JP2010155856A - Osteogenesis promoting and bone resorption inhibiting agent - Google Patents
Osteogenesis promoting and bone resorption inhibiting agent Download PDFInfo
- Publication number
- JP2010155856A JP2010155856A JP2010046910A JP2010046910A JP2010155856A JP 2010155856 A JP2010155856 A JP 2010155856A JP 2010046910 A JP2010046910 A JP 2010046910A JP 2010046910 A JP2010046910 A JP 2010046910A JP 2010155856 A JP2010155856 A JP 2010155856A
- Authority
- JP
- Japan
- Prior art keywords
- bone
- bone resorption
- plasminogen
- prothrombin
- transferrin
- 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.)
- Granted
Links
- 208000006386 Bone Resorption Diseases 0.000 title claims abstract description 50
- 230000024279 bone resorption Effects 0.000 title claims abstract description 50
- 230000011164 ossification Effects 0.000 title claims abstract description 47
- 230000001737 promoting effect Effects 0.000 title claims abstract description 24
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 16
- 102000013566 Plasminogen Human genes 0.000 claims abstract description 36
- 108010051456 Plasminogen Proteins 0.000 claims abstract description 36
- 108090000901 Transferrin Proteins 0.000 claims abstract description 34
- 102000004338 Transferrin Human genes 0.000 claims abstract description 34
- 239000012581 transferrin Substances 0.000 claims abstract description 34
- 108010015742 Cytochrome P-450 Enzyme System Proteins 0.000 claims abstract description 32
- 102000002004 Cytochrome P-450 Enzyme System Human genes 0.000 claims abstract description 32
- 108010088842 Fibrinolysin Proteins 0.000 claims abstract description 29
- 229940012957 plasmin Drugs 0.000 claims abstract description 29
- 239000004480 active ingredient Substances 0.000 claims abstract description 19
- 235000013305 food Nutrition 0.000 claims abstract description 14
- 239000003814 drug Substances 0.000 claims abstract description 10
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 4
- 229940078581 Bone resorption inhibitor Drugs 0.000 claims description 15
- 230000001629 suppression Effects 0.000 claims description 10
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- 235000013361 beverage Nutrition 0.000 claims description 4
- 229940079593 drug Drugs 0.000 abstract description 6
- 230000005764 inhibitory process Effects 0.000 abstract 1
- 108010094028 Prothrombin Proteins 0.000 description 43
- 102100027378 Prothrombin Human genes 0.000 description 42
- 229940039716 prothrombin Drugs 0.000 description 42
- 108090000190 Thrombin Proteins 0.000 description 26
- 229960004072 thrombin Drugs 0.000 description 26
- 210000000988 bone and bone Anatomy 0.000 description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 18
- 239000000203 mixture Substances 0.000 description 15
- 235000013336 milk Nutrition 0.000 description 12
- 210000004080 milk Anatomy 0.000 description 12
- 239000008267 milk Substances 0.000 description 12
- 235000018102 proteins Nutrition 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 108090000623 proteins and genes Proteins 0.000 description 12
- 239000002994 raw material Substances 0.000 description 11
- 239000000126 substance Substances 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 10
- 238000009472 formulation Methods 0.000 description 10
- 230000009471 action Effects 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 210000000963 osteoblast Anatomy 0.000 description 9
- 239000011780 sodium chloride Substances 0.000 description 9
- 238000012360 testing method Methods 0.000 description 8
- 208000020084 Bone disease Diseases 0.000 description 7
- 208000001132 Osteoporosis Diseases 0.000 description 7
- 210000002997 osteoclast Anatomy 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 238000005728 strengthening Methods 0.000 description 7
- 241000283690 Bos taurus Species 0.000 description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 5
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 229960005069 calcium Drugs 0.000 description 5
- 229910052791 calcium Inorganic materials 0.000 description 5
- 239000011575 calcium Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000002244 precipitate Substances 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- 108010046377 Whey Proteins Proteins 0.000 description 4
- 102000007544 Whey Proteins Human genes 0.000 description 4
- LZCZIHQBSCVGRD-UHFFFAOYSA-N benzenecarboximidamide;hydron;chloride Chemical compound [Cl-].NC(=[NH2+])C1=CC=CC=C1 LZCZIHQBSCVGRD-UHFFFAOYSA-N 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 208000008035 Back Pain Diseases 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- 239000004365 Protease Substances 0.000 description 3
- 235000015895 biscuits Nutrition 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 229910052742 iron Inorganic materials 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 235000020183 skimmed milk Nutrition 0.000 description 3
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010014173 Factor X Proteins 0.000 description 2
- 102000009123 Fibrin Human genes 0.000 description 2
- 108010073385 Fibrin Proteins 0.000 description 2
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 2
- 206010017076 Fracture Diseases 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 108010001014 Plasminogen Activators Proteins 0.000 description 2
- 102000001938 Plasminogen Activators Human genes 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- 229920002684 Sepharose Polymers 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 229930003448 Vitamin K Natural products 0.000 description 2
- 239000005862 Whey Substances 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical class N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 229910052788 barium Inorganic materials 0.000 description 2
- DSAJWYNOEDNPEQ-UHFFFAOYSA-N barium atom Chemical compound [Ba] DSAJWYNOEDNPEQ-UHFFFAOYSA-N 0.000 description 2
- 229940006612 barium citrate Drugs 0.000 description 2
- PAVWOHWZXOQYDB-UHFFFAOYSA-H barium(2+);2-hydroxypropane-1,2,3-tricarboxylate Chemical compound [Ba+2].[Ba+2].[Ba+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O PAVWOHWZXOQYDB-UHFFFAOYSA-H 0.000 description 2
- 210000002798 bone marrow cell Anatomy 0.000 description 2
- 210000002805 bone matrix Anatomy 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 description 2
- 235000013351 cheese Nutrition 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 210000001612 chondrocyte Anatomy 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 210000003278 egg shell Anatomy 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 229950003499 fibrin Drugs 0.000 description 2
- DWYMPOCYEZONEA-UHFFFAOYSA-L fluoridophosphate Chemical compound [O-]P([O-])(F)=O DWYMPOCYEZONEA-UHFFFAOYSA-L 0.000 description 2
- 235000013350 formula milk Nutrition 0.000 description 2
- 235000015110 jellies Nutrition 0.000 description 2
- 239000008274 jelly Substances 0.000 description 2
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 2
- 229940127126 plasminogen activator Drugs 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 235000019419 proteases Nutrition 0.000 description 2
- 235000020185 raw untreated milk Nutrition 0.000 description 2
- 238000007634 remodeling Methods 0.000 description 2
- 238000001223 reverse osmosis Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000012064 sodium phosphate buffer Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000009469 supplementation Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 235000019168 vitamin K Nutrition 0.000 description 2
- 239000011712 vitamin K Substances 0.000 description 2
- 150000003721 vitamin K derivatives Chemical class 0.000 description 2
- 229940046010 vitamin k Drugs 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 235000021119 whey protein Nutrition 0.000 description 2
- 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
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- WOVKYSAHUYNSMH-RRKCRQDMSA-N 5-bromodeoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-RRKCRQDMSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 102000055006 Calcitonin Human genes 0.000 description 1
- 108060001064 Calcitonin Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 108010084457 Cathepsins Proteins 0.000 description 1
- 102000005600 Cathepsins Human genes 0.000 description 1
- 108090000317 Chymotrypsin Proteins 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 206010072132 Fracture pain Diseases 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 108010051815 Glutamyl endopeptidase Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000798109 Homo sapiens Melanotransferrin Proteins 0.000 description 1
- 102000008133 Iron-Binding Proteins Human genes 0.000 description 1
- 108010035210 Iron-Binding Proteins Proteins 0.000 description 1
- 108060005987 Kallikrein Proteins 0.000 description 1
- 102000001399 Kallikrein Human genes 0.000 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 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
- 102100032239 Melanotransferrin Human genes 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 108090001109 Thermolysin Proteins 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000007059 acute toxicity Effects 0.000 description 1
- 231100000403 acute toxicity Toxicity 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- OFHCOWSQAMBJIW-AVJTYSNKSA-N alfacalcidol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)C[C@H](O)C1=C OFHCOWSQAMBJIW-AVJTYSNKSA-N 0.000 description 1
- 239000003957 anion exchange resin Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 description 1
- 229910001626 barium chloride Inorganic materials 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 229940019700 blood coagulation factors Drugs 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 108010009896 bone resorption factor Proteins 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 1
- 229960004015 calcitonin Drugs 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 description 1
- 239000001527 calcium lactate Substances 0.000 description 1
- 229960002401 calcium lactate Drugs 0.000 description 1
- 235000011086 calcium lactate Nutrition 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229960002376 chymotrypsin Drugs 0.000 description 1
- 235000013353 coffee beverage Nutrition 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 238000000909 electrodialysis Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 239000006167 equilibration buffer Substances 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 235000014058 juice drink Nutrition 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000011034 membrane dialysis Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 235000020124 milk-based beverage Nutrition 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 229940127293 prostanoid Drugs 0.000 description 1
- 150000003814 prostanoids Chemical class 0.000 description 1
- 230000002787 reinforcement Effects 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
- 235000013580 sausages Nutrition 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 229960001322 trypsin Drugs 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Non-Alcoholic Beverages (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
本発明は、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を有効成分とする骨形成促進及び骨吸収抑制剤に関する。また、本発明は、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を配合した骨形成促進及び骨吸収抑制用飲食品、医薬又は飼料に関する。 The present invention relates to a bone formation promotion and bone resorption inhibitor comprising as an active ingredient any one or more of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin. The present invention also relates to a food / beverage product, pharmaceutical or feed for promoting bone formation and suppressing bone resorption, which contains at least one of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin.
近年、高齢者人口の増加に伴い、骨粗鬆症、骨折、腰痛等の各種骨疾患が増加する傾向にある。骨組織においては、絶えず骨形成と骨吸収が営まれており、若い時には骨形成と骨吸収のバランスが保たれているが、加齢に伴い種々の原因でそのバランスが骨吸収に傾く(アンカップリング)。そして、この状態が長期間続くと骨組織が脆くなり、骨粗鬆症、骨折、腰痛等の各種骨疾患を生じることになる。このアンカップリングを防止することができれば、骨粗鬆症、骨折、腰痛等の各種骨疾患を予防することができると考えられている。 In recent years, various bone diseases such as osteoporosis, fractures, and back pain tend to increase as the elderly population increases. In bone tissue, bone formation and bone resorption are constantly carried out, and the balance between bone formation and bone resorption is maintained when young, but the balance tends to bone resorption due to various causes (aging) Coupling). If this state continues for a long period of time, the bone tissue becomes brittle, and various bone diseases such as osteoporosis, fractures, and back pain occur. If this uncoupling can be prevented, it is considered that various bone diseases such as osteoporosis, fracture, and back pain can be prevented.
従来より、アンカップリングを防止し、各種骨疾患を予防あるいは治療する方法として、(1)食餌によるカルシウム補給、(2)軽い運動、(3)日光浴、(4)薬物治療等が行われている。食餌によるカルシウム補給には、炭酸カルシウム、リン酸カルシウム等のカルシウム塩や卵殻、魚骨粉等の天然カルシウム剤が使用されている。しかし、これらは必ずしも経口摂取に適している素材であるとはいえない。
軽い運動はジョギングや散歩等が良いとされるが、体が弱っている場合は軽い運動も厄介なものであり、まして寝たきりの老人では殆ど運動できない。日光浴は活性型ビタミンD3の補給という点では良いとされているが、これだけでは不充分である。薬物投与には、1α−ヒドロキシビタミンD3やカルシトニン製剤等が使用されており、骨粗鬆症の治療には有効であるということが知られている。しかし、これらの物質は医薬そのものであり、食品素材としても使用可能なものではない。
Conventionally, as methods for preventing uncoupling and preventing or treating various bone diseases, (1) dietary calcium supplementation, (2) light exercise, (3) sunbathing, (4) drug treatment, and the like have been performed. Yes. For calcium supplementation with diet, calcium salts such as calcium carbonate and calcium phosphate, and natural calcium preparations such as eggshell and fish bone powder are used. However, these are not necessarily materials suitable for oral intake.
Light exercise is good for jogging or taking a walk, but if the body is weak, light exercise is troublesome, and even a bedridden elderly person can hardly exercise. Sunbathing is considered good in terms of supplementing active vitamin D 3 , but it is not enough. For drug administration, 1α-hydroxyvitamin D 3 , calcitonin preparation and the like are used, and it is known that they are effective for the treatment of osteoporosis. However, these substances are medicines themselves and cannot be used as food materials.
一方、本発明者らは、食品素材として使用可能な骨強化作用や骨吸収抑制作用を有する物質を得るために、乳清タンパク質中に存在する骨強化及び骨吸収抑制因子を探索し続けてきた。その結果、逆浸透膜や電気透析等の処理により、乳清タンパク質の水溶性画分から乳清由来の塩を除去したタンパク質及びペプチド混合物に骨強化作用があることを見出した (例えば、特許文献1参照。)。そして、このタンパク質及びペプチド混合物の水溶液をエタノール処理、加熱処理、加塩処理、限外濾過膜処理して得られる画分に骨芽細胞増殖促進作用及び骨強化作用があることを見出した(例えば、特許文献2及び3参照。)。また、乳中に微量にしか存在しない塩基性タンパク質に骨芽細胞増殖促進作用、骨強化作用及び骨吸収抑制作用があることを見出した(例えば、特許文献4参照。)。 On the other hand, the present inventors have continued to search for bone strengthening and bone resorption inhibiting factors present in whey protein in order to obtain a substance having bone strengthening action and bone resorption inhibiting action that can be used as a food material. . As a result, it was found that a protein and peptide mixture obtained by removing whey-derived salts from a water-soluble fraction of whey protein by treatment such as reverse osmosis membrane or electrodialysis has a bone strengthening action (for example, Patent Document 1). reference.). And, it was found that the fraction obtained by subjecting the aqueous solution of the protein and peptide mixture to ethanol treatment, heat treatment, salt treatment, and ultrafiltration membrane treatment has an osteoblast proliferation promoting action and a bone strengthening action (for example, (See Patent Documents 2 and 3.) Moreover, it discovered that the basic protein which exists only in trace amount in milk has an osteoblast proliferation promotion effect, a bone reinforcement effect | action, and a bone resorption suppression effect (for example, refer patent document 4).
本発明者らは、更に探索を進め、骨形成促進及び骨吸収抑制作用を有する成分の分離精製を試み、その物質を同定したところ、既知物質であるプロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンであることを確認した。そしてこれらの物質を骨形成促進及び骨吸収抑制剤として利用することができることを見出し、本発明を完成するに至った。
プロトロンビンは、分子量約7.2kDaのタンパク質であり、血液凝固II因子とも呼ばれる。ビタミンK依存性血液凝固因子の1つであり、ビタミンK存在下で第X因子によりトロンビンに分解される。生じたトロンビンはフィブリノーゲンをフィブリンへと分解し、フィブリンが架橋することによって血液が凝固する。プロトロンビンは、血液を始め、全身組織中に広く分布していることが知られており、ヒトやウシでは乳中にも存在していることが報告されている。
プロトロンビンやトロンビンの骨強化及び骨吸収抑制等の骨への作用についてはこれまで報告されていない。
The present inventors further advanced the search, tried to separate and purify a component having an action of promoting bone formation and suppressing bone resorption, and identified the substance. Prothrombin, cytochrome P-450, plasminogen, which are known substances, It was confirmed to be transferrin, thrombin and plasmin. And it discovered that these substances could be utilized as a bone formation promotion and bone resorption inhibitor, and came to complete this invention.
Prothrombin is a protein having a molecular weight of about 7.2 kDa and is also called blood coagulation factor II. It is one of vitamin K-dependent blood coagulation factors, and is decomposed into thrombin by factor X in the presence of vitamin K. The generated thrombin breaks fibrinogen into fibrin, and the blood is coagulated by fibrin cross-linking. Prothrombin is known to be widely distributed in whole body tissues including blood, and it has been reported that it is also present in milk in humans and cows.
The effects of prothrombin and thrombin on bone such as bone strengthening and bone resorption suppression have not been reported so far.
チトクロームP-450(CytochromeP-450)は、還元型で一酸化炭素を結合して450nm付近にソーレー吸収帯を示す一群のプロトヘムタンパク質の総称である。機能的には、各種のステロイドホルモン、胆汁酸、プロスタノイドの合成・分解反応、脂肪酸のω酸化、ビタミンDの活性化反応、無数の外来薬物や体内に取り込まれた環境汚染物質等の酸化的解毒反応に関与している。 Cytochrome P-450 (Cytochrome P-450) is a generic name for a group of protoheme proteins that bind to carbon monoxide in a reduced form and show a Soret absorption band near 450 nm. Functionally, various steroid hormones, bile acids, prostanoid synthesis / decomposition reactions, fatty acid ω oxidation, vitamin D activation reactions, oxidative effects of numerous foreign drugs and environmental pollutants taken into the body Involved in detoxification reactions.
プラスミノーゲン(Plasminogen)は、プラスミンの前駆体であり、動物血漿中に存在し、プラスミノーゲンアクチベーターによって分子内のArg-Val結合が切断されて活性なプラスミンとなる。分子量約91,000の一本鎖の糖タンパク質であり、プラスミン等のプロテアーゼによってN末端側のペプチドが切断されて分子量約82,000の修飾プラスミノーゲンとなる。 Plasminogen is a precursor of plasmin, exists in animal plasma, and plasminogen activator cleaves the Arg-Val bond in the molecule into active plasmin. It is a single-chain glycoprotein having a molecular weight of about 91,000, and a peptide on the N-terminal side is cleaved by a protease such as plasmin to form a modified plasminogen having a molecular weight of about 82,000.
トランスフェリン(Transferrin)は、血中の輸送鉄と結合する分子量約75,000のタンパク質で、鉄結合性グロブリンとも呼ばれる。幼若赤血球はトランスフェリンと結合した鉄に対する受容体を有しており、血清鉄がヘモグロビンの合成に利用されるためにはトランスフェリンと結合していなければならない。トランスフェリンについては膜結合型トランスフェリン様タンパク質が軟骨細胞の形成を促進すると開示されているが、軟骨細胞の形成促進であり、骨形成促進とはメカニズムが全く異なっている (例えば、特許文献5及び6参照。)。 Transferrin is a protein with a molecular weight of about 75,000 that binds to transported iron in the blood, and is also called iron-binding globulin. Immature erythrocytes have receptors for iron bound to transferrin, and serum iron must be bound to transferrin in order to be used for hemoglobin synthesis. Regarding transferrin, it has been disclosed that a membrane-bound transferrin-like protein promotes the formation of chondrocytes, but it is the promotion of chondrocyte formation, and the mechanism is completely different from the promotion of bone formation (for example, Patent Documents 5 and 6). reference.).
プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれについても骨形成を促進したり、骨吸収を抑制する働きについて記載した文献は見られない。 There is no literature describing the action of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin on promoting bone formation or inhibiting bone resorption.
本発明は、食品素材としても使用可能な、長期的に経口摂取することができ、安全な物質であって、骨形成促進及び骨吸収抑制作用を有する、骨形成促進及び骨吸収抑制剤を得ることを課題とする。また、本発明は、骨形成促進及び骨吸収抑制用飲食品、医薬品又は飼料を得ることを課題とする。 The present invention provides a bone formation promotion and bone resorption inhibitor that can be used as a food material, can be taken orally for a long period of time, is a safe substance, and has a bone formation promotion and bone resorption suppression action. This is the issue. Moreover, this invention makes it a subject to obtain the food-drinks, pharmaceuticals, or feed for bone formation promotion and bone resorption suppression.
本発明者らは、これらの問題点を鑑み、広く食品素材に含まれている骨強化作用を示す物質について、鋭意、探索を進め、骨芽細胞増殖促進作用及び骨吸収抑制作用を有するタンパク質について、骨形成促進及び骨吸収抑制作用を有する成分の分離精製を試み、その物質を同定したところ、既知物質であるプロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンであることを確認した。そしてこれらの物質を骨芽細胞増殖促進及び骨吸収抑制剤として利用することができることを見出し、本発明を完成するに至った。
すなわち、本発明は、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を有効成分とする骨形成促進及び骨吸収抑制剤に関する。
また、本発明は、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を配合した骨形成促進及び骨吸収抑制用飲食品、医薬又は飼料に関する。
In view of these problems, the present inventors have eagerly and continuously searched for substances that have a bone strengthening action widely contained in food materials, and have a protein having an osteoblast proliferation promoting action and a bone resorption inhibiting action. Attempts were made to isolate and purify components that have bone formation promotion and bone resorption suppression effects, and the substances were identified and confirmed to be known substances prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin. did. And it discovered that these substances can be utilized as an osteoblast proliferation promotion and a bone resorption inhibitor, and came to complete this invention.
