Chemical Stability of Pentostatin (NSC-218321), a Cytotoxic and Immunosuppressant Agent

Abstract

Pentostatin, an unusual nucleoside of natural origin, has been used for the treatment of hairy cell leukemia, as an immunosuppressant agent, and as an inhibitor of adenosine deaminase. The studies of the physicochemical properties and solution stability of pentostatin (1) are important to the development of a parenteral formulation for extensive preclinical and clinical testing. Pentostatin displayed apparent pK _a values at 25 ± 0.1°C and ionic strength of 0.15 M of 2.03 ± 0.03 and 5.57 ± 0.14 (spectrophotometric) and 5.50 ± 0.02 (potentiometric) for N₁ and the amidine nitrogen in the seven-membered ring, respectively, which are the most likely protonation sites. The rates of degradation of pentostatin were determined as a function of pH, buffer concentration, and temperature. In the pH range 1.0–4.0, pentostatin undergoes acid-catalyzed glycosidic cleavage leading to the formation of the base compound, 2, and 2-deoxyribose. A carbonium ion mechanism in which C–N bond cleavage was the rate-determining step was consistent with the data. In the pH range 6.5–10.5, the imine bond at C₅ position in pentostatin is hydrolyzed to form the corresponding formamide. Pentostatin hydrolysis in this pH range was independent of pH. At pH >11, pentostatin decomposes to nonchromophoric products probably through multiple-step base-catalyzed hydrolytic mechanisms. Pentostatin appears to be quite stable after reconstitution of a lyophilized experimental dosage form. Care must be taken if pentostatin is extensively diluted with 5% dextrose in water, as pentostatin stability is compromised at pH values less than 5.