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Enzymatic Estimation and Metabolism of 1-ß-D-Arabinofuranosylcytosine in Man¹

McGill University Cancer Research Unit, Montreal, Quebec, Canada

An enzymatic method for the estimation of the concentration of 1-ß-D-arabinofuranosylcytosine (ara-C) in biological fluid has been developed. The method consists of the phosphorylation of ara-C with ATP--³²P in the presence of deoxycytidine kinase and Mg⁺⁺ and subsequent isolation of the 5'-monophosphate of ara-C (ara-CMP--³²P) by thin-layer chromatography on diethylaminoethyl-cellulose. This method requires only 3 hr to complete and can detect concentrations of ara-C in the plasma as low as 2.5 M. With this enzymatic method the half-life of ara-C in the plasma was estimated to be 15 min in a patient with acute leukemia who received a continuous i.v. infusion of ara-C.

Since for most nucleoside analogs the active inhibitor is a phosphorylated derivative, the enzymatic method can be used as an additional procedure for the screening of potential antitumor nucleosides.

¹ Supported by the National Cancer Institute of Canada.

² Recipient of USPHS International Fellowship 1F05 TWO 1731-01. Present address: Laboratory of Molecular Embryology, Naples, Italy.

Received 9/28/71. Accepted 11/ 4/71.