1995 Fiscal Year Final Research Report Summary
Development of Assay Methods to Measure Salmon Calcitonin with Highly Sensitivity : Determination of the Bioavailability of Nasal Salmon Calcitonin.
| Project/Area Number |
06557136
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
医薬分子機能学
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| Research Institution | Setsunan University |
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Principal Investigator |
KOIDA Masao Setsunan University Faculty of Pharmaceutical Sciences Professor, 薬学部, 教授 (80039651)
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| Co-Investigator(Kenkyū-buntansha) |
OHKUBO Satoru Sandoz Pharmaceuticals, LTD.Tsukuba research center Research Scientist, 筑波総合研究所, 研究員
KOHNO Takeyuki Setsunan University Faculty of Pharmaceutical Sciences Associate Professor, 薬学部, 講師 (50178224)
NAKAMUTA Hiromichi Setsunan University Faculty of Pharmaceutical Sciences Assistant Professor, 薬学部, 助手 (70164275)
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| Project Period (FY) |
1994 – 1995
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| Keywords | calcitonin / osteoporosis / osteomalacia / osteoclast / bone absorption / enzyme immunoassay |
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| Research Abstract |
Salmon calcitonin (SCT), one of the standard peptide therapeutics for diffuse bone diseases (i.e.osteoporosis and Paget's disease), has been clinically applied by the parenteral injections. Lately the nasal application formula tends to replace the parenterals especially for chronic use in senile and postmenopausal osteoporosis, due to a lower incidence of annoying side effects (vomiting, nausea and flushing) with the ease for self-administration. Not only clinically but also experimentally, however, no direct study has existed so far about the pharmacokinetic property of nasal SCT given in the therapeutic doses, mainly due to the lack of a rliable assay method of picomolar level of SCT.In this study, it was attempted to develop assay methods to measure SCT with highly sensitivity to determine the bioavailability of the nasal SCT.
1. Development of highly sensitive assay of SCT based on the physiological activity. (1) Cell suspensions containing osteoclasts were prepared from long bones of neonatal rats and (2) seeded onto the devitalized slices (100 mum in thickness) of bovine femur, (3) under the scanning electron microscopy and light microscopy to measure the numbers and areas of excavated lacunae (pits) and finally to evaluate the inhibitory effect of SCT on the pit formation. The detection limit of SCT achieved was less than 1 pM.
2. A highly sensitive non-competitive enzyme immunoassay method (hetero-two-site enzyme immunoassay) for SCT has been developed.
3. The application of the hetero-two-site enzyme immunoassay has enable us to directly estimate the bioavailability of nasal SCT at therapeutic levels in rats.
4. Highly sensitive and less-time consuming enzyme immunoassay (immune complex transfer two-site enzyme immunoassay) for SCT has been developed.
5. In preliminary experiments, the immune complex transfer two-site enzyme immunoassay has been applicable to measure SCT concentration in serum after intranasal administration of SCT to human.
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