Carbodine, the carbocyclic analog of cytidine, was found to possess significant antiviral activity against influenza virus types A0/PR-8/34 and A2/Aichi/2/68 (Hong Kong) in vitro. The compound selectively inhibited PR-8 influenza virus-induced cytopathogenic effects in Madin-Darby canine kidney and inhibited Hong Kong influenza virus replication in primary rhesus monkey kidney cell cultures. The 50% minimum inhibitory concentration for inhibition of human influenza type A viruses by carbodine was approximately 2.6 microgram/ml (i.e., in the range of antiviral potency of ribavirin, but less potent than amantadine hydrochloride in concomitant assays). The fact that carbodine is metabolized to carbodine triphosphate in mammalian cells makes interference with the viral ribonucleic acid-dependent ribonucleic acid polymerase reaction a likely possibility for its principal mode of action. The carbocyclic analogs of uridine (the deamination product of carbodine), 2'-deoxycytidine, 3'-deoxycytidine, N,N-dimethylcytidine, N-methylcytidine, and some related carbocyclic analogs of pyrimidine nucleosides were inactive against PR-8 influenza virus in vitro. The combination of carbodine plus tetrahydrouridine was no more effective in vitro than carbodine alone, thus indirectly indicating that deamination of carbodine probably did not occur to a significant degree during the cell culture experiments. Although reproducibly active in vitro, carbodine did not exhibit any efficacy against lethal influenza virus infections in mice when administered by either the intraperitoneal or intranasal routes up to dose-limiting toxic levels.