Discovery and analysis of a series of C2-symmetric HIV-1 proteinase inhibitors derived from penicillin.
In order to identify a suitable peptide substrate for human immunodeficiency virus-1 (HIV-1) proteinase, a range of peptides from various cleavage sites within the gag-pol polyprotein were assayed by HPLC for specific cleavage. The peptide with the optimal combination of favorable kinetics and good solubility was based on the N-terminus cleavage site of HIV-1 proteinase (KQGTVSFNF*PQIT). The HPLC assay, using the above peptide, was developed into a rapid isocratic method in order to analyze inhibition kinetics. An assay suitable for high-throughput screening was developed using a radioactively labeled peptide with the same sequence, coupled to a solid phase. Using this assay, a C2-symmetric HIV-1 proteinase inhibitor derived from penicillin was discovered during random screening of a compound library. A chemical synthesis program developed this structure into a series of potent inhibitors. The lead structures were highly selective for HIV-1 proteinase with good antiviral activity in vitro against HIV and no cytotoxicity. The HPLC assay was used to demonstrate that these compounds are competitive tight-binding inhibitors of HIV-1 proteinase.
Gray
N M
NM
Department of Virology, Glaxo Group Research Ltd., Greenford, Middlesex, United Kingdom.
Cameron
J M
JM
Cammack
N
N
Cobley
K N
KN
Holmes
D S
DS
Humber
D C
DC
Orr
D C
DC
Penn
C R
CR
Potter
R
R
Madar
S
S
D000595:Amino Acid Sequence; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D017320:HIV Protease Inhibitors / Q000032:analysis* / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D010406:Penicillins / Q000032:analysis* / Q000494:pharmacology
D017320:HIV Protease Inhibitors; D010406:Penicillins