Although the main role of 1, 25-dihydroxyvitamin D3 (1,25-(OH)₂D3) is to regulate calcium homeostasis, the valuable therapeutic applications of this compound has led to the search of new 1,25-(OH)₂D3-Vitamin D receptor (VDR) ligands with less side effects. In this work we have characterized seven 1,25-(OH)₂D3 derivatives (ZK136607, ZK161422, ZK157202, ZK159222, ZK168492, ZK191732 and ZK168289). ZK157202 is an agonist that gives a pattern similar to that of 1,25-(OH)₂D3 or ZK161422 in limited trypsin digestion assays, is able to recruit p160 and DRIP205 coactivators, is as potent as 1,25-(OH)₂D3 to stimulate VDRES-dependent transcription in HeLa cells and acts as a superagonist in HEK 293T cells. This compound is also more potent than the natural ligand to transrepress the activation of the RAR2 promoter by retinoic acid (RA) and the response of the collagenase promoter to 4-12-O-tetradecanoylphorbol 13-acetate (TPA). ZK136607, ZK168492, ZK191732 and ZK168289 have a profile similar to that of the partial antagonist ZK159222. They induce an antagonistic-type proteolytic pattern, do not recruit classical coactivators and have little transactivation potency. However, they act in a cell context-dependent manner because they lack activity in HeLa cells while presenting some agonistic activity in HEK 293T cells, or vice versa. Furthermore, some of these compounds have a "dissociated" activity: they cannot transactivate but they are as potent as 1,25-(OH)₂D3 in transrepression assays. Together our results demonstrate the existence of novel VDR ligands with variable biological functions and "dissociated" activity. They should represent useful tools for studying VDR function and could have therapeutic utility.