The in vitro evaluation of nucleoside analogues as probes for use in the noninvasive diagnosis of herpes simplex encephalitis.
1-(2-Deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodouracil (FIAU) and the 5-iodo (FIRU), 5-bromo (FBrRU), 5-chloro (FClRU) and 5-fluoro (FFRU) analogues of 1-(2-deoxy-2-fluoro-beta-D-ribofuranosyl)uracil were examined in four in vitro tests designed to evaluate their potential as radiopharmaceuticals in a non-invasive diagnostic test for herpes simplex encephalitis (HSE). The tests involved, (a) cellular uptake studies--which showed that all five nucleosides are selectively trapped in rabbit kidney cells infected with herpes simplex virus, type I, which codes for its own thymidine kinase; (b) incubation studies--which showed that, in human platelets, FIAU, FIRU and FFRU are not degraded by thymidine phosphorylase (an enzyme which catalyzes the glycosidic bond cleavage of physiological nucleosides); (c) a transport study, using mouse erythrocytes--which indicated that all five nucleosides have good affinity for a NBMPR-sensitive nucleoside transporter, with inhibition constants (Ki values in the range of 0.177-0.41; and (d) determination of octanol/water partition coefficients--which (log P = -0.14 to -1.67) indicated that FIAU is the most lipophilic of the five compounds studied, and may therefore cross the blood-brain barrier most readily. We conclude from the results that FIAU is the most promising compound for further evaluation in HSE animal models. The combination of tests described in this study provide a useful screening protocol for evaluation of other nucleoside analogues of potential use in a diagnostic test for HSE.
Tovell
D R
DR
Faculty of Medicine, University of Alberta, Edmonton, Canada.
Samuel
J
J
Mercer
J R
JR
Misra
H K
HK
Xu
L
L
Wiebe
L I
LI
Tyrrell
D L
DL
Knaus
E E
EE
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology; D001921:Brain / Q000000981:diagnostic imaging; D002478:Cells, Cultured; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D004660:Encephalitis / Q000175:diagnosis* / Q000000981:diagnostic imaging; D004912:Erythrocytes / Q000378:metabolism; D006561:Herpes Simplex / Q000175:diagnosis* / Q000000981:diagnostic imaging; D007457:Iodine Radioisotopes; D007668:Kidney / Q000378:metabolism; D009705:Nucleosides; D011817:Rabbits; D011877:Radionuclide Imaging; D018139:Simplexvirus; D012995:Solubility; D013939:Thymidine Phosphorylase / Q000378:metabolism
D007457:Iodine Radioisotopes; D009705:Nucleosides; D013939:Thymidine Phosphorylase