Isolation and identification of 1α‐hydroxy‐3‐epi‐vitamin D 3 , a potent suppressor of parathyroid hormone secretion

Since our original demonstration of the metabolism of 1α,25(OH) 2 D 3 into 1α,25(OH) 2 ‐3‐epi‐D 3 in human keratinocytes, there have been several reports indicating that epimerization of the 3 hydroxyl group of vitamin D compounds is a common metabolic process. Recent studies reported the metabolism of 25OHD 3 and 24( R ),25(OH) 2 D 3 into their respective C‐3 epimers, indicating that the presence of 1α hydroxyl group is not necessary for the 3‐epimerization of vitamin D compounds. To determine whether the presence of a 25 hydroxyl group is required for 3‐epimerization of vitamin D compounds, we investigated the metabolism of 1αOHD 3 , a non‐25 hydroxylated vitamin D compound, in rat osteosarcoma cells (ROS 17/2.8). We noted metabolism of 1αOHD 3 into a less polar metabolite which was unequivocally identified as 1αOH‐3‐epi‐D 3 using the techniques of HPLC, GC/MS, and 1 H‐NMR analysis. We also identified 1αOH‐3‐epi‐D 3 as a circulating metabolite in rats treated with pharmacological concentrations of 1αOHD 3 . Thus, these results indicated that the presence of a 25 hydroxyl group is not required for 3‐epimerization of vitamin D compounds. Furthermore, the results from the same studies also provided evidence to indicate that 1αOH‐3‐epi‐D 3 , like 1αOHD 3 , is hydroxylated at C‐25. We then evaluated the biological activities of 1αOH‐3‐epi‐D 3. Treatment of normal rats every other day for 7 days with 2.5 nmol/kg of 1αOH‐3‐epi‐D 3 did not raise serum calcium, while the same dose of 1αOHD 3 increased serum calcium by 3.39 ± 0.52 mg/dl. Interestingly, in the same rats which received 1αOH‐3‐epi‐D 3 we also noted a reduction in circulating PTH levels by 65 ± 7%. This ability of 1αOH‐3‐epi‐D 3 to suppress PTH levels in normal rats without altering serum calcium was further tested in rats with reduced renal function. The results indicated that the ED50 of 1αOH‐3‐epi‐D 3 for suppression of PTH was only slightly higher than that of 1α,25(OH) 2 D 3 , but that the threshold dose of the development of hypercalcemia (total serum Ca > 10.5 mg/dl) was nearly 80 times higher. These findings indicate that 1αOH‐3‐epi‐D 3 is a highly selective vitamin D analog with tremendous potential for treatment of secondary hyperparathyroidism in chronic renal failure patients. © 2005 Wiley‐Liss, Inc.