title: Metabolism of 20-hydroxyvitamin D3 by mouse liver microsomes.
abstract: 20-Hydroxyvitamin D3 [20(OH)D3], the major product of CYP11A1 action on vitamin D3, is biologically active and like 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] can inhibit proliferation and promote differentiation of a range of cells, and has anti-inflammatory properties. However, unlike 1,25(OH)2D3, it does not cause toxic hypercalcemia at high doses and is therefore a good candidate for therapeutic use to treat hyperproliferative and autoimmune disorders. In this study we analyzed the ability of mouse liver microsomes to metabolize 20(OH)D3. The two major products were identified from authentic standards as 20,24-dihydroxyvitamin D3 [20,24(OH)2D3] and 20,25-dihydroxyvitamin D3 [20,25(OH)2D3]. The reactions for synthesis of these two products from 20(OH)D3 displayed similar Km values suggesting that they were catalyzed by the same cytochrome P450. Some minor metabolites were produced by reactions with higher Km values for 20(OH)D3. Some metabolites gave mass spectra suggesting that they were the result of hydroxylation followed by dehydrogenation. One product had an increase in the wavelength for maximum absorbance from 263nm seen for 20(OH)D3, to 290nm, suggesting a new double bond was interacting with the vitamin D-triene chromophore. The two major products, 20,24(OH)2D3 and 20,25(OH)2D3 have both previously been shown to have higher potency for inhibition of colony formation by melanoma cells than 20(OH)D3, thus it appears that metabolism of 20(OH)D3 by mouse liver microsomes can generate products with enhanced activity.
authors.affiliation: School of Chemistry and Biochemistry, The University of Western Australia, Crawley, WA 6009, Australia.
authors.forename: Chloe Y S
authors.initials: CY
authors.lastname: Cheng
authors.affiliation: Department of Pathology and Laboratory Medicine, Center for Adult Cancer Research, University of Tennessee Health Science Center, Memphis, TN, USA.
authors.forename: Andrzej T
authors.initials: AT
authors.lastname: Slominski
authors.affiliation: School of Chemistry and Biochemistry, The University of Western Australia, Crawley, WA 6009, Australia. Electronic address: robert.tuckey@uwa.edu.au.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Tuckey
mesh_terms: D000818:Animals; D002112:Calcifediol / Q000031:analogs & derivatives* / Q000378:metabolism; D005260:Female; D006900:Hydroxylation; D008810:Mice, Inbred C57BL; D008862:Microsomes, Liver / Q000378:metabolism*
chemical_list: C480634:20-hydroxyvitamin D3; D002112:Calcifediol
keywords: 20-Hydroxyvitamin D3; Cytochrome P450; Hydroxylation; Liver; Microsomes