Quantification of 2'-fluoro-2'-deoxyuridine and 2'-fluoro-2'-deoxycytidine in DNA and RNA isolated from rats and woodchucks using LC/MS/MS.
Apatmers are synthesized using 2'-fluoropyrimdines in place of normal pyrmidines to stabilize them against enzymatic degradation, and thereby improve their therapeutic efficacy. Despite this stabilizing effect, the apatmers can still be degraded by nucleases in the blood. Primer template extension studies have demonstrated that mammalian DNA polymerases can incorporate these 2'-fluoropyrimidines into growing strands of DNA. The toxicologic effects of these compounds have been examined in rats and woodchucks, animals known to be susceptible to the toxic effects of other modified pyrimidines. Whether these nucleosides can be incorporated into DNA in vivo has not been established. These studies report the development of methodologies and the results of studies designed to determine if and to what extent 2'-fluoropyrimidines are incorporated into tissue DNA following long-term treatment. Rats were dosed intravenously with either 2'-fluorouridine (2'-FU) or 2'-fluorocytidine (2'-FC) at doses of 5, 50, and 500 mg/kg/day for 90 days. Woodchucks were dosed intravenously with either 2'-FU or 2'-FC at doses of 0.75 or 7.5 mg/kg/day for 90 days. The amounts of 2'-FU or 2'-FC in DNA and RNA were quantified using newly developed LC/MS/MS methodologies. Administration of 2'-FU to rats and woodchucks resulted in incorporation of the compound into DNA from liver, spleen, testis, muscle, and kidney. Incorporation also occurred in RNA from rat liver (only tissue examined). Similarly, administration of 2'-FC to rats and woodchucks resulted in incorporation into liver DNA (only tissue examined). These data demonstrate that 2'-fluoropyrimidines are incorporated into DNA and RNA of various tissues of rats and woodchucks following long-term administration.
Richardson
Frank C
FC
OSI Pharmaceuticals, Inc., Boulder, Colorado 80301, USA.
Zhang
Chenghong
C
Lehrman
S Russ
SR
Koc
Hasan
H
Swenberg
James A
JA
Richardson
Katherine A
KA
Bendele
Raymond A
RA
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000096:biosynthesis / Q000737:chemistry*; D003841:Deoxycytidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000032:analysis* / Q000378:metabolism; D005467:Floxuridine / Q000008:administration & dosage / Q000032:analysis* / Q000378:metabolism; D006868:Hydrolysis; D008297:Male; D008392:Marmota; D009928:Organ Specificity; D012313:RNA / Q000096:biosynthesis / Q000737:chemistry*; D051381:Rats; D011916:Rats, Inbred F344; D021241:Spectrometry, Mass, Electrospray Ionization
D005467:Floxuridine; D003841:Deoxycytidine; D012313:RNA; D004247:DNA; C007251:2'-fluoro-2'-deoxycytidine