Enhanced clearance of topoisomerase I inhibitors from human colon cancer cells by glucuronidation

As part of a program to identify novel mechanisms of resistance to topoisomerase I (topo 1) inhibitors, the cellular pharmacology of 7-ethyl-10-hydroxycamptothecin (SN-38), the active metabolite of clinically used irinotecan (CPT-11) and NU/ICRF 505, an anthraquinone-tyrosine conjugate, has been investigated in two human colorectal cancer (CRC) cell lines. Two novel metabolites of NU/ICRF 505 (M I and M2) and a single metabolite of SN-38 (M 1) were detected by high performance liquid chromatography in the culture medium of HT29 cells but were absent in HCT116 cells. Identities of all three metabolites were established by a combination of biochemical and physicochemical techniques. M I of SN-38 was the C10-(beta)-glucuronide of the parent lactone while M I of NU/ICRF 505 was the C4-O-glucuronide and M2 the tyrosine-O-glucuronide. both of the parent compound. Drug transport studies revealed that by 24 hr HT29 cells had effectively cleared 82.5% of NU/ICRF 505 (10 muM) into the culture medium as the two glucuronides. In contrast, intracellular concentrations of NU/ICRF 505 were maintained in HCT116 cells in the absence of glucuronidation at a level 550 times greater than in HT29 cells. HT29 cells cleared 40.9% of SN-38 (I muM) as the glucuronide to the culture medium, while the parent drug was maintained at a level 2-fold greater in HCT116 cells. Enhanced drug clearance due to glucuronidation may contribute to intrinsic drug resistance of human CRC. (C) 2002 Elsevier Science Inc. All rights reserved.