyl-L-alaninep-nitroanilide(AAPA) 溶液を使用した。測定にはOPHを0.01 U/mL、0.005 U/mL、0.001 U/mLに調製して使用した。96ウェルマイクロプレートの各wellにOPH、AAPA、試料溶液を混合添加し、37°Cに設定したインキュベーター内で4時間反応させた反応液の405nmにおける吸光度をマイクロプレートリーダーで測定した。OPHの酵素活性は1時間当たりの吸光度変化量(反応速度)を求めた。同時にreference(Ref)として試料無添加時の反応速度を求め、下式に従ってRefの反応速度を100%とした時の活性増強作用を算出した。OPH活性増強作用ネガティブコントロールにはエピガロカテキンガレート(EGCg)を使用した。
≪2≫ Method
Acylamino-acid releasing enzyme (AARE) was used as OPH, and N-acetyl-L-alaninep-nitroanilide (AAPA) solution was used as a reaction substrate for OPH. For the measurement, OPH was adjusted to 0.01 U / mL, 0.005 U / mL, and 0.001 U / mL. The OPH, AAPA, and sample solution were mixed and added to each well of the 96-well microplate, and the absorbance at 405 nm of the reaction solution reacted for 4 hours in an incubator set at 37 ° C was measured with a microplate reader. The amount of change in absorbance per hour (reaction rate) was determined for the enzyme activity of OPH. At the same time, the reaction rate when no sample was added was determined as reference (Ref), and the activity enhancing action when the Ref reaction rate was 100% was calculated according to the following formula. Epigallocatechin gallate (EGCg) was used as a negative control for enhancing OPH activity.
≪3≫結果
図8(a)に測定結果を示す。図8(b)はグラフで示したものである。図示するように、黒ガリンガル抽出液はOPH活性を試料無添加時(Ref)よりも約2倍増強させる作用がみられた。
<効果>
<< 3 >> Results FIG. 8A shows the measurement results. FIG. 8B is a graph. As shown in the figure, the black galling extract showed an effect of enhancing the OPH activity by about 2 times compared to when no sample was added (Ref).
<Effect>
本実施形態により、黒ガリンガル抽出物を有効成分とするOPH活性増強剤などを提供することができる。 According to this embodiment, it is possible to provide an OPH activity enhancer containing black galling extract as an active ingredient.
Claims (9)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2016105996A JP6411403B2 (en) | 2016-05-27 | 2016-05-27 | Protein saccharification reaction inhibitor |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2016105996A JP6411403B2 (en) | 2016-05-27 | 2016-05-27 | Protein saccharification reaction inhibitor |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2017210452A JP2017210452A (en) | 2017-11-30 |
JP6411403B2 true JP6411403B2 (en) | 2018-10-24 |
Family
ID=60475225
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016105996A Active JP6411403B2 (en) | 2016-05-27 | 2016-05-27 | Protein saccharification reaction inhibitor |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP6411403B2 (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP6462755B2 (en) * | 2017-04-04 | 2019-01-30 | 丸善製薬株式会社 | Brain function improving agent and food and drink for improving brain function |
JP2019187354A (en) * | 2018-04-27 | 2019-10-31 | 株式会社アンチエイジングコミュニケーション | OPH activity enhancer |
JP7229513B2 (en) * | 2018-10-16 | 2023-02-28 | 丸善製薬株式会社 | Brain function improving agent and food and drink for improving brain function |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP5804391B2 (en) * | 2012-03-08 | 2015-11-04 | 株式会社アップウェル | Anti-glycation agent |
JP2015168668A (en) * | 2014-03-10 | 2015-09-28 | 株式会社ダイセル | Anti-saccharification agent containing equol |
JP6490342B2 (en) * | 2014-03-12 | 2019-03-27 | 株式会社再春館製薬所 | Anti-skin aging agent containing Shiranui Chrysanthemum extract |
JP6359850B2 (en) * | 2014-03-24 | 2018-07-18 | 株式会社コーセー | Anti-glycation agent |
JP2016003193A (en) * | 2014-06-16 | 2016-01-12 | 株式会社ダイセル | Anti-glycation agent comprising phytoene and/or phytofluene |
JP2016088933A (en) * | 2014-11-01 | 2016-05-23 | 育宏 南 | Age production inhibitor and application thereof |
-
2016
- 2016-05-27 JP JP2016105996A patent/JP6411403B2/en active Active
Also Published As
Publication number | Publication date |
---|---|
JP2017210452A (en) | 2017-11-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5538611B2 (en) | Maillard reaction inhibitor | |
KR101841255B1 (en) | Composition for improving skin wrinkle and enhancing elasticity | |
JP4286513B2 (en) | Anti-aging composition | |
JP6411403B2 (en) | Protein saccharification reaction inhibitor | |
JP2006124350A (en) | Nmf production promoter | |
JPH08259431A (en) | Maillard reaction inhibitor and dermal preparation for external use containing the same | |
JP2016193857A (en) | Skin cosmetic, and food and drink | |
JP2015027998A (en) | Moisturizer, skin barrier function activator, tight junction formation accelerator, trpv4 expression enhancer, intracellular calcium concentration increaser, intracellular calcium concentration increaser, lipid synthesis accelerator, blood flow improver and periocular darkness ameliorator | |
JP2018188375A (en) | Pentosidine production inhibitor | |
JP6159918B2 (en) | Hyaluronidase inhibitors and methods for inhibiting hyaluronidase activity | |
JP6122200B1 (en) | Anti-glycation composition | |
JP3827581B2 (en) | Skin cosmetics and beauty food and drink | |
JP4247091B2 (en) | Skin anti-aging agent | |
KR20190047582A (en) | Two Step-sequential Enzyme treated extract of Plant originated materials, its functional cosmetic composition comprising the same and method producing the same | |
WO2008035583A1 (en) | Moisturizing agent, cell-activating agent, antioxidant agent, protease activity enhancing agent, anti-aging agent, skin-whitening agent, anti-inflammatory agent, and neutral fat accumulation-inhibiting agent | |
JP4201091B1 (en) | Moisturizer, anti-aging agent, antioxidant and external preparation for skin | |
JP2006298800A (en) | Polyphenol having action of fibroblast activation and food supplement and cosmetic containing the same | |
JP2014208613A (en) | Agent for increasing ceramide and hyaluronic acid content of skin | |
KR20180114308A (en) | Method for producing microparticle of natural dye with increased stability using lecithin | |
JP6544503B2 (en) | Cosmetic composition and functional food containing collagenase inhibitor and collagenase inhibitor | |
WO2015002279A1 (en) | Composition for promoting glutathione production | |
JP2011032177A (en) | Inhibitor of kit cleavage | |
JP7076062B2 (en) | External skin agents, anti-glycation agents and collagenase inhibitors | |
JP5294847B2 (en) | Moisturizer, whitening agent and slimming agent | |
JP2019182863A (en) | Skin firmness or moisture improving composition |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A80 | Written request to apply exceptions to lack of novelty of invention |
Free format text: JAPANESE INTERMEDIATE CODE: A80 Effective date: 20160622 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20171006 |
|
A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20171006 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20180131 |
|
A975 | Report on accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A971005 Effective date: 20180301 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20180613 |
|
RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20180622 |
|
RD04 | Notification of resignation of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7424 Effective date: 20180705 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20180806 |
|
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: 20180828 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20180926 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6411403 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |