CN1500096A - 乳中的osteoprotegerin - Google Patents
乳中的osteoprotegerin Download PDFInfo
- Publication number
- CN1500096A CN1500096A CNA028078640A CN02807864A CN1500096A CN 1500096 A CN1500096 A CN 1500096A CN A028078640 A CNA028078640 A CN A028078640A CN 02807864 A CN02807864 A CN 02807864A CN 1500096 A CN1500096 A CN 1500096A
- Authority
- CN
- China
- Prior art keywords
- bone
- osteoprotegerin
- opg
- purposes
- composition
- 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.)
- Pending
Links
- 102000008108 Osteoprotegerin Human genes 0.000 title claims abstract description 87
- 108010035042 Osteoprotegerin Proteins 0.000 title claims abstract description 87
- XXUPLYBCNPLTIW-UHFFFAOYSA-N octadec-7-ynoic acid Chemical compound CCCCCCCCCCC#CCCCCCC(O)=O XXUPLYBCNPLTIW-UHFFFAOYSA-N 0.000 title claims abstract description 87
- 235000013336 milk Nutrition 0.000 title claims abstract description 8
- 210000004080 milk Anatomy 0.000 title claims abstract description 8
- 239000008267 milk Substances 0.000 title claims abstract description 8
- 238000002360 preparation method Methods 0.000 claims abstract description 23
- 239000000203 mixture Substances 0.000 claims abstract description 17
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 8
- 210000000988 bone and bone Anatomy 0.000 claims description 40
- 201000010099 disease Diseases 0.000 claims description 20
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 20
- 239000000463 material Substances 0.000 claims description 16
- 208000001132 Osteoporosis Diseases 0.000 claims description 15
- 235000013305 food Nutrition 0.000 claims description 13
- 210000000481 breast Anatomy 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 10
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 9
- 229920001184 polypeptide Polymers 0.000 claims description 6
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 6
- 206010065687 Bone loss Diseases 0.000 claims description 5
- 230000006907 apoptotic process Effects 0.000 claims description 5
- 235000020247 cow milk Nutrition 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 238000012546 transfer Methods 0.000 claims description 5
- 239000000654 additive Substances 0.000 claims description 4
- 230000000996 additive effect Effects 0.000 claims description 4
- 230000013595 glycosylation Effects 0.000 claims description 4
- 238000006206 glycosylation reaction Methods 0.000 claims description 4
- 208000037147 Hypercalcaemia Diseases 0.000 claims description 3
- 206010031252 Osteomyelitis Diseases 0.000 claims description 3
- 230000008859 change Effects 0.000 claims description 3
- 230000000148 hypercalcaemia Effects 0.000 claims description 3
- 208000030915 hypercalcemia disease Diseases 0.000 claims description 3
- 208000015181 infectious disease Diseases 0.000 claims description 3
- 210000000214 mouth Anatomy 0.000 claims description 3
- 201000008482 osteoarthritis Diseases 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 3
- 206010020751 Hypersensitivity Diseases 0.000 claims description 2
- 206010031264 Osteonecrosis Diseases 0.000 claims description 2
- 208000006399 Premature Obstetric Labor Diseases 0.000 claims description 2
- 206010036600 Premature labour Diseases 0.000 claims description 2
- 208000026935 allergic disease Diseases 0.000 claims description 2
- 230000007815 allergy Effects 0.000 claims description 2
- 230000001363 autoimmune Effects 0.000 claims description 2
- 230000004663 cell proliferation Effects 0.000 claims description 2
- 235000013351 cheese Nutrition 0.000 claims description 2
- 235000015140 cultured milk Nutrition 0.000 claims description 2
- 235000013350 formula milk Nutrition 0.000 claims description 2
- 235000015243 ice cream Nutrition 0.000 claims description 2
- 230000036039 immunity Effects 0.000 claims description 2
- 230000002757 inflammatory effect Effects 0.000 claims description 2
- 208000018773 low birth weight Diseases 0.000 claims description 2
- 231100000533 low birth weight Toxicity 0.000 claims description 2
- 230000003239 periodontal effect Effects 0.000 claims description 2
- 208000026440 premature labor Diseases 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 210000002345 respiratory system Anatomy 0.000 claims description 2
- 230000009885 systemic effect Effects 0.000 claims description 2
- 235000013618 yogurt Nutrition 0.000 claims description 2
- 230000036737 immune function Effects 0.000 abstract description 3
- 230000004097 bone metabolism Effects 0.000 abstract description 2
- 241000283690 Bos taurus Species 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 35
- 235000020256 human milk Nutrition 0.000 description 19
- 210000004251 human milk Anatomy 0.000 description 19
