PROJECT SUMMARY / ABSTRACT The HIV-1 (Human Immunodeficiency Virus) is a member of the retroviral family which contains a single-stranded RNA genome and is the major etiological agent involved in the development of acquired immunodeficiency syndrome or AIDS. The WHO now estimates that in 2022 over 40 million people worldwide are infected. Development of antiretroviral therapy (ART) provided much progress over the past several decades. Continual emergence of drug resistance HIV variants and side effects of life-long therapy necessitates the development of new agents and long-acting therapies to increase patient compliance. Developing PrEP prophylactic and combination therapies would be highly beneficial as well as new approaches to reduce HIV reservoirs. Successful drug targets include HIV reverse transcriptase (RT), HIV protease, integration, viral entry, attachment, and capsid. Drugs targeting RT remain a cornerstone of AIDS therapy and are divided into two classes: nucleoside inhibitors (NRTIs) and non-nucleoside inhibitors (NNRTIs). NNRTIs with improved safety, pharmacological, drug resistance profiles, and dosing regimens are still needed. Building on the discovery of potent novel lead compounds, using computational and structure-guided design, the PI and an established set of collaborators, have used lead optimization to develop several new classes of