JP4300324B2 - Age-related eye disease ameliorating agent, and pharmaceutical composition, food and drink, cosmetics and feed containing the same - Google Patents
Age-related eye disease ameliorating agent, and pharmaceutical composition, food and drink, cosmetics and feed containing the same Download PDFInfo
- Publication number
- JP4300324B2 JP4300324B2 JP2005033913A JP2005033913A JP4300324B2 JP 4300324 B2 JP4300324 B2 JP 4300324B2 JP 2005033913 A JP2005033913 A JP 2005033913A JP 2005033913 A JP2005033913 A JP 2005033913A JP 4300324 B2 JP4300324 B2 JP 4300324B2
- Authority
- JP
- Japan
- Prior art keywords
- age
- lactoferrin
- granules
- related eye
- eye disease
- 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
- 208000030533 eye disease Diseases 0.000 title description 57
- 239000003795 chemical substances by application Substances 0.000 title description 36
- 239000008194 pharmaceutical composition Substances 0.000 title description 16
- 239000002537 cosmetic Substances 0.000 title description 8
- 235000013305 food Nutrition 0.000 title description 6
- 210000004561 lacrimal apparatus Anatomy 0.000 claims description 34
- 208000002780 macular degeneration Diseases 0.000 claims description 17
- 239000004480 active ingredient Substances 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 6
- 208000002177 Cataract Diseases 0.000 claims description 5
- 208000003464 asthenopia Diseases 0.000 claims description 5
- 208000010412 Glaucoma Diseases 0.000 claims description 4
- 201000010041 presbyopia Diseases 0.000 claims description 4
- 230000003248 secreting effect Effects 0.000 claims description 3
- 238000009472 formulation Methods 0.000 claims 2
- 235000021242 lactoferrin Nutrition 0.000 description 73
- 102000010445 Lactoferrin Human genes 0.000 description 71
- 108010063045 Lactoferrin Proteins 0.000 description 71
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 description 71
- 229940078795 lactoferrin Drugs 0.000 description 71
- 241000700159 Rattus Species 0.000 description 50
- 239000008187 granular material Substances 0.000 description 50
- 230000032683 aging Effects 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 22
- 206010064930 age-related macular degeneration Diseases 0.000 description 15
- 201000004569 Blindness Diseases 0.000 description 13
- 241001465754 Metazoa Species 0.000 description 12
- 239000003814 drug Substances 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 238000011282 treatment Methods 0.000 description 10
- -1 food-drinks Substances 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 239000000126 substance Substances 0.000 description 9
- 230000005540 biological transmission Effects 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 description 8
- 206010061218 Inflammation Diseases 0.000 description 7
- 230000037396 body weight Effects 0.000 description 7
- 229940079593 drug Drugs 0.000 description 7
- 230000037406 food intake Effects 0.000 description 7
- 230000004054 inflammatory process Effects 0.000 description 7
- 208000003556 Dry Eye Syndromes Diseases 0.000 description 6
- 206010013774 Dry eye Diseases 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 230000002265 prevention Effects 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 5
- 230000033115 angiogenesis Effects 0.000 description 5
- 230000003110 anti-inflammatory effect Effects 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 239000001768 carboxy methyl cellulose Substances 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 238000001000 micrograph Methods 0.000 description 5
- 229950003937 tolonium Drugs 0.000 description 5
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 5
- JKQXZKUSFCKOGQ-JLGXGRJMSA-N (3R,3'R)-beta,beta-carotene-3,3'-diol Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C[C@@H](O)CC1(C)C JKQXZKUSFCKOGQ-JLGXGRJMSA-N 0.000 description 4
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 4
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- JKQXZKUSFCKOGQ-LQFQNGICSA-N Z-zeaxanthin Natural products C([C@H](O)CC=1C)C(C)(C)C=1C=CC(C)=CC=CC(C)=CC=CC=C(C)C=CC=C(C)C=CC1=C(C)C[C@@H](O)CC1(C)C JKQXZKUSFCKOGQ-LQFQNGICSA-N 0.000 description 4
- QOPRSMDTRDMBNK-RNUUUQFGSA-N Zeaxanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCC(O)C1(C)C)C=CC=C(/C)C=CC2=C(C)CC(O)CC2(C)C QOPRSMDTRDMBNK-RNUUUQFGSA-N 0.000 description 4
- JKQXZKUSFCKOGQ-LOFNIBRQSA-N all-trans-Zeaxanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2=C(C)CC(O)CC2(C)C JKQXZKUSFCKOGQ-LOFNIBRQSA-N 0.000 description 4
- 235000014113 dietary fatty acids Nutrition 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 239000000194 fatty acid Substances 0.000 description 4
- 229930195729 fatty acid Natural products 0.000 description 4
- 235000012680 lutein Nutrition 0.000 description 4
- 229960005375 lutein Drugs 0.000 description 4
- 239000001656 lutein Substances 0.000 description 4
- KBPHJBAIARWVSC-RGZFRNHPSA-N lutein Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\[C@H]1C(C)=C[C@H](O)CC1(C)C KBPHJBAIARWVSC-RGZFRNHPSA-N 0.000 description 4
- ORAKUVXRZWMARG-WZLJTJAWSA-N lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C ORAKUVXRZWMARG-WZLJTJAWSA-N 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 230000003716 rejuvenation Effects 0.000 description 4
- 239000004094 surface-active agent Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 description 4
- 235000010930 zeaxanthin Nutrition 0.000 description 4
- 229940043269 zeaxanthin Drugs 0.000 description 4
- 239000001775 zeaxanthin Substances 0.000 description 4
- 241000283690 Bos taurus Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 208000007135 Retinal Neovascularization Diseases 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 3
- 239000011230 binding agent Substances 0.000 description 3
- 230000007850 degeneration Effects 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 230000004438 eyesight Effects 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 239000000314 lubricant Substances 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 238000000879 optical micrograph Methods 0.000 description 3
- 230000001575 pathological effect Effects 0.000 description 3
- 230000003449 preventive effect Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 3
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 230000004393 visual impairment Effects 0.000 description 3
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 2
- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 2
- 241000283086 Equidae Species 0.000 description 2
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 2
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 208000034038 Pathologic Neovascularization Diseases 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 2
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 2
- 229930003427 Vitamin E Natural products 0.000 description 2
- 208000000208 Wet Macular Degeneration Diseases 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 2
- 239000002870 angiogenesis inducing agent Substances 0.000 description 2
- 230000003712 anti-aging effect Effects 0.000 description 2
- 235000013793 astaxanthin Nutrition 0.000 description 2
- 229940022405 astaxanthin Drugs 0.000 description 2
- 239000001168 astaxanthin Substances 0.000 description 2
- MQZIGYBFDRPAKN-ZWAPEEGVSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-ZWAPEEGVSA-N 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 230000023555 blood coagulation Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 2
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 2
- 235000021466 carotenoid Nutrition 0.000 description 2
- 150000001747 carotenoids Chemical class 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 235000010980 cellulose Nutrition 0.000 description 2
- 208000037976 chronic inflammation Diseases 0.000 description 2
- 230000006020 chronic inflammation Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000003889 eye drop Substances 0.000 description 2
- 229940012356 eye drops Drugs 0.000 description 2
- 229940126864 fibroblast growth factor Drugs 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 239000001341 hydroxy propyl starch Substances 0.000 description 2
- 235000013828 hydroxypropyl starch Nutrition 0.000 description 2
- 230000002452 interceptive effect Effects 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- GLDOVTGHNKAZLK-UHFFFAOYSA-N octadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCCCO GLDOVTGHNKAZLK-UHFFFAOYSA-N 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 210000001525 retina Anatomy 0.000 description 2
- 230000002207 retinal effect Effects 0.000 description 2
- 210000004739 secretory vesicle Anatomy 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 235000000891 standard diet Nutrition 0.000 description 2
- 230000000451 tissue damage Effects 0.000 description 2
- 231100000827 tissue damage Toxicity 0.000 description 2
- 230000009261 transgenic effect Effects 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 230000004304 visual acuity Effects 0.000 description 2
