Metabolism of 20-hydroxyvitamin D3 and 20,23-dihydroxyvitamin D3 by rat and human CYP24A1.
CYP11A1 hydroxylates vitamin D3 producing 20S-hydroxyvitamin D3 [20(OH)D3] and 20S,23-dihydroxyvitamin D3 [20,23(OH)2D3] as the major and most characterized metabolites. Both display immuno-regulatory and anti-cancer properties while being non-calcemic. A previous study indicated 20(OH)D3 can be metabolized by rat CYP24A1 to products including 20S,24-dihydroxyvitamin D3 [20,24(OH)2D3] and 20S,25-dihydroxyvitamin D3, with both producing greater inhibition of melanoma colony formation than 20(OH)D3. The aim of this study was to characterize the ability of rat and human CYP24A1 to metabolize 20(OH)D3 and 20,23(OH)2D3. Both isoforms metabolized 20(OH)D3 to the same dihydroxyvitamin D species with no secondary metabolites being observed. Hydroxylation at C24 produced both enantiomers of 20,24(OH)2D3. For rat CYP24A1 the preferred initial site of hydroxylation was at C24 whereas the human enzyme preferred C25. 20,23(OH)2D3 was initially metabolized to 20S,23,24-trihydroxyvitamin D3 and 20S,23,25-trihydroxyvitamin D3 by rat and human CYP24A1 as determined by NMR, with both isoforms showing a preference for initial hydroxylation at C25. CYP24A1 was able to further oxidize these metabolites in a series of reactions which included the cleavage of C23-C24 bond, as indicated by high resolution mass spectrometry of the products, analogous to the catabolism of 1,25(OH)2D3 via the C24-oxidation pathway. Similar catalytic efficiencies were observed for the metabolism of 20(OH)D3 and 20,23(OH)2D3 by human CYP24A1 and were lower than for the metabolism of 1,25(OH)2D3. We conclude that rat and human CYP24A1 metabolizes 20(OH)D3 producing only dihydroxyvitamin D3 species as products which retain biological activity, whereas 20,23(OH)2D3 undergoes multiple oxidations which include cleavage of the side chain.
School of Chemistry and Biochemistry, The University of Western Australia, Crawley, WA 6009, Australia.
Elaine W
EW
Tieu
Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, TN, USA.
Wei
W
Li
Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, TN, USA; Department of Pharmaceutical Sciences, School of Pharmacy, South College, Knoxville, TN, USA.
Jianjun
J
Chen
Department of Pathology and Laboratory Medicine, Center for Cancer Research, University of Tennessee Health Science Center, Memphis, TN, USA.
Tae-Kang
TK
Kim
Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, TN, USA.
Dejian
D
Ma
Department of Pathology and Laboratory Medicine, Center for Cancer Research, University of Tennessee Health Science Center, Memphis, TN, USA; Department of Dermatology, University of Alabama Birmingham, Birmingham, AL, USA.
Andrzej T
AT
Slominski
School of Chemistry and Biochemistry, The University of Western Australia, Crawley, WA 6009, Australia. Electronic address: robert.tuckey@uwa.edu.au.
Robert C
RC
Tuckey
D000818:Animals; D002112:Calcifediol / Q000031:analogs & derivatives* / Q000378:metabolism; D004100:Dihydroxycholecalciferols / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D010084:Oxidation-Reduction; D051381:Rats; D014807:Vitamin D / Q000031:analogs & derivatives / Q000378:metabolism; D065668:Vitamin D3 24-Hydroxylase / Q000378:metabolism*
C547379:20,23-dihydroxyvitamin D3; C480634:20-hydroxyvitamin D3; D004100:Dihydroxycholecalciferols; C118756:dihydroxy-vitamin D3; D014807:Vitamin D; C585549:CYP24A1 protein, human; C585551:Cyp24a1 protein, rat; D065668:Vitamin D3 24-Hydroxylase; D002112:Calcifediol
20,23-Dihydroxyvitamin D3; 20-Hydroxyvitamin D3; CYP24A1; Cytochrome P450; Phospholipid vesicles; Vitamin D3