JPH0228118A - Production of immunologically active peptide - Google Patents
Production of immunologically active peptideInfo
- Publication number
- JPH0228118A JPH0228118A JP63178447A JP17844788A JPH0228118A JP H0228118 A JPH0228118 A JP H0228118A JP 63178447 A JP63178447 A JP 63178447A JP 17844788 A JP17844788 A JP 17844788A JP H0228118 A JPH0228118 A JP H0228118A
- Authority
- JP
- Japan
- Prior art keywords
- active peptide
- immunologically active
- acid
- animal
- bone
- 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
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 29
- 238000004519 manufacturing process Methods 0.000 title claims description 5
- 210000000988 bone and bone Anatomy 0.000 claims abstract description 19
- 241001465754 Metazoa Species 0.000 claims abstract description 10
- 210000001185 bone marrow Anatomy 0.000 claims abstract description 7
- 239000011260 aqueous acid Substances 0.000 claims description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract description 22
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 abstract description 13
- 239000000243 solution Substances 0.000 abstract description 11
- 241000283690 Bos taurus Species 0.000 abstract description 6
- 239000002253 acid Substances 0.000 abstract description 6
- 208000001132 Osteoporosis Diseases 0.000 abstract description 5
- 239000007864 aqueous solution Substances 0.000 abstract description 5
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 abstract description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 abstract description 4
- 206010039984 Senile osteoporosis Diseases 0.000 abstract description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 abstract description 4
- 230000032683 aging Effects 0.000 abstract description 3
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 abstract description 3
- 229910052500 inorganic mineral Inorganic materials 0.000 abstract description 3
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 abstract description 3
- 239000011707 mineral Substances 0.000 abstract description 3
- 150000007524 organic acids Chemical class 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract description 3
- 210000001519 tissue Anatomy 0.000 abstract description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 abstract description 2
- 241000287828 Gallus gallus Species 0.000 abstract description 2
- 241000124008 Mammalia Species 0.000 abstract description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 abstract description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 2
- 230000007246 mechanism Effects 0.000 abstract description 2
- 230000002503 metabolic effect Effects 0.000 abstract description 2
- 210000005036 nerve Anatomy 0.000 abstract description 2
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract description 2
- 241000282898 Sus scrofa Species 0.000 abstract 2
- 206010057178 Osteoarthropathies Diseases 0.000 abstract 1
- 201000010099 disease Diseases 0.000 abstract 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract 1
- 238000010790 dilution Methods 0.000 description 14
- 239000012895 dilution Substances 0.000 description 14
- 210000002966 serum Anatomy 0.000 description 11
- 102000004196 processed proteins & peptides Human genes 0.000 description 10
- 239000000284 extract Substances 0.000 description 9
- 229960000583 acetic acid Drugs 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 238000003127 radioimmunoassay Methods 0.000 description 6
- 239000000427 antigen Substances 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 235000015278 beef Nutrition 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- NVEXXUGCBSXDLS-LNEXRSTESA-N (4S)-4-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[[(2S)-1-[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-6-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[2-[[(2S)-2-amino-1-hydroxy-3-(4-hydroxyphenyl)propylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxyethylidene]amino]-1-hydroxy-3-phenylpropylidene]amino]-1-hydroxy-4-methylpentylidene]amino]-5-carbamimidamido-1-hydroxypentylidene]amino]-5-carbamimidamido-1-hydroxypentylidene]amino]-1,5-dihydroxy-5-iminopentylidene]amino]-1-hydroxy-3-phenylpropylidene]amino]-1-hydroxyhexylidene]amino]-1-hydroxy-3-methylbutylidene]amino]-1-hydroxy-3-methylbutylidene]amino]-1,3-dihydroxybutylidene]amino]-5-carbamimidamido-1-hydroxypentylidene]amino]-1,3-dihydroxypropylidene]amino]-1,5-dihydroxy-5-iminopentylidene]amino]-4-carboxy-1-hydroxybutylidene]amino]-3-carboxypropanoyl]pyrrolidin-2-yl]-hydroxymethylidene]amino]-1,4-dihydroxy-4-iminobutylidene]amino]-1-hydroxypropylidene]amino]-1-hydroxy-3-(4-hydroxyphenyl)propylidene]amino]-1,3-dihydroxypropylidene]amino]-1-hydroxyethylidene]amino]-5-[(2S)-1-[(2S)-1-[(2S)-3-carboxy-1-[(1S)-1-carboxyethyl]imino-1-hydroxypropan-2-yl]imino-1-hydroxy-3-phenylpropan-2-yl]imino-1-hydroxy-4-methylpentan-2-yl]imino-5-hydroxypentanoic