Isolation, identification and biological activity of 24R,25-dihydroxy-3-epi-vitamin D3: a novel metabolite of 24R,25-dihydroxyvitamin D3 produced in rat osteosarcoma cells (UMR 106).
We recently identified 1alpha,25-dihydroxy-3-epi-vitamin D3 [1alpha,25(OH)2-3-epi-D3] as a metabolite of 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] produced in rat osteosarcoma cells (UMR 106). We now report the isolation of 24R,25-dihydroxy-3-epi-vitamin D3 [24R,25(OH)2-3-epi-D3] as a metabolite of 24R,25-dihydroxyvitamin D3 [24R,25(OH)2D3] by high-performance liquid chromatography (HPLC) with chiral column and its structure assignment by proton nuclear magnetic resonance (1H-NMR) and liquid chromatography-mass spectrometry (LC-MS) analysis. We also demonstrated the production of 24R,25(OH)2-3-epi-D, in two other cell lines [human colon carcinoma cells (Caco-2) and porcine kidney cells (LLC-PK1)] which were previously shown to convert 1alpha,25(OH)2D3 into 1alpha,25(OH)2-3-epi-D3. It can be seen that the production of 24R,25(OH)2- 3-epi-D3 from 24R,25(OH)2D3 is lower than that of 1alpha,25(OH)2-3-epi-D3 from 1alpha,25(OH)2D3 in all the cells studied. 24R,25(OH)2-3-epi-D3 was found to be inactive in terms of its ability to bind to the vitamin D receptor (VDR), in inhibiting proliferation and in inducing differentiation of human promyelocytic leukemia cells (HL-60). Thus, our study indicates that the C-3 epimerization pathway is common to both 1alpha,25(OH)2D3 and 24R,25(OH)2D3 and may play an important role in modulating the concentration and the biological activity of these two major vitamin D3 metabolites in target tissues.
Kamao
M
M
Department of Hygienic Sciences, Kobe Pharmaceutical University, Japan.
Tatematsu
S
S
Reddy
G S
GS
Hatakeyama
S
S
Sugiura
M
M
Ohashi
N
N
Kubodera
N
N
Okano
T
T
D015650:24,25-Dihydroxyvitamin D 3 / Q000378:metabolism*; D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology; D000818:Animals; D001859:Bone Neoplasms; D018938:Caco-2 Cells; D002851:Chromatography, High Pressure Liquid; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007536:Isomerism; D009682:Magnetic Resonance Spectroscopy; D012516:Osteosarcoma; D013552:Swine; D014407:Tumor Cells, Cultured; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology
D020533:Angiogenesis Inhibitors; C118756:dihydroxy-vitamin D3; D014807:Vitamin D; D015650:24,25-Dihydroxyvitamin D 3