The synthetic 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) analog 20-epi-22-oxa-24a,26a,27a-tri-homo-1,25-(OH)2vitamin D3 (KH1060) is considerably more potent than its cognate hormone. The mechanism of action of KH1060 includes interaction with the vitamin D receptor (VDR). We previously showed that KH1060 increases VDR stability in ROS 17/2.8 osteoblastic cells by inducing a specific conformational change in the VDR. KH1060 is metabolized, both in vivo and in vitro, into several stable products. In the present study, we investigated whether these metabolites might contribute to the increased biological activity of KH1060. We found that the potencies of two of these metabolites, 24a-OH-KH1060 and 26-OH-KH1060, were similar to that of 1,25-(OH)2D3 in inducing osteocalcin production by the osteoblast cell line ROS 17/2.8. This report further showed that these metabolites had the same effects as KH1060 on VDR: they increased VDR stability in ROS 17/2.8 cells, while limited proteolytic analysis revealed that they caused a conformational change in the VDR, resulting in an increased resistance against proteolytic cleavage. Furthermore, as shown in gel mobility shift assays, both compounds clearly induced VDR binding to vitamin D response elements. Together, these results show that the potent in vitro activity of KH1060 is not only directed by the effects on the VDR conformation/stabilization of the analog itself, but also by certain of its long-lived metabolites, and emphasizes the importance of detailed knowledge of the metabolism of synthetic hormonal analogs. Copyright (C) 2000 Elsevier Science Inc.

KH1060, Metabolites, Receptor conformation, Receptor stability, Vitamin D analogs, Vitamin D receptor,
Biochemical Pharmacology
Department of Internal Medicine

van den Bemd, G.J.C.M, Dilworth, F.J, Makin, H.L.J, Prahl, C, Deluca, H.F, Jones, G, … van Leeuwen, J.P.T.M. (2000). Contribution of several metabolites of the vitamin D analog 20-epi-22-oxa-24a,26a,27a-tri-homo-1,25-(OH)2 vitamin D3 (KH 1060) to the overall biological activity of KH1060 by a shared mechanism of action. Biochemical Pharmacology, 59(6), 621–627. doi:10.1016/S0006-2952(99)00371-8