title: Lack of contribution of dihydrofluorouracil and alpha-fluoro-beta-alanine to the cytotoxicity of 5'-deoxy-5-fluorouridine on human keratinocytes.
abstract: Capecitabine (Xeloda) is a very active oral fluoropyrimidine (colon and breast cancers) whose clinical use is complicated by the presence of hand-foot syndrome (HFS). This cutaneous toxicity is less frequently encountered with other oral fluoropyrimidines containing a dihydropyrimidine dehydrogenase (DPD) inhibitor. The HFS is thus attributed to the presence of the main 5-fluorouracil (5-FU) metabolites, dihydrofluorouracil (5-FUH2) and alpha-fluoro-beta-alanine (FBAL), but without strong pharmacological arguments. The aim of the present study was to closely examine this latter hypothesis. Capecitabine generates 5'-deoxyflourouridine (5'-DFUR) which is transformed into 5-FU at the cellular target site through the intermediary of thymidine phosphorylase (TP). The cytotoxic effects (MTT test, 4-day exposure) of 5'-DFUR, 5-FU, 5-FUH2 and FBAL were tested against the spontaneously immortalized human keratinocyte cell line (HaCaT) and the human cancer colon cell line WiDr as a control. Mean IC50s on HaCaT and WiDr were, respectively, 1.3 and 10 microM for 5'-DFUR, 0.2 and 3.3 microM for 5-FU, 13.4 and 560 microM for 5-FUH2, and greater than 650 and 6500 microM for FBAL. The respective 5'-DFUR IC50s values were not different when cells were exposed to 5'-DFUR alone or in combination with 5-FU, 5-FUH2 and FBAL in both cell lines, the relative proportion of each drug reflecting known pharmacokinetic data for capecitabine (5'-DFUR 12.4%, 5-FUH2 6.4%, 5-FU 1.2% and FBAL 80%). This latter finding demonstrates the relative lack of significant cytotoxic activity of 5-FUH2 and FBAL on human keratinocytes. TP activity was particularly high in HaCaT cells and DPD activity was very low in both cell lines. These data strongly suggest that the presence of 5-FU metabolites does not play a major role in the HFS generated by capecitabine and that it can probably be attributed to particularly high TP activity in keratinocytes. This observation may have important clinical consequences such as a possible local pharmacological inhibition of TP for controlling HFS.
authors.affiliation: Centre Antoine-Lacassagne, Nice, France; Pharmacy School, Marseille, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Fischel
authors.forename: Patricia
authors.initials: P
authors.lastname: Formento
authors.forename: Joseph
authors.initials: J
authors.lastname: Ciccolini
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Etienne-Grimaldi
authors.forename: G
authors.initials: G
authors.lastname: Milano
mesh_terms: D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000633:toxicity*; D045744:Cell Line, Tumor; D042943:Dihydrouracil Dehydrogenase (NADP) / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D005467:Floxuridine / Q000493:pharmacokinetics / Q000633:toxicity*; D005472:Fluorouracil / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000633:toxicity*; D005533:Foot Dermatoses / Q000139:chemically induced; D006229:Hand Dermatoses / Q000139:chemically induced; D006801:Humans; D020128:Inhibitory Concentration 50; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D013577:Syndrome; D013939:Thymidine Phosphorylase / Q000378:metabolism; D015091:beta-Alanine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000633:toxicity*
chemical_list: D000970:Antineoplastic Agents; D005467:Floxuridine; D015091:beta-Alanine; C032348:alpha-fluoro-beta-alanine; C019389:5-fluorodihydrouracil; D042943:Dihydrouracil Dehydrogenase (NADP); D013939:Thymidine Phosphorylase; D005472:Fluorouracil; C025034:doxifluridine