acid Chemical compound CC(C)C[C@H](\N=C(/O)[C@H](CCC(O)=O)\N=C(/O)C\N=C(/O)[C@H](CO)\N=C(/O)[C@H](Cc1ccc(O)cc1)\N=C(/O)[C@H](C)\N=C(/O)[C@H](CC(O)=N)\N=C(/O)[C@@H]1CCCN1C(=O)[C@H](CC(O)=O)\N=C(/O)[C@H](CCC(O)=O)\N=C(/O)[C@H](CCC(O)=N)\N=C(/O)[C@H](CO)\N=C(/O)[C@H](CCCNC(N)=N)\N=C(/O)[C@@H](\N=C(/O)[C@@H](\N=C(/O)[C@@H](\N=C(/O)[C@H](CCCCN)\N=C(/O)[C@H](Cc1ccccc1)\N=C(/O)[C@H](CCC(O)=N)\N=C(/O)[C@H](CCCNC(N)=N)\N=C(/O)[C@H](CCCNC(N)=N)\N=C(/O)[C@H](CC(C)C)\N=C(/O)[C@H](Cc1ccccc1)\N=C(/O)C\N=C(/O)C\N=C(/O)[C@@H](N)Cc1ccc(O)cc1)C(C)C)C(C)C)[C@@H](C)O)C(\O)=N\[C@@H](Cc1ccccc1)C(\O)=N\[C@@H](CC(O)=O)C(\O)=N\[C@@H](C)C(O)=O NVEXXUGCBSXDLS-LNEXRSTESA-N 0.000 description 4
- 108090001053 Gastrin releasing peptide Proteins 0.000 description 4
- 102400000236 Leumorphin Human genes 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 108010074732 preproenkephalin Proteins 0.000 description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 3
- 102100036519 Gastrin-releasing peptide Human genes 0.000 description 3
- 239000011575 calcium Substances 0.000 description 3
- 229910052791 calcium Inorganic materials 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 230000005965 immune activity Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- HFDKKNHCYWNNNQ-YOGANYHLSA-N 75976-10-2 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@@H](NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](C)N)C(C)C)[C@@H](C)O)C1=CC=C(O)C=C1 HFDKKNHCYWNNNQ-YOGANYHLSA-N 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 206010068975 Bone atrophy Diseases 0.000 description 1
- 208000020084 Bone disease Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000252233 Cyprinus carpio Species 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
- 241000196324 Embryophyta Species 0.000 description 1
- 102400001368 Epidermal growth factor Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 102400001370 Galanin Human genes 0.000 description 1
- 101800002068 Galanin Proteins 0.000 description 1
- 102000004862 Gastrin releasing peptide Human genes 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- VXKCPBPQEKKERH-IUCAKERBSA-N Gly-Arg-Pro Chemical compound NC(N)=NCCC[C@H](NC(=O)CN)C(=O)N1CCC[C@H]1C(O)=O VXKCPBPQEKKERH-IUCAKERBSA-N 0.000 description 1
- 208000012659 Joint disease Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102000018886 Pancreatic Polypeptide Human genes 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 102000005157 Somatostatin Human genes 0.000 description 1
- 108010056088 Somatostatin Proteins 0.000 description 1
- 101000983124 Sus scrofa Pancreatic prohormone precursor Proteins 0.000 description 1
- 241000473945 Theria <moth genus> Species 0.000 description 1
- 241000186514 Warburgia ugandensis Species 0.000 description 1
- 230000009102 absorption Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000035508 accumulation Effects 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- VDQQXEISLMTGAB-UHFFFAOYSA-N chloramine T Chemical compound [Na+].CC1=CC=C(S(=O)(=O)[N-]Cl)C=C1 VDQQXEISLMTGAB-UHFFFAOYSA-N 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 108010074605 gamma-Globulins Proteins 0.000 description 1
- PUBCCFNQJQKCNC-XKNFJVFFSA-N gastrin-releasingpeptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)CNC(=O)[C@H](C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)C(C)C)[C@@H](C)O)C(C)C)[C@@H](C)O)C(C)C)C1=CNC=N1 PUBCCFNQJQKCNC-XKNFJVFFSA-N 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- SLZIZIJTGAYEKK-CIJSCKBQSA-N molport-023-220-247 Chemical compound C([C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)CN)[C@@H](C)O)C1=CNC=N1 SLZIZIJTGAYEKK-CIJSCKBQSA-N 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 230000009711 regulatory function Effects 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 1
- 229960000553 somatostatin Drugs 0.000 description 1
- 230000002227 vasoactive effect Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は、免疫活性ペプチドの製造方法に関する。[Detailed description of the invention] [Industrial application field] The present invention relates to a method for producing an immunoactive peptide.
