Stability of the new prodrug 9-aminocamptothecin glucuronide (9ACG) in the presence of human serum albumin

Abstract

9-aminocamptothecin glucuronide (9ACG) is a new water-soluble prodrug of 9-aminocamptothecin (9AC) that is a substrate for beta-glucuronidase and displays potent antitumor activity against human tumor xenografts. The lactone ring of camptothecins (CPTs) is required for antitumor activity but spontaneously opens under physiological conditions to an inactive carboxy form. The carboxy form of many CPTs, including 9AC, preferentially binds to human serum albumin (HSA), which further reduces the equilibrium amount of active lactone and greatly decreases antitumor efficacy. In this study, we examined the hypothesis that the unique structure of 9ACG might alter prodrug interaction with HSA and increase 9ACG lactone stability as compared with 9AC. HPLC analysis revealed that HSA did not affect the equilibrium level of 9ACG lactone whereas both CPT lactone and 9AC lactone were greatly reduced in the presence of HSA as compared to their equilibrium levels in PBS. Similar results were found in human serum and whole blood. The lactone ring of 9ACG also opened more slowly (t(1/2)=50 min) as compared with 9AC (t(1/2)=20 min) in the presence of HSA. Both 9ACG lactone and 9ACG carboxy bound HSA with similar affinities (K(D) approximately 4.5 x 10(-5)M(-1)). Binding of 9ACG to HSA reduced prodrug toxicity to cancer cells by about 10-fold in vitro. Injection of HSA into nude mice prolonged the half-life of 9ACG by about 3-fold, indicating that albumin-bound 9ACG lactone may act as a depot of active prodrug in vivo. Our results suggests that in contrast to CPT and 9AC, HSA does not appear to adversely affect 9ACG and may enhance the selective antitumor activity of 9ACG in tumors that contain beta-glucuronidase.