- 238000000034 method Methods 0.000 description 12
- 239000000523 sample Substances 0.000 description 11
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 9
- 238000011534 incubation Methods 0.000 description 9
- 210000002997 osteoclast Anatomy 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 238000003752 polymerase chain reaction Methods 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 6
- 238000003757 reverse transcription PCR Methods 0.000 description 6
- 102000011632 Caseins Human genes 0.000 description 5
- 108010076119 Caseins Proteins 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 5
- 239000005862 Whey Substances 0.000 description 5
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 5
- 235000021240 caseins Nutrition 0.000 description 5
- 230000001413 cellular effect Effects 0.000 description 5
- 239000000470 constituent Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 210000002919 epithelial cell Anatomy 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 102000007544 Whey Proteins Human genes 0.000 description 4
- 108010046377 Whey Proteins Proteins 0.000 description 4
- 239000013543 active substance Substances 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 230000033228 biological regulation Effects 0.000 description 4
- 210000000845 cartilage Anatomy 0.000 description 4
- 239000005018 casein Substances 0.000 description 4
- 230000030833 cell death Effects 0.000 description 4
- 210000004443 dendritic cell Anatomy 0.000 description 4
- 239000003446 ligand Substances 0.000 description 4
- 210000005075 mammary gland Anatomy 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 238000010839 reverse transcription Methods 0.000 description 4
- 208000006386 Bone Resorption Diseases 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 108010025832 RANK Ligand Proteins 0.000 description 3
- 239000013614 RNA sample Substances 0.000 description 3
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 3
- 102100024568 Tumor necrosis factor ligand superfamily member 11 Human genes 0.000 description 3
- 241000235013 Yarrowia Species 0.000 description 3
- 241000235015 Yarrowia lipolytica Species 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000024279 bone resorption Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 230000006651 lactation Effects 0.000 description 3
- 230000006798 recombination Effects 0.000 description 3
- 238000005215 recombination Methods 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 230000009182 swimming Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 102000010170 Death domains Human genes 0.000 description 2
- 108050001718 Death domains Proteins 0.000 description 2
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 2
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 2
- 101000830565 Homo sapiens Tumor necrosis factor ligand superfamily member 10 Proteins 0.000 description 2
- 101000798130 Homo sapiens Tumor necrosis factor receptor superfamily member 11B Proteins 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102000003982 Parathyroid hormone Human genes 0.000 description 2
- 108090000445 Parathyroid hormone Proteins 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 102000009618 Transforming Growth Factors Human genes 0.000 description 2
- 108010009583 Transforming Growth Factors Proteins 0.000 description 2
- 102000018594 Tumour necrosis factor Human genes 0.000 description 2
- 108050007852 Tumour necrosis factor Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 230000037182 bone density Effects 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 210000001612 chondrocyte Anatomy 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000000539 dimer Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 229940126864 fibroblast growth factor Drugs 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 102000052781 human TNFRSF11B Human genes 0.000 description 2
- 102000044949 human TNFSF10 Human genes 0.000 description 2
- 208000026278 immune system disease Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 210000002751 lymph Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 230000001582 osteoblastic effect Effects 0.000 description 2
- 210000004409 osteocyte Anatomy 0.000 description 2
- 210000004663 osteoprogenitor cell Anatomy 0.000 description 2
- 239000000199 parathyroid hormone Substances 0.000 description 2
- 229960001319 parathyroid hormone Drugs 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 229940108461 rennet Drugs 0.000 description 2
- 108010058314 rennet Proteins 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 108091092562 ribozyme Proteins 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 230000001131 transforming effect Effects 0.000 description 2
