Metabolism of the vitamin D3 derivative (24R)‐hydroxycalcidiol by human promyelocytic leukemia cells (HL‐60): Isolation and identification of (5Z) and (5E)‐(24R)‐19‐nor‐10‐oxo‐24‐hydroxycalcidiol

Seiichi ISHIZUKA, Toshimitsu MATSUI, Yoshinobu NAKAO, Takuo FUJITA, Helmut REICHEL, Anthony W. NORMAN

Research output: Contribution to journalArticlepeer-review

Abstract

Human promyelocytic leukemia cells (HL‐60 cells) incubated with (24R)‐hydroxy[26,27‐methyl‐3H]calcidiol (0.2 μCi) or non‐radioactive (24R)‐hydroxycalcidiol (370 μg) produced significant quantities of two new vitamin D3 (calciol) metabolites. The metabolites were isolated from HL‐60 cell culture media by methanol/chloroform extraction and a series of chromatographic procedures. The two new metabolites were identified as (5Z)‐ and (5E)‐(24R)‐19‐nor‐10‐oxo‐24‐hydroxycalcidiol by HPLC analysis, ultraviolet absorption spectrophotometry, mass spectrometry and Fourier‐transfrom infrared spectrophotometry. According to the isolation and purification procedures, the total amounts of 3.04 μg (5Z)‐(24R)‐19‐nor‐10‐oxo‐24‐hydroxycalcidiol (λmax= 310 nm, ɛ= 17070 M−1 cm−1) and 8.89 μg (5E)‐(24R)‐19‐nor‐10‐oxo‐24‐hydroxycalcidiol (λmax= 312 nm, e= 24500 M−1 cm−1) were calculated, assuming an M1 of 418. The activity of 19‐nor‐10‐oxo‐(24R)‐hydroxycalcidiol to promote HL‐60 cell differentiation was higher than the activity of the precursor (24R)‐hydroxycalcidiol suggesting a possible biological action of this metabolite in HL‐60 cells.

Original languageEnglish
Pages (from-to)475-483
Number of pages9
JournalEuropean Journal of Biochemistry
Volume170
Issue number1-2
DOIs
StatePublished - Dec 1987
Externally publishedYes

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