- 235000019155 vitamin A Nutrition 0.000 description 2
- 239000011719 vitamin A Substances 0.000 description 2
- 235000019165 vitamin E Nutrition 0.000 description 2
- 229940046009 vitamin E Drugs 0.000 description 2
- 239000011709 vitamin E Substances 0.000 description 2
- 229940045997 vitamin a Drugs 0.000 description 2
- 239000001993 wax Substances 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- PZNPLUBHRSSFHT-RRHRGVEJSA-N 1-hexadecanoyl-2-octadecanoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[C@@H](COP([O-])(=O)OCC[N+](C)(C)C)COC(=O)CCCCCCCCCCCCCCC PZNPLUBHRSSFHT-RRHRGVEJSA-N 0.000 description 1
- RFVNOJDQRGSOEL-UHFFFAOYSA-N 2-hydroxyethyl octadecanoate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCCO RFVNOJDQRGSOEL-UHFFFAOYSA-N 0.000 description 1
- HIQIXEFWDLTDED-UHFFFAOYSA-N 4-hydroxy-1-piperidin-4-ylpyrrolidin-2-one Chemical compound O=C1CC(O)CN1C1CCNCC1 HIQIXEFWDLTDED-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 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 1
- 102000009840 Angiopoietins Human genes 0.000 description 1
- 108010009906 Angiopoietins Proteins 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- 101000798100 Bos taurus Lactotransferrin Proteins 0.000 description 1
- 108010074051 C-Reactive Protein Proteins 0.000 description 1
- 102100032752 C-reactive protein Human genes 0.000 description 1
- 241000282832 Camelidae Species 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000725101 Clea Species 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 208000028399 Critical Illness Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines 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
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 208000009386 Experimental Arthritis Diseases 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 102000052396 Hephaestin Human genes 0.000 description 1
- 108700038053 Hephaestin Proteins 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010065973 Iron Overload Diseases 0.000 description 1
- 102000008133 Iron-Binding Proteins Human genes 0.000 description 1
- 108010035210 Iron-Binding Proteins Proteins 0.000 description 1
- 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 1
- 239000004166 Lanolin Substances 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 239000004264 Petrolatum Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241000320126 Pseudomugilidae Species 0.000 description 1
- 101710132082 Pyrimidine/purine nucleoside phosphorylase Proteins 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- FKNQFGJONOIPTF-UHFFFAOYSA-N Sodium cation Chemical compound [Na+] FKNQFGJONOIPTF-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 102100031372 Thymidine phosphorylase Human genes 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 125000005907 alkyl ester group Chemical group 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
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000002917 arthritic effect Effects 0.000 description 1
- 210000000544 articulatio talocruralis Anatomy 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 201000009310 astigmatism Diseases 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000036765 blood level Effects 0.000 description 1
- 229940072440 bovine lactoferrin Drugs 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- ACTIUHUUMQJHFO-UPTCCGCDSA-N coenzyme Q10 Chemical compound COC1=C(OC)C(=O)C(C\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UPTCCGCDSA-N 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 208000021921 corneal disease Diseases 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 235000013350 formula milk Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 239000008172 hydrogenated vegetable oil Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229940039717 lanolin Drugs 0.000 description 1
- 235000019388 lanolin Nutrition 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- HCWCAKKEBCNQJP-UHFFFAOYSA-N magnesium orthosilicate Chemical compound [Mg+2].[Mg+2].[O-][Si]([O-])([O-])[O-] HCWCAKKEBCNQJP-UHFFFAOYSA-N 0.000 description 1
- 239000000391 magnesium silicate Substances 0.000 description 1
- 229910052919 magnesium silicate Inorganic materials 0.000 description 1
- 235000019792 magnesium silicate Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 208000031225 myocardial ischemia Diseases 0.000 description 1
- GOQYKNQRPGWPLP-UHFFFAOYSA-N n-heptadecyl alcohol Natural products CCCCCCCCCCCCCCCCCO GOQYKNQRPGWPLP-UHFFFAOYSA-N 0.000 description 1
- 230000014399 negative regulation of angiogenesis Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 229940066842 petrolatum Drugs 0.000 description 1
- 235000019271 petrolatum Nutrition 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920001223 polyethylene glycol 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
- 230000002980 postoperative effect Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 235000013594 poultry meat Nutrition 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000020341 sensory perception of pain Effects 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 229910001415 sodium ion Inorganic materials 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000008347 soybean phospholipid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Images
Landscapes
- Fodder In General (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Cosmetics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
本発明は、加齢により機能低下または組織損傷した眼を修復・治療し、視力低下または失明を予防するための加齢性眼疾患改善剤、特に、これまで不可避の自然現象と考えられてきた眼の加齢変性（眼の老化現象）を遅らせ、または若返らせ、最終的には眼疾患の治癒をもたらす加齢性眼疾患改善剤に関する。さらに、本発明は、この改善剤を含む医薬組成物、飲食品、化粧料および飼料に関する。 The present invention is considered to be an aging eye disease ameliorating agent for repairing / treating an eye whose function has deteriorated or tissue damage due to aging, and to prevent visual loss or blindness, particularly an inevitable natural phenomenon so far. The present invention relates to an agent for improving age-related eye diseases that delays or rejuvenates eye age-related degeneration (eye aging phenomenon), and ultimately leads to healing of eye diseases. Furthermore, this invention relates to the pharmaceutical composition, food-drinks, cosmetics, and feed containing this improving agent.
医学の進歩に伴う平均寿命の延長にもかかわらず、先進国における失明率は増大の一途である。視覚は、生活の質（ＱＯＬ）のなかで最重要ファクターであり、寿命が長くなっても、失明しては幸福な人生とはいえない。 Despite the increase in life expectancy associated with medical advances, blindness rates in developed countries continue to increase. Vision is the most important factor in quality of life (QOL), and even if the lifespan is long, it cannot be said that it is a happy life if it loses sight.
米国を始めとする先進諸国は失明を克服するための研究に躍起であるが、それに加え、近年、失明にも増して社会的な関心をひくようになったのが、視力の質（Ｑｕａｌｉｔｙ ｏｆ Ｖｉｓｉｏｎ： ＱＯＶ）である。すなわち、従来は単に見えればよいと考えていた人々も、「もっと快適にものを見たい」、「コンタクトレンズなしでものを見たい」、「老眼鏡はかけたくない」等と考えるようになってきている。 Advanced countries such as the United States are eager to research to overcome blindness, but in addition to this, the quality of vision (Quality of (Vision: QOV). In other words, people who previously thought that they should simply be able to see are now thinking that they want to see things more comfortably, want to see things without contact lenses, and do not want to wear reading glasses. ing.
また、ドライアイ患者も増加している。ドライアイになると、眼が乾くために長時間眼を開けておくことができず、ドライアイは眼の疲れの最大原因になっている。こうした眼の疲れに対しては、従来は眼を休める程度の対応で充分な改善効果も見込めたが、近年のコンピューター技術の発達やインターネットの普及によって、仕事中に眼を休めることすらできず、眼の休養による充分な改善効果も見込めなくなってきている。 The number of dry eye patients is also increasing. When it comes to dry eyes, it is impossible to keep the eyes open for a long time because the eyes are dry, and dry eyes are the biggest cause of eye fatigue. With regard to such eye fatigue, it has been expected that a sufficient improvement effect can be expected by taking measures to rest the eyes, but due to the recent development of computer technology and the spread of the Internet, it is not possible to rest the eyes during work. It is no longer possible to expect sufficient improvement from eye rest.
このような、加齢性網膜黄斑変性症、緑内障、白内障、ドライアイ、老視といった眼疾患の多くが、加齢によるものである。したがって、視力の低下防止（失明対策）またはＱＯＶの向上を図るためには、未だ発展途上の分野ではあるが、加齢を防止するためのアンチエイジング医学が非常に重要である。眼疾患の原因が加齢によるものであれば、加齢そのものに干渉することによって眼の病気を発症させないようにできないものかという観点から、眼の加齢を遅らせる、あるいは安全に修復することができる抗加齢物質、若返り物質が世界的に希求されている。 Many of these eye diseases such as age-related retinal macular degeneration, glaucoma, cataract, dry eye and presbyopia are caused by aging. Therefore, anti-aging medicine for preventing aging is very important in order to prevent a decrease in visual acuity (measures for blindness) or improve QOV, although it is still a developing field. If the cause of the eye disease is due to aging, it may be possible to delay the aging of the eye or restore it safely from the viewpoint of preventing the development of an eye disease by interfering with the aging itself. Anti-aging substances that can be rejuvenated and rejuvenating substances are in demand worldwide.