That is, the present invention relates to a bone formation promotion and bone resorption inhibitor comprising as an active ingredient any one or more of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin.
The present invention also relates to a food / beverage product, pharmaceutical or feed for promoting bone formation and suppressing bone resorption, which contains at least one of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin.
本発明のプロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を有効成分とする骨形成促進及び骨吸収抑制剤は、これを経口投与することにより、骨粗鬆症等の各種骨疾患の予防や改善に有用である。また、この有効成分を飲食品、医薬、飼料等に配合すると骨形成を促進し、骨吸収を抑制して、骨粗鬆症等の各種骨疾患を予防し、改善する効果を奏する。 The bone formation promoting and bone resorption inhibitor comprising any one or more of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin, and plasmin of the present invention as an osteoporosis, etc. It is useful for the prevention and improvement of various bone diseases. In addition, when this active ingredient is blended in foods, drinks, medicines, feeds, etc., bone formation is promoted, bone resorption is suppressed, and various bone diseases such as osteoporosis are prevented and improved.
破骨細胞は造血幹細胞から発生して海綿骨表面に存在し、骨を溶解する細胞である。破骨細胞が骨基質を溶解し(骨吸収)、その後、骨芽細胞が骨基質を合成することによって、骨の形成や成長(モデリング)、代謝(リモデリング)が起こると考えられている。本発明は、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を経口投与することによって、骨芽細胞による骨形成を促進し、破骨細胞による骨吸収が顕著に抑制されることを見出したものである。 Osteoclasts are cells that originate from hematopoietic stem cells and are present on the surface of cancellous bone and lyse bone. It is believed that osteoclasts dissolve bone matrix (bone resorption), and then osteoblasts synthesize bone matrix, resulting in bone formation and growth (modeling) and metabolism (remodeling). The present invention promotes bone formation by osteoblasts by orally administering at least one of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin, and bone resorption by osteoclasts It has been found that it is remarkably suppressed.
プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンに見出された骨形成促進作用と骨吸収抑制作用を用いることによって、骨のモデリング及びリモデリングの両方がバランスよく働くことが期待できる。よって本発明は、骨形成促進作用及び骨吸収抑制作用のいずれをも有する骨形成促進及び骨吸収抑制剤、飲食品、医薬又は飼料を提供するものである。 Expected to balance both bone modeling and remodeling by using the bone formation-promoting and bone resorption-inhibiting effects found in prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin it can. Therefore, this invention provides the bone formation promotion and bone resorption inhibitor, food / beverage products, medicine, or feed which have both a bone formation promotion effect and a bone resorption suppression effect.
本発明で有効成分として使用するプロトロンビン、チトクロームP-450、プラスミノーゲン、及びトランスフェリンはいずれも、ウシ、水牛、ヒト、ブタ、ヒツジ、ヤギ、ウマ等の乳中に微量に存在しており、乳から分離抽出が可能である。乳成分としては、生乳、脱脂乳、又はホエーから分離したタンパク質画分、あるいは生乳から分離した乳中細胞画分が利用できる。プロトロンビン、チトクロームP-450、プラスミノーゲン、及びトランスフェリンは、健康な家畜の血液から調製することもできる。プロトロンビン、チトクロームP-450、プラスミノーゲン、及びトランスフェリンはいずれも市販されており、この市販品を用いることも可能である。また、遺伝子工学的に作製したリコンビナント品を用いてもよい。 Prothrombin, cytochrome P-450, plasminogen, and transferrin used as active ingredients in the present invention are all present in trace amounts in milk such as cows, buffalo, humans, pigs, sheep, goats, horses, Separation and extraction from milk is possible. As the milk component, a protein fraction separated from raw milk, skim milk or whey, or a cell fraction in milk separated from raw milk can be used. Prothrombin, cytochrome P-450, plasminogen, and transferrin can also be prepared from healthy domestic blood. Prothrombin, cytochrome P-450, plasminogen, and transferrin are all commercially available, and these commercially available products can also be used. Moreover, you may use the recombinant goods produced by genetic engineering.
さらに、プロトロンビン、チトクロームP-450、プラスミノーゲン、及びトランスフェリンをトリプシン、パンクレアチン、キモトリプシン、ペプシン、パパイン、カリクレイン、カテプシン、サーモライシン、V8プロテアーゼ等のタンパク質分解酵素により分解したものを用いても良い。例えば、プロトロンビン分解物は、上記のプロトロンビンをタンパク質分解酵素で限定分解したペプチド混合物である。プロトロンビンをファクターX等の酵素で分解したトロンビンを用いても良い。また、プラスミノーゲンをプラスミノーゲンアクチベーター等の酵素で分解したプラスミンを用いても良い。トロンビン、プラスミンもプロトロンビン、プラスミノーゲンと同様に市販されており、この市販のものを用いることもできる。 Further, prothrombin, cytochrome P-450, plasminogen, and transferrin may be decomposed with a protease such as trypsin, pancreatin, chymotrypsin, pepsin, papain, kallikrein, cathepsin, thermolysin, or V8 protease. For example, the prothrombin degradation product is a peptide mixture obtained by limited degradation of the above prothrombin with a proteolytic enzyme. Thrombin obtained by decomposing prothrombin with an enzyme such as Factor X may be used. Alternatively, plasmin obtained by decomposing plasminogen with an enzyme such as plasminogen activator may be used. Thrombin and plasmin are also commercially available in the same manner as prothrombin and plasminogen, and these commercially available products can also be used.
乳からの調製については、例えば、新鮮な牛乳を陰イオン交換樹脂に接触させてプロトロンビン、チトクロームP-450、プラスミノーゲン、及びトランスフェリンを含む画分を吸着させ、塩化ナトリウムの濃度を徐々に高めて溶出した後、ゲル濾過クロマトグラフィーによりプロトロンビン、チトクロームP-450、プラスミノーゲン、及びトランスフェリンをそれぞれ得ることができる。
血液からの調製については、クエン酸血漿をクエン酸バリウムと反応させ、不溶性バリウムに吸着したプロトロンビン、チトクロームP-450、プラスミノーゲン、及びトランスフェリンの沈殿をそれぞれ回収する。さらにイオン交換クロマトグラフィーにより純度を高めることができる。
For preparation from milk, for example, fresh milk is contacted with an anion exchange resin to adsorb fractions containing prothrombin, cytochrome P-450, plasminogen, and transferrin, and gradually increase the concentration of sodium chloride. After elution, prothrombin, cytochrome P-450, plasminogen, and transferrin can be obtained by gel filtration chromatography.
