4-Hydroxy-1α,25-Dihydroxyvitamin D 3 : Synthesis and Structure-Function Study.

The active vitamin D metabolites, 25-hydroxyvitamin D 3 (25D 3 ) and 1,25-dihydroxyvitamin D 3 (1,25D 3 ), are produced by successive hydroxylation steps and play key roles in several cellular processes. However, alternative metabolic pathways exist, and among them, the 4-hydroxylation of 25D 3 is a major one. This study aims to investigate the structure-activity relationships of 4-hydroxy derivatives of 1,25D 3 . Structural analysis indicates that 1,4α,25(OH) 3 D 3 and 1,4β,25(OH) 3 D 3 maintain the anchoring hydrogen bonds of 1,25D 3 and form additional interactions, stabilizing the active conformation of VDR. In addition, 1,4α,25D 3 and 1,4β,25D 3 are as potent as 1,25D 3 in regulating the expression of VDR target genes in rat intestinal epithelial cells and in the mouse kidney. Moreover, these two 4-hydroxy derivatives promote hypercalcemia in mice at a dose similar to that of the parent compound.