眼疾患の中でもとりわけ高齢者に多発し、失明に至る疾患として世界的な課題とされているのは、加齢性網膜黄斑変性症（Ａｇｅ−ｒｅｌａｔｅｄ Ｍａｃｕｌａｒ Ｄｅｇｅｎｅｒａｔｉｏｎ： ＡＭＤ）である。ＡＭＤは、６５歳以上の高齢者に多く見られ、失明の原因となる病気であり、厚生労働省の特定疾患に指定されている。この病気にかかると、視野がぼやけ、線が歪んで見え、視野の中央部にいくつかの黒点が現れる。日本では、推定患者数が１万５千人と比較的少ないが、２００４年の米国では初期のＡＭＤが１，５００万人、失明の危険がある重症患者が１６０万人と推定されほど高齢者に多発する眼疾患である。しかも、ＡＭＤは白内障と違って治療法が確立されておらず、現状では有効な治療手段がない。疫学調査ではルテインおよびゼアキサンチンを含む黄緑野菜を多く摂取する結果、これらカロチノイドの血中濃度が高い人は発症リスクが低いことがわかっているが、ルテインおよびゼアキサンチンを摂取してもＡＤＭの発症および進行に影響がないことが大規模な二重盲検試験によって明らかにされている。 Among eye diseases, Age-related Macular Degeneration (AMD) is a world-wide problem that frequently occurs in the elderly and leads to blindness. AMD is a disease that is frequently seen in elderly people over the age of 65 and causes blindness, and is designated as a specific disease by the Ministry of Health, Labor and Welfare. When this illness occurs, the field of view is blurred, the lines appear distorted, and several black spots appear in the center of the field of view. In Japan, the estimated number of patients is relatively small at 15,000, but in the US in 2004, the early AMD was estimated at 15 million, and the number of critically ill patients at risk of blindness was estimated at 1.6 million. It is a frequent eye disease. Moreover, unlike cataracts, AMD has not established a treatment method, and currently there is no effective treatment means. Epidemiological studies have shown that people who have high blood levels of these carotenoids have low risk of developing as a result of ingesting large amounts of yellow-green vegetables containing lutein and zeaxanthin. However, even if lutein and zeaxanthin are ingested, the occurrence of ADM and Large double-blind trials have shown no effect on progression.
ＡＭＤは、その約９割を占める萎縮型と網膜の血管新生を伴う滲出型とに分類される。このうち失明のリスクが高い滲出型では、診断半年後から１年半後に重篤な視力減退が起こり、失明が多発するが、現在、その進行をくい止め、また確実に失明を阻止する治療手段はない。ＡＭＤの危険因子は、年齢、家族歴、青い目、喫煙、血管障害、黄緑野菜の摂取不足、強度の光線に長時間暴露されること等であるが、中でも、年齢は発症と最も強い相関があり、血族に患者が存在する家族歴も１０〜２０％の相関があるとされている。また、虚血性心疾患、高血圧症等の心血管病との高い相関も注目されている。滲出型ＡＭＤにより失明に至る過程には、網膜の炎症および炎症部位を栄養する血管新生が関わっている。そのため、網膜の血管新生を抑制すれば、滲出型ＡＭＤの進行を阻止し、失明を防止できるものと考えられる。 AMD is classified into an atrophic type, which accounts for about 90%, and an exudative type with retinal neovascularization. Of these, the exudative type, which has a high risk of blindness, causes severe visual loss and occurs frequently after one and a half years from the diagnosis, but currently there are no treatments that can stop the progression and reliably prevent blindness. Absent. Risk factors for AMD include age, family history, blue eyes, smoking, vascular disorders, inadequate intake of yellow-green vegetables, prolonged exposure to intense light, etc. Among them, age has the strongest correlation with onset There is also a 10-20% correlation between the family history of patients in the family. In addition, a high correlation with cardiovascular diseases such as ischemic heart disease and hypertension has attracted attention. The process leading to blindness by wet AMD involves inflammation of the retina and angiogenesis that nourishes the site of inflammation. Therefore, it is considered that if the retinal neovascularization is suppressed, the progression of wet AMD can be prevented and blindness can be prevented.
血管新生には、生理的なものと、癌、炎症等で見られる病的なものとがある。生理的血管新生、とりわけ胎生期の血管新生は、炎症、出血、血液凝固を伴なわず、血管内皮細胞に特異的に作用する血管新生因子（ＶＥＧＦ）やアンジオポエチン等の因子によって整然と制御されたプロセスである。それとは対照的に、癌、炎症等で見られる病的な血管新生には、線維芽細胞成長因子（ＦＧＦ）やチミジンフォスフォリラーゼ等の内皮細胞に対して非特異的な因子が関与する。しかも、病的な血管新生は、炎症、出血、血液凝固等を必ず伴っており、これが血管新生像を複雑なものとしている。病的な新生血管は、破綻・出血して血栓を生じ、血管から血漿成分がしみ出して周囲の組織にフィブリノイド変性を起こす。このような過程を経て網膜の神経細胞が破壊され、失明に至るのである。 There are two types of neovascularization: physiological ones and pathological ones found in cancer, inflammation and the like. Physiological angiogenesis, especially embryonic angiogenesis, is an orderly controlled process by factors such as angiogenic factors (VEGF) and angiopoietin that act specifically on vascular endothelial cells without inflammation, bleeding, or blood clotting. It is. In contrast, pathological angiogenesis seen in cancer, inflammation, etc. involves factors that are not specific for endothelial cells such as fibroblast growth factor (FGF) and thymidine phosphorylase. In addition, pathological angiogenesis is always accompanied by inflammation, bleeding, blood coagulation, and the like, which complicates angiogenesis. Pathological new blood vessels rupture and bleed to form blood clots, and plasma components ooze out from the blood vessels, causing fibrinoid degeneration in the surrounding tissues. Through these processes, neurons in the retina are destroyed, leading to blindness.
したがって、ＡＭＤを予防または治療するには、少なくとも、（１）安全であること、（２）加齢変性した眼組織を若返らせること、（３）網膜の血管新生を阻止すること、および（４）抗炎症作用があること、の４条件を備えた薬剤が必要である。最近、ＡＭＤ予防・治療薬として血管新生因子（ＶＥＧＦ）の遺伝子発現に干渉するリボ核酸断片（ＲＮＡｉ）が特に注目されている。既に米国ではＲＮＡｉを注射してＡＭＤを治療することを目指し、バイオテクノロジーのベンチャー企業が数社も創立され、熾烈な開発競争を展開している。米国食品薬品局は、医薬品としての承認申請が行われた場合、最優先で審査することを公表している。ＡＭＤの予防・治療薬は緊急度が高いからである。 Thus, to prevent or treat AMD, at least (1) be safe, (2) rejuvenate age-degenerated ocular tissue, (3) prevent retinal neovascularization, and (4 ) Drugs with 4 conditions of having anti-inflammatory action are necessary. Recently, a ribonucleic acid fragment (RNAi) that interferes with gene expression of angiogenic factor (VEGF) has attracted particular attention as a preventive / therapeutic agent for AMD. Already in the United States, several biotechnology venture companies have been established with the aim of treating RNA by injecting RNAi, and fierce development competition is taking place. The US Food and Drug Administration has announced that if a drug application is submitted, it will be reviewed with the highest priority. This is because the preventive and therapeutic drugs for AMD are highly urgent.
加齢性眼疾患の予防・治療に従来用いられてきた製品は、ルテイン、ゼアキサンチン、アスタキサンチンなどのカロチノイド、視力に関係するビタミンＡ、抗酸化作用があるアスコルビン酸、ビタミンＥ、コエンザイムＱ１０等々を有効成分として含んでいる。 Products that have been used for the prevention and treatment of age-related eye diseases include carotenoids such as lutein, zeaxanthin, and astaxanthin, vitamin A related to visual acuity, ascorbic acid with antioxidant action, vitamin E, coenzyme Q 10 and the like. Contains as an active ingredient.
ラクトフェリン（以下、ＬＦということがある）は、哺乳動物の乳汁中に含まれる可溶性の鉄結合性の糖タンパク質である。その分子量は、ウシで８６，０００、ヒトで８８，０００である。ラクトフェリンは、溶液中から鉄イオンを奪うことによって、抗菌活性を示すことがわかっており（非特許文献１）、従来、健康食品等の成分として利用されてきた。 Lactoferrin (hereinafter sometimes referred to as LF) is a soluble iron-binding glycoprotein contained in mammalian milk. Its molecular weight is 86,000 for cattle and 88,000 for humans. Lactoferrin has been shown to exhibit antibacterial activity by depriving iron ions from solution (Non-Patent Document 1) and has been conventionally used as a component of health foods and the like.
前記４条件のうち、安全性に関しては、牛乳から抽出したラクトフェリンが高い安全性を示すことが知られている（特許文献１）。すなわち、ラクトフェリンは、雌雄のビーグル犬およびラットに５ｇ／ｋｇを１回経口投与、または２ｇ／ｋｇを１２週間連続経口投与しても何らの毒性も示さない。既に育児用調製粉乳、健康食品およびヨーグルト、飲料などに添加されており、不特定多数の人に摂取されて久しいにもかかわらず、ラクトフェリンの安全性に懸念を抱かせる情報は一切報告されていない。 Among the above four conditions, it is known that lactoferrin extracted from milk shows high safety regarding safety (Patent Document 1). That is, lactoferrin does not show any toxicity when 5 g / kg is orally administered to male and female beagle dogs and rats once or 2 g / kg is orally administered continuously for 12 weeks. Already added to infant formulas, health foods and yogurt, beverages, etc., no information has been reported that raises concerns about the safety of lactoferrin even though it has been consumed by an unspecified number of people. .