近年、社会の高齢化に伴い、加齢による各種骨関節疾患
に随伴した、いわゆる老化性骨粗鬆症（オステオポロー
ゼ）、すなわち骨萎縮を起こすことが知られている。In recent years, with the aging of society, it has been known that so-called senile osteoporosis (osteoporosis), that is, bone atrophy, is associated with various bone and joint diseases due to aging.
生骨髄カルシウムについて、老化性骨粗鬆症に対する有
効性が指摘されている（天井および均温、新薬と臨床、
↓ユ、１（１９８４））。しかしながら、カルシウムの
吸収、蓄積、代謝、排泄等の経路は、複雑であり生体内
では、これらが各種ホルモン、ペプチドにより調節をう
けていることが、示唆されている。It has been pointed out that raw bone marrow calcium is effective against senile osteoporosis (ceiling and isotherm, new drugs and clinical studies,
↓Yu, 1 (1984)). However, the pathways of absorption, accumulation, metabolism, excretion, etc. of calcium are complex, and it has been suggested that these are regulated in vivo by various hormones and peptides.
現在、多種多様の生理活性ペプチドが見出だされ、それ
らの細胞内カルシウム調節に対する作用がン主目されて
いる。At present, a wide variety of physiologically active peptides have been discovered, and their effects on intracellular calcium regulation are the focus of attention.
しかしながら、骨および骨髄中の生理活性ペプチドの存
在については、全く検討されておらず、現在なお不明の
ままである。However, the presence of bioactive peptides in bone and bone marrow has not been investigated at all and remains unclear at present.
天然補助食品の効果を検討するうえにもその存在の有無
の検索は、重要な基礎的な情報を提供する物と考える。We believe that searching for the presence or absence of natural supplements provides important basic information when examining the effects of natural supplements.
本発明は、骨および骨髄の微粉砕物中に含まれる各種免
疫活性ペプチド成分の製造をその目的とするものである
。The object of the present invention is to produce various immunoactive peptide components contained in finely ground bone and bone marrow.
即ち本発明は、鳥獣骨または骨髄を含有する粉砕物を酸
水溶液と接触させ、酸水溶液中に免疫活性ペプチドを抽
出することを特徴とする、免疫活性ペプチドの製造方法
をその要旨とするものである。That is, the gist of the present invention is a method for producing an immunoactive peptide, which is characterized by contacting a pulverized product containing bird and animal bones or bone marrow with an aqueous acid solution, and extracting the immunoactive peptide into the aqueous acid solution. be.
鳥獣骨または骨髄を含有する粉砕物における、鳥獣とし
ては、鶏、豚、猪、牛等を挙げることができる。Examples of the birds and animals in the crushed material containing bird and animal bones or bone marrow include chickens, pigs, boars, and cows.
鳥獣骨または骨髄を含有する粉砕物の製造方法としては
、たとえば、鳥獣の頭部と四肢を除去した鳥獣骨を、約
２°Ｃ以下の温度下で粗粉砕し、液体アンモニウム冷却
下で凍結し、粉砕したのち、脱窒素化し、マイクロ波お
よび熱風等により乾燥する方法等を挙げることができる
。As a method for producing a crushed product containing bird and animal bones or bone marrow, for example, bird and animal bones from which the head and limbs have been removed are coarsely ground at a temperature of about 2°C or less, and then frozen under liquid ammonium cooling. Examples include a method of pulverizing, denitrifying, and drying using microwaves, hot air, or the like.
