title: Glucuronidation of 7-ethyl-10-hydroxycamptothecin (SN-38), an active metabolite of irinotecan (CPT-11), by human UGT1A1 variants, G71R, P229Q, and Y486D.
abstract: 7-Ethyl-10-hydroxycamptothecin (SN-38), an active metabolite of antitumor agent irinotecan (CPT-11), is conjugated and detoxified to SN-38-glucuronide by UDP-glucuronosyltransferase (UGT) 1A1. Genetic polymorphisms in UGT1A1 are thought to contribute to severe diarrhea and/or leukopenia caused by CPT-11. In this regard, it has been reported that polymorphisms in the promoter region could affect the CPT-11 pharmacokinetics and interindividual variation of toxicity. However, little information is available on the influence of UGT1A1 polymorphisms in the coding region on the SN-38 glucuronidation activity. In the present study, wild-type (WT) and three variant (G71R, P229Q, and Y486D) cDNAs of human UGT1A1s were transiently expressed in COS-1 cells, and the kinetic parameters of these UGT1A1s were determined for SN-38 glucuronidation. A partially reduced UGT1A1 protein expression was observed in COS-1 cells for G71R and Y486D. WT UGT1A1 catalyzed SN-38 glucuronidation with an apparent K(m) value of 11.5 microM, whereas those of G71R, P229Q, and Y486D were 14.0, 18.0, and 63.5 microM, respectively. The SN-38 glucuronidation efficiency ratio (V(max)/K(m)) normalized for the level of expression was 1.4, 0.66 (47% of WT), 0.73 (52%), and 0.07 (5%) microl/min/mg of protein for WT, G71R, P229Q, and Y486D, respectively. Thus, the SN-38 glucuronidation activity of Y486D was drastically reduced, whereas the reduction in the G71R and P229Q activities was fractional. The decreased SN-38 glucuronidation efficiency ratio of G71R and P229Q could be critical in combination with other polymorphisms in the UGT1A1 gene.
authors.affiliation: Division of Environmental Chemistry, National Institute of Health Sciences, Tokyo, Japan.
authors.forename: Hideto
authors.initials: H
authors.lastname: Jinno
authors.forename: Toshiko
authors.initials: T
authors.lastname: Tanaka-Kagawa
authors.forename: Nobumitsu
authors.initials: N
authors.lastname: Hanioka
authors.forename: Mayumi
authors.initials: M
authors.lastname: Saeki
authors.forename: Seiichi
authors.initials: S
authors.lastname: Ishida
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Nishimura
authors.forename: Masanori
authors.initials: M
authors.lastname: Ando
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Saito
authors.forename: Shogo
authors.initials: S
authors.lastname: Ozawa
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Sawada
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics*; D000818:Animals; D001120:Arginine / Q000235:genetics; D001224:Aspartic Acid / Q000235:genetics; D019556:COS Cells; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000378:metabolism*; D002522:Chlorocebus aethiops; D014644:Genetic Variation; D020719:Glucuronides / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000235:genetics* / Q000378:metabolism*; D005973:Glutamine / Q000235:genetics; D005998:Glycine / Q000235:genetics; D006801:Humans; D000077146:Irinotecan; D011392:Proline / Q000235:genetics; D014443:Tyrosine / Q000235:genetics; D000099148:UGT1A1 Enzyme
chemical_list: D001120:Arginine; D001224:Aspartic Acid; D002166:Camptothecin; D020719:Glucuronides; D014453:Glucuronosyltransferase; D005973:Glutamine; D005998:Glycine; D000077146:Irinotecan; D011392:Proline; D014443:Tyrosine; D000099148:UGT1A1 Enzyme