また、血管新生の阻害効果については、津田等（日本国立がんセンター）により、ラクトフェリンが化学発ガン剤による発ガンを抑制し、移植ガンの転移を抑制するのは、ガン細胞により誘導される血管新生を阻害するのも一因であると報告されている（非特許文献２）。 Regarding the angiogenesis inhibitory effect, it is induced by cancer cells that lactoferrin suppresses carcinogenesis by chemical carcinogens and suppresses metastasis of transplanted cancer by Tsuda et al. (Japan National Cancer Center). Inhibition of angiogenesis has also been reported to be a cause (Non-patent Document 2).
さらに、抗炎症作用については、ラクトフェリンを慢性炎症の病態モデルであるアジュバント関節炎ラットに経口投与すると、足関節の腫脹を抑制し、細菌リポ多糖を投与した際の血中ＴＮＦ−α上昇を抑え、抗炎症サイトカインであるＩＬ−１０を上昇させるという報告があり（非特許文献３）、ラクトフェリンは慢性炎症に対し消炎作用を呈するものと考えられる。 Furthermore, for anti-inflammatory action, when lactoferrin is orally administered to adjuvant arthritic rats, which are chronic inflammation pathological models, it suppresses swelling of the ankle joint, suppresses blood TNF-α elevation when bacterial lipopolysaccharide is administered, There is a report that IL-10 which is an anti-inflammatory cytokine is increased (Non-patent Document 3), and lactoferrin is considered to exhibit an anti-inflammatory action against chronic inflammation.
一方、ラクトフェリンは涙液に多量に含まれるにもかかわらず、その眼に対する作用については充分に明らかになっていない。すなわち、このタンパク質が持つ本来の機能からラクトフェリンが感染防御、抗炎症作用を有することが想像されるものの、眼に対する具体的な薬理効果としては、角膜障害治療作用（特許文献２）および術後乱視予防作用（特許文献３）以外については報告されていない。すなわち、ラクトフェリンが、加齢性眼疾患の予防または治療に有効であることを具体的に示すデータは報告されておらず、ラクトフェリンが加齢性眼疾患を改善することは従来まったく知られていなかった。 On the other hand, despite the fact that lactoferrin is contained in a large amount in tears, its action on the eye has not been fully clarified. That is, although lactoferrin is supposed to have infection protection and anti-inflammatory action from the original functions of this protein, specific pharmacological effects on the eyes include corneal disorder treatment (Patent Document 2) and postoperative astigmatism. No report has been made other than the preventive action (Patent Document 3). In other words, no data specifically showing that lactoferrin is effective in preventing or treating age-related eye diseases has been reported, and it has not been known at all that lactoferrin improves age-related eye diseases. It was.
本発明は、眼精疲労、加齢性網膜黄斑変性症、緑内障、白内障、ドライアイ、老視等の、加齢に起因または関連する眼疾患（加齢性眼疾患ともいう）を予防または治療するために有効で、安全性の高い薬剤を提供すること、特に、加齢の過程に干渉することによってこれらの眼疾患を発症させない、加齢による機能低下または組織傷害を阻止する、または加齢減少を逆転させて若返らせることができる、眼疾患の予防または治療用薬剤を提供することを目的とする。さらに、本発明は、そのような薬剤を配合した加齢性眼疾患予防または治療用の医薬組成物、飲食品、化粧料および飼料を提供することを目的とする。 The present invention prevents or treats eye diseases caused by or related to aging (also referred to as age-related eye diseases) such as eye strain, age-related macular degeneration, glaucoma, cataract, dry eye, presbyopia, etc. To provide drugs that are effective and safe to do, in particular, do not cause these eye diseases by interfering with the aging process, prevent functional deterioration or tissue damage due to aging, or aging An object of the present invention is to provide a drug for preventing or treating eye diseases, which can reverse the decrease and rejuvenate. Furthermore, an object of this invention is to provide the pharmaceutical composition, food-drinks, cosmetics, and feed for age-related eye disease prevention or treatment which mix | blended such a chemical | medical agent.
本発明者らは、前記の課題を解決するために、広く天然素材に含まれている加齢性眼疾患予防または治療効果を有する可能性のある物質について、鋭意、探索を進めていたところ、ラクトフェリンが、加齢性眼疾患を予防・治療することができる、すなわち、老化した眼を若返らせる作用があること、そしてラクトフェリンが前記４条件をすべて満たす物質であることを見出し、本発明を完成するに至った。 In order to solve the above-mentioned problems, the present inventors have been diligently searching for substances that may have the effect of preventing or treating age-related eye diseases that are widely included in natural materials. Discovered that lactoferrin can prevent and treat age-related eye diseases, that is, has the effect of rejuvenating aging eyes, and that lactoferrin is a substance that satisfies all of the above four conditions, thereby completing the present invention. It came to do.
すなわち、本発明は、
〔１〕ラクトフェリンを有効成分として含有する加齢性眼疾患改善剤；
〔２〕加齢性眼疾患が、眼精疲労、加齢性網膜黄斑変性症、緑内障、白内障、ドライアイ、老視のいずれか１以上である、前記〔１〕記載の加齢性眼疾患改善剤；
〔３〕前記〔１〕または〔２〕記載の加齢性眼疾患改善剤を含む医薬組成物；
〔４〕前記〔１〕または〔２〕記載の加齢性眼疾患改善剤を含む飲食品；
〔５〕前記〔１〕または〔２〕記載の加齢性眼疾患改善剤を含む化粧料；
〔６〕前記〔１〕または〔２〕記載の加齢性眼疾患改善剤を含む飼料；
を提供する。
That is, the present invention
[1] Age-related eye disease ameliorating agent containing lactoferrin as an active ingredient;
[2] Age-related eye disease according to [1], wherein the age-related eye disease is one or more of eye strain, age-related macular degeneration, glaucoma, cataract, dry eye, and presbyopia Improver;
[3] A pharmaceutical composition comprising the age-related eye disease ameliorating agent according to [1] or [2];
[4] A food or drink containing the aging eye disease ameliorating agent according to [1] or [2];
[5] A cosmetic comprising the aging eye disease ameliorating agent according to [1] or [2];
[6] A feed containing the age-related eye disease ameliorating agent according to [1] or [2];
I will provide a.
本発明によりラクトフェリンを有効成分とする加齢性眼疾患改善剤、ならびにこれを有効成分として含有する加齢性眼疾患改善用医薬組成物、加齢性眼疾患改善用飲食品、加齢性眼疾患改善用化粧料、および加齢性眼疾患改善用飼料（以下、加齢性眼疾患改善剤等ということがある）が提供される。本発明の加齢性眼疾患改善剤等は、加齢性眼疾患の発症予防および発症後の治療に有用である。 Age-related eye disease ameliorating agent comprising lactoferrin as an active ingredient according to the present invention, a pharmaceutical composition for improving age-related eye disease containing the same as an active ingredient, food and drink for improving age-related eye disease, and aging eye A disease improving cosmetic and an age-related eye disease improving feed (hereinafter sometimes referred to as an age-related eye disease improving agent) are provided. The agent for improving age-related eye diseases of the present invention is useful for preventing the onset of age-related eye diseases and treating them after onset.
本発明の加齢性眼疾患改善剤等は、加齢性眼疾患の予防・治療に従来用いられてきた化合物と比べ、安全性が高く、確実に加齢性眼疾患を改善する特徴がある。また、ラクトフェリンは乳汁、粘液や好中球の成分であることから、加齢性眼疾患改善剤等は、他の薬物の生物活性を損なうことがなく、それらとの併用が可能である。 The agent for improving age-related eye diseases of the present invention is characterized by higher safety and surely improving age-related eye diseases compared to compounds conventionally used for the prevention and treatment of age-related eye diseases. . In addition, since lactoferrin is a component of milk, mucus, and neutrophils, age-related eye disease ameliorating agents and the like can be used in combination with them without impairing the biological activity of other drugs.
本発明の加齢性眼疾患改善剤は、ラクトフェリンを有効成分として含有することを特徴とする。本発明において使用されるラクトフェリンとしては、ヒトおよび種々の動物、例えば、ウシ、ウマ、ブタ、ヒツジ、ヤギ、ラクダ等から得られる天然のラクトフェリン分子（鉄イオンの有無またはその含量、由来する生物種などを問わない）、遺伝子工学技術により改変されたラクトフェリン遺伝子に基づいて産生される組換え型ラクトフェリン、トランスジェニック動物が泌乳するラクトフェリンなどのラクトフェリンのほか、ラクトフェリンの活性フラグメントなどの機能的等価物のいずれであってもよい。したがって、本発明に関して用語「ラクトフェリン」は、特に示さない限り、これらの種々のラクトフェリンをも包含する意味で用いられる。 The agent for improving age-related eye diseases of the present invention is characterized by containing lactoferrin as an active ingredient. The lactoferrin used in the present invention includes natural lactoferrin molecules obtained from humans and various animals, such as cows, horses, pigs, sheep, goats, camels, etc. (presence or absence of iron ions or their contents, derived biological species). Functional lactoferrin such as lactoferrin produced by transgenic animals, lactoferrin produced by transgenic animals, and functional equivalents of lactoferrin, etc. Either may be sufficient. Therefore, the term “lactoferrin” in the context of the present invention is used to encompass these various lactoferrins unless otherwise indicated.
