The circulating hormonal form of human parathyroid hormone (hPTH-1-37) has been assessed in vitro as well as in vivo in the ovariectomized rat, a model for postmenopausal osteoporosis. In vitro, hPTH-1-37 induces a dose-dependent cAMP formation and increases vitality as well as alkaline phosphatase activity in UMR106 osteosarcoma cells. Differentiation and proliferation of osteoclasts in rat bone marrow-derived stem cell preparations are decreased. Daily hPTH-1-37 s.c. administration in ovariectomized rats for 60 days results in augmented formation of new bone, in amplified femural bone density, and in thickening of the calvaria.

01 Feb 1998·Arzneimittel-Forschung

A new immunoenzymometric assay for bioactive N-terminal human parathyroid hormone fragments and its application in pharmacokinetic studies in dogs.

Article

Author: Mägerlein, M ; Neidlein, R ; Forssmann, W G ; Adermann, K ; Müller-Beckmann, B ; Stein, K ; Hock, D

Advances in the treatment of clinical disorders of mineral in homeostatis and metabolic bone disease with intact parathyroid hormone 1-84 or one of the biologically active N-terminal fragments require a precise and sensitive measurement in serum. Therefore, a two-site immunoenzymometric assay for the quantitative determination of bioactive hPTH-1-37 (human parathyroid hormone) at picomolar concentrations was developed. Monoclonal antibodies (mAB) against hPTH-1-37 were raised by hybridoma cells in serum-free cell culture. Furthermore, sequence-specific polyclonal antibodies were obtained by immunisation of rabbits using multiple antigenic peptides (MAP) representing the conspicuous regions of the primary structure of hPTH-1-37. The polyclonal and monoclonal antibodies were characterised by epitope mapping. The combination of a monoclonal antibody (13C63/5) recognising hPTH fragment 16-24 with a polyclonal antibody (k2) showing a predominant binding sequence at hPTH-1-5 led to a sandwich assay specific for N-terminally intact and therefore biologically active hPTH. The validated assay ranging from 4 to 1000 pmol/l was applied to pharmacokinetic studies of hPTH-1-37. After s.c. administration of 30 mu g/kg in 5 beagles, the maximum serum concentrations of hPTH-1-37 ranging at 2139 +/- 857 pmol/l were observed 45 min after the injection. Clearance of the peptide calculated from the exponential disappearance curve was 32.0 +/- 9.1 ml/min/kg with a mean t1/2 of 37 +/- 10 min.

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Indication	Highest Phase	Country/Location	Organization	Date
Hypoparathyroidism	Phase 1	DE	IPF PharmaCeuticals GmbH	-
Psoriasis	Phase 1	DE	PHARIS Biotec GmbH	-

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