- 230000032258 transport Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- JYGRAOYMDDFOSM-FQJIPJFPSA-N (4s)-4-[[(2s)-4-carboxy-2-[[(2s)-3-carboxy-2-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]propanoyl]amino]butanoyl]amino]-5-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-5-oxopentano Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN JYGRAOYMDDFOSM-FQJIPJFPSA-N 0.000 description 1
- KZMAWJRXKGLWGS-UHFFFAOYSA-N 2-chloro-n-[4-(4-methoxyphenyl)-1,3-thiazol-2-yl]-n-(3-methoxypropyl)acetamide Chemical compound S1C(N(C(=O)CCl)CCCOC)=NC(C=2C=CC(OC)=CC=2)=C1 KZMAWJRXKGLWGS-UHFFFAOYSA-N 0.000 description 1
- XMTQQYYKAHVGBJ-UHFFFAOYSA-N 3-(3,4-DICHLOROPHENYL)-1,1-DIMETHYLUREA Chemical compound CN(C)C(=O)NC1=CC=C(Cl)C(Cl)=C1 XMTQQYYKAHVGBJ-UHFFFAOYSA-N 0.000 description 1
- QRXMUCSWCMTJGU-UHFFFAOYSA-N 5-bromo-4-chloro-3-indolyl phosphate Chemical compound C1=C(Br)C(Cl)=C2C(OP(O)(=O)O)=CNC2=C1 QRXMUCSWCMTJGU-UHFFFAOYSA-N 0.000 description 1
- 206010000599 Acromegaly Diseases 0.000 description 1
- 208000034309 Bacterial disease carrier Diseases 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 102000055006 Calcitonin Human genes 0.000 description 1
- 108060001064 Calcitonin Proteins 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 208000014311 Cushing syndrome Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000007260 Deoxyribonuclease I Human genes 0.000 description 1
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 208000001730 Familial dysautonomia Diseases 0.000 description 1
- 108060003393 Granulin Proteins 0.000 description 1
- 206010020365 Homocystinuria Diseases 0.000 description 1
- 201000002980 Hyperparathyroidism Diseases 0.000 description 1
- 206010020850 Hyperthyroidism Diseases 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- 206010027294 Menkes' syndrome Diseases 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 241000713869 Moloney murine leukemia virus Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 230000004988 N-glycosylation Effects 0.000 description 1
- 108010057466 NF-kappa B Proteins 0.000 description 1
- 102000003945 NF-kappa B Human genes 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 201000000023 Osteosclerosis Diseases 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 201000001638 Riley-Day syndrome Diseases 0.000 description 1
- 206010039984 Senile osteoporosis Diseases 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 108010017842 Telomerase Proteins 0.000 description 1
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000003143 atherosclerotic effect Effects 0.000 description 1
- 235000008452 baby food Nutrition 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 210000002805 bone matrix Anatomy 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229940021722 caseins Drugs 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 230000007541 cellular toxicity Effects 0.000 description 1
- 238000010909 chemical acidification Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000001728 clone cell Anatomy 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 210000003022 colostrum Anatomy 0.000 description 1
- 235000021277 colostrum Nutrition 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 108010053500 delicious peptide Proteins 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000007882 dietary composition Nutrition 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- KAKKHKRHCKCAGH-UHFFFAOYSA-L disodium;(4-nitrophenyl) phosphate;hexahydrate Chemical compound O.O.O.O.O.O.[Na+].[Na+].[O-][N+](=O)C1=CC=C(OP([O-])([O-])=O)C=C1 KAKKHKRHCKCAGH-UHFFFAOYSA-L 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000000416 exudates and transudate Anatomy 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000011132 hemopoiesis Effects 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000031891 intestinal absorption Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000009245 menopause Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 235000021239 milk protein Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 230000001599 osteoclastic effect Effects 0.000 description 1
- 102000013415 peroxidase activity proteins Human genes 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 210000004224 pleura Anatomy 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 201000009395 primary hyperaldosteronism Diseases 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008521 reorganization Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 108010048734 sclerotin Proteins 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000009772 tissue formation Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70578—NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/19—Dairy proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/06—Antiabortive agents; Labour repressants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/12—Drugs for disorders of the metabolism for electrolyte homeostasis