本発明においては、これらのラクトフェリンの１種または２種以上を適宜選択して用いることができる。ラクトフェリンは、公知の物質であって、市販されているものであるが、それを製造するには、公知の方法、例えばスルホン化担体を用いてラクトフェリンを精製する方法（特開平３−１０９４００号公報）を工業的に有利に利用することができる。 In the present invention, one or more of these lactoferrins can be appropriately selected and used. Lactoferrin is a known substance and is commercially available. To produce it, a known method, for example, a method of purifying lactoferrin using a sulfonated carrier (JP-A-3-109400). ) Can be used industrially advantageously.
本発明の加齢性眼疾患改善剤は、ラクトフェリンを唯一の必須成分とするが、所望により、製薬または食品業界で公知の種々の成分などを含んでいてもよい。例えば、ビタミンＡ、ルテイン、ゼアキサンチン、アスタキサンチン、ビタミンＣ、ビタミンＥ等、従来から視力低下の回復に有効な作用を持つと考えられている成分を含むことができる。これらの他の有効成分をさらに含有させることによって、一層の加齢性眼疾患改善作用を期待できる。なお、本発明に関して加齢性眼疾患の「改善」は、予防および治療（軽快、治癒を含む）を包含する意味で用いられる。したがって、本発明の加齢性眼疾患改善剤は、加齢性眼疾患の予防または治療のいずれか一方または両方を目的とすることができる。 The agent for improving age-related eye diseases of the present invention contains lactoferrin as the only essential component, but may contain various components known in the pharmaceutical or food industry as desired. For example, ingredients such as vitamin A, lutein, zeaxanthin, astaxanthin, vitamin C, vitamin E, and the like that have been conventionally considered to have an effective action for recovery of vision loss can be included. By further containing these other active ingredients, a further aging eye disease improving effect can be expected. In the present invention, “amelioration” of age-related eye diseases is used in the meaning including prevention and treatment (including remission and cure). Therefore, the agent for improving age-related eye diseases of the present invention can be aimed at either or both of prevention or treatment of age-related eye diseases.
本発明の加齢性眼疾患改善剤は、添加剤として、製薬産業において日常的に使用されている賦形剤、崩壊剤、滑沢剤、結合剤、界面活性剤、流動性促進剤、着色剤、香料等を適宜添加して所望の剤型の医薬組成物とすることができる。本発明の加齢性眼疾患改善剤または医薬組成物の投与経路は、公知のいずれの経路であってもよく、例えば点眼、経口、経皮、注射、経腸、直腸内等の任意の経路を選択することができる。好ましくは点眼および経口投与である。 The aging eye disease ameliorating agent of the present invention includes, as additives, excipients, disintegrating agents, lubricants, binders, surfactants, fluidity promoters, colorings that are routinely used in the pharmaceutical industry. Agents, fragrances and the like can be added as appropriate to obtain a pharmaceutical composition of a desired dosage form. The administration route of the aging ophthalmic disease improving agent or pharmaceutical composition of the present invention may be any known route, for example, any route such as eye drops, oral, transdermal, injection, enteral, rectal, etc. Can be selected. Instillation and oral administration are preferred.
例えば、粉末剤、顆粒剤、錠剤、カプセル剤等の経口剤は、澱粉、乳糖、白糖、マンニット、カルボキシメチルセルロース、コーンスターチ、無機塩類等を用いて常法によって製剤化される。この種の製剤には、前記賦形剤の他に、結合剤、崩壊剤、界面活性剤、滑沢剤、流動性促進剤、着色料、香料等を適宜使用することができる。より具体的には、結合剤としては、例えば、澱粉、デキストリン、アラビアガム末、ゼラチン、ヒドロキシプロピルスターチ、カルボキシメチルセルロースナトリウム、メチルセルロース、結晶性セルロース、エチルセルロース、ポリビニルピロリドンが挙げられる。また、崩壊剤としては、例えば、澱粉、ヒドロキシプロピルスターチ、カルボキシメチルセルロースナトリウム、架橋カルボキシメチルセルロースナトリウム、結晶性セルロース、カルボキシメチルセルロース等が挙げられる。界面活性剤としては、大豆レシチン、蔗糖脂肪酸エステル等が、滑沢剤としては、タルク、ロウ、蔗糖脂肪酸エステル、水素添加植物油等が、流動性促進剤としては、無水ケイ酸、乾燥水酸化アルミニウム、ケイ酸マグネシウム等が、それぞれ挙げられる。 For example, oral preparations such as powders, granules, tablets and capsules are formulated by conventional methods using starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, inorganic salts and the like. In this type of preparation, in addition to the above-mentioned excipients, binders, disintegrants, surfactants, lubricants, fluidity promoters, colorants, fragrances, and the like can be appropriately used. More specifically, examples of the binder include starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch, sodium carboxymethylcellulose, methylcellulose, crystalline cellulose, ethylcellulose, and polyvinylpyrrolidone. Moreover, as a disintegrating agent, starch, hydroxypropyl starch, sodium carboxymethylcellulose, bridge | crosslinking sodium carboxymethylcellulose, crystalline cellulose, carboxymethylcellulose etc. are mentioned, for example. Surfactants include soy lecithin, sucrose fatty acid esters, lubricants include talc, wax, sucrose fatty acid esters, hydrogenated vegetable oils, etc., and fluidity promoters include anhydrous silicic acid and dry aluminum hydroxide. And magnesium silicate.
また、経口投与のほか、本発明の加齢性眼疾患改善剤、またはそれを含む医薬組成物または化粧料は、点眼等の方法で局所適用することによっても、加齢性眼疾患予防または治療効果を発揮する。局所適用剤とするには、その使用目的に応じて、通常用いられる公知の成分に配合することによって、液剤、固形剤、半固形剤等の各種剤形に調製することが可能であり、好ましい組成物としては、軟膏、ゲル、クリーム、スプレー剤、貼付剤、ローション、粉末等が挙げられる。例えば、本発明の加齢性眼疾患改善剤を、ワセリン等の炭化水素、ステアリルアルコール、ミリスチン酸イソプロピル等の高級脂肪酸低級アルキルエステル、ラノリン等の動物性油脂、グリセリン等の多価アルコール、グリセリン脂肪酸エステル、モノステアリン酸ポリエチレングリコール等の界面活性剤、無機塩、ロウ、樹脂、水、パラオキシ安息香酸メチル、パラオキシ安息香酸ブチル等の保存料等の１以上の成分と混合することによって、加齢性眼疾患改善用医薬組成物または化粧料を製造することができる。 In addition to oral administration, the age-related eye disease ameliorating agent of the present invention, or a pharmaceutical composition or cosmetic containing the same can be applied or applied locally by methods such as eye drops to prevent or treat age-related eye diseases. Demonstrate the effect. In order to obtain a topical application agent, it can be prepared into various dosage forms such as a liquid agent, a solid agent, a semi-solid agent and the like by blending with commonly known components depending on the purpose of use. Examples of the composition include ointments, gels, creams, sprays, patches, lotions, and powders. For example, the age-related eye disease ameliorating agent of the present invention includes hydrocarbons such as petrolatum, stearyl alcohol, higher fatty acid lower alkyl esters such as isopropyl myristate, animal fats and oils such as lanolin, polyhydric alcohols such as glycerin, glycerin fatty acids By mixing with one or more components such as ester, surfactant such as polyethylene glycol monostearate, inorganic salt, wax, resin, water, methyl paraoxybenzoate, butyl paraoxybenzoate, etc. A pharmaceutical composition for improving eye diseases or a cosmetic can be produced.
このような各種製剤の製造方法は、当業者には充分公知である。なお、本発明に関して「医薬組成物」という場合、人間に対して適用するもののほか、獣医学的に動物に対して適用するもの（獣医薬）をも含む。 Methods for producing such various preparations are well known to those skilled in the art. The term “pharmaceutical composition” in the context of the present invention includes not only those applied to humans but also those applied to animals veterinarily (veterinary medicine).
