Camptothecin derivatives as anti-hiv agents and methods of identifying agents that disrupt vif self-association

Abstract

The present invention relates to the use of camptothecin derivatives as anti-HIV agents that disrupt self-association of the viral infectivity factor (Vif) found in HIV and other retroviruses. The present invention also relates to methods of identifying agents that disrupt VIf self-association and methods of using these agents, including methods of treating or preventing HIV infection.

1 . A method for treating or preventing HIV infection or AIDS in a patient, said method comprising:
 administering to a patient in need of such treatment or prevention a therapeutically effective amount of a compound of Formula (I):   
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof, 
         wherein: 
         Q is selected from NH, O, and S; 
         R 20a  and R 20b  are individually selected from hydrogen, hydroxy, and C 1-6 alkyl; 
         R 21  is selected from hydrogen, —NHC(═O)(CH 2 ) p NR 23 R 24 , and —(CH 2 ) p NR 23 R 24 ; 
         p is 0, 1, 2, 3, or 4; 
         R 22  is selected from hydrogen and hydroxyl; 
         R 23  and R 24  are individually selected from hydrogen and C 1-6 alkyl; and 
         R 25  and R 26  are individually selected from hydrogen and —NO 2 . 
       
     
     
         2 - 11 . (canceled) 
     
     
         12 . The method according to  claim 1 , wherein the compound of Formula (I) or pharmaceutically acceptable salt thereof is administered with a pharmaceutically acceptable carrier. 
     
     
         13 . The method according to  claim 1 , further comprising:
 administering a therapeutically effective amount of at least one other agent for treating HIV selected from the group consisting of HIV reverse transcriptase inhibitors, non-nucleoside HIV reverse transcriptase inhibitors, HIV protease inhibitors, HIV fusion inhibitors, HIV attachment inhibitors, CCR5 inhibitors, CXCR4 inhibitors, HIV budding or maturation inhibitors, and HIV integrase inhibitors.   
     
     
         14 . The method according to  claim 17 , wherein said contacting step is effective to inhibit A method for inhibiting infectivity of a lentivirus in the cell. 
     
     
         15 . (canceled) 
     
     
         16 . The method according to  claim 14 , wherein the lentivirus is selected from the group consisting of HIV-1 and HIV-2. 
     
     
         17 . A method for inhibiting Vif self-association in a cell, said method comprising:
 contacting a cell with an inhibitory-effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein said compound of Formula (I) comprises:   
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof, 
         wherein: 
         Q is selected from NH, O, and S; 
         R 20a  and R 20b  are individually selected from hydrogen, hydroxy, and C 1-6 alkyl; 
         R 21  is selected from hydrogen, —NHC(═O)(CH 2 ) p NR 23 R 24 , and —(CH 2 ) p NR 23 R 24    
         p is 0, 1, 2, 3, or 4; 
         R 22  is selected from hydrogen and hydroxyl; 
         R 23  and R 24  are individually selected from hydrogen and C 1-6 alkyl; and 
         R 25  and R 26  are individually selected from hydrogen and —NO 2 . 
       
     
     
         18 . The method according to  claim 17 , wherein said compound of Formula (I) or pharmaceutically acceptable salt thereof is administered with a pharmaceutically acceptable carrier,
 wherein said compound of Formula (I) or pharmaceutically acceptable salt thereof is effective to disrupt or inhibit multimerization of Vif in the cell, thereby inhibiting Vif self-association in the cell.   
     
     
         19 . (canceled) 
     
     
         20 . A method of identifying an agent that disrupts Vif self-association, said method comprising:
 providing a Vif:Vif complex comprising a first Vif protein or fragment associated with a second Vif protein or fragment;   contacting the Vif:Vif complex with a test agent under conditions effective to generate a detectable signal when the Vif:Vif complex is disrupted; and   detecting the detectable signal to determine whether or not the test agent disrupts the Vif:Vif complex,   wherein disruption of the Vif:Vif complex by the test agent identifies an agent that disrupts Vif self-association.   
     
     
         21 . (canceled) 
     
     
         22 . The method according to  claim 20 , wherein the test agent is from a library of small molecule compounds. 
     
     
         23 . The method according to  claim 20 , wherein the contacting step comprises incubating the Vif:Vif complex with one type of test agent or more than one type of test agent. 
     
     
         24 . (canceled) 
     
     
         25 . The method according to  claim 20 , wherein the detectable signal is detected using a detection technique selected from the group consisting of fluorimetry, microscopy, spectrophotometry, computer-aided visualization, and the like, or combinations thereof. 
     
     
         26 . The method according to  claim 25 , wherein the detectable signal is selected from the group consisting of a fluorescent signal, a phosphorescent signal, a luminescent signal, an absorbent signal, and a chromogenic signal. 
     
     
         27 . The method according to  claim 26 , wherein the fluorescent signal is detectable by its fluorescence properties selected from the group consisting of fluorescence resonance energy transfer (FRET), fluorescence emission intensity, and fluorescence lifetime (FL). 
     
     
         28 . The method according to  claim 20 , wherein the Vif:Vif complex is provided with a first detection moiety attached to the first Vif protein or fragment and a second detection moiety attached to the second Vif protein or fragment
 wherein the first detection moiety and the second detection moiety comprise a fluorescence resonance energy transfer (FRET) pair, wherein the first detection moiety is a FRET donor and the second detection moiety is a FRET acceptor.   
     
     
         29 . (canceled) 
     
     
         30 . (canceled) 
     
     
         31 . The method according to  claim 28 , wherein the FRET donor and the FRET acceptor comprise a fluorophore pair selected from the group consisting of EGFP-REACh2, GFP-YFP, EGFP-YFP, EGFP-REACh2, CFP-YFP, CFP-dsRED, BFP-GFP, GFP or YFP-dsRED, Cy3-Cy5, Alexa488-Alexa555, Alexa488-Cy3, FITC-Rhodamine (TRITC), YFP-TRITC or Cy3, and the like. 
     
     
         32 . The method according to  claim 20 , wherein the Vif:Vif complex is provided in a host cell co-transfected with a first plasmid encoding the first Vif protein or fragment and a second plasmid encoding the second Vif protein or fragment. 
     
     
         33 . (canceled) 
     
     
         34 . (canceled) 
     
     
         35 . The method according to  claim 32 , wherein the host cell is stably or transiently co-transfected with the first and second plasmids. 
     
     
         36 - 50 . (canceled) 
     
     
         51 . A method for treating or preventing HIV infection or AIDS in a patient, said method comprising:
 identifying an agent that disrupts Vif self-association by performing the method according to  claim 20 ; and   administering to a patient in need of such treatment or prevention a therapeutically effective amount of the agent.   
     
     
         52 - 55 . (canceled)