- A61P3/14—Drugs for disorders of the metabolism for electrolyte homeostasis for calcium homeostasis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physical Education & Sports Medicine (AREA)
- Rheumatology (AREA)
- Immunology (AREA)
- Zoology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Hematology (AREA)
- Endocrinology (AREA)
- Diabetes (AREA)
- Reproductive Health (AREA)
- Molecular Biology (AREA)
- Pulmonology (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Animal Husbandry (AREA)
- Toxicology (AREA)
- Cell Biology (AREA)
- Urology & Nephrology (AREA)
Abstract
本发明涉及可从乳源,特别是人乳和牛乳中获得的osteoprotegerin。本发明还涉及所述成分在制备可以食用的制剂和/或药物组合物中的用途,特别是所述制剂/组合物在预防或治疗与骨代谢和免疫功能相关的疾病中的用途。
Description
本发明涉及可从乳源,特别是人乳和牛乳中获得的osteoprotegerin。本发明还涉及其在制备可以食用的制剂和/或药物组合物中的用途,特别是所述制剂/组合物在预防或治疗与骨代谢和免疫功能相关的疾病中的用途。
在哺乳动物中,骨骼提供了对身体的支撑,并且由矿物质、胶原蛋白和非胶原蛋白的基质、和细胞成分组成。其生长和维持受多种不同因素的控制,包括其组成细胞类型的调控和相互作用,所述细胞类型即形成软骨的软骨细胞,合成并且沉积骨基质的成骨细胞,以及负责骨物质吸收的破骨细胞。
软骨细胞源于间充质细胞,并且产生软骨内骨形成所需要的起始软骨模板。促进骨组织形成的成骨细胞源于间充质骨祖细胞,并且位于骨表面,在这里它们合成、转运并且排列所述基质蛋白,另一方面,负责骨吸收的破骨细胞源于存在于造血骨髓中的粒细胞-单核细胞前体。破骨细胞和成骨细胞的作用是密切联系的,例如,在破骨细胞介导的吸收过程中,所述蛋白因子专门起着信号传导分子的作用,由破骨细胞启动骨更新。相反,成骨细胞可以通过表达可溶性或膜结合调节剂影响破骨细胞功能。因此,正常骨重建取决于骨形成和骨吸收的相反功能之间的明确平衡,所述功能是由各种相应的细胞类型产生的。
诸如成纤维细胞生长因子(FGF)和转化生长因子(TGF)-β的生长因子储存在骨细胞外基质中,并且能在分泌时刺激骨祖细胞的局部释放。然后,诸如骨形态发生蛋白(BMPs)和甲状旁腺素(PTH)的因子能影响所述祖细胞发育成成骨细胞,骨形成细胞、其最终分化和功能,是通过所述细胞与骨基质蛋白的相互作用调控的。
在衰老期间,个体会经历骨质的逐渐损失,这种现象被称为解偶联,它被认为是由于破骨细胞的活性超过了成骨细胞的活性所导致的。当这种解偶联持续较长时间时,会有越来越多的骨物质被破坏/吸收,这种状况被称为骨质疏松结果。
除了取决于年龄的现象之外,骨损失还可能由钙或激素缺乏引起,或由会导致多种不同疾病的状况引起,如骨质疏松症、高钙血症、骨佩吉氏病,由于骨关节炎或类风湿关节炎或骨髓炎所导致的骨损失等。降低了的骨密度,通常会导致机械强度减弱,并且增加了骨折的可能性。
现有用于骨质疏松和/或相关的骨疾病的方法包括给需要的个体服用钙。最近,业已设计出用于治疗上述疾病状况的试剂,包括刺激和/或抑制骨细胞,如激素、降钙素、胰岛素样生长因子或osteoprotegerin(OPG)。所述制剂通常是通过重组方法制备的,并且业已配制/制备成galenic形式,以便相应的物质能够以活性形式达到靶,即骨骼。
WO 00/24771披露了编码osteoprotegerin样蛋白的核酸及其在诸如治疗骨质疏松方面的用途。这种多肽是通过重组方法合成的,然后进行配制,以便与希望的给药途径相容。推荐静脉内、真皮内、皮下、口服(例如吸入)、经皮(局部)、经粘膜和直肠内给药途径。
一般,对于特定的物质来说,要找到合适的galenic形式要花费很长时间并且是很烦琐的,因为用于这一目的的成分必须与所述活性物质相容,并且还必须提供对人体内不同条件的足够的保护作用。不过,由于刺激骨骼生长的试剂是局部合成的-骨组织内/骨组织上-给予这种物质是困难的。通常,必须设计出胶囊,它有助于使所述物质通过胃肠道,而又不会受到胃肠道中占主导地位的不利环境条件的破坏。不过,这种给药途径还是具有某些缺陷,因为所述物质必须通过肝脏,并且在体液中运输然后才能达到骨骼。另外,它通常会导致较少数量的活性生物学物质达到靶组织。
因此,本发明的问题是提供一种给予个体活性物质的方法,以便所述物质能在所述个体的特定靶组织中起作用。
因此,业已通过提供可通过乳获得的osteoprotegerin解决了上述问题。
在附图中,
图1表示在哺乳期的不同阶段,人乳中osteoprotegerin的浓度;
图2表示在还原条件下使用10%SDS-凝胶进行的人乳级份的蛋白印迹分析。用购自R&D Systems的生物素化的抗OPG多克隆抗体BAF805和链亲和素-碱性磷酸酶(SAPP)显示OPG带;
图3表示整合到Yarrowia转化体中的基因组DNA中的质粒的限制图谱;
图4表示人乳细胞和人乳腺上皮细胞MCF-7的RT-PCR分析;泳道1和2:β-肌动蛋白(预期的带大小:460bp);泳道3和4:OPG(预期的带大小:603bp);1.人乳细胞;2.MCF-7;3.人乳细胞;4.MCF-7;
图5表示一种实验的结果,其中,本发明的OPG能抑制TRAIL-诱导的Jurkat细胞的凋亡。
Osteoprotegerin(OPG)又被称为破骨细胞发生抑制因子(OCIF)和TNF-受体样分子1(TR1)最近被报导为受体的肿瘤坏死因子家族(TNFR)的成员。它能在体外和体内抑制破骨细胞发育,并且提高骨密度(骨硬化病)。在正常的小鼠胚胎中,业已将OPG定位于发育中骨骼的软骨原基和小肠内。
不过,与TNF受体家族的其他成员不同,OPG不具有跨膜结构域。另外,还可以证实OPG也是细胞毒性配体TRAIL(TNF-相关的细胞凋亡诱导配体)的受体,并且与滤泡树突细胞来源的受体-1相同。因此,推测它能调控细胞死亡,并且在淋巴样组织的形成和免疫应答调控方面发挥重要作用。实际上,业已证实缺乏OPG的动物具有发育不足的淋巴组织。
在导致本发明的研究中,业已惊奇地发现,除了在骨组织中存在osteoprotegerin之外,它还存在于人类乳中。其结果是,在哺乳期间,母亲显然以能够在胃肠道中存活的形式给新生儿提供所述物质。随后发现,由乳腺细胞生产的OPG明显不同于从其他来源分离的OPG,这些不同表现在其稳定性和/或抗降解能力方面。
并不希望受任何理论的约束,目前认为,在乳腺细胞中所产生的对蛋白的特异的糖基化模式,使得所述多肽在遇到酸性胃液和/或在肠道中遇到的碱性环境中更稳定,以便在肠道吸收和运输到骨组织时,所述活性结构域保持完整,并且能够发挥其生物学活性。
本发明的OPG,即可以从乳源中获得的形式具有SEQ ID.No.1所示的多肽序列,并且分别具有大约80,130和200kDa的大小,其大小不同于通过重组方法获得的大小(55kDa)。
本发明的OPG可以包含在可食用的制剂中,所述制剂可以是食品物质,如乳、酸乳、凝乳、奶酪、发酵乳、乳基发酵食品、冰淇淋、谷基发酵食品、乳基粉末、婴儿配方以及宠物食品。同样,本发明的OPG还可以掺入肠道或药物组合物中,例如,选自下列一组:溶液、干燥的口服添加剂、液体口服添加剂、干燥的管装饲料或液体管装饲料。
实际上,由于本发明的OPG是稳定的,没有必要将所述活性化合物制成特定的galenic形式,以便避免它受到在胃肠道和体液中占主导地位的不同的和潜在的不利条件的破坏。
根据本发明的另一方面,还提供了来自乳的osteoprotegerin在制备可以食用的制剂中的用途,如制备食品物质或肠道组合物或药物组合物。
可以将本发明的osteoprotegerin和上文详细说明的可食用制剂用于治疗和/或预防骨重建疾病。
最常见的骨骼疾病是骨量减少,这是一种一般与骨量低于正常水平的任何疾病相关的状况。这种状况可能是由骨合成的速度降低或骨分解速度的提高或同时由这两者所导致的。骨量减少的最常见的形式是原发性骨质疏松,又被称作绝经后和老年骨质疏松。这种形式的骨质疏松是随着年龄增加骨普遍损失的后果,并且通常是在具有正常的骨骼形成速度下,骨吸收速度增加的结果。另一种形式的骨质疏松包括内分泌骨质疏松(甲状腺机能亢进、甲状旁腺机能亢进、柯兴氏综合征和肢端肥大症),遗传性和先天性骨质疏松(骨发育不全,高胱氨酸尿、Menkes′综合征,和Riley-Day综合征)和由于四肢固定而导致的骨质疏松。
最近,业已认识到人群中的骨质疏松同样与动脉钙化的高发病率相关,它是很多动脉粥样硬化病变的一种成分。
因此,如上文所述的食品可以被很好地用于预防分别与骨量减少或骨质疏松相关的症状和/或骨结构变化的发生或减轻这种症状。可以理解的是,可以将所述活性物质以足以实现所需生物学反应的量添加到所述食品物质中。由于业已发现OPG本身是母乳的一种成分,乳基产品本身就很好地适合将所述物质输送到人体。
在另一方面,为了分别治疗严重的骨量减少或骨质疏松病例,优选方案可能是一种药物组合物,该组合物包括较高含量的本发明的osteoprotegerin,该含量足以阻止或者甚至是逆转有关疾病过程。所述组合物可以含有本发明的OPG作为唯一的活性成分,其优点是没有必要涉及所述物质的主要配方。因此,在本发明范围内,可以简单地压制一种由任选补充了载体或香味剂的“OPG-粉末”组成的片剂,不过,当需要将本发明的OPG与其他活性物质一起配制时,应当考虑所述额外物质的性质以及在胃肠道中降解这些物质的可靠性。以剂量单位形式配制的本发明的OPG,使得主治医师能更好地控制所述活性化合物的每日或每周的剂量。
还可将本发明的osteoprotegerin用于预防骨佩吉氏病,骨髓炎,导致骨损失的骨感染性病变,高钙血症,骨坏死,由于骨关节炎或类风湿关节炎所导致的骨损失,牙周骨损失和/或溶骨性转移的发生和/或治疗所述疾病。
