Human immunodeficiency virus type 1 (HIV-1) protease is essential for viral replication and has proved a very effective target for antiviral drugs to treat AIDS. But, the long term effectiveness of current AIDS therapy is confronted by the major challenge of rapid development of drug-resistant HIV. Multiple mutations accumulate in the protease in response to inhibitor therapy and produce resistance. The overall goal is to develop new antiviral protease inhibitors and therapeutic strategies to overcome the problem of drug- resistance. The overarching hypothesis is that improved knowledge of the molecular mechanisms for HIV resistance to protease inhibitors will aid in development of potent new therapeutic agents to combat drug resistant virus. Our kinetic and structural analysis has shown that protease mutations can produce inhibitor resistance by several mechanisms, including lower affinity for inhibitor due to mutations that alter the inhibitor binding site, and altered protease stability due to mutations that alter the dimer interface. The central design strategy is that inhibitors with improved polar interactions with conserved regions of HIV protease will be potent drugs for resistant HIV. Our analysis of HIV protease-inhibitor structures has demonstrated the importance of the conserved set of hydrogen bond interactions between main chain atoms of peptide analogs and the protease backbone atoms. Our crystallographic analysis shows that the earlier clinical inhibitors have fewer of these polar interactions and high affinity is achieved by van der Waals interactions with protease side chains, leading to sensitivity to mutations in the binding site. The new antiviral inhibitor darunavir (TMC114;UIC-94017) was designed and confirmed to include more hydrogen bonds with protease main chain atoms. Darunavir showed fewer changes than other clinical inhibitors on the structures and activities of mutant proteases, resulting in high potency, excellent resistance profile, and approval for AIDS salvage therapy in June 2006. Our design strategy was further verified by analysis of the antiviral inhibitors GRL-06579A and GRL-98065. The appearance of new resistance mutations, diverse mechanisms of resistance and adverse side effects of drugs necessitate the development of new non-peptide inhibitors to expand the repertoire and potency of antiviral agents for resistant HIV. This research integrates in vitro analysis of the crystal structures and enzymatic activities of HIV protease mutants, chemical synthesis, and antiviral studies in HIV-infected cells to design novel protease inhibitors in the combat against resistant HIV.

Public Health Relevance

The effectiveness of drugs that target HIV-1 protease for HIV/AIDS therapy is greatly reduced by drug resistance due to mutant proteases. New antiviral agents are being developed to overcome drug resistance with the aid of analysis of the structure and activity of mutants of HIV protease.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
Project #
Application #
Study Section
AIDS Discovery and Development of Therapeutics Study Section (ADDT)
Program Officer
Hagan, Ann A
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Georgia State University
Schools of Arts and Sciences
United States
Zip Code
Gerlits, Oksana; Keen, David A; Blakeley, Matthew P et al. (2017) Room Temperature Neutron Crystallography of Drug Resistant HIV-1 Protease Uncovers Limitations of X-ray Structural Analysis at 100 K. J Med Chem 60:2018-2025
Ghosh, Arun K; Sean Fyvie, W; Brindisi, Margherita et al. (2017) Design, synthesis, X-ray studies, and biological evaluation of novel macrocyclic HIV-1 protease inhibitors involving the P1'-P2' ligands. Bioorg Med Chem Lett 27:4925-4931
Ghosh, Arun K; Rao, Kalapala Venkateswara; Nyalapatla, Prasanth R et al. (2017) Design and Development of Highly Potent HIV-1 Protease Inhibitors with a Crown-Like Oxotricyclic Core as the P2-Ligand To Combat Multidrug-Resistant HIV Variants. J Med Chem 60:4267-4278
Ghosh, Arun K; Brindisi, Margherita; Nyalapatla, Prasanth R et al. (2017) Design of novel HIV-1 protease inhibitors incorporating isophthalamide-derived P2-P3 ligands: Synthesis, biological evaluation and X-ray structural studies of inhibitor-HIV-1 protease complex. Bioorg Med Chem 25:5114-5127
Ghosh, Arun K; Osswald, Heather L; Glauninger, Kristof et al. (2016) Probing Lipophilic Adamantyl Group as the P1-Ligand for HIV-1 Protease Inhibitors: Design, Synthesis, Protein X-ray Structural Studies, and Biological Evaluation. J Med Chem 59:6826-37
Gerlits, Oksana; Wymore, Troy; Das, Amit et al. (2016) Long-Range Electrostatics-Induced Two-Proton Transfer Captured by Neutron Crystallography in an Enzyme Catalytic Site. Angew Chem Int Ed Engl 55:4924-7
Park, Joon H; Sayer, Jane M; Aniana, Annie et al. (2016) Binding of Clinical Inhibitors to a Model Precursor of a Rationally Selected Multidrug Resistant HIV-1 Protease Is Significantly Weaker Than That to the Released Mature Enzyme. Biochemistry 55:2390-400
Shen, ChenHsiang; Yu, Xiaxia; Harrison, Robert W et al. (2016) Automated prediction of HIV drug resistance from genotype data. BMC Bioinformatics 17 Suppl 8:278
Agniswamy, Johnson; Louis, John M; Shen, Chen-Hsiang et al. (2015) Substituted Bis-THF Protease Inhibitors with Improved Potency against Highly Resistant Mature HIV-1 Protease PR20. J Med Chem 58:5088-95
Ghosh, Arun K; Yu, Xufen; Osswald, Heather L et al. (2015) Structure-based design of potent HIV-1 protease inhibitors with modified P1-biphenyl ligands: synthesis, biological evaluation, and enzyme-inhibitor X-ray structural studies. J Med Chem 58:5334-43

Showing the most recent 10 out of 80 publications