Cellular pharmacology ofd-3-azido-3-deoxy-myo-inositol, an inhibitor of phosphatidylinositol signaling having antiproliferative activity


d-3-Azido-3-deoxy-myo-inositol (3AMI) is an inhibitor of the growth of v-sis-transformed NIH 3T3 cells but not of wild-type NIH 3T3 cells, whose effects may be mediated through the phosphatidylinositol-3′-kinase pathway. We studied some properties of the cellular pharmacology of 3AMI using high-specific-activity [3H]-3AMI. The uptake of [3H]-3AMI by wild-type NIH 3T3 and v-sis NIH 3T3 cells was similar. [3H]-3AMI was a substrate for phosphatidylinositol synthetase, with the maximal velocity (Vmax) being 1.0 nmol min−1 mg−1 and the Michaelis constant (K m) being 23 mM. Corresponding values obtained for [3H]-myo-inositol as a substrate were 5.5 nmol min−1 mg−1 and 3.2 mM. [3H]-3AMI was incorporated into the cellular inositol lipids of v-sis NIH 3T3 cells to a similar extent as that observed for [3H]-myo-inositol but was not incorporated into the inositol lipids of wild-type NIH 3T3 cells. The [3H]-3AMI incorporated by the v-sis NIH 3T3 cells was present in the phosphatidylinositol and phosphatidylinositol phosphate fractions but not in bisphosphorylated phosphatidylinositol.myo-Inositol antagonized the growth-inhibitory effects of 3AMI. The v-sis NIH 3T3 cells were found to be more sensitive than the wild-type NIH 3T3 cells to growth inhibition (without 3AMI) caused by the removal ofmyo-inositol from the medium. The results of the study suggest that 3AMI is an antimetabolite ofmyo-inositol. The relative sensitivity of v-sis NIH 3T3 and some other cells to 3AMI may be a reflection of increasedmyo-inositol requirements for the growth of these cells as compared with wild-type NIH 3T3 cells.


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