Screen for inhibitors of filovirus and uses therefor

Abstract

The invention provides methods to identify agents useful to prevent, inhibit or treat viral infections, e.g., filovirus infections, as well as compositions having one or more agents to prevent, inhibit or treat viral infection.

1 . A method to identify one or more agents that inhibit Ebola virus infection, comprising:
 a) contacting a host cell, one or more agents and recombinant infectious, biologically contained Ebola virus, the genome of which contains a deletion of sequences corresponding to Ebola virus VP30 sequences, which deletion is effective to prevent expression of functional VP30 upon infection of a cell with the recombinant virus; and   b) identifying one or more agents that inhibit viral infection without substantially decreasing host cell viability.   
     
     
         2 . The method of  claim 1  wherein the host cell is contacted with the virus before or after the one or more agents. 
     
     
         3 . The method of  claim 1  wherein the host cell is contacted with the virus and the agents simultaneously. 
     
     
         4 . The method of  claim 1  wherein the identified agent inhibits the infectivity of the virus by at least 90%. 
     
     
         5 . The method of  claim 1  wherein the detected agent has an IC 50  of less than about 10.0 μM or a CC 50  of more than about 0.1 μM. 
     
     
         6 . (canceled) 
     
     
         7 . A method to identify inhibitors of  filovirus  glycoprotein receptor binding or fusion, comprising:
 a) contacting a host cell with one or more agents and a recombinant replication incompetent pseudotyped rhabdovirus comprising  filovirus  glycoprotein and a mutant negative sense rhabdovirus genome which lacks sequences for a rhabdovirus glycoprotein but comprises a sequence for a reporter protein; and   b) identifying at least one agent that inhibits reporter protein levels or expression in the host cell.   
     
     
         8 . The method of  claim 7  wherein the rhabdovirus is VSV. 
     
     
         9 . The method of  claim 7  wherein the  filovirus  glycoprotein is a Marburg virus glycoprotein or an  Ebolavirus  glycoprotein. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 7  wherein the reporter protein levels or expression in the presence of the one or more agents is compared to reporter protein levels or expression in the absence of the agent(s). 
     
     
         12 . The method of  claim 7  wherein at least one agent inhibits  filovirus  glycoprotein binding. 
     
     
         13 . The method of  claim 7  wherein at least one agent inhibits  filovirus  glycoprotein fusion. 
     
     
         14 . (canceled) 
     
     
         15 . The method of  claim 7  wherein the host cell is a mammalian cell. 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 1  wherein the at least one agent contacted with the cell is a triphenylethylene, an inhibitor of calcium-independent phospholipase A 2  and/or of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of PGE 2  synthase, a steroid, dopamine antagonist, anticholinergic or an Hsp90 inhibitor. 
     
     
         18 . The method of  claim 1  wherein the at least one agent is a compound of formula (I)-(XIII). 
     
     
         19 - 21 . (canceled) 
     
     
         22 . A method to prevent, inhibit or treat  filovirus  infection in a mammal, comprising administering to the mammal an effective amount of a compound of formula (I)-(XIII) or an effective amount of a pharmaceutical composition comprising an agent, wherein the composition comprises a calcium channel blocker, a tetranortriterpenoid, a sigma receptor agonist, triphenylethylene, an inhibitor of calcium-independent phospholipase A 2 , an inhibitor of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of PGE 2  synthase, a steroid, adopamine antagonist, an inhibitor of Hsp90, or an anticholinergic. 
     
     
         23 . (canceled) 
     
     
         24 . The method of  claim 22  wherein the mammal is a human. 
     
     
         25 . The method of  claim 22  wherein the agent is bepridil hydrochloride, clomiphene citrate, benzotropin mesylate, 7-deacetoxy-3-deacetyl-7-oxokhivorin, 1,2alpha-epoxy-7-deacetoxy-7-oxodihxdrogedunin, epoxygedunin, 1,3-dideacetyl-7-deacetoxy-7-oxokhivorin, gedunin, tamoxifen citrate, fluspirilene, raloxifene hydrochloride, bromoenol lactone, cortexolone maleate, (R,R)-cis,diethyl tetrahydro-2,8-chrysenediol, MK-866, L-687, 384 hydrochloride, cycloheximide, gedunol, dihydrogedunin, 3beta-acetoxydeoxodihydrogedunin, 3alpha-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 3beta-hydroxydeoxodihydrogedunin, deacetoxy-7-oxogedunin, 1,2alpha-epoxydeacetoxydihydrogedunin, 3beta-hydroxydeoxydesacetoxy-7-oxogedunin, tridesacetoxykhivorin, 1,3-dideacetylkhivorin, heudelottin C, geldanamycin, 17-allylamino-17-demethoxygeldanamycin, 4-[4-(2,3-dihydro-1,4-benzodioxin-6-yl)-5-methyl-1H-pyrazol-3-yl]-6-ethyl-1,3-benzenediol, 6-phenylimidazo[2,1-b]-1,3,4-thiadiazole-2-sulfonamide, geldanamycin, 17-Allylamino-17-demethoxygeldanamycin, 4-[4-(2,3-Dihydro-1,4-benzodioxin-6-yl)-5-methyl-1H-pyrazol-3-yl]-6-ethyl-1,3-benzenediol, 6-Phenylimidazo[2,1-b]-1,3,4-thiadiazole-2-sulfonamide, or any combination thereof. 
     
     
         26 . The method of  claim 22  wherein the agent is orally, intravenously or subcutaneously administered. 
     
     
         27 - 30 . (canceled) 
     
     
         31 . The method of  claim 7  wherein the at least one agent contacted with the cell is a triphenylethylene, an inhibitor of calcium-independent phospholipase A 2  and/or of magnesium-dependent phosphatidate phosphohydrolase, an inhibitor of PGE 2  synthase, a steroid, dopamine antagonist, anticholinergic or an Hsp90 inhibitor. 
     
     
         32 . The method of  claim 7  wherein the at least one agent is a compound of formula (I)-(XII).