また、本発明の加齢性眼疾患改善剤は、経口投与するために、栄養剤や飲食品に添加して、飲食品または飼料としてヒトまたはヒト以外の対象動物に摂取させることもできる。このような飲食品または飼料の製造方法も当業者には公知である。 Moreover, in order to administer the age-related eye disease improvement agent of this invention orally, it can also be added to a nutrient or food-drinks, and can also be made to ingest in humans or a non-human target animal as food-drinks or feed. Such a method for producing a food or drink or feed is also known to those skilled in the art.
本発明の加齢性眼疾患改善剤またはそれを含む医薬組成物、飲食品、化粧料もしくは飼料を与えるべき対象となるのは、ヒトをはじめとして、ヒト以外の哺乳類、鳥類などの種々の動物であり、例えば、ウシ、ウマ、ブタ、ヒツジ、ヤギ、ニワトリ等の家畜、家禽類やイヌ、ネコ等のペット類が挙げられる。 The subject to be given the age-related eye disease ameliorating agent of the present invention or a pharmaceutical composition, food / beverage product, cosmetics or feed containing the same is not limited to humans, but also various animals such as mammals other than humans and birds. Examples thereof include livestock such as cows, horses, pigs, sheep, goats and chickens, and pets such as poultry, dogs and cats.
本発明の加齢性眼疾患改善剤の経口投与による有効量は、その製剤形態、投与方法、使用目的、および投与される対象の種類、年齢、体重、病状等によって異なり、それらに応じて各々に適した量で投与することができる。哺乳類に対して投与する場合、好ましくは体重１ｋｇあたり２ｍｇ／日以上とすることができる。ヒトに投与する場合、一般的には、有効成分量として、１日あたり８０ｍｇ〜１６０ｍｇ、例えば、成人１人当たり１日１２０ｍｇ以上の量であることができる。このような１日あたりの用量を一度にまたは分割して、本発明の加齢性眼疾患改善剤による加齢性眼疾患の予防、治療または状態の改善が必要とされている対象に対し、投与することができる。 The effective amount of the age-related eye disease ameliorating agent of the present invention by oral administration varies depending on the preparation form, administration method, purpose of use, and type, age, weight, medical condition, etc. of the subject to be administered, and depending on them, respectively. Can be administered in an amount suitable for the dose. When administered to a mammal, the dose can be preferably 2 mg / day or more per kg of body weight. When administered to humans, in general, the amount of the active ingredient may be 80 mg to 160 mg per day, for example, 120 mg or more per day per adult. For a subject in need of prevention, treatment or amelioration of age-related eye diseases with the agent for improving age-related eye diseases of the present invention by dividing such daily dose at once or divided, Can be administered.
本発明の加齢性眼疾患改善剤の点眼による有効量は、製剤形態、使用目的等により異なるが、適用する組成物全量を基準として、好ましくは０．００１〜２重量％となるように、ラクトフェリンを配合すればよい。 The effective amount of the aging eye disease ameliorating agent of the present invention by instillation varies depending on the preparation form, purpose of use, etc., but is preferably 0.001 to 2% by weight based on the total amount of the composition to be applied. What is necessary is just to mix | blend lactoferrin.
本発明を実施例により詳細に説明するが、本発明はそれに限定されるものではない。 The present invention will be described in detail by way of examples, but the present invention is not limited thereto.
実施例１：加齢性眼疾患改善剤の製造
ナトリウムイオン吸着型スルホン酸陽イオン交換樹脂４００ｇを充填したカラム（直径５ｃｍ×高さ３０ｃｍ）を脱イオン水で十分に洗浄した後、このカラムに未殺菌脱脂乳４０Ｌ（ｐＨ６．７）を流速２５ｍＬ／ｍｉｎで通液した。通液後、カラムを脱イオン水で十分洗浄し、２．０Ｍ塩化ナトリウムを含む０．０２Ｍ炭酸緩衝液（ｐＨ７．０）で溶出した。ラクトフェリンを含有する溶出画分をＳ−Ｓｅｐｈａｒｏｓｅ ＦＦカラムに吸着させ、脱イオン水で十分洗浄し、１０ｍＭリン酸緩衝液（ｐＨ７．０）で平衡化した後、０〜２．０Ｍ塩化ナトリウムの濃度勾配を溶出し、ラクトフェリンを含む画分を回収した。そして、その画分をゲル濾過クロマトグラフィーで処理し、凍結乾燥してラクトフェリン３．５ｇを得た。なお、このようにして得られたラクトフェリンの純度は９３％であり、そのまま加齢性眼疾患改善剤として使用可能である。
Example 1: Manufacture of an agent for improving age-related eye diseases A column (
実施例２：ラクトフェリンを含有する加齢性眼疾患改善医薬組成物
株式会社ＮＲＬファーマ製造の腸溶性ラクトフェリン粉末（純ラクトフェリンとして１０％を含有）を本発明の医薬組成物として使用した。
Example 2: Pharmaceutical composition for improving age-related eye diseases containing lactoferrin Enteric lactoferrin powder (containing 10% as pure lactoferrin) manufactured by NRL Pharma Co. , Ltd. was used as the pharmaceutical composition of the present invention.
試験例１：ラットにおける眼の加齢現象に及ぼす効果
ラットを実験動物として用い、眼の加齢現象に及ぼす効果を調べるために、生後１年８ヶ月〜２年半のウイスター系雌ラット（老齢）１８頭を無作為に２群に分け、一群を、ラクトフェリンを投与していない群（「老齢対照群」または「対照群」ともいう；ｎ＝９）、他方を、ラクトフェリンを投与した群（「老齢ラクトフェリン（ＬＦ）摂取群」または「老齢ＬＦ摂取群」ともいう；ｎ＝９）として４週間飼育し、涙腺を光学顕微鏡と透過電子顕微鏡で観察した。なお、ラットの寿命は約２年半であるから、実験に使用したラットは、人の７０〜８０歳代に相当する。
Test Example 1: Effects on the aging phenomenon of the eyes in rats In order to investigate the effects on the aging phenomenon of the eyes using rats as experimental animals, Wistar female rats (aged ages 1 to 8 months to 2 and a half years old) ) 18 were randomly divided into two groups, one group not receiving lactoferrin (also referred to as “old age control group” or “control group”; n = 9), and the other group receiving lactoferrin ( Also referred to as an “aged lactoferrin (LF) ingestion group” or “aged LF ingestion group”; n = 9), the lacrimal glands were observed with an optical microscope and a transmission electron microscope. In addition, since the lifetime of a rat is about two and a half years, the rat used for experiment is equivalent to a person's 70-80 generations.
両群とも飼料と飲料水は自由摂取とし、対照群には粉末のラット用標準飼料（日本クレア、ＣＥ−２）を与え、ＬＦ摂取群には対照群と同じラット用標準飼料に実施例２記載の医薬組成物粉末を４％添加した飼料（飼料中のＬＦ含量：０．４％）を与え、それぞれ４週間飼育した。同時に、若齢ラット群として、成熟期に達した５頭のラット（１０週齢若齢ラット）もＣＥ−２粉末を与えて４週間飼育した。 In both groups, feed and drinking water were allowed to be freely consumed, powdered standard diet for rats (Clea Japan, CE-2) was given to the control group, and the same standard diet for rats as the control group was given to the LF intake group. A feed (4% LF content in the feed) to which 4% of the described pharmaceutical composition powder was added was fed and bred for 4 weeks each. At the same time, as a group of young rats, 5 rats (10-week-old young rats) that reached maturity were also fed with CE-2 powder and bred for 4 weeks.
実験開始４週間後に各動物を屠殺し、それぞれの標本について涙腺摘出直前の体重および摘出直後の涙腺（左側）の重量を計測した。また、今回の光学顕微鏡および電子顕微鏡による観察には、対照群から６頭、ＬＦ摂取群から５頭をそれぞれ無作為に抽出した標本を用いた。それらの標本を用いて、常法にしたがって固定、脱水、包埋を行った後、準超薄トルイジン青染色切片ならびに超薄電子染色切片を作製し、光学顕微鏡および透過電子顕微鏡で観察した。なお、今回は老齢ラットにおけるラクトフェリン投与実験の評価を行う際に、成熟期に達した直後の若齢ラット（１０週齢若齢ラット）涙腺細胞の構造を参考とした。 Four weeks after the start of the experiment, each animal was sacrificed, and the body weight immediately before lacrimal gland extraction and the weight of the lacrimal gland (left side) just after excision were measured for each specimen. In addition, in this observation with an optical microscope and an electron microscope, specimens obtained by randomly extracting 6 animals from the control group and 5 animals from the LF intake group were used. Using these specimens, fixation, dehydration, and embedding were performed according to a conventional method, and then a semi-ultra thin toluidine blue stained section and an ultra thin electron stained section were prepared and observed with an optical microscope and a transmission electron microscope. In addition, this time, when evaluating the lactoferrin administration experiment in an old rat, the structure of the lacrimal gland cell of the young rat (10-week-old young rat) immediately after reaching the mature stage was referred.