业已发现OPG是肿瘤坏死因子相关配体(TRAIL)的一种受体,它在与其含有死亡结构域的受体结合之后能诱导细胞凋亡。据推测它能调控细胞死亡,并且在淋巴组织形成和免疫应答调控方面发挥重要作用。另外,OPG是RANKL的引诱受体(NF-κB的受体激活物的配体),业已报导它是激活的T细胞的产物。RANKL受体在成熟树突细胞上的连接,能增强树突细胞的存活,另外,RANKL与其受体的结合,能增强T-细胞生长和树突细胞功能。
因此,本发明提供了可以从人和/或牛乳中获得的osteoprotegerin在生产可食用制剂,如饮食组合物或肠道组合物,或生产药物中的用途,所述可食用制剂和药物分别用于导致免疫组织的正常发育,用于导致正常的免疫功能,甚至用于预防和/或治疗免疫系统疾病。
本发明考虑的免疫系统疾病包括过敏、自身免疫、脓毒症、癌症、炎性肠病、系统性自身免疫状况、心血管疾病和皮肤、口腔、胃肠道、泌尿生殖道或呼吸道的免疫病理学状况。
另外,本发明的osteoprotegerin同样可应用于调控细胞增殖和凋亡,用于促进口腔耐受性,调节感染过程和新生儿的细菌集群。特别是对于新生儿来说,上述疾病可能在很大程度上与早产和/或低出生体重相关,因此,在这种情况下,可以通过婴儿食品的方式给所述婴儿简单地服用osteoprotegerin。
可以理解的是,任何年龄的个体都可以成为接受治疗的个体,不过,婴儿和老年人是主要考虑对象,因为他们本身需要外源osteoprotegerin。特别是诸如新生儿的个体,其骨物质和/或免疫系统的发育需要osteoprotegerin,因此,在这种情况下,可以有利地给予本发明的所述化合物和/或食品物质和/或药物组合物。
不过,应当理解的是,本发明还可应用于成年人,以便预防上述任何疾病的发生。还应当理解的是,除了人之外,要治疗的个体还可以是动物,如宠物,其中,将本发明的OPG掺入宠物食品。
本发明的OPG可以从乳源中获得,所述乳源来自哺乳动物,特别是来自人或牛乳或初乳。人乳OPG具有380个氨基酸的氨基酸序列,与用作分子量标准的蛋白标记(BioRad)相比,其分子量大约为80、130和200kDa。它具有4个N-糖基化位点,并且能够以单体形式存在,并且通过Cys379形成S-S键,从而形成二聚体。
本发明的OPG可以从诸如人或牛乳的乳源中分离。不过,应当理解的是,本发明的OPG可以通过重组方法在合适的细胞中制备,产生存在于“乳-OPG”中的糖基化模式。用于表达的优选细胞是乳腺细胞,因为预计这种细胞能产生相同的或基本上相同的糖基化模式。
适用于表达本发明OPG的合适的细胞,可以通过用合适的方法永生化而获得,如SV40载体或端粒酶基因,并且用含有编码OPG多肽的核苷酸序列的表达载体转化。当感兴趣的多肽是分泌的多肽时,可以通过从上清液中分离获得这种多肽,或者通过收集所述细胞,并且从所述细胞中分离所述多肽。对于连续生产来说,优选从上清液中分离。下面的实施例对本发明进行说明,但本发明并不局限于这些实施例。
实施例
人乳和人血清样品
在无菌条件下,通过吸乳泵或偶然通过人工挤压从健康母亲体内采集人乳样品(10-60ml),直到产后17天。将所述人乳挤入无菌的50ml离心管中,并且在收集之后2小时内处理,在离心(200×g,10分钟)之后,马上取出细胞沉淀,并且处理,以便提取RNA。其余的乳在-20℃下冷冻。人血清样品是从健康供体获得的,并且在-20℃下保存。
人乳的分级分离
通过高速离心从全乳中提取乳脂。取出上层乳脂层,用水洗涤,并且将洗涤过的乳脂放在-20℃下保存直到使用。乳清和酪蛋白的分离是通过对脱脂乳进行粗制凝乳酶处理或化学酸化(用盐酸)诱导酪蛋白凝固而实现的。对处理过的乳进行离心,然后分别从非溶解地的粗制凝乳酶酪蛋白中分离甜乳清,并且从酸性酪蛋白中分离酸性乳清。最后,通过超速离心制备可溶性乳蛋白(超速离心的乳清)和不可溶性胶束酪蛋白。在-20℃下冷冻所有酪蛋白和乳清级份,直到使用。
人乳腺细胞系
MCf-7(美国典型培养物保藏中心(ATCC),Manassas,VA.,HTB-22)是一种源于乳腺癌胸膜渗出液的人乳腺细胞系,它保留了分化的乳腺上皮细胞的若干特征。在添加了lO%胎牛血清(FCS,Amimed Bioconcept)的DMEM(Amimed Bioconcept,Allschwill,SwitZ6rland)中培养所述细胞,并且在37℃下,在含有5% CO2的潮湿气体中保持。每周更换2-3次培养基。在达到铺满之后,在37℃下,用胰蛋白酶/EDTA(GibcoBRL)分离细胞。然后制备所述细胞进行RNA提取。
蛋白印迹分析
用Laemmli还原样品缓冲液将乳样品稀释l/25倍,并且煮沸5分钟。通过10%SDS-PAGE分离所述蛋白,并且转移到硝酸纤维素膜(BioRad)上。用生物素化的多克隆抗人OPG(BAF805,O.2μg/ml;R&D systems)和链亲和素碱性磷酸酶(Pieroe)探测所述印迹。用碱性磷酸酶底物BCIP/NBT(Zymed Laboratofies)观察免疫反应性。将预先染色的蛋白标记用作分子量标准物(BioRad)。将以25ng/泳道的用量加样的重组人OPG(R&D systems)用作阳性对照。
通过人乳腺乳细胞和人乳腺上皮细胞表达OPG
在逆转录之后接着进行PCR,在来自产后18天的单身母亲的全人乳细胞群中,以及在人乳腺上皮细胞系MCF-7中扩增OPG转录物。通过Trizol方法(Gibco-BRL)从所述细胞中提取总RNA。简单地讲,将Trizol(5-10×106细胞使用lml)添加到细胞沉淀中,用移液管吸入和排出若干次,并且转移到一个Eppendorf试管中。添加氯仿(每lml Trizol添加O.2m1),并且将该试管温育5分钟,然后在4℃下以12,000×g的速度离心15分钟。用等体积的异丙醇使RNA沉淀,并且以12,000×g的速度离心10分钟。用70%的乙醇洗涤沉淀,然后重新悬浮在无菌的去离子水中。在-20℃下保存RNA直到使用。
用不含RNA酶的DNase I处理RNA样品,以便消除基因组DNA的污染。通过在光谱仪中测定合适稀释度(100-200倍)在260曲和280nm下的吸收值,对RNA进行定量。按以下方法计算RNA浓度(μg/ml):在A260×稀释倍数×40mg/ml下的吸收值。基本上不含蛋白的总RNA样品的A260/A280的比值应当为1.8-2.2。
用莫洛尼鼠白血病病毒逆转录酶(Perkin Elmer)对RNA进行逆转录。将RNA样品(0.5μg总RNA),0.5个单位的RNA酶抑制剂,1mM每一种dNTP,0.5nmol/ml特异性3’引物,5mM MgCl2和1.25个单位的逆转录酶放入含有由生产商提供的酶缓冲液的总体积为10μl反应混合物中温育。在42℃下,将该反应混合物温育30分钟,然后在95℃下加热5分钟。然后在热循环仪(Biolabo,Scientific Instruments,ChatelSt Denis,Switzerland)上用Gold DNA聚合酶(Perkin Elmer)扩增所述逆转录产物。在50μl的总体积中进行聚合酶链式反应(PCR)。在PCR缓冲液中使用10μl的逆转录产物,2mM MgCl2,5μM每一种dNTP,0.2nmol/ml OPG-特异性反义
ACTAGTTATAAGCAGCTTATTTTTACTG,
和有义GGAGGCATTCTTCAGGTTTGCTG引物,和1.25个单位的DNA聚合酶。在95℃下进行10分钟的最初变性步骤,然后通过在94℃下变性45秒,在60℃下退火1分钟,并且在72℃下延伸1分30秒然后在72℃下进行7分钟的延伸,共进行35轮,以便扩增样品。用β-肌动蛋白作为阳性对照对所有样品进行RT-PCR。将RT-PCR产物的样品加样到用1×TAE缓冲液配制的1.2%的琼脂糖凝胶(含有溴化乙锭)上,并且通过在150伏电压下电泳1小时分离。在UV光下观察RT-PCR产物。通过与DNA大小标记(Boehringer Mannheim)比较,确定所述带的正确大小。
人OPG的ELI SA-(夹心酶免疫测定)
通过ELISA测定存在于乳和不同乳级份中OPG的浓度。为此,通过在4℃下温育过夜,将抗OPG的单克隆抗体(MAB805,1μg/ml;R&DSystems,UK)包被到96孔平板(Nunc)上。然后用由PBS配制的0.05%的Tween-20洗涤平板2次。通过在室温下用由PBS配制的2%的牛血清白蛋白(BSA)将所述平板再温育2小时抑制非特异性结合。在室温下,在PBS-BSA中将样品或标准浓度的重组OPG(0.119-121.5ng/ml;R&DSystems)温育3小时。然后用PBS-Tween洗涤平板4次,然后添加生物素标记过的抗人OPG多克隆抗体(BAF805,0.5μg/ml;R&D Systems),在室温下再温育1小时。再经过4次洗涤之后,添加链亲和素-过氧化物酶(SAAP,0.5μg/ml,Kirkegaard % Perry KPL)在室温下温育1小时。洗涤平板4次,并且添加底物TMB过氧化物酶(KPL)。覆盖平板,并且遮光温育5分钟。通过添加1N HCl终止所述酶促反应。在450nm下用ELISA读数器(Dynex Technologies)读出吸光度。检测限为大约30pg/ml。
人乳OPG的生物学活性
OPG是肿瘤坏死因子相关配体的受体(TRAIL),它在与它的含有死亡结构域的受体DR4和DR5结合之后,能诱导细胞凋亡。开发了一种生物测定方法,其中,可以测定人乳OPG阻断TRAIL-诱导的所述细胞的凋亡的能力。
就此而言,将Jurkat细胞、克隆E6-1(ATCC)维持在由供应商ATCC改进的含有10% FCS的RPMI 1640培养基中(37℃,5% CO2)。以5×104细胞/孔的密度将细胞接种到96孔平板(Nunc)上。在存在2μg/ml增强蛋白的条件下向每一个孔中添加各种浓度的可溶性重组人TRAIL(0-20ng/ml),所述增强子蛋白是一种抗体,它能与可溶性重组人TRAIL反应,并因此增强其活性(Alexis,Laufelfingen,CH)。某些孔还装有50ng/ml重组人OPG(R&D Systems),人乳样品(HM;1/80最终稀释度;在产后第一天或第九天采集),和/或20μg/ml抗-OPG单克隆抗体(MAB805,R&DSystems)。在37℃下温育平板16小时。在培养的最后6小时,通过添加3H-胸苷(1μCi/孔)测定细胞存活力。
在培养基对照中,在超过5ng/ml的浓度下观察到了TRAIL-诱导的细胞增殖的抑制。不过,HM样品能阻止这种抑制作用。这种作用显然是由于OPG的存在所导致的,因为抗OPG单克隆抗体能逆转这种作用。
结果如图5所示
蛋白印迹分析
OPG在细胞中是以55kDa的单体形式合成的,不过,在分泌到细胞外之后,转化成大约110kDa的二硫键连接的二聚体。在乳中,检测到大约80,130和200kDa的带。
人乳中的OPG浓度
通过ELISA检查了在哺乳的前17天的不同时间,10位哺乳母亲的乳样品中的OPG含量。在哺乳的前1-3天,其浓度提高到最大值,然后降低。OPG在乳中的浓度为大约50ng/ml-大约2μg/ml(图1)。
