In vitro metabolism of the vitamin D analog, 22-oxacalcitriol, using cultured osteosarcoma, hepatoma, and keratinocyte cell lines.
Using four cultured cell models representing liver, keratinocyte, and osteoblast, we have demonstrated that the vitamin D analog, 22-oxacalcitriol is degraded into a variety of hydroxylated and side chain truncated metabolites. Four of these metabolic products have been rigorously identified by high pressure liquid chromatography, diode array spectrophotometry, and gas chromatography-mass spectrometry analysis as 24-hydroxylated and 26-hydroxylated derivatives as well as the cleaved molecules, hexanor-1alpha,20-dihydroxyvitamin D3 and hexanor-20-oxo-1alpha-hydroxyvitamin D3. Comparison with chemically synthesized standards has revealed the stereochemistry of the biological products. Although differences exist in the amounts of products formed with the different cell types, it is apparent that 22-oxacalcitriol is subject to metabolism by both vitamin D-inducible and noninducible enzymes. Time course studies suggest that the truncated 20-alcohol is derived from a side chain hydroxylated molecule via a hemiacetal intermediate and the 20-oxo derivative is likely formed from the 20-alcohol. Biological activity measurements of the metabolites identified in our studies are consistent with the view that these are catabolites and that the biological activity of 22-oxacalcitriol is due to the parent compound. These results are also consistent with recent findings of others that the biliary excretory form of 22-oxacalcitriol is a glucuronide ester of the truncated 20-alcohol.
Masuda
S
S
Department of Biochemistry, Queen's University, Kingston, Ontario, K7L 3N6 Canada.
Byford
V
V
Kremer
R
R
Makin
H L
HL
Kubodera
N
N
Nishii
Y
Y
Okazaki
A
A
Okano
T
T
Kobayashi
T
T
Jones
G
G
D000818:Animals; D002117:Calcitriol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D006133:Growth Substances / Q000737:chemistry; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D012516:Osteosarcoma / Q000378:metabolism*; D051381:Rats; D018167:Receptors, Calcitriol / Q000378:metabolism; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D006133:Growth Substances; D018167:Receptors, Calcitriol; D002117:Calcitriol; C051883:maxacalcitol