また、食肉処理工場において抜管した牛骨を約２０゛Ｃ
に冷却し、出口孔径６．５胴のチョンパーにより、牛骨
細片とし、これの約１００部に対し５５部の氷水を加え
、さらに出口孔径約２．５ ｍｍのチゴッパーにより細
片化し、次いでこれの約１５５部に氷水約４５部を加え
て摺動磨砕機（たとえばスーパーマスコロイダ−ＭＫＺ
８１．５−４０、増幸産業株式会社製造）を通過させ、
さらに磨砕機（プーラ−社製造）に、冷却下に通過させ
て粉砕する等の方法（特願昭５５−６７５９１号明細書
、実施例）を挙げることができる。In addition, extubated beef bones at a meat processing plant are heated to approximately 20°C.
The beef bones were cooled to 100 mL, cut into small pieces using a chomper with an outlet hole diameter of 6.5 mm, and 55 parts of ice water was added to about 100 parts of the beef bones. Approximately 45 parts of ice water is added to approximately 155 parts of this, and a sliding mill (for example, Super Mascolloid-MKZ) is used.
81.5-40, manufactured by Masuko Sangyo Co., Ltd.),
Further, a method of pulverizing the material by passing it through a grinder (manufactured by Puller Co., Ltd.) under cooling (Japanese Patent Application No. 55-67591, Examples) can be mentioned.
免疫活性ペプチドを抽出する酸水溶液としては、鉱酸水
溶液および有機酸水溶液を挙げることができる。Examples of the acid aqueous solution for extracting immunoactive peptides include mineral acid aqueous solutions and organic acid aqueous solutions.
鉱酸としては、塩酸、硫酸、燐酸等を挙げることができ
る。Examples of mineral acids include hydrochloric acid, sulfuric acid, and phosphoric acid.
有機酸としては、蟻酸、酢酸、プロピオン酸、酪酸等を
挙げることができる。これらの酸は、分岐していても差
支えない。Examples of organic acids include formic acid, acetic acid, propionic acid, butyric acid, and the like. These acids may be branched.
次に、実施例により本発明をさらに説明するが、本発明
は、かかる実施例に限定されるものではない。Next, the present invention will be further explained with reference to examples, but the present invention is not limited to these examples.
実施例１
成牛の頭部と四肢とを除去した新鮮な牛骨を２℃の温度
下で粗粉砕し、液体アンモニウム冷却下に凍結（約−１
００”Ｃ）粉砕したのち、マイクロ波および熱風（約９
０°Ｃ１５分間）乾燥した。Example 1 Fresh beef bones from which the head and limbs of an adult cow were removed were coarsely ground at a temperature of 2°C, and frozen under liquid ammonium cooling (approximately -1
00”C) After crushing, microwave and hot air (approx.
Dry at 0°C for 15 minutes).
収量は乾燥重量比で約３３％であった。得られた乾燥物
は、第１表に示す組成を有していた。The yield was about 33% on a dry weight basis. The obtained dried product had the composition shown in Table 1.
第 １ 表
（重量％）
上記により得られた牛骨及び骨髄乾燥物１ｇに、０、Ｉ
Ｎ塩酸の１０ｍｆｆ１を加え、ｐｏｌｙｔｒｏｎ（Ｐｏ
ｌｙＬｒｏｎ 社製造）にてホモジナイズした後、４
０００ｒｐｍにて３０分間遠心分離し、上滑を分離した
。沈渣は、０．ＩＮ塩酸の１０＋ｙ／２を添加し、十分
撹拌したのち遠心分離し、その上清をさきの上清と合せ
、凍結乾燥し、これを塩酸抽出物とした。Table 1 (% by weight) 0, I
Add 10 mff1 of N-hydrochloric acid and
After homogenizing with lyLron Co., Ltd.),
The mixture was centrifuged at 000 rpm for 30 minutes, and the supernatant was separated. The sediment is 0. 10+y/2 of IN hydrochloric acid was added, thoroughly stirred, and then centrifuged. The supernatant was combined with the previous supernatant and freeze-dried to obtain a hydrochloric acid extract.