乳OPG的细胞来源
RT-PCR分析发现,本发明的OPG可能存在于人乳细胞和乳腺上皮细胞中。在两种类型的未处理过的细胞中证实了OPG mRNA的组成型表达(图4)。
将人乳OPG克隆到酵母中
通过离心(200×g,10分钟)从人乳(产后18天)中分离细胞。用TRIzol(Life Technologies,Basel,Switzerland)从所述细胞沉淀中提取总RNA,用DNAseI处理,并且按照生产商推荐的方法在RNeasy旋转柱(Qiagen,Basel)上进一步纯化。按照生产商提供的方法(RocheDiagnostics,Rotkreuz),使用TitanTMOne试管RT-PCR系统,由该总RNA扩增编码成熟形式的OPG的PCR产物。
使用OPG特异性反义引物
CC
GGCCTCTTCGGCCGCCAAGCGA
GAAACGTTTCCTCCAAAGTACC,和有义引物ACTAGTTATAAGCAGCTTATTTTTACTG,由该cDNA扩增了一个1174bp PCR片段。对该PCR产物进行SfiI-SpeI消化,凝胶纯化,并且将所得到的1156bp片段连接到SfiI-XbaI消化过的和SAP-处理过的pINA1267上,形成pNFF270。然后通过转化将质粒pNFF270导入酵母Yarrowia lipolytica中。图3表示整合到Yarrowia转化体的基因组DNA中的所述质粒的限制图谱,而SEQ ID.No.1表示由该OPG质粒编码的蛋白。
pGEM-T OPG克隆的序列如图6所示。成熟的OPG用黑体表示,并且翻译。在公开的OPG/OCIF序列上,成熟OPG的242号氨基酸残基是Ala-残基(A),而分析过的所有pGEM-T OPG克隆编码位于该位置上的Asp-残基(D)。将该克隆的SfiI-SpeI OPG片段转移到SfiI-XbaI消化过的pINA1267上。所得到的质粒具有图3所示的限制图谱。将单拷贝的该质粒整合到Yarrowia转化体的基因组DNA上。在图4中示出了由该质粒编码的蛋白。成熟的OPG用粗体字母表示。通过转化将质粒pNFF270导入Yarrowia lipolytica中。所得到的转化体能将一种与OPG-特异性抗体交叉反应的蛋白分泌到培养基中,而具有空的表达载体的Y.lipolytica转化体不能分泌这种蛋白。
Claims (14)
1.可以从人或牛乳或初乳中获得的osteoprotegerin。
2.如权利要求1的osteoprotegerin,其中,所述osteoprotegerin具有能产生具有大约80,130和200kDa的分子量的多肽的糖基化模式。
3.一种食品物质,含有如权利要求1或2中任意一项的osteoprotegerin。
4.选自下列一组的食品物质:乳、酸乳、凝乳、奶酪、发酵乳、乳基发酵制品、冰淇淋、谷基发酵制品、乳基粉末、婴儿配方和宠物食品。
5.一种肠道组合物或药物组合物,含有权利要求1或2中任意一项的osteoprotegerin。
6.如权利要求5的肠道或药物组合物,该组合物选自下列一组:溶液、干燥的口服添加剂、液体口服添加剂、干燥的管装饲料或液体管装饲料。
7.权利要求1或2的osteoprotegerin在生产可以食用的制剂中的用途。
8.如权利要求7的用途,其中,所述可以食用的制剂是权利要求3或4的食品物质或权利要求5或6的组合物。
9.权利要求1或2的osteoprotegerin在制备用于治疗与骨重建相关的疾病的如权利要求3-6中任意一项的物质或组合物中的用途。
10.如权利要求9的用途,其中,所述疾病是骨质疏松症、骨佩吉氏病、骨髓炎、导致骨损失的骨感染性病变、高钙血症、骨量减少、骨坏死、由骨关节炎或类风湿关节炎导致的骨损失、牙周骨损失和/或溶骨性转移。
11.权利要求1或2的osteoprotegerin在制备用于治疗和/或预防免疫疾病的如权利要求3-6中任意一项的物质或组合物中的用途。
12.如权利要求11的用途,其中,所述疾病是过敏、自身免疫、炎性肠病、系统性自身免疫状况、细胞增殖和细胞凋亡失调以及皮肤、口腔、胃肠道、泌尿生殖道或呼吸道的免疫病理学状况。
13.如权利要求9-12中任意一项的用途,其中,所述疾病与早产和/或低出生体重相关。
14.权利要求1或2的osteoprotegerin在制备用于骨物质和/或免疫系统发育的如权利要求3-6中任意一项的物质或组合物中的用途。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP01108414 | 2001-04-03 | ||
EP01108414.2 | 2001-04-03 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN1500096A true CN1500096A (zh) | 2004-05-26 |
Family
ID=8177040
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNA028078640A Pending CN1500096A (zh) | 2001-04-03 | 2002-03-15 | 乳中的osteoprotegerin |
Country Status (18)
Country | Link |
---|---|
US (2) | US7749960B2 (zh) |
EP (2) | EP1757619B1 (zh) |
JP (1) | JP4921687B2 (zh) |
CN (1) | CN1500096A (zh) |
AT (1) | ATE458004T1 (zh) |
AU (1) | AU2002253133B2 (zh) |
BR (1) | BR0208554A (zh) |
CA (1) | CA2442694C (zh) |
DE (1) | DE60235418D1 (zh) |
ES (1) | ES2337716T3 (zh) |
IL (1) | IL158152A0 (zh) |
MX (1) | MXPA03008895A (zh) |
PL (1) | PL366451A1 (zh) |
PT (1) | PT1757619E (zh) |
RU (1) | RU2324705C2 (zh) |
UA (1) | UA84831C2 (zh) |
WO (1) | WO2002081521A2 (zh) |
ZA (1) | ZA200308490B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109803681A (zh) * | 2016-08-19 | 2019-05-24 | 新加坡保健服务集团有限公司 | 用于治疗免疫病症的免疫抑制组合物 |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7943328B1 (en) | 2006-03-03 | 2011-05-17 | Prometheus Laboratories Inc. | Method and system for assisting in diagnosing irritable bowel syndrome |
GB0604460D0 (en) | 2006-03-06 | 2006-04-12 | Isis Innovation | Treatment |
US9878003B2 (en) | 2006-03-06 | 2018-01-30 | The University Of Manchester | Method of treating bone disorders using TSG-6 |
US20080085524A1 (en) * | 2006-08-15 | 2008-04-10 | Prometheus Laboratories Inc. | Methods for diagnosing irritable bowel syndrome |
US20100094560A1 (en) * | 2006-08-15 | 2010-04-15 | Prometheus Laboratories Inc. | Methods for diagnosing irritable bowel syndrome |
WO2008078588A1 (ja) * | 2006-12-25 | 2008-07-03 | National University Corporation Tokyo Medical And Dental University | 関節軟骨の変性を治療又は予防するための医薬及び方法 |
AU2022318574B2 (en) | 2021-07-30 | 2024-03-21 | Helaina, Inc. | Methods and compositions for protein synthesis and secretion |
Family Cites Families (48)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB124309A (en) | 1918-04-16 | 1919-03-27 | John Page Croft | An Improved Coffee Preparation. |
FR2039157A7 (en) | 1969-04-14 | 1971-01-15 | Marlan Sl | Liquid coffee concentrate |
DE3821374A1 (de) | 1988-06-24 | 1990-01-18 | Nolde Sylvia | Kaffee-bonbons |
US4946701A (en) | 1989-08-04 | 1990-08-07 | Procter & Gamble | Beverages |
US5182926A (en) | 1991-09-16 | 1993-02-02 | Nestec S.A. | Recovery of aroma gases |
EP0652889A4 (en) * | 1992-06-15 | 1997-05-07 | Gene Pharming Europ Bv | PRODUCTION OF RECOMBINANT POLYPEPTIDES BY BOVINE SPECIES AND TRANSGENIC PROCESSES. |
GB2285578A (en) | 1993-12-22 | 1995-07-19 | Leilani Lea | Beverages containing added guaranine |
JP3112637B2 (ja) | 1994-09-30 | 2000-11-27 | 雪印乳業株式会社 | 骨強化剤 |
JPH08165249A (ja) | 1994-12-14 | 1996-06-25 | Snow Brand Milk Prod Co Ltd | 骨強化作用を有する組成物 |
