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

Jeffrey Cummings, Gary Boyd, Brian T. Ethell, Janet S. Macpherson, Brian Burchell, John F. Smyth, Duncan I. Jodrell

Research output: Contribution to journalArticle

Abstract

As part of a program to identify novel mechanisms of resistance to topoisomerase I (topo I) 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 (M1 and M2) and a single metabolite of SN-38 (M1) 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. M1 of SN-38 was the C10-(β)-glucuronide of the parent lactone while M1 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 μM) 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 (1 μM) 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.
Original languageEnglish
Pages (from-to)607-613
Number of pages7
JournalBiochemical Pharmacology
Volume63
Issue number4
DOIs
Publication statusPublished - Feb 2002
Externally publishedYes

Fingerprint

irinotecan
Topoisomerase I Inhibitors
Colonic Neoplasms
Glucuronides
Cells
HT29 Cells
Metabolites
HCT116 Cells
Culture Media
Pharmaceutical Preparations
Colorectal Neoplasms
High performance liquid chromatography
Lactones
Drug Resistance
Tyrosine

Keywords

  • SN-38
  • NU/ICRF 505
  • drug uptake
  • Glucuronidation
  • drug clearance
  • human colon cancer cells

Cite this

Cummings, J., Boyd, G., Ethell, B. T., Macpherson, J. S., Burchell, B., Smyth, J. F., & Jodrell, D. I. (2002). Enhanced clearance of topoisomerase I inhibitors from human colon cancer cells by glucuronidation. Biochemical Pharmacology, 63(4), 607-613. https://doi.org/10.1016/S0006-2952(01)00812-7
Cummings, Jeffrey ; Boyd, Gary ; Ethell, Brian T. ; Macpherson, Janet S. ; Burchell, Brian ; Smyth, John F. ; Jodrell, Duncan I. . / Enhanced clearance of topoisomerase I inhibitors from human colon cancer cells by glucuronidation. In: Biochemical Pharmacology. 2002 ; Vol. 63, No. 4. pp. 607-613.
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Cummings, J, Boyd, G, Ethell, BT, Macpherson, JS, Burchell, B, Smyth, JF & Jodrell, DI 2002, 'Enhanced clearance of topoisomerase I inhibitors from human colon cancer cells by glucuronidation' Biochemical Pharmacology, vol. 63, no. 4, pp. 607-613. https://doi.org/10.1016/S0006-2952(01)00812-7

Enhanced clearance of topoisomerase I inhibitors from human colon cancer cells by glucuronidation. / Cummings, Jeffrey; Boyd, Gary; Ethell, Brian T. ; Macpherson, Janet S. ; Burchell, Brian; Smyth, John F. ; Jodrell, Duncan I. .

In: Biochemical Pharmacology, Vol. 63, No. 4, 02.2002, p. 607-613.

Research output: Contribution to journalArticle

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T1 - Enhanced clearance of topoisomerase I inhibitors from human colon cancer cells by glucuronidation

AU - Cummings, Jeffrey

AU - Boyd, Gary

AU - Ethell, Brian T.

AU - Macpherson, Janet S.

AU - Burchell, Brian

AU - Smyth, John F.

AU - Jodrell, Duncan I.

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AB - As part of a program to identify novel mechanisms of resistance to topoisomerase I (topo I) 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 (M1 and M2) and a single metabolite of SN-38 (M1) 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. M1 of SN-38 was the C10-(β)-glucuronide of the parent lactone while M1 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 μM) 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 (1 μM) 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.

KW - SN-38

KW - NU/ICRF 505

KW - drug uptake

KW - Glucuronidation

KW - drug clearance

KW - human colon cancer cells

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DO - 10.1016/S0006-2952(01)00812-7

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