実施例２
ビーフインド（三菱化成食品株式会社販売、登録商標）
Ｉｇに、０．１Ｍ酢酸１０ｍ１を加え、ｐｏｌｙｔｒｏ
ｎによりホモジナイズし、熱湯浴中にて１０分間煮沸し
た。直ちに水冷した後氷酢酸を加え、ＩＭ濃度に調整し
た。実施例１と同様に遠心分離し、上清を分離した。沈
渣は、１Ｍ酢酸１０ｍｊ２に懸濁し、再び同様に遠心分
離し、上清を分離した。これをさきの上清と合せ凍結乾
燥し、熱酢酸抽出物とした。Example 2 Beef India (Sold by Mitsubishi Kasei Foods Co., Ltd., registered trademark)
Add 10ml of 0.1M acetic acid to Ig, and
The mixture was homogenized by n and boiled for 10 minutes in a hot water bath. After immediately cooling with water, glacial acetic acid was added to adjust the concentration to IM. Centrifugation was performed in the same manner as in Example 1, and the supernatant was separated. The precipitate was suspended in 10 mj2 of 1M acetic acid, centrifuged again in the same manner, and the supernatant was separated. This was combined with the previous supernatant and lyophilized to obtain a hot acetic acid extract.
■）ラジオイムノアッセイ（ＲＩＡ）法検索する活性ペ
プチドとして、
Ｖａｓｏａｃｔｉｖｅ １ｎｔｅｓｔｉｎａｌＰｏｌ
ｙｐｅｐｔｉｄｅ （ＶＩＰ）
Ｐｅｐｔｉｄｅ ｈｆｓｔｉｄｉｎｅｉｓｏｌｅｕｃ
ｆｎｅ （ＰＨＩ）
Ｐａｎｃｒｅａｔｉｃ Ｐｏ１ｙｐｅｐｔｉｄｅ（ｐ
ｐ） ガストリン放出ペプチド（ＣＡＲＰ）Ｍｅｔｈ
ｉｏｎｉｎｅ−ｅｎｋｅｐｈａｌｉｎＡｒｇ−Ｇａｙ−
Ｌｅｕ （Ｍｅｔ−Ｅｎｋ８）ロイモルフィン
ガラニン
ソマトスフチン
ｅｐｉｄｅｒｍａｌ ｇｒｏｗｔｈ
ｆ ａ ｃ ｔ ｏ ｒ （ＥＧＦ）
を選定した。■) Radioimmunoassay (RIA) method Vasoactive 1ntestinalPol is used as an active peptide to search.
ypeptide (VIP) Peptide hfstidineisoleuc
fne (PHI) Pancreatic polypeptide (p
p) Gastrin releasing peptide (CARP) Meth
ionine-enkephalinArg-Gay-
Leu (Met-Enk8) leumorphin galanin somatosfutin epidermal growth factor (EGF) was selected.
上記活性ペプチドのそれぞれに特異的なＲＩＡの確立に
、それぞれ、抗ブタＶＩＰ血清Ｒ５０２、抗ブタＰＨ１
（１−１５）血清Ｒ８４０３、抗ヒトｐｐ血清ＲＯ２１
２、抗ブタＧＲＰ血清Ｒ６９０２、抗ＭｅＬ−Ｅｎｋ８
血清ＲＯ１７１、抗ブタロイモルフイン血清ＲＯＯ１４
、抗ブタガラニン血清Ｒ１９８５、抗ソマトスタフチン
血清０ＡＬ２７２および抗ヒトＥＧＦ血清ＴＫＲ１０２
をそれぞれ用いた。For establishment of RIA specific for each of the above active peptides, anti-pig VIP serum R502, anti-pig PH1, respectively, were used.
(1-15) Serum R8403, anti-human pp serum RO21
2. Anti-pig GRP serum R6902, anti-MeL-Enk8
Serum RO171, anti-porcine leumorphin serum ROO14
, anti-porcine galanin serum R1985, anti-somatostafutin serum 0AL272 and anti-human EGF serum TKR102.
were used respectively.
いずれの抗血清も、ＲＩＡ系に用いた標準抗原に特異的
であり、既知の類似ペプチドとは交差反応しないことを
確認している。It has been confirmed that all antisera are specific to the standard antigen used in the RIA system and do not cross-react with known similar peptides.