IL117175A (en) * | 1995-02-20 | 2005-11-20 | Sankyo Co | Osteoclastogenesis inhibitory factor protein |
US6369027B1 (en) | 1995-12-22 | 2002-04-09 | Amgen Inc. | Osteoprotegerin |
US6613544B1 (en) * | 1995-12-22 | 2003-09-02 | Amgen Inc. | Osteoprotegerin |
JP3578538B2 (ja) | 1996-01-09 | 2004-10-20 | 雪印乳業株式会社 | 離乳食 |
JP3613488B2 (ja) | 1996-01-17 | 2005-01-26 | 雪印乳業株式会社 | 栄養補給用組成物 |
JP2974604B2 (ja) * | 1996-01-23 | 1999-11-10 | 雪印乳業株式会社 | 塩基性タンパク質組成物、塩基性ペプチド組成物及びその利用 |
JP3061364B2 (ja) | 1996-04-26 | 2000-07-10 | 雪印乳業株式会社 | 乳児用調製乳 |
JPH1057071A (ja) | 1996-08-19 | 1998-03-03 | Snow Brand Milk Prod Co Ltd | 新規dna及びそれを用いた蛋白質の製造方法 |
EP0826308B1 (en) | 1996-08-26 | 2000-12-27 | Societe Des Produits Nestle S.A. | Coffee extraction process and product |
EP0951546A2 (en) | 1996-12-20 | 1999-10-27 | The Board Of Regents, The University Of Texas System | Compositions and methods of use for osteoclast inhibitory factors |
RU2238949C2 (ru) | 1997-04-15 | 2004-10-27 | Санкио Компани Лимитед | Белок, специфически связывающийся с ингибирующим остеокластогенез фактором (ocif) (варианты), днк его кодирующая (варианты), днк в качестве зонда (варианты), способ получения белка (варианты), способ скрининга вещества (варианты), антитело (варианты), способ получения поликлонального антитела, гибридома (варианты), способ получения моноклонального антитела, способ измерения ocif-связывающего белка, фармацевтическая композиция (варианты) и лекарственное средство (варианты) |
JP2002514079A (ja) | 1997-05-01 | 2002-05-14 | アムジエン・インコーポレーテツド | キメラopgポリペプチド |
AU7705098A (en) | 1997-05-29 | 1998-12-30 | Human Genome Sciences, Inc. | Human tumor necrosis factor receptor-like protein 8 |
US6346388B1 (en) | 1997-08-13 | 2002-02-12 | Smithkline Beecham Corporation | Method of identifying agonist and antagonists for tumor necrosis related receptors TR1 and TR2 |
ATE328281T1 (de) * | 1997-09-24 | 2006-06-15 | Sankyo Co | Methode zur diagnose von abnormalem knochenstoffwechsel |
US6194151B1 (en) | 1997-09-26 | 2001-02-27 | Millenium Pharmaceuticals, Inc. | Molecules of the TNF receptor superfamily and uses therefor |
JP3933273B2 (ja) | 1997-09-30 | 2007-06-20 | 雪印乳業株式会社 | 家畜飼料 |
US6297022B1 (en) | 1997-10-08 | 2001-10-02 | Smithkline Beecham Corporation | Method of identifying agonists and antagonists for tumor necrosis related receptor TR1 |
US6087555A (en) | 1997-10-15 | 2000-07-11 | Amgen Inc. | Mice lacking expression of osteoprotegerin |
US5997929A (en) | 1997-11-03 | 1999-12-07 | Nestec S.A. | Extraction process |
AU1535699A (en) | 1997-11-24 | 1999-06-15 | Apotech S.A. | Novel receptors opg-2 |
JPH11155420A (ja) | 1997-12-02 | 1999-06-15 | Snow Brand Milk Prod Co Ltd | トランスジェニック動物 |
CA2316545A1 (en) | 1997-12-29 | 1999-07-08 | Regeneron Pharmaceuticals, Inc. | Novel nucleic acid and polypeptide with homology to the tnf-receptors |
US6093436A (en) | 1998-02-04 | 2000-07-25 | Nestec S.A. | Beverage antioxidant system |
US6149957A (en) | 1998-04-09 | 2000-11-21 | Nestec S.A. | Aroma recovery process |
US6790823B1 (en) * | 1998-04-23 | 2004-09-14 | Amgen Inc. | Compositions and methods for the prevention and treatment of cardiovascular diseases |
JP4082782B2 (ja) | 1998-04-30 | 2008-04-30 | 雪印乳業株式会社 | 歯周病予防及び改善剤 |
WO1999061038A1 (en) | 1998-05-29 | 1999-12-02 | Adams Food Ltd. | Composition having therapeutic and/or nutritionally active substituent |
JP2000086697A (ja) | 1998-09-10 | 2000-03-28 | Snow Brand Milk Prod Co Ltd | 新規蛋白質、その製造法及び用途 |
AU754971B2 (en) | 1998-09-15 | 2002-11-28 | Pharmexa A/S | Method for down-regulating osteoprotegerin ligand activity |
JP4097335B2 (ja) * | 1998-09-30 | 2008-06-11 | 生化学工業株式会社 | 食品添加物 |
JP2000102390A (ja) | 1998-09-30 | 2000-04-11 | Snow Brand Milk Prod Co Ltd | Dna及びそれを用いた蛋白質の製造方法 |
AU6006099A (en) | 1998-10-09 | 2000-05-01 | Sankyo Company Limited | Preventives or remedies for cachexia |
US6916907B1 (en) * | 1998-10-23 | 2005-07-12 | Curagen Corporation | Nucleic acids encoding osteoprotegern-like proteins and methods of using same |
EP1127578A4 (en) * | 1998-10-28 | 2004-12-15 | Sankyo Co | MEDICINE FOR BONE METABOLIC DEVIATIONS |
AU765510B2 (en) | 1999-01-28 | 2003-09-18 | Societe Des Produits Nestle S.A. | Aromatised soluble creamer powder |
US6413558B1 (en) | 1999-07-19 | 2002-07-02 | The Proctor & Gamble Co. | Compositions, kits, and methods for providing and maintaining energy and metal alertness |
AU6946300A (en) * | 1999-08-30 | 2001-03-26 | Mayo Foundation For Medical Education And Research | Use of dna encoding osteoprotegerin to prevent or inhibit metabolic bone disorders |
RU2268069C2 (ru) * | 2000-04-18 | 2006-01-20 | Сосьете Де Продюи Нестле С.А. | Питательные модули |
-
2002
