A New High-throughput Analyzing System of Ligand Induced Cofactor Recruitment on Vitamin D Receptor; Synthesis and Biological Evaluation of 19-Norvitamin D3 Derivatives

Abstract

Synthesis of 25-hydroxy-19-norvitamin D3 derivatives as prohormone type agents for anti-prostate diseases was accomplished utilizing Julia-type olefination. The compounds showed potent antiproliferative activity on an immortalized normal prostate cell line, PZ-HPV-7. We demonstrated that 25-hydroxy-19-norvitamin D3 was hydroxylated to form 1?,25-dihydroxy-19-norvitamin D3 by human 1?-hydroxylase (CYP27B1) in a cell-free system. Moreover, we have developed a novel rapid analyzing system of ligand induced cofactor recruitment on vitamin D receptor (VDR) using fluorophore labeled coactivator (CoA) and HCHO fixing of the complex. Recruitment of CoA by 19-norvitamin D3 derivatives was analyzed using this system in a high-throughput manner. Efficiency of recruiting of CoA would explain discrepancy between strong biological activity and weak VDR binding affinity of ligands.

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