For preparation from blood, citrate plasma is reacted with barium citrate and the precipitates of prothrombin, cytochrome P-450, plasminogen and transferrin adsorbed on insoluble barium are recovered. Furthermore, the purity can be increased by ion exchange chromatography.
本発明の骨形成促進及び骨吸収抑制剤は、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を有効成分とする。また、このプロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンのいずれか1種以上を、牛乳、乳飲料、コーヒー飲料、ジュース、ゼリー、ビスケット、パン、麺、ソーセージ等の飲食品に配合しても良いし、錠剤や粉末等の医薬としても良い。さらに、塩化カルシウム、炭酸カルシウム、乳酸カルシウム、卵殻、牛乳由来のカルシウム等の吸収性が良好なカルシウム剤を併用することにより、骨強化作用を一層高めることもできる。 The bone formation promotion and bone resorption inhibitor of the present invention contains at least one of prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin as an active ingredient. In addition, any one or more of these prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin are used in foods and drinks such as milk, milk drinks, coffee drinks, juice, jelly, biscuits, bread, noodles and sausages. It may be blended in or as a medicine such as a tablet or powder. Furthermore, the bone strengthening action can be further enhanced by using a calcium agent with good absorbability such as calcium chloride, calcium carbonate, calcium lactate, eggshell and milk-derived calcium.
本発明の骨形成促進及び骨吸収抑制剤の投与量は、有効成分、年齢、治療効果及び病態等により異なるが、成人の場合、1日当たり 1ng〜100mgを数回に分けて経口的に摂取すれば良い。このように、本発明の骨形成促進及び骨吸収抑制剤を摂取することにより、骨粗鬆症等の各種骨疾患を予防又は改善することができる。なお、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンは、元来、血漿や乳由来の成分であり、ラットにおける急性毒性は認められなかった。また、これらの有効成分を飼料に含有させて、家畜や家禽等の骨形成を促進したり、骨吸収を抑制したりすることもできる。 The dose of the bone formation promoting and bone resorption inhibitor of the present invention varies depending on the active ingredient, age, therapeutic effect and disease state, etc. In the case of adults, 1 ng to 100 mg per day should be taken orally in several divided doses. It ’s fine. Thus, various bone diseases such as osteoporosis can be prevented or improved by taking the bone formation promoting and bone resorption inhibitor of the present invention. Prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin were originally derived from plasma and milk, and no acute toxicity was observed in rats. Moreover, these active ingredients can be contained in feed to promote bone formation of livestock, poultry, etc., or to suppress bone resorption.
以下に実施例及び試験例を示し、本発明についてより詳細に説明するが、これらは単に例示するのみであり、本発明はこれらによって何ら限定されるものではない。 EXAMPLES Examples and test examples are shown below, and the present invention will be described in more detail. However, these are merely illustrative, and the present invention is not limited by these.
既知の方法(Hashimoto, N., T. Morita, and S. Iwanaga, J. Biochem. (Tokyo), vol.97, p.1347-1355, 1985)に準じ、ウシ血漿プロトロンビンを調製した。すなわち、0.01 Mになるようにクエン酸ナトリウムを加え、遠心分離によりウシ血漿を得た。10lのウシ血漿に0.1 Mになるようにクエン酸バリウムをゆっくりと加え、緩やかに1時間撹拌した後、4,000rpmで10分間遠心分離した。得られた沈殿を0.01 M塩化バリウム、1mMベンザミジン塩酸、0.02%アジ化ナトリウムを含む、0.1 M塩化ナトリウム溶液で洗浄し、遠心分離した。この洗浄操作を2回繰り返した。バリウム沈殿を40%飽和硫安1lで懸濁し、1 mMになるようにジイソプロリルフォスフォロフルオリデート(DFP)を加えて、1晩撹拌した。5,000rpmで30分間、遠心分離を行ない、上清を得た。この上清に67%飽和度になるように飽和硫安を加え、5,000rpm、30分間の遠心分離により、沈殿を回収した。この沈殿を0.2 M塩化ナトリウム、1 mM DFP、1 mMベンザミジン塩酸を含む0.05 Mリン酸ナトリウム緩衝液(pH 6.0)100〜150mlで溶解し、同じ緩衝液20lに対して1晩透析を行なった。不溶成分を遠心分離で除いた後、DEAE-セファデックスA50クロマトグラフィーに供した。0.2 Mの塩化ナトリウム、1 mMベンザミジン塩酸を含む0.05 Mリン酸ナトリウム緩衝液(pH 6.0)で平衡化したDEAE−セファデックスA-50カラム(200×100 mm)に供し、塩化ナトリウム濃度を 600 mMまで次第に高め、吸着したタンパク質を溶出した。プロトロンビンは塩化ナトリウム濃度が約 300 mMとなったところで溶出した。これを透析により脱塩し、次いで、0.1 Mの塩化ナトリウム、1 mMベンザミジン塩酸を含む50mMトリス−塩酸緩衝液(pH 6.3)で平衡化したブルーセファロースCL-6Bカラム (80×200mm)に供し、平衡化緩衝液で洗浄した後、塩化ナトリウム濃度を4 Mとした緩衝液で吸着したタンパク質を溶出した。これを透析により脱塩し、凍結乾燥して本発明の有効成分であるプロトロンビン1.0 gを得た。このプロトロンビン1.0gを乳糖9.0 gと混合し、顆粒状に成型して本発明の骨形成促進及び骨吸収抑制剤を得た。
なお、チトクロームP-450、プラスミノーゲン、及びトランスフェリンについても上記プロトロンビンと同様の方法によりウシ血漿より調製することが可能である。
Bovine plasma prothrombin was prepared according to a known method (Hashimoto, N., T. Morita, and S. Iwanaga, J. Biochem. (Tokyo), vol. 97, p. 1347-1355, 1985). That is, sodium citrate was added to 0.01 M, and bovine plasma was obtained by centrifugation. Barium citrate was slowly added to 10 l of bovine plasma so that the concentration was 0.1 M, and the mixture was gently stirred for 1 hour and then centrifuged at 4,000 rpm for 10 minutes. The obtained precipitate was washed with a 0.1 M sodium chloride solution containing 0.01 M barium chloride, 1 mM benzamidine hydrochloride and 0.02% sodium azide, and centrifuged. This washing operation was repeated twice. The barium precipitate was suspended in 1 l of 40% saturated ammonium sulfate, diisoprolyl phosphorofluoridate (DFP) was added to 1 mM, and the mixture was stirred overnight. Centrifugation was performed at 5,000 rpm for 30 minutes to obtain a supernatant. Saturated ammonium sulfate was added to this supernatant to 67% saturation, and the precipitate was recovered by centrifugation at 5,000 rpm for 30 minutes. This precipitate was dissolved in 100 to 150 ml of 0.05 M sodium phosphate buffer (pH 6.0) containing 0.2 M sodium chloride, 1 mM DFP and 1 mM benzamidine hydrochloride, and dialyzed overnight against 20 l of the same buffer. Insoluble components were removed by centrifugation, and then subjected to DEAE-Sephadex A50 chromatography. The sample was applied to a DEAE-Sephadex A-50 column (200 x 100 mm) equilibrated with 0.05 M sodium phosphate buffer (pH 6.0) containing 0.2 M sodium chloride and 1 mM benzamidine hydrochloride, and the sodium chloride concentration was 600 mM. And the adsorbed protein was eluted. Prothrombin was eluted when the sodium chloride concentration reached about 300 mM. This was desalted by dialysis and then applied to a Blue Sepharose CL-6B column (80 × 200 mm) equilibrated with 50 mM Tris-HCl buffer (pH 6.3) containing 0.1 M sodium chloride and 1 mM benzamidine hydrochloride. After washing with the equilibration buffer, the adsorbed protein was eluted with a buffer with a sodium chloride concentration of 4 M. This was desalted by dialysis and freeze-dried to obtain 1.0 g of prothrombin, which is an active ingredient of the present invention. 1.0 g of this prothrombin was mixed with 9.0 g of lactose and molded into granules to obtain the bone formation promotion and bone resorption inhibitor of the present invention.
