The human immunodeficiency virus type 1 (HIV-l) protease is an attractive target for antiviral therapy because of its small size (99 residues), its homology to other viral and cellular proteases, in particular the aspartic proteinases, and its presumed essential role in HIV-l replication. This proposal aims at the development of specific HIV protease inhibitors capable of preventing viral replication with the hope that these compounds may prove therapeutically useful. The Group will express HIV-l protease in both bacterial and mammalian systems in order to produce adequate amounts of purified protein for structural and mechanistic studies. The chemical syntheses of HIV-2 protease will provide a source of this enzyme for comparison and the preparation of isotopically labelled analogs for NMR studies. Based on the suggested analogy with aspartic proteinases, a variety of dipeptide analogs of the transition state variety will be incorporated into known substrate sequences for the proteases. The affinities of the observed inhibitors will be optimized by a variation of the Geysen procedure for epitope mapping HIV-l function DNA clones will be constructed with deletions or mutations in the protease region to determine if protease is essential for viral replication and to investigate its role in the virus life style. Inhibitors of the HIV protease will be tested in vivo for their ability to inhibit virus multiplication in a number of different lymphoid and monocytoid cultures. Models of the protease will be built based on the available information from sequence homology, 2-D NMR studies, structure-activity studies, and X-ray studies (presuming sufficient protein can be obtained and crystallized).

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AI027302-03
Application #
3547219
Study Section
Special Emphasis Panel (SRC (34))
Project Start
1988-09-30
Project End
1993-08-31
Budget Start
1990-09-01
Budget End
1991-08-31
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Ho, C M; Marshall, G R (1995) DBMAKER: a set of programs to generate three-dimensional databases based upon user-specified criteria. J Comput Aided Mol Des 9:65-86
Oprea, T I; Waller, C L; Marshall, G R (1994) Three-dimensional quantitative structure-activity relationship of human immunodeficiency virus (I) protease inhibitors. 2. Predictive power using limited exploration of alternate binding modes. J Med Chem 37:2206-15
Spearman, P; Wang, J J; Vander Heyden, N et al. (1994) Identification of human immunodeficiency virus type 1 Gag protein domains essential to membrane binding and particle assembly. J Virol 68:3232-42
Marinier, A; Toth, M V; Houseman, K et al. (1994) HIV-1 protease inhibitors: ketomethylene isosteres with unusually high affinity compared with hydroxyethylene isostere analogs. Bioorg Med Chem 2:919-25
Waller, C L; Oprea, T I; Giolitti, A et al. (1993) Three-dimensional QSAR of human immunodeficiency virus (I) protease inhibitors. 1. A CoMFA study employing experimentally-determined alignment rules. J Med Chem 36:4152-60
Gu, R; Westervelt, P; Ratner, L (1993) Role of HIV-1 envelope V3 loop cleavage in cell tropism. AIDS Res Hum Retroviruses 9:1007-15
Ratner, L; Vander Heyden, N; Garcia, J et al. (1991) Formation of noninfectious HIV-1 virus particles lacking a full-length envelope protein. AIDS Res Hum Retroviruses 7:287-94
Ratner, L; vander Heyden, N; Dedera, D (1991) Inhibition of HIV and SIV infectivity by blockade of alpha-glucosidase activity. Virology 181:180-92
Swain, A L; Miller, M M; Green, J et al. (1990) X-ray crystallographic structure of a complex between a synthetic protease of human immunodeficiency virus 1 and a substrate-based hydroxyethylamine inhibitor. Proc Natl Acad Sci U S A 87:8805-9
Bryant, M; Ratner, L (1990) Myristoylation-dependent replication and assembly of human immunodeficiency virus 1. Proc Natl Acad Sci U S A 87:523-7

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