title: Chemical synthesis of 20S-hydroxyvitamin D3, which shows antiproliferative activity.
abstract: 20S-hydroxyvitamin D3 (20S-(OH)D3), an in vitro product of vitamin D3 metabolism by the cytochrome P450scc, was recently isolated, identified and shown to possess antiproliferative activity without inducing hypercalcemia. The enzymatic production of 20S-(OH)D3 is tedious, expensive, and cannot meet the requirements for extensive chemical and biological studies. Here we report for the first time the chemical synthesis of 20S-(OH)D3 which exhibited biological properties characteristic of the P450scc-generated compound. Specifically, it was hydroxylated to 20,23-dihydroxyvitamin D3 and 17,20-dihydroxyvitamin D3 by P450scc and was converted to 1alpha,20-dihydroxyvitamin D3 by CYP27B1. It inhibited proliferation of human epidermal keratinocytes with lower potency than 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) in normal epidermal human keratinocytes, but with equal potency in immortalized HaCaT keratinocytes. It also stimulated VDR gene expression with similar potency to 1,25(OH)2D3, and stimulated involucrin (a marker of differentiation) and CYP24 gene expression, showing a lower potency for the latter gene than 1,25(OH)2D3. Testing performed with hamster melanoma cells demonstrated a dose-dependent inhibition of cell proliferation and colony forming capabilities similar or more pronounced than those of 1,25(OH)2D3. Thus, we have developed a chemical method for the synthesis of 20S-(OH)D3, which will allow the preparation of a series of 20S-(OH)D3 analogs to study structure-activity relationships to further optimize this class of compound for therapeutic use.
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, TN 38163, USA. wli@uthsc.edu
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Jianjun
authors.initials: J
authors.lastname: Chen
authors.forename: Zorica
authors.initials: Z
authors.lastname: Janjetovic
authors.forename: Tae-Kang
authors.initials: TK
authors.lastname: Kim
authors.forename: Trevor
authors.initials: T
authors.lastname: Sweatman
authors.forename: Yan
authors.initials: Y
authors.lastname: Lu
authors.forename: Jordan
authors.initials: J
authors.lastname: Zjawiony
authors.forename: Robert C
authors.initials: RC
authors.lastname: Tuckey
authors.forename: Duane
authors.initials: D
authors.lastname: Miller
authors.forename: Andrzej
authors.initials: A
authors.lastname: Slominski
mesh_terms: D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase / Q000378:metabolism; D000818:Animals; D002112:Calcifediol / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002762:Cholecalciferol / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D006224:Cricetinae; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects; D008958:Models, Molecular; D008968:Molecular Conformation
chemical_list: C480634:20-hydroxyvitamin D3; D002762:Cholecalciferol; D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase; D002786:Cholesterol Side-Chain Cleavage Enzyme; D002112:Calcifediol