Note that cytochrome P-450, plasminogen, and transferrin can also be prepared from bovine plasma by the same method as for prothrombin.
牛乳からプロトロンビンを含む画分を調製した。陽イオン交換樹脂のSP セファロース HP(アマシャムバイオサイエンス社製)400 gを充填したカラム(直径5 cm×高さ30 cm)を脱イオン水で十分洗浄した後、このカラムに未殺菌脱脂乳40l(pH 6.7)を流速25 ml/minで通液した。通液後、このカラムを脱イオン水で十分洗浄し、1.5M塩化ナトリウムを含む 0.02M炭酸緩衝液(pH 7.0)で樹脂に吸着したタンパク質画分を溶出した。そして、この溶出液を逆浸透(RO)膜により脱塩して、濃縮した後、凍結乾燥してタンパク質画分粉末 21 gを得、本発明の骨形成促進及び骨吸収抑制剤とした。なお、このタンパク質画分粉末には、プロトロンビンが0.01重量%含まれていた。 A fraction containing prothrombin was prepared from milk. A column (5 cm in diameter x 30 cm in height) packed with 400 g of the cation exchange resin SP Sepharose HP (Amersham Biosciences) was thoroughly washed with deionized water, and then 40 l of non-sterilized skim milk ( pH 6.7) was passed at a flow rate of 25 ml / min. After passing through the column, the column was thoroughly washed with deionized water, and the protein fraction adsorbed on the resin was eluted with 0.02 M carbonate buffer (pH 7.0) containing 1.5 M sodium chloride. This eluate was desalted with a reverse osmosis (RO) membrane, concentrated, and then lyophilized to obtain 21 g of protein fraction powder, which was used as the bone formation promotion and bone resorption inhibitor of the present invention. This protein fraction powder contained 0.01% by weight of prothrombin.
[試験例1]
プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、トロンビン及びプラスミンについて骨芽細胞増殖作用を調べた。試験に供した各有効成分はいずれも市販のものを用いた。試験に供した各有効成分の由来及び各有効成分の最終濃度は表1記載の通りである。
マウス骨芽細胞MC3T3-E1細胞を10%牛胎児血清を含むα−MEM培地(Flow Laboratories社製) で、2×104/mlの細胞数で96穴プレートに播種し、5% CO2存在下、37℃で24時間培養し、試験用培養細胞とした。そして、培地を牛胎児血清を含まないα−MEM培地に交換し、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、プラスミン及びトロンビンを表1記載のそれぞれの最終濃度となるよう培地に添加して、37℃で18時間培養した。これにブロモデオキシウリジン(BrdU)を用いたセルプロロフィレーションアッセイキット(アマシャムバイオサイエンス社製)で細胞増殖活性を調べた。対照として、有効成分無添加のものを用い、有効成分無添加のものの細胞増殖活性を 100%としたときのそれぞれの細胞増殖活性 (%) で表した。その結果を表1に示す。
[Test Example 1]
Prothrombin, cytochrome P-450, plasminogen, transferrin, thrombin and plasmin were examined for osteoblast proliferation. Each active ingredient used in the test was commercially available. The origin of each active ingredient subjected to the test and the final concentration of each active ingredient are shown in Table 1.
Mouse osteoblasts MC3T3-E1 cells are seeded in 96-well plates in α-MEM medium (Flow Laboratories) containing 10% fetal bovine serum at a cell count of 2 × 10 4 / ml and 5% CO 2 is present Then, the cells were cultured at 37 ° C. for 24 hours to obtain cultured cells for testing. Then, the medium is replaced with an α-MEM medium not containing fetal bovine serum, and prothrombin, cytochrome P-450, plasminogen, transferrin, plasmin and thrombin are added to the medium so as to have respective final concentrations shown in Table 1. And then cultured at 37 ° C. for 18 hours. Cell proliferation activity was examined with a cell prolophylation assay kit (Amersham Bioscience) using bromodeoxyuridine (BrdU). As a control, an active ingredient-free product was used, and the cell-growth activity (%) when the active component-free product was defined as 100%. The results are shown in Table 1.
プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、プラスミン及びトロンビンを添加したものはいずれも有効成分無添加のものに比べ細胞増殖活性が増加しており、骨芽細胞増殖作用を有することが判った。 Prothrombin, cytochrome P-450, plasminogen, transferrin, plasmin, and thrombin all have an increased cell proliferative activity compared to those without any active ingredient, and are found to have osteoblast proliferative activity. It was.
[試験例2]
プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、プラスミン及びトロンビンについて骨吸収抑制作用を調べた。試験に供した各有効成分はいずれも市販のものを用いた。試験に供した各有効成分の由来及び各有効成分の最終濃度は表2記載の通りである。
生後10〜20日齢のICR系マウスの長管骨を摘出し、軟組織を除去した後、5%牛胎児血清を含むα−MEM(Flow Laboratories社製)溶液中で骨を機械的に細切し、破骨細胞を含む全骨髄細胞を得た。この破骨細胞を含む全骨髄細胞を約2×106 細胞になるように象牙片の上に撒き込み、5%牛胎児血清を含むα−MEM溶液でスポットした。2時間後、プロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、プラスミン及びトロンビンを、表2記載のそれぞれの最終濃度となるように、5%牛胎児血清を含むα−MEM溶液を加え、5日間培養し、既存の破骨細胞の骨吸収抑制活性を調べた。
骨吸収抑制作用の評価は、培養後、象牙片上の細胞を剥がしてヘマトキシリン染色し、PIASLA-555により画像解析して骨吸収窩(ピット) の数を測定した。(瀬野悍二ら,研究テーマ別動物培養細胞マニュアル,pp.199-200, 1993)。すなわち、ピット数が少ないということは、破骨細胞の活性が低下して骨吸収活性が抑制されたことを意味している。また、ピットアッセイで骨吸収抑制効果が示された物質は、動物実験でも骨吸収の抑制効果が示されており(Tobaら、Bone, vol.27, p.403-408, 2000)、一般的にピットアッセイは骨吸収抑制効果を調べる上で適した実験系である。対照として、有効成分無添加のものを用い、有効成分無添加のものの骨吸収活性を100%としたときのそれぞれの骨吸収活性 (%) で表した。その結果を表2に示す。
[Test Example 2]
Bone resorption inhibitory effects of prothrombin, cytochrome P-450, plasminogen, transferrin, plasmin and thrombin were examined. Each active ingredient used in the test was commercially available. The origin of each active ingredient subjected to the test and the final concentration of each active ingredient are shown in Table 2.