涙腺の平均重量に関する所見
老齢ラットの対照群（９匹）における涙腺重量平均値は、０．１５ｇであり、ＬＦ摂取群（９匹）の涙腺重量平均値は、０．１８ｇであった。また、それぞれの個体における涙腺の対体重比を算出し、さらに各群における対体重比の算術平均を求めた結果、老齢対照群の対体重比平均値は０．０４５％であり、老齢ＬＦ摂取群においては０．０４９％であった。この実験では両群の間には統計学的有意差は認められなかったものの、以上のように涙腺の重量平均値および対体重比平均値ともに老齢ＬＦ摂取群が老齢対照群に比して僅かに大きい値となっていた。
Observation on Average Lacrimal Gland Weight The average lacrimal gland weight in the control group (9 animals) of the old rats was 0.15 g, and the lacrimal gland weight average value in the LF intake group (9 animals) was 0.18 g. In addition, the ratio of the lacrimal gland to body weight in each individual was calculated, and the arithmetic average of the body weight ratio in each group was calculated. As a result, the average value of body weight ratio in the elderly control group was 0.045%, and the elderly LF was ingested. In the group it was 0.049%. In this experiment, no statistically significant difference was found between the two groups, but as described above, both the weight average value of the lacrimal gland and the average value of body weight ratio were slightly higher in the elderly LF intake group than in the old control group. It was a large value.
涙腺細胞内の構造に関する光学顕微鏡的所見
エポキシ樹脂包埋の準超薄切片にトルイジン青染色を施して観察を行った結果、老齢ラット対照群の涙腺細胞内は、ほぼ均一の大きさでトルイジン青に均質に染色される顆粒状構造物（記載の便宜上、この顆粒を「Ａ顆粒」とする）によって占められている状態が多く認められた（図１；（ａ）は１００倍、（ｂ）は２５０倍）。また、それらの顆粒と同等ないしはやや大きい顆粒であってトルイジン青に淡染する明調顆粒も僅かに観察された。
Light microscopic observations on the structure of lacrimal gland cells Toluidine blue staining was performed on a semi-thin section of epoxy resin-embedded semi-thin sections. (FIG. 1; (a) is 100 times, (b). FIG. 1; (a) is 100 times larger). Is 250 times). In addition, a slight amount of light-colored granules that were equivalent to or slightly larger than those granules and lightly dyed toluidine blue were also observed.
老齢ＬＦ摂取群の涙腺細胞内には、老齢対照群で多く観察されるＡ顆粒もみられるが、中等度の大きさのトルイジン青に淡染する明調顆粒のほか、特に大型の明調顆粒が多数観察された（この種のトルイジン青に淡染する顆粒を「Ｂ顆粒」とする）（図２および３；図２（ａ）、（ｂ）および図３（ａ）は１００倍、図３（ｂ）は２５０倍）。また、Ｂ顆粒は大きいものほど不規則な外形を示す傾向があった。さらに拡大して観察を行うと、Ｂ顆粒内部の染色性は、近傍にみられる導管内部を満たしている構造物の染色性と類似した状態が認められた（図２（ｂ））。いずれにしても老齢ＬＦ摂取群においては、Ａ顆粒やＢ顆粒のほか、種々の染色性と大きさを示す顆粒状構造物が多数混在しているのが特徴的であった。 In the lacrimal gland cells of the aged LF intake group, A granules, which are often observed in the aged control group, are also observed, but in addition to the light-colored granules that are lightly stained with medium-sized toluidine blue, particularly large light-colored granules are present. Many observed (granules that lightly dye this type of toluidine blue are referred to as “B granules”) (FIGS. 2 and 3; FIGS. 2 (a), (b) and 3 (a) are 100 times larger, FIG. (B) is 250 times). In addition, the larger the B granules, the more apt to show irregular shapes. When further observing, the stainability inside the B granule was found to be similar to the stainability of the structure filling the inside of the conduit seen in the vicinity (FIG. 2 (b)). In any case, in the aged LF intake group, in addition to A granules and B granules, a large number of granular structures having various staining properties and sizes are characteristically mixed.
若齢ラットにおける涙腺細胞内の顆粒状構造物の分布および形態は、老齢のＬＦ摂取群と極めて類似した特徴を有していた（図４；（ａ）は１００倍、（ｂ）は２５０倍）。若齢ラットにおけるＢ顆粒の出現度を基準（＋＋）として、老齢ラットの涙腺細胞内に認められるＢ顆粒の出現度を評価した結果を表１に示す。 The distribution and morphology of granular structures in lacrimal gland cells in young rats had characteristics very similar to those in the elderly LF intake group (FIG. 4; (a) was 100 times, (b) was 250 times. ). Table 1 shows the results of evaluating the degree of appearance of B granules found in lacrimal gland cells of old rats, using the degree of appearance of B granules in young rats as a reference (++).
＊ 若齢ラットにおけるＢ顆粒の出現度を基準（＋＋）として、老齢ラットの各個体におけるＢ顆粒の出現状態を示す。（＋＋）は若齢ラットにおけるＢ顆粒の出現度と同程度、（＋＋＋）はそれより高い出現度を示し、（＋）はやや低く、（−）はかなり低い状態であることを示している。 * Appearance state of B granules in each individual of old rats is shown with reference to the degree of appearance of B granules in young rats (++). (++) is similar to the appearance of B granules in young rats, (++) is higher, (+) is slightly lower, (-) is much lower .
老齢対照群においても一部の個体（個体番号Ｃ−９）では、若齢ラットと同程度のＢ顆粒の出現度を示す個体も認められたが、他の個体は＋ないし−となっており、Ａ顆粒が主体をなしていることがわかる。一方、ＬＦ摂取老齢ラット群では、いずれの個体も＋＋あるいは＋＋＋となっており、若齢ラットにおけるＢ顆粒の出現度と同等、あるいはそれ以上の結果を示している。 Even in the old age control group, some individuals (individual number C-9) showed the appearance of B granules at the same level as young rats, but other individuals were + or-. It can be seen that the A granule is the main component. On the other hand, in the LF-incorporated elderly rat group, all individuals are ++ or ++, and the results are equal to or higher than the appearance degree of B granules in young rats.
涙腺細胞内の構造に関する透過電子顕微鏡的所見
対照老齢ラットの涙腺細胞内にみられる顆粒の主体をなすＡ顆粒の内部は、電子密度がやや高く、ほぼ均一な大きさを示す微細粒子によって満たされていた（図５；（ａ）は２，９００倍、（ｂ）は５，８００倍）。一方、ＬＦ摂取老齢ラットに多数認められる大型のＢ顆粒の内部には、電子密度の高い羽毛状の構造物とそれらの構造物の間に介在するコロイド状の物質が観察された（図６および図７（ａ）；図６（ａ）は２，９００倍、図６（ｂ）は５，８００倍、図７（ａ）は７，３００倍）。また、これらの顆粒内にみられる羽毛状の構造物とコロイド状の物質は、その近傍の導管内にも認められた（図７（ｂ）；７，３００倍）。中等度の大きさのＢ顆粒は、互いに融合して不規則な外形をした大型のＢ顆粒になると思われる像も観察された。
Transmission electron microscopic observation on the structure of lacrimal gland cells The inside of granule A, which is the main granule found in lacrimal gland cells of aged rats, is filled with fine particles with slightly higher electron density and almost uniform size. (FIG. 5; (a) is 2,900 times, (b) is 5,800 times). On the other hand, inside the large B granules observed in many LF-ingested aged rats, feather-like structures with high electron density and colloidal substances interposed between these structures were observed (FIG. 6 and FIG. 6). FIG. 7 (a); FIG. 6 (a) is 2,900 times, FIG. 6 (b) is 5,800 times, and FIG. 7 (a) is 7,300 times. Moreover, the feather-like structure and colloidal substance which are seen in these granules were recognized also in the nearby conduit (FIG. 7 (b); 7,300 times). An image was also observed in which moderately sized B granules fused to each other to form large B granules having an irregular shape.
ラクトフェリンを投与していない若齢ラットの涙腺細胞内にみられる顆粒の形態や顆粒内ならびに導管内の微細構造は、老齢ＬＦ摂取群のものと極めて類似した特徴を有していた（図８および図９；図８（ａ）は２，９００倍、図８（ｂ）、図９（ａ）および（ｂ）は７，３００倍）。 The granule morphology and intragranular and intraductal microstructure found in the lacrimal gland cells of young rats not administered lactoferrin had characteristics very similar to those in the aged LF intake group (Fig. 8 and FIG. 9: FIG. 8 (a) is 2,900 times, FIG. 8 (b), FIGS. 9 (a) and 9 (b) are 7,300 times).
