A NEW SERIES OF VITAMIN-D ANALOGS IS HIGHLY-ACTIVE FOR CLONAL INHIBITION, DIFFERENTIATION, AND INDUCTION OF WAF1 IN MYELOID-LEUKEMIA

The active form of vitamin D-3 [1 alpha,25-dihydroxyvitamin-D-3 (1 alp ha,25(OH)(2)D-3)] modulates the proliferation and differentiation of h ematopoietic cells. Analogs of 1 alpha,25(OH)(2)D-3 that have greater potency may have the potential as adjuvant therapy for high-risk patie nts in remission for acute myelogenous leukemia (AML) and myelodysplas tic syndromes. A new generation of 11 analogs of 1 alpha,25(OH)(2)D-3 has been synthesized, and we examined their effects on the human leuke mic cell line HL-60. This cell line provides a sensitive monitor of ac tivity of the 1 alpha,25(OH)(2)D-3 analogs. All the compounds were pot ent, producing a 50% clonal inhibition (ED(50)) in the range of 10(-8) to 10(-11) mol/L; nine of the 11 analogs had ED(50s) gt concentration s that were at least 10-fold lower than those for the parental 1,25(OH )(2)D-3. The most active compound [cmpd LA, (22R)-1 alpha,25-(OH)(2)-1 6,22,23-triene-D-3] had an ED(50) of 2 x 10(-11) mol/L; it was also te sted on clonogenic cells from patients with AML, and it achieved an ED S, of approximately 6 x 10(-11) mol/L, while 1 alpha,25(OH)(2)D-3 prod uced an ED(50) Of approximately 10(-8) mol/L on the same population of cells. Five different cell surface markers were examined on HL-60 cel ls exposed to the 1 alpha,25(OH)(2)D-3 analogs: HLA-DR and CD11b were induced by all of the compounds; CD13 was induced by six of the 12 com pounds, including 1,25(OH)(2)D-3; CD14 was strongly induced by all com pounds; and CD38 was induced rather weakly by nine of 12 analogs. WAF1 /CIP1/p21, a cyclin-dependent kinase inhibitor (CDKI), which is import ant in blocking the cell cycle, was examined by Western blot and was f ound to be induced by all of the compounds, suggesting a possible mech anism by which these analogs inhibit leukemic growth. The induction of WAF1 occurred at concentrations of vitamin D analogs as low as 10(-10 ) mol/L. This structure-function study showed that a new series of 1 a lpha,25(OH)(2)D-3 analogs was active in clonal inhibition, as well as induction of differentiation and WAF1 expression of HL-60 cells. The k ey structural motifs included C-16 double bond, double and/or triple b onds in the side chain, lengthening of the side chain, 20-epi-conforma tion of the side chain, replacement of six hydrogens at the end of the side chain with fluorines, and the removal of C-19. Consideration sho uld be given to further in vivo testing of toxicity and efficacy to mo ve toward a clinical trial, especially in a setting of minimal residua l disease. (C) 1996 by The American Society of Hematology.