- 2002-03-15 ES ES06124157T patent/ES2337716T3/es not_active Expired - Lifetime
- 2002-03-15 PT PT06124157T patent/PT1757619E/pt unknown
- 2002-03-15 AU AU2002253133A patent/AU2002253133B2/en not_active Ceased
- 2002-03-15 JP JP2002579906A patent/JP4921687B2/ja not_active Expired - Fee Related
- 2002-03-15 PL PL02366451A patent/PL366451A1/xx not_active Application Discontinuation
- 2002-03-15 EP EP06124157A patent/EP1757619B1/en not_active Expired - Lifetime
- 2002-03-15 EP EP02722228A patent/EP1377610A2/en not_active Ceased
- 2002-03-15 CA CA2442694A patent/CA2442694C/en not_active Expired - Fee Related
- 2002-03-15 BR BR0208554-2A patent/BR0208554A/pt not_active Application Discontinuation
- 2002-03-15 MX MXPA03008895A patent/MXPA03008895A/es unknown
- 2002-03-15 RU RU2003132059/13A patent/RU2324705C2/ru not_active IP Right Cessation
- 2002-03-15 CN CNA028078640A patent/CN1500096A/zh active Pending
- 2002-03-15 WO PCT/EP2002/002912 patent/WO2002081521A2/en not_active Application Discontinuation
- 2002-03-15 DE DE60235418T patent/DE60235418D1/de not_active Expired - Lifetime
- 2002-03-15 IL IL15815202A patent/IL158152A0/xx unknown
- 2002-03-15 AT AT06124157T patent/ATE458004T1/de not_active IP Right Cessation
- 2002-03-21 UA UA2003098661A patent/UA84831C2/ru unknown
-
2003
- 2003-10-02 US US10/676,358 patent/US7749960B2/en not_active Expired - Lifetime
- 2003-10-30 ZA ZA200308490A patent/ZA200308490B/en unknown
-
2005
- 2005-06-02 US US11/144,236 patent/US7524815B2/en not_active Expired - Fee Related
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109803681A (zh) * | 2016-08-19 | 2019-05-24 | 新加坡保健服务集团有限公司 | 用于治疗免疫病症的免疫抑制组合物 |
CN109803681B (zh) * | 2016-08-19 | 2023-12-12 | 新加坡保健服务集团有限公司 | 用于治疗免疫病症的免疫抑制组合物 |
Also Published As
Publication number | Publication date |
---|---|
PT1757619E (pt) | 2010-03-25 |
EP1757619A3 (en) | 2007-03-14 |
JP2004537980A (ja) | 2004-12-24 |
ATE458004T1 (de) | 2010-03-15 |
CA2442694C (en) | 2012-03-06 |
BR0208554A (pt) | 2004-03-02 |
EP1757619B1 (en) | 2010-02-17 |
US7524815B2 (en) | 2009-04-28 |
ZA200308490B (en) | 2005-01-31 |
EP1757619A2 (en) | 2007-02-28 |
MXPA03008895A (es) | 2005-03-07 |
RU2003132059A (ru) | 2005-05-10 |
IL158152A0 (en) | 2004-03-28 |
PL366451A1 (en) | 2005-02-07 |
CA2442694A1 (en) | 2002-10-17 |
US20040137074A1 (en) | 2004-07-15 |
WO2002081521A8 (en) | 2003-10-23 |
RU2324705C2 (ru) | 2008-05-20 |
WO2002081521A2 (en) | 2002-10-17 |
WO2002081521A3 (en) | 2003-08-28 |
UA84831C2 (ru) | 2008-12-10 |
AU2002253133B2 (en) | 2008-02-28 |
JP4921687B2 (ja) | 2012-04-25 |
US20050288219A1 (en) | 2005-12-29 |
EP1377610A2 (en) | 2004-01-07 |
US7749960B2 (en) | 2010-07-06 |
ES2337716T3 (es) | 2010-04-28 |
DE60235418D1 (de) | 2010-04-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Xu et al. | Postnatal adaptation of the gastrointestinal tract in neonatal pigs: a possible role of milk-borne growth factors | |
US7524815B2 (en) | Osteoprotegerin in milk | |
JP4740531B2 (ja) | 骨吸収抑制剤 | |
AU754676B2 (en) | Food-induced antisecretory proteins in egg yolk | |
AU2002253133A1 (en) | Osteoprotegerin in milk | |
Iskandar et al. | Gene prolactine receptor (PRLR) and signal transducer and activator of transcription 5 (STAT5) on milk production | |
CN100358919C (zh) | 哺乳动物促乳素变体 | |
US6967090B2 (en) | Nucleic acid and protein sequences of bovine epidermal growth factor | |
Maeda et al. | Gene expression of lymphocyte prolactin receptor was suppressed in lactating mothers | |
ARITON et al. | IMPACT OF A2 MILK ON HUMAN HEALTH AND THE DAIRY INDUSTRY-A REVIEW. | |
Colditz | Metabolic effects of host defence responses during gastrointestinal parasitism in sheep | |
CN101868475A (zh) | 骨吸收抑制用食品原材料 | |
Yadav | Isolation and Characterization of Genetic Variants of Beta-Casein Protein (A1/A2) and Study Their Impact on Early Precipitation of Osteoporosis | |
JP5120971B2 (ja) | 骨吸収抑制剤 | |
Kobayashi et al. | Expression of stanniocalcin-1 in gastrointestinal tracts of neonatal and mature rats | |
CN111657397A (zh) | 一种基于直链淀粉提升山羊生长性能的饲料 | |
JP2772499B2 (ja) | 豚用飼料およびそれを用いて豚を飼育する方法 | |
JP5379884B2 (ja) | 骨吸収抑制剤 | |
CN116671583A (zh) | 低胆固醇鸡蛋饲料及其制备方法 | |
Hwang et al. | Effect of IGF-I rich fraction from bovine colostral whey on murine immunity | |
Wimmers | Articles in PresS. Am J Physiol Regul Integr Comp Physiol (March 16, 2016). doi: 10.1152/ajpregu. 00215.2015 | |
Sanglikar | Genes in IGF pathway and their association with feed efficiency | |
JP2018052846A (ja) | 心房性ナトリウム利尿ペプチド分泌促進用組成物 | |
JPH0764739B2 (ja) | 骨成分改善組成物 | |
AU2002257450A1 (en) | Nucleic acid and protein sequences of bovine epidermal growth factor |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
WD01 | Invention patent application deemed withdrawn after publication |