After removing the long bones of 10-20 days old ICR mice, removing soft tissues, mechanically chopping bones in α-MEM (Flow Laboratories) solution containing 5% fetal calf serum And whole bone marrow cells containing osteoclasts were obtained. The whole bone marrow cells containing osteoclasts were seeded on ivory pieces so as to be about 2 × 10 6 cells, and spotted with an α-MEM solution containing 5% fetal calf serum. Two hours later, prothrombin, cytochrome P-450, plasminogen, transferrin, plasmin and thrombin were added to an α-MEM solution containing 5% fetal bovine serum so that the final concentrations shown in Table 2 were obtained. After culturing for one day, the bone resorption inhibitory activity of existing osteoclasts was examined.
To evaluate the bone resorption inhibitory action, after culturing, the cells on the ivory pieces were peeled off and stained with hematoxylin, and image analysis was performed with PIASLA-555 to measure the number of bone resorption pits. (Kenji Seno et al., Animal Culture Cell Manual by Research Theme, pp.199-200, 1993). That is, the small number of pits means that the activity of osteoclasts was reduced and the bone resorption activity was suppressed. In addition, substances that have been shown to inhibit bone resorption by pit assay have also been shown to inhibit bone resorption in animal experiments (Toba et al., Bone, vol.27, p.403-408, 2000). The pit assay is a suitable experimental system for examining the bone resorption inhibitory effect. As a control, an active ingredient-free additive was used, and the bone-resorbing activity (%) of each additive-free additive was defined as 100%. The results are shown in Table 2.
本発明の有効成分であるプロトロンビン、チトクロームP-450、プラスミノーゲン、トランスフェリン、プラスミン及びトロンビンを添加したものはいずれも、有効成分無添加のものに比べ骨吸収活性が抑制されており、顕著な骨吸収抑制作用を有することが判った。 Any of the active ingredients of the present invention to which prothrombin, cytochrome P-450, plasminogen, transferrin, plasmin and thrombin are added has suppressed bone resorption activity compared to those without any active ingredient, and is remarkable. It was found to have a bone resorption inhibitory action.
表3に示した配合で原料を混合した後、容器に充填し、加熱殺菌して、骨形成促進及び骨吸収抑制用果汁飲料を製造した。 After mixing the raw materials with the formulation shown in Table 3, the container was filled and sterilized by heating to produce a juice drink for promoting bone formation and suppressing bone resorption.
表4に示した配合で原料を混合した後、打錠機により加圧成型して、錠剤型の骨形成促進及び骨吸収抑制剤を製造した。 After mixing the raw materials with the formulation shown in Table 4, the tablet was pressed by a tableting machine to produce a tablet-type bone formation promoting and bone resorption inhibitor.
表5に示した配合で市販のプラスミノーゲンを用いて原料を混合し、ドウを作成して成型した後、焙焼して、骨形成促進及び骨吸収抑制用ビスケットを製造した。 The ingredients shown in Table 5 were mixed using commercially available plasminogen, the dough was created and molded, and then baked to produce bone formation promotion and bone resorption suppression biscuits.
表6に示した配合で市販のプラスミンを用いて原料を混合し、骨形成促進及び骨吸収抑制用ビスケットを製造した。 The raw materials were mixed using commercially available plasmin with the formulation shown in Table 6 to produce bone formation promotion and bone resorption suppression biscuits.
表7に示した配合で原料を混合した後、容器に充填し、加熱殺菌して、骨形成促進及び骨吸収抑制用ゼリーを製造した。 After mixing the raw materials with the formulation shown in Table 7, the container was filled and sterilized by heating to produce a jelly for promoting bone formation and suppressing bone resorption.
表8に示した配合で市販のプロトロンビンを用いて原料を混合した後、85℃で乳化して、骨形成促進及び骨吸収抑制用プロセスチーズを製造した。 The raw materials were mixed using commercially available prothrombin with the formulation shown in Table 8, and then emulsified at 85 ° C. to produce a process cheese for promoting bone formation and suppressing bone resorption.
表9に示した配合で市販のトロンビンを用いて原料を混合し、骨形成促進及び骨吸収抑制用プロセスチーズを製造した。 The raw materials were mixed using commercially available thrombin with the formulation shown in Table 9 to produce a process cheese for promoting bone formation and suppressing bone resorption.
12重量%還元脱脂乳を90℃で20分間加熱殺菌した後、ラクトバチルス・アシドフィルス(Lactobacillus acidophilus)及びストレプトコッカス・サーモフィルス(Streptococcus thermophilus) をそれぞれ接種し、2種類のスターターカルチャーを得て両者を等量混合した。そして、表10に示した配合で原料を混合した後、発酵させて、骨形成促進及び骨吸収抑制用ヨーグルトを製造した。 After 12% by weight reduced skim milk was sterilized by heating at 90 ° C for 20 minutes, Lactobacillus acidophilus and Streptococcus thermophilus were inoculated to obtain two types of starter cultures, etc. The amount was mixed. And after mixing a raw material with the mixing | blending shown in Table 10, it was made to ferment and the yoghurt for bone formation promotion and bone resorption suppression was manufactured.
表11に示した配合で実施例1で得られたプロトロンビンを用いて原料を混合し、骨形成促進及び骨吸収抑制用乳児用調製粉乳を製造した。 The raw materials were mixed using the prothrombin obtained in Example 1 with the formulation shown in Table 11 to produce infant formula for promoting bone formation and suppressing bone resorption.
表12に示した配合で市販のトロンビンを用いて原料を混合し、骨形成促進及び骨吸収抑制用乳児用調製粉乳を製造した。 The raw materials were mixed using commercially available thrombin with the formulation shown in Table 12 to produce infant formula for promoting bone formation and suppressing bone resorption.
表13に示した配合で市販のプラスミノーゲンを用いて原料を混合して、骨形成促進及び骨吸収抑制用イヌ飼育用飼料(ドッグフード)を製造した。 Raw materials were mixed using commercially available plasminogen with the formulation shown in Table 13 to produce a dog breeding feed for promoting bone formation and suppressing bone resorption (dog food).
