Assay for screening for an anti-viral agent

Abstract

An antiviral agent capable of disrupting the association of two viral proteins required for DNA replication in herpesviruses. The agents disrupt the association of UL8 and POL in HSV-1 or the association of equivalent homologues of these proteins in other herpesviruses (for example UL 102 and UL54 in HCMV) Suitable agents are peptides which mimic the C-terminal or C-proximal portion of UL8 (or its homologues) for example the peptide IELVFTGVLAGVWGEGGKFV. Peptidomimetic compounds of such peptides are also suitable anti-viral agents. An assay to test for agents capable of disrupting association of POL and UL8 (or homologues thereof) is also described.

1 - 14 . (canceled)  
     
     
         15 . An in vitro assay method to determine the ability of a substance to inhibit the association of UL8 and POL wherein “UL8” is defined as UL8 of HSV-1 or the homologues thereof in other herpesviruses and “POL” is defined as POL of HSV-1 or the homologues thereof in other herpesviruses, said method comprising: 
 i) providing a first viral component consisting of UL8;    ii) providing a second viral component consisting of POL;    iii) exposing said first viral component to said substance followed by addition of the second viral component, or exposing said second viral component to said substance followed by addition of the said first viral component, or exposing the first viral component to said second viral component followed by said substance;    iv) washing to remove any second viral component and/or substance not associated with the first viral component; and    v) detecting the presence, and optionally determining the amount, of associated first and second viral components.    
     
     
         16 . The assay method as claimed in  claim 15  wherein UL8 of HSV-1 is the first viral component and POL of HSV-1 is the second viral component.  
     
     
         17 . The assay method as claimed in  claim 15  wherein one of UL102 of HCMV is the first viral component and UL54 of HCMV is the second viral component.  
     
     
         18 . The assay method as claimed in  claim 15  wherein one of said first and second viral components is localised on a surface.  
     
     
         19 . The assay method as claimed in  claim 15  wherein an antibody is used to detect association of said viral components.  
     
     
         20 . (canceled)  
     
     
         21 . (canceled)  
     
     
         22 . (canceled)  
     
     
         23 . (canceled)  
     
     
         24 . (canceled)  
     
     
         25 . (canceled)  
     
     
         26 . (canceled)  
     
     
         27 . (canceled)  
     
     
         28 . An in vitro assay method to determine the ability of a substance to inhibit the association of POL and a polypeptide selected from the group consisting of:  
       
         
           
                 
                 
                 
                 
               
                     
                 
                   a) 
                   VFTGVLAGVWGEGGKFVYPFDDKMSFLFA; 
                   (SEQ ID NO: 5) 
                     
                 
                     
                 
                   b) 
                   IELVFTGVLAGVWGEGGKFV; 
                   (SEQ ID NO: 7) 
                 
                     
                 
                   c) 
                   DEWVRSLAVDAQHAAKRVASEGLRFFRLNA; 
                   (SEQ ID NO: 11) 
                 
                   and 
                 
                     
                 
                   d) 
                   TWLEERDEWVRSLAVDAQHAARRVAS. 
                   (SEQ ID NO: 12) 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       wherein “POL” is defined as POL of HSV-1 or the homologues thereof in other herpesviruses, said method comprising: 
 i) providing said polypeptide;  
 ii) exposing said polypeptide to said substance followed by addition of POL, or exposing POL to said substance followed by addition of the said polypeptide, or exposing POL to said polypeptide followed by said substance;  
 iii) washing to remove any POL and/or substance not associated with the polypeptide; and  
 v) detecting the presence, and optionally determining the amount, of associated polypeptide and POL.