２）標識抗原：
標識抗原は、標準抗原に対応するペプチドをクロラミン
Ｔ法により ＋！５ｉを導入し、５ｅｐｈａｄｅｘＧ
１０カラム（ＩＸ３０ｃｍ）または５ＥＰ−ＰＡＫＣ
１Ｂカートリッジ（ＷａｔｅｒｓＡｓｓｏｃｔａｔｅｓ
）により精製した。2) Labeled antigen: Labeled antigen is obtained by preparing a peptide corresponding to a standard antigen using the chloramine T method +! Introduced 5i and 5ephadexG
10 columns (IX30cm) or 5EP-PAKC
1B Cartridge (Waters Associates)
).
３）標準希釈液：
標準希釈液は、０．０２５Ｍ ＥＤＴＡ、０．１４Ｍ
ＮａＣ１，０，１４Ｍ 塩化ナトリウムおよび０
．５％ウシ血清アルブミンを含む０． ＯＩ Ｍ リ
ン酸緩衝液を用いた。3) Standard dilution solution: Standard dilution solution is 0.025M EDTA, 0.14M
NaCl 1,0,14M Sodium chloride and 0
．． 0.0 with 5% bovine serum albumin. OIM phosphate buffer was used.
なお、ソマトスタチンＲＩＡ系では、７０ｍＭＥＤＴＡ
、０．１％ゼラチンを含む０．１ Ｍヒスチジン緩衝液
（ｐＨ６，０）を用いた。In addition, in the somatostatin RIA system, 70mMEDTA
, 0.1 M histidine buffer (pH 6,0) containing 0.1% gelatin was used.
４）ＲＩＡ測定： ｔＡは、二重測定で行い、ガラス試験管を用いた。4) RIA measurement: tA was performed in duplicate and using glass test tubes.
標準希釈液（０，４ｎｆり、標準ペプチド（０，１ｍ１
）および抗血清溶液（０，１ｎｆ）および標準抗原溶液
（５，０００〜１０．ＯＯＯｃｐｍ１０．１ ｍｌ、
０．１ ｍｌ）をガラス試験管に分注、混和させた。Standard dilution solution (0.4nf), standard peptide (0.1ml
) and antiserum solution (0.1nf) and standard antigen solution (5,000-10.00cpm 10.1 ml,
0.1 ml) was dispensed into a glass test tube and mixed.
次いで、４ ’Ｃで８時間反応させた後、正常家兎血清
（１００倍希釈、０．１ｎｆ抗家兎ガンマグロブリン血
清（２０倍希釈、０．１ｍｊ２）、１０％ポリエチレン
グリコール液（ポリエチレングリコール６０００．０．
１ｎｆ）を順次添加、混和し、さらに４°Ｃに３時間反
応後、遠心分離（３，０００ｒｐｍ、３０分、４’Ｃ）
ｌ、、上清を吸引除去し、沈渣の放射活性をガンマカウ
ンタで測定した。Next, after reacting at 4'C for 8 hours, normal rabbit serum (100-fold dilution, 0.1nf anti-rabbit gamma globulin serum (20-fold dilution, 0.1mj2), 10% polyethylene glycol solution (polyethylene glycol 6000) .0.
1nf) was added in sequence, mixed, and further incubated at 4°C for 3 hours, followed by centrifugation (3,000 rpm, 30 minutes, 4'C).
l. The supernatant was removed by suction, and the radioactivity of the precipitate was measured using a gamma counter.
５）活性ペプチドの検索：
４）に記載した方法により、実施例１および２により得
られた牛骨及び骨髄粗抽出物の活性ペプチドを検索した
。5) Search for active peptides: The crude bovine bone and bone marrow extracts obtained in Examples 1 and 2 were searched for active peptides by the method described in 4).
実施例１および２により得られた牛骨及び骨髄粗抽出物
（０，１ｇ）に標準希釈液（１，０ｍｊ２）を加え、十
分混和したのち不溶物を遠心分離（４゜０００ｒｐｍ、
３０分、４゛Ｃ）にて除去した。得られた抽出液を倍々
または４倍希釈した希釈系列について測定を行った。The standard dilution solution (1.0 mj2) was added to the crude bovine bone and bone marrow extracts (0.1 g) obtained in Examples 1 and 2, and after mixing thoroughly, the insoluble matter was centrifuged (4°000 rpm,
It was removed for 30 minutes at 4°C. Measurements were performed on a dilution series in which the obtained extract was diluted twofold or fourfold.