本発明の医薬組成物を経口摂取した老齢ラット（ＬＦ摂取老齢ラット群）においては、これを投与しなかった老齢ラット（対照老齢ラット群）に比して、涙腺細胞内に種々の大きさや構造を有する顆粒が豊富に認められ、それらの顆粒が互いに融合して大型化していくと思われる状態も観察された。また、大型のＢ顆粒内にみられる構造物（顆粒内構造物）は、近傍の導管内に認められる構造物と形態的に極めて類似していることから、大型のＢ顆粒は導管に排出される段階に近い状態にまで成熟した分泌顆粒であると考えられる。 In the aged rat (LF-ingested aged rat group) orally ingested the pharmaceutical composition of the present invention, various sizes and structures in the lacrimal gland cells as compared to the aged rat (control aged rat group) that was not administered. There were also abundant granules having a size, and a state in which these granules seemed to be fused and enlarged was also observed. In addition, since the structure (intragranular structure) found in the large B granule is very similar in form to the structure found in the nearby conduit, the large B granule is discharged into the conduit. It is thought that it is a secretory granule that has matured to a state close to that stage.
以上の所見によって、比較的均一な大きさと構造を示す顆粒（Ａ顆粒）によって占められている対照老齢ラットの涙腺細胞に比べて、ラクトフェリンを投与した老齢ラットにおいては、分泌顆粒が旺盛に形成されている可能性が示唆された。 Based on the above findings, secretory granules were actively formed in the aged rats administered with lactoferrin as compared to the lacrimal gland cells of the control aged rats occupied by granules (A granules) showing relatively uniform size and structure. The possibility is suggested.
すなわち、大型のＢ顆粒がラクトフェリンを投与した老齢ラットの涙腺細胞内において極めて豊富に観察されるのは、ラクトフェリンの投与により、涙腺細胞の分泌機能が亢進していることを示す形態的特徴であると考えられる。 That is, it is a morphological feature that the lacrimal gland cell secretion function is enhanced by the administration of lactoferrin that large B granules are observed in abundantly in the lacrimal gland cells of old rats administered with lactoferrin. it is conceivable that.
また、ラクトフェリンを投与していない若齢ラットの涙腺細胞における観察でも、ラクトフェリンを投与した老齢ラットとほぼ同様な所見が認められることから、ＬＦ摂取老齢ラット群にみられる構造的特徴は、ラクトフェリン投与の影響で生じた異常所見である可能性はないものと理解される。また、涙腺の重量平均値ならびに対体重比平均値ともに両群の間には統計学的有意差は認められなかったことから、ラクトフェリン投与が特に老齢ラットにおける涙腺の重量増加を促すように作用したのではなく、分泌機能の亢進に作用した可能性があると思われる。すなわち、加齢によって変性した組織の若返り活性をラクトフェリンが有することが確認された。 In addition, the observation in the lacrimal gland cells of young rats not administered lactoferrin shows almost the same findings as the old rats administered lactoferrin. It is understood that there is no possibility of abnormal findings caused by In addition, there was no statistically significant difference between the two groups in both the weight average value of the lacrimal gland and the mean value of body weight ratio, so lactoferrin administration acted to promote an increase in the weight of the lacrimal gland especially in aged rats It seems that it may have acted on the enhancement of secretory function. That is, it was confirmed that lactoferrin has the rejuvenation activity of tissues denatured by aging.
なお、通常のＬＦ投与実験では、ＬＦを２〜４％の割合で混和した飼料が多用されているが、本実験ではその１／１０に相当する正味０．２〜０．４％のＬＦを投与したにも関わらず、ＬＦの効果を認めることができた。これは、本実験で用いた医薬組成物が腸溶性製剤であることに起因する可能性も高い。本実験から腸溶性粉末ラクトフェリンの経口摂取により、老齢ラットにおける涙腺細胞の分泌機能が亢進すると思われる形態的特徴が認められた。 In a normal LF administration experiment, feed mixed with LF at a ratio of 2 to 4% is frequently used. In this experiment, a net 0.2 to 0.4% LF corresponding to 1/10 of the feed is used. Despite the administration, the effect of LF could be recognized. This is highly likely due to the fact that the pharmaceutical composition used in this experiment is an enteric preparation. From this experiment, morphological features that the ingestion of enteric powdered lactoferrin is thought to enhance the secretory function of lacrimal gland cells in aged rats were observed.
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005033913A JP4300324B2 (en) | 2005-02-10 | 2005-02-10 | Age-related eye disease ameliorating agent, and pharmaceutical composition, food and drink, cosmetics and feed containing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005033913A JP4300324B2 (en) | 2005-02-10 | 2005-02-10 | Age-related eye disease ameliorating agent, and pharmaceutical composition, food and drink, cosmetics and feed containing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2006219415A JP2006219415A (en) | 2006-08-24 |
| JP4300324B2 true JP4300324B2 (en) | 2009-07-22 |
Family
ID=36981960
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2005033913A Active JP4300324B2 (en) | 2005-02-10 | 2005-02-10 | Age-related eye disease ameliorating agent, and pharmaceutical composition, food and drink, cosmetics and feed containing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4300324B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007031403A (en) * | 2005-07-29 | 2007-02-08 | Morinaga Milk Ind Co Ltd | Lactic acid production inhibitor |
| JP5302546B2 (en) * | 2008-02-12 | 2013-10-02 | 雪印メグミルク株式会社 | Cataract preventive agent |
-
2005
- 2005-02-10 JP JP2005033913A patent/JP4300324B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| JP2006219415A (en) | 2006-08-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2618832B1 (en) | Composition comprising sod, lutein and zeaxanthin | |
| AU2012209285B2 (en) | Methods and compositions for treating, reducing or preventing deterioration of the visual system of animals | |
| CA2979112C (en) | Eye health composition | |
| JP6638092B2 (en) | Use of pyrroloquinoline quinone, its derivatives and / or salts in dry syndrome and pharmaceutical compositions | |
| KR20190034638A (en) | Composition containing a benzoate compound and tannic acid for treating central nervous system disorders | |
| JP5865242B2 (en) | Composition for treating eye strain | |
| JP2015523333A (en) | Cosmetic, pharmaceutical and food composition containing a crushed or extracted fish eyeball | |
| JP4300324B2 (en) | Age-related eye disease ameliorating agent, and pharmaceutical composition, food and drink, cosmetics and feed containing the same | |
| JP6869367B2 (en) | Composition for prevention and treatment of diseases related to deterioration of Bruch membrane function containing sea cucumber extract as an active ingredient | |
| KR102080864B1 (en) | Functional feed composition for pet using microalgae having antiobecity and antidiabetes | |
| CN117580573A (en) | L-ergothioneine for the relief and prevention of age-related vision deterioration | |
| ES2940767T3 (en) | Nutraceutical ophthalmic composition for the treatment of retinal pathologies with a neovascular component | |
| EP2709643B1 (en) | Compositions comprising extracts or materials derived from palm oil vegetation liguor for inhibition of vision loss due to macular degeneration | |
| KR20230032154A (en) | Exosomes as the composition for the treatment and prevention of dry eye syndrome | |
| EP4096414A1 (en) | Undenatured type ii collagen in food and beverage applications and uses thereof | |
| WO2021195068A1 (en) | Undenatured type ii collagen in animal food and treats | |
| WO2022040026A1 (en) | Method for improving inflammation, joint health, joint mobility, and joint comfort in healthy mammals | |
| JP2008505097A (en) | Compounds and methods for treating diabetic vascular disease | |
| US10183054B2 (en) | Compositions comprising extracts or materials derived from palm oil vegetation liquor for inhibition of vision loss due to angiogenesis and method of preparation there | |
| TWI789123B (en) | Using Growth Factors to Treat Arthritis | |
| RU2806096C2 (en) | Nutriceutical ophthalmological composition for treatment of retinal pathologies with a neovascular component | |
| TW202321280A (en) | Use of growth factor to treat arthritis being applied to preparation of a composition for treating the arthritis | |
| KR101751617B1 (en) | A composition comprising peptide extracts of ogye and method thereof | |
| Hung et al. | Dragon fruit (Hylocereus undatus (Haw.) Britt.) powders attenuate ulcerative disease symptoms via dextran sulfate sodium-induced mouse model | |
| KR20200042745A (en) | Composition for protecting neuronal cells comprising ginsenoside Rb2 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20071219 |
|
| RD01 | Notification of change of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7426 Effective date: 20080125 |
|
| A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20080430 |
|
| A975 | Report on accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A971005 Effective date: 20080522 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20080610 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20080805 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20081021 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20081217 |
|
| 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: 20090303 |
|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20090401 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20090401 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120501 Year of fee payment: 3 |
|
| R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 Ref document number: 4300324 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130501 Year of fee payment: 4 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20140501 Year of fee payment: 5 |
|
| 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 |
|
| 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 |
|
| 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 |
|
| 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 |