in cv 1 monkey kidney cells using a methyl cellulose disk overlay assay as described by schroeder et al j med chem 24 pp 1078 1983 1981 the inhibition of virus replication was determined quantitatively by measuring the amount of virus produced by infected cells during a single cell cycle of infection in the presence of the compound tested cv 1 cells were infected with 20 pfu cell of herpes simplex virus one hour post infection cells were rinsed with modified eagle 39 s medium and exposed for 30 min to a medium composed of 1 anti herpes rabbit antiserum to neutralize unpenetrated virus the cells were rinsed once with medium and then cultured with the test compound a baseline used for measuring net virus production was obtained by harvesting and freezing one sample which was untreated 4 hours after infection the remaining samples were collected and frozen 18 20 hours post infection virus was titrated in cv 1 cells by a plague assay as described by hughes et al j virol 16 pp 275 283 975 in studies designed to prevent the anti viral activity of the chelators fecl 3 at various concentrations was added to medium containing a fixed concentration of the test compounds and the anti vital assay performed as described above the partition equilibrium values for octanol and phosphate buffered saline were determined for certain of the test compounds the compounds were added in nitrogen saturated methanol at the same concentration to three acid washed test tubes and the alcohol evaporated leaving a thin film of compounds coating the interior of the tube next 5 ml each of degassed phosphate buffered saline ph 7 4 and n octanol were added the tubes were sealed with teflon caps under.