活性ペプチド成分の含有量は、各ＲＩＡ系における粗抽
出物の希釈曲線から評価し、希釈各点について、得られ
た免疫活性の平均値を牛骨及び骨髄乾燥物あたりに換算
し、測定値とした。なお、標準曲線ならびに希釈曲線は
ＩｏｇｉＬ−１ｏｇ変換し、直線化した。The content of active peptide components is evaluated from the dilution curve of the crude extract in each RIA system, and the average value of the obtained immune activity for each dilution point is converted to the amount per dried bovine bone and bone marrow, and the measured value and did. In addition, the standard curve and dilution curve were converted into IogiL-1og and linearized.
６）結果： 実施例１で得られた塩酸抽出物のＶＩＰ、ＰＰ。6) Results: VIP, PP of the hydrochloric acid extract obtained in Example 1.
Ｍｅｔ−Ｅｎｋ８およびＰＨ１について、標準曲線なら
びに希釈曲線を作図し、第１図に示した。Standard curves and dilution curves were drawn for Met-Enk8 and PH1 and shown in FIG.
ＶＩＰとＭｅｔ−Ｅｎｋ８とが、標準ペプチドと平行し
た。VIP and Met-Enk8 were paralleled with standard peptides.
実施例１で得られた塩酸抽出物（収率２８％）中の免疫
活性を、第２表に示した。The immunoactivity in the hydrochloric acid extract (yield 28%) obtained in Example 1 is shown in Table 2.
第 ２ 表 ｐ ｍ ｏ Ｉ ／ ｇ乾燥物 実施例２で得られた酢酸抽出物のｖｒｐ、ｐｐ。Table 2 p m / g dry matter vrp, pp of the acetic acid extract obtained in Example 2.
Ｍｅｔ−Ｅｎｋ８．ＥＧＦ、ＧＲＰおよびロイモルフィ
ンについて標準曲線ならびに希釈曲線を作図し、第２図
に示した。Met-Enk8. Standard curves and dilution curves were constructed for EGF, GRP and leumorphin and are shown in FIG.
ＶＩＰ、ＰＰ、Ｍｅｔ−Ｅｎｋ８．ＧＲＰおよびロイモ
ルフインが標準ペプチドと平行した。VIP, PP, Met-Enk8. GRP and leumorphin were paralleled with standard peptides.
実施例２で得られた酢酸抽出物（収率２４％）中の免疫
活性を、第３表に示した。The immunoactivity in the acetic acid extract (yield 24%) obtained in Example 2 is shown in Table 3.
第３表
哺乳動物などの動物組機とくに神経および消化管組織に
分布し存在するもので、生体の様々な調節機能を行って
いると考えられていることから、加齢による各種骨関節
疾患に随伴した、いわゆる老化性骨粗■症（オステオボ
ローゼ）の患者に、本発明の免疫活性ペプチタドを投与
することにより、生体の骨代謝機構に有効に作用するも
のと考えられる。Table 3: These substances are distributed and present in the tissues of animals such as mammals, especially the nerves and gastrointestinal tract, and are thought to perform various regulatory functions in the living body. It is believed that by administering the immunoactive peptide of the present invention to a patient suffering from so-called senile osteoporosis (osteoborosis), it will effectively act on the bone metabolic mechanism of the body.
