Nelfinavir conjugates and antibodies useful in immunoassay

Abstract

Activated haptens useful for generating immunogens to HIV protease inhibitors, immunogens useful for producing antibodies to HIV protease inhibitors, and antibodies and labeled conjugates useful in immunoassays for the HIV protease inhibitor saquinavir. The novel haptens feature an activated functionality at the central, non-terminal hydroxyl group. Also described are monoclonal antibodies specific for saquinavir having less than 10% cross-reactivity with lopinavir, nelfinavir, amprenavir, ritonavir, and indinavir, and a murine hybridoma producing said antibodies.

1 . A compound having the structure  
     
       
         
         
             
             
         
       
     
     wherein 
 Y is O, S or NH,  
 m is 0 or 1,  
 L is a linker comprising from 0 to 40 carbon atoms arranged in a straight chain or a branched chain, saturated or unsaturated, and containing up to two ring structures and 0-20 heteroatoms, with the proviso that not more than two heteroatoms may be linked in sequence, and  
 A is an activated functionality selected from the group consisting of esters, isocyanates, isothiocyanates, thiols, imidoesters, anhydrides, maleimides, thiolactones, diazonium groups and aldehydes.  
 
   
   
       2 . The compound of  claim 1  wherein Y is O and m is 1.  
   
   
       3 . The compound of  claim 1  wherein Y is O, m is 1, and the first atom in L adjacent to C═Y is N.  
   
   
       4 . The compound of  claim 1  wherein Y is S, m is 1, and the first atom is L adjacent to C═Y is N.  
   
   
       5 . The compound of  claim 1  wherein Y is NH and m is 1.  
   
   
       6 . The compound of  claim 1  wherein m is 0.  
   
   
       7 . The compound Oc-(succinimido-oxycarbonyl-butyryl-aminocaproyl)-nelfinavir.  
   
   
       8 . The compound Oc-[4′-(succinimido-oxycarbonyl)-benzoyl-aminocaproyl]-nelfinavir.  
   
   
       9 . The compound Oc-(succinimido-oxycarbonyl-propylamino-co-glycyl-glycyl-glycyl-carbonyl)-nelfinavir.  
   
   
       10 . The compound Oar-MEM-Oc-(succinimido-oxycarbonyl-methyl)-nelfinavir.  
   
   
       11 . A compound having the structure  
     
       
         
         
             
             
         
       
     
     wherein 
 Y is O, S, or NH,  
 m is 0 or 1,  
 L is a linker comprising 0 to 40 carbon atoms arranged in a straight chain or a branched chain, saturated or unsaturated, and further comprising up to two ring structures and 0-20 heteroatoms, with the proviso that not more than two heteroatoms are linked in sequence,  
 Z is a moiety selected from the group consisting of —CONH—, —NHCO—, —NHCONH—, —NHCSNH—, —OCONH—, —NHOCO—, —S—, —NH(C═NH)—, —N═N—, —NH—, and  
                     
 P is selected from the group consisting of polypeptides, polysaccharides and synthetic polymers, and  
 n is a number from 1 to 50 per 50 kilodaltons molecular weight of P.  
 
   
   
       12 . The compound of  claim 11  wherein P is an aminated dextran.  
   
   
       13 . The compound of  claim 11  wherein P is bovine serum albumin.  
   
   
       14 . The compound of  claim 11  wherein P is keyhole limpet hemocyanin.  
   
   
       15 . The compound of  claim 11  wherein P is Limulus polyphemus hemocyanin.  
   
   
       16 . The compound of  claim 11  wherein P is bovine thyroglobulin.  
   
   
       17 . The compound Oc-(succinimido-oxycarbonyl-butyryl-aminocaproyl)-nelfinavir conjugate with KLH.  
   
   
       18 . The compound Oc-[4′-(succinimido-oxycarbonyl)-benzoyl-aminocaproyl]-nelfinavir conjugate with BSA.  
   
   
       19 . A compound having the structure  
     
       
         
         
             
             
         
       
     
     wherein 
 Y is O, S, or NH,  
 m is 0 or 1,  
 L is a linker comprising 0 to 40 carbon atoms arranged in a straight chain or a branched chain saturated or unsaturated, and further comprising up to two ring structures and 0-20 heteroatoms, with the proviso that not more than two heteroatoms are linked in sequence,  
 Z is a moiety selected from the group consisting of —CONH—, —NHCO—, —NHCONH—,  
 —NHCSNH—, —OCONH—, —NHOCO—, —S—, —NH(C═NH)—, —N═N—, —NH—, and  
                     
 Q is selected from the group consisting of non-isotopic labels and biotin,  
 and n is a number from 1 to 50 per 50 kilodaltons molecular weight of Q.  
 
   
   
       20 . The compound of  claim 19  wherein Q is biotin.  
   
   
       21 . An antibody generated in response to a compound having the structure:  
     
       
         
         
             
             
         
       
     
     wherein 
 Y is O, S, or NH,  
 m is 0 or 1,  
 L is a linker comprising 0 to 40 carbon atoms arranged in a straight chain or a branched chain, saturated or unsaturated, and further comprising up to two ring structures and 0-20 heteroatoms, with the proviso that not more than two heteratoms are linked in sequence,  
 Z is a moiety selected from the group consisting of —CONH—, —NHCO—, —NHCONH—, —NHCSNH—, —OCONH—, —NHCO—, —S—, —NH(C═NH)—, —N═N—, —NH—, and  
                     
 P is selected from the group consisting of polypeptides, a polysaccharides, and synthetic polymers,  
 and n is a number from 1 to 50 kilodaltons molecular weight of P.  
 
   
   
       22 . (canceled)  
   
   
       23 . A monoclonal antibody specific for nelfinavir having less than 10% cross-reactivity with saquinavir, indinavir, amprenavir, ritonavir and lopinavir.  
   
   
       24 . Murine hybridoma NEL 5.4.1 having ATCC No. PTA-4475.