表14に示した配合で市販のプラスミンを用いて原料を混合して、骨形成促進及び骨吸収抑制用イヌ飼育用飼料(ドッグフード)を製造した。 The raw materials were mixed using commercially available plasmin with the formulation shown in Table 14 to produce a dog breeding feed for promoting bone formation and suppressing bone resorption (dog food).
Claims (3)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2010046910A JP5188523B2 (en) | 2010-03-03 | 2010-03-03 | Bone formation promoter and bone resorption inhibitor |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2010046910A JP5188523B2 (en) | 2010-03-03 | 2010-03-03 | Bone formation promoter and bone resorption inhibitor |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2003340804A Division JP4740532B2 (en) | 2003-09-30 | 2003-09-30 | Bone formation promoter and bone resorption inhibitor |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012141624A Division JP2012197313A (en) | 2012-06-25 | 2012-06-25 | Agent for promoting bone formation and inhibiting bone resorption |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2010155856A true JP2010155856A (en) | 2010-07-15 |
JP5188523B2 JP5188523B2 (en) | 2013-04-24 |
Family
ID=42574027
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2010046910A Expired - Fee Related JP5188523B2 (en) | 2010-03-03 | 2010-03-03 | Bone formation promoter and bone resorption inhibitor |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP5188523B2 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109925507A (en) * | 2017-12-15 | 2019-06-25 | 深圳瑞健生命科学研究院有限公司 | A kind of method and drug prevented or treat osteoarthritis |
JP2020502102A (en) * | 2016-12-15 | 2020-01-23 | タレンゲン インターナショナル リミテッドTalengen International Limited | Drug for preventing and treating osteoporosis and use thereof |
JP2020524161A (en) * | 2017-06-19 | 2020-08-13 | タレンゲン インターナショナル リミテッドTalengen International Limited | Method and drug for regulating and controlling GLP-1/GLP-1R |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0399022A (en) * | 1989-09-12 | 1991-04-24 | Teijin Ltd | Medicine of bone cacochymia and evaluation of bone metabolism state |
JPH05279266A (en) * | 1992-04-02 | 1993-10-26 | Imuno Japan:Kk | Medicine composition for treating and preventing periodontosis |
JPH08256684A (en) * | 1995-03-27 | 1996-10-08 | Snow Brand Milk Prod Co Ltd | Cheese and its production |
JPH10512284A (en) * | 1995-08-09 | 1998-11-24 | ベーリンガー マンハイム ゲゼルシャフト ミット ベシュレンクテル ハフツング | Use of plasminogen activator for local formation of bone tissue |
JP2000281587A (en) * | 1999-03-30 | 2000-10-10 | Snow Brand Milk Prod Co Ltd | Bone resoprtion suppressor |
-
2010
- 2010-03-03 JP JP2010046910A patent/JP5188523B2/en not_active Expired - Fee Related
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0399022A (en) * | 1989-09-12 | 1991-04-24 | Teijin Ltd | Medicine of bone cacochymia and evaluation of bone metabolism state |
JPH05279266A (en) * | 1992-04-02 | 1993-10-26 | Imuno Japan:Kk | Medicine composition for treating and preventing periodontosis |
JPH08256684A (en) * | 1995-03-27 | 1996-10-08 | Snow Brand Milk Prod Co Ltd | Cheese and its production |
JPH10512284A (en) * | 1995-08-09 | 1998-11-24 | ベーリンガー マンハイム ゲゼルシャフト ミット ベシュレンクテル ハフツング | Use of plasminogen activator for local formation of bone tissue |
JP2000281587A (en) * | 1999-03-30 | 2000-10-10 | Snow Brand Milk Prod Co Ltd | Bone resoprtion suppressor |
Non-Patent Citations (1)
Title |
---|
TSUNOI, M. ET AL, EXP. CELL RES., vol. 153, no. 1, JPN6012020764, 1984, pages 240 - 244, ISSN: 0002207630 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2020502102A (en) * | 2016-12-15 | 2020-01-23 | タレンゲン インターナショナル リミテッドTalengen International Limited | Drug for preventing and treating osteoporosis and use thereof |
JP7531080B2 (en) | 2016-12-15 | 2024-08-09 | タレンゲン インターナショナル リミテッド | Drug for preventing and treating osteoporosis and use thereof |
JP2020524161A (en) * | 2017-06-19 | 2020-08-13 | タレンゲン インターナショナル リミテッドTalengen International Limited | Method and drug for regulating and controlling GLP-1/GLP-1R |
JP7335609B2 (en) | 2017-06-19 | 2023-08-30 | タレンゲン インターナショナル リミテッド | Methods and Agents for Regulating GLP-1/GLP-1R |
US11938172B2 (en) | 2017-06-19 | 2024-03-26 | Talengen International Limited | Method for regulating and controlling GLP-1/GLP-1R and drug |
CN109925507A (en) * | 2017-12-15 | 2019-06-25 | 深圳瑞健生命科学研究院有限公司 | A kind of method and drug prevented or treat osteoarthritis |
Also Published As
Publication number | Publication date |
---|---|
JP5188523B2 (en) | 2013-04-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP3929088B2 (en) | Bone formation promoter and bone resorption inhibitor | |
JP4536341B2 (en) | Bone formation promoter | |
TWI586362B (en) | Novel protein material | |
JP5188523B2 (en) | Bone formation promoter and bone resorption inhibitor | |
JP6124420B2 (en) | Fermented milk and method for producing the same | |
KR101170852B1 (en) | Agent for promoting osteogenesis and/or inhibiting bone resorption | |
JPWO2014020679A1 (en) | Fermented milk and method for producing the same | |
JP6124418B2 (en) | Beverage and method for producing the same | |
JP4740532B2 (en) | Bone formation promoter and bone resorption inhibitor | |
JP3073439B2 (en) | Bone formation promotion and bone resorption inhibitor | |
JP2012197313A (en) | Agent for promoting bone formation and inhibiting bone resorption | |
JP6203723B2 (en) | Protein material for bone disease prevention or treatment and method for producing the same | |
JP6357265B2 (en) | Protein material for bone disease prevention or treatment | |
JP6357266B2 (en) | Protein material for bone disease prevention or treatment | |
JP6562957B2 (en) | Fermented milk and method for producing the same | |
JP6562956B2 (en) | Beverage and method for producing the same | |
JP6208129B2 (en) | Beverage and method for producing the same | |
JP2017123860A (en) | Fermented milk and method for manufacturing the same | |
JP2016152809A (en) | Beverage and manufacturing method therefor |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A712 Effective date: 20110721 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110801 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20120424 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20120625 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20121017 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20121210 |
|
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: 20130115 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20130122 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20160201 Year of fee payment: 3 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5188523 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 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 |
|
LAPS | Cancellation because of no payment of annual fees |