第１図は、実施例１で得られた活性ペプチドの免疫活性
希釈曲線および標準曲線であり、第２図は、実施例２で
得られた活性ペプチドの免疫活性希釈曲線および標準曲
線である。
ｐｍｏｌ／ｇ乾燥物
〔発明の効果〕FIG. 1 shows the immunoreactivity dilution curve and standard curve of the active peptide obtained in Example 1, and FIG. 2 shows the immunoreactivity dilution curve and standard curve of the active peptide obtained in Example 2. pmol/g dry matter [Effects of the invention]
Claims (1)
接触させ、酸水溶液中に免疫活性ペプチドを抽出するこ
とを特徴とする免疫活性ペプチドの製造方法。(1) A method for producing an immunoactive peptide, which comprises contacting a pulverized product containing bird and animal bones or bone marrow with an aqueous acid solution, and extracting the immunoactive peptide into the aqueous acid solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63178447A JPH0228118A (en) | 1988-07-18 | 1988-07-18 | Production of immunologically active peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63178447A JPH0228118A (en) | 1988-07-18 | 1988-07-18 | Production of immunologically active peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0228118A true JPH0228118A (en) | 1990-01-30 |
Family
ID=16048680
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63178447A Pending JPH0228118A (en) | 1988-07-18 | 1988-07-18 | Production of immunologically active peptide |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0228118A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107874062A (en) * | 2017-11-14 | 2018-04-06 | 王清秀 | FOS ox bone marrow peptide compound powder and its application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61268700A (en) * | 1985-05-24 | 1986-11-28 | Dai Ichi Pure Chem Co Ltd | Novel physiologically active peptide |
-
1988
- 1988-07-18 JP JP63178447A patent/JPH0228118A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61268700A (en) * | 1985-05-24 | 1986-11-28 | Dai Ichi Pure Chem Co Ltd | Novel physiologically active peptide |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107874062A (en) * | 2017-11-14 | 2018-04-06 | 王清秀 | FOS ox bone marrow peptide compound powder and its application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bühler et al. | Small intestinal morphology in eight-day-old calves fed colostrum for different durations or only milk replacer and treated with long-R3-insulin-like growth factor I and growth hormone | |
| Houseknecht et al. | Effect of dietary energy source and level on serum growth hormone, insulin-like growth factor 1, growth and body composition in beef heifers | |
| Yambayamba et al. | Hormonal status, metabolic changes, and resting metabolic rate in beef heifers undergoing compensatory growth | |
| Moughan et al. | Perchloric and trichloroacetic acids as precipitants of protein in endogenous ileal digesta from the rat | |
| Moughan et al. | Composition of nitrogen-containing fractions in digesta from the distal ileum of pigs fed a protein-free diet | |
| Hendricksen et al. | The voluntary intake, digestibility and retention time by cattle and sheep of stem and leaf fractions of a tropical legume (Lablab purpureus) | |
| Tatara et al. | Effects of combined maternal administration with alpha-ketoglutarate (AKG) and β-hydroxy-β-methylbutyrate (HMB) on prenatal programming of skeletal properties in the offspring | |
| Schaudies et al. | EGF content in the gastrointestinal tract of rats: effect of age and fasting/feeding | |
| Braude et al. | Artificial rearing of pigs: 2.* The time course of milk protein digestion and proteolytic enzyme secretion in the 28-day-old pig | |
| Płoszaj et al. | Polyamines in goat's colostrum and milk | |
| Salter et al. | Digestibilities of nitrogen compounds in rumen bacteria and in other components of digesta in the small intestine of the young steer | |
| Duehlmeier et al. | Distribution patterns of the glucose transporters GLUT4 and GLUT1 in skeletal muscles of rats (Rattus norvegicus), pigs (Sus scrofa), cows (Bos taurus), adult goats, goat kids (Capra hircus), and camels (Camelus dromedarius) | |
| GOFF et al. | Parathyroid hormone-related peptide content of bovine milk and calf blood assessed by radioimmunoassay and bioassay | |
| Lapierre et al. | Effect of human growth hormone-releasing factor and (or) thyrotropin-releasing factor on growth, carcass composition, diet digestibility, nutrient balance, and plasma constituents in dairy calves | |
| 岸田光代 et al. | Development of radioimmunoassay for eel growth hormone. | |
| Anast et al. | Calcitonin | |
| Dinusson | The Effects of Stilbestrol, Testosterone, and Thyroid Alteration on the Growth and Fattening of Beef Heifers | |
| JPH0228118A (en) | Production of immunologically active peptide | |
| Steele et al. | Regulation of somatomedin production, release, and mechanism of action | |
| Herington | Ontogenesis of hepatic growth hormone receptors in the rat | |
| Knopf | The role of host hormones in controlling survival and development of Schistosoma mansoni | |
| Grimes et al. | Effect of short-term fasting/refeeding on epidermal growth factor content in the gastrointestinal tract of suckling rats | |
| Bunnett et al. | Immunocytochemical localization of gastric inhibitory peptide and glucagon in the alimentary tract of ruminants | |
| Galvin et al. | Effects of purified porcine relaxin on the uterus of gilts during early pregnancy | |
| Koldovský et al. | The developing gastrointestinal tract and milk-borne epidermal growth factor |