Reduced albumin binding promotes the stability and activity of topotecan in human blood.

Topotecan, a semisynthetic water-soluble analogue of camptothecin, is the first topoisomerase I targeting anticancer agent to enter comparative phase III clinical trials. Here we elucidate the biophysical factors underlying the markedly improved bloodstream stability and cytotoxic activity of topotecan relative to camptothecin. Each agent contains an alpha-hydroxy-delta-lactone ring that hydrolyzes under physiological pH to yield a biologically-inactive carboxylate form. In human plasma, camptothecin lactone converts rapidly and completely to its carboxylate form due to a 200-fold binding preference by serum albumin (HSA) for the latter [Mi, Z., & Burke, T.G. (1994) Biochemistry 33, 10540-12545]. Time-resolved fluorescence anisotropy measurements reveal that neither topotecan lactone nor carboxylate associates with HSA, thereby resulting in a significantly higher level of lactone stability in plasma for topotecan (t1/2 = 23.1 min, percent lactone at equilibrium of 17.6) relative to camptothecin (t1/2 = 10.6 min, percent lactone at equilibrium of < 0.2). Moreover, studies with HL-60 human promyelocytic leukemia cells reveal that a physiologically-relevant level (40 mg/mL) of HSA dramatically attenuates the cytotoxic activity of camptothecin in excess of 2600-fold (for a 72 h exposure, the IC50 value of 1.5 nM in the absence of HSA increased to 4 microM in the presence of HSA). The activities of other clinically relevant anticancer analogues, 9-aminocamptothecin and SN-38, were also strongly modulated by the presence of 40 mg/mL HSA. In marked contrast, the presence of HSA effected no change on the cytotoxic activity of topotecan (IC50 = 12 nM both in the absence and in presence of HSA).(ABSTRACT TRUNCATED AT 250 WORDS)