Figure 1.
Effect of M2 expression and amantadine on yeast growth.
Yeast strains containing an empty plasmid (A) or plasmid bearing WT M2 (B), S31N M2 (C) or V27A M2 (D) were distributed into 96-well plates and their growth was measured over time following transfer at 0h to medium containing galactose and addition of the indicated concentrations of amantadine.
Figure 2.
Effect of a spiroadamantane amine on the growth of yeast expressing WT and mutated M2.
Yeast strains containing the indicated plasmids were distributed in 96-well plates in medium containing galactose and exposed to the indicated concentrations of the spiroadamantane amine (structure shown) for 40 h.
Figure 3.
(A) Yeast expressing WT M2 were exposed to 1,440 screening chemicals (M2+ Drugs) with 8 untreated wells per plate as negative controls (M2, n = 144). Each plate also contained 8 positive controls not treated with drugs (Empty plasmid, n = 144). Growth was measured after 40 h and the % growth restoration was calculated as described in Materials and Methods. The arrow points to the single active compound found in this batch. (B) the active compound found in A was retested at different concentrations against yeast expressing WT M2 or bearing the empty plasmid and growth was measured after 40 h.
Figure 4.
Structures, names and EC50 of active compounds found in the screen.
Figure 5.
Activity of hexamethylene amiloride and selected triazine analogs in the TEVC assay.
Each point is the mean and standard deviation of five to eight oocytes.
Figure 6.
Antiviral activity and cytotoxicity of selected compounds.
The effect of the compounds on influenza A/Udorn/72 WT virus replication was evaluated by plaque formation in MDCK cells. Cytotoxicity towards MDCK cells was assayed after 48 h drug exposure with the MTT tetrazolium dye assay as described [55].