Inhibition of Hiv-1 Replication by Disruption of the Processing of the Viral Capsid-Spacer Peptide 1 Protein

Abstract

Inhibition of HIV-1 replication by disrupting the processing of the viral Gag capsid (CA) protein (p24) from the CA-spacer peptide 1 (SP1) protein precursor (p25) is disclosed. Amino acid sequences containing a mutation in the Gag p25 protein, with the mutation resulting in a decrease in the inhibition of processing of p25 to p24 by dimethylsuccinyl betulinic acid or dimethylsuccinyl betulin, polynucleotides encoding such mutated sequences and antibodies that selectively bind such mutated sequences are also included. Methods of inhibiting, inhibitory compounds and methods of discovering inhibitory compounds that target proteolytic processing of the HIV Gag protein are included. In one embodiment, such compounds inhibit the interaction of the HIV protease enzyme with Gag by binding to Gag rather than to the protease enzyme. In another embodiment, viruses or recombinant proteins that contain mutations in the region of the Gag proteolytic cleavage site can be used in screening assays to identify compounds that target proteolytic processing.

1 - 5 . (canceled) 
     
     
         6 . A method of treating HIV-1-infection in a patient, comprising administering a compound that inhibits processing of the viral Gag p25 protein (CA-SP1) to p24 (CA), wherein said compound binds to a polypeptide encoded by a polynucleotide having a sequence at least about 70% identical to a sequence selected from the group consisting of:
 (a) about nucleotides 1243-1435 of SEQ ID NO: 18;   (b) about nucleotides 1729-1920 of SEQ ID NO: 19;   (c) about nucleotides 1344-1435 of SEQ ID NO: 18;   (d) about nucleotides 1828-1920 of SEQ ID NO: 19;   (e) about nucleotides 1370-1413 of SEQ ID NO: 18;   (f) about nucleotides 1857-1899 of SEQ ID NO: 19   (g) about nucleotides 1372-1419 of SEQ ID NO: 18;   (h) about nucleotides 1858-1905 of SEQ ID NO: 19;   (i) about nucleotides 1372-1434 of SEQ ID NO: 18; and   (j) about nucleotides 1858-1920 of SEQ ID NO: 19.   
     
     
         7 - 52 . (canceled) 
     
     
         53 . A recombinant non-HIV-1 retrovirus the replication of which is inhibited by 3-O-(3′,3′-dimethylsuccinyl)betulinic acid (DSB). 
     
     
         54 - 64 . (canceled) 
     
     
         65 . An animal model of lentivirus infection comprising a suitable non-human animal host infected with the recombinant non-HIV-1 retrovirus of  claim 53 . 
     
     
         66 - 89 . (canceled) 
     
     
         90 . An isolated polypeptide from the HIV CA-SP1 polypeptide which contains a mutation that results in a decrease in inhibition of processing of p25 by 3-O-(3′,3′-dimethylsuccinyl)betulinic acid. 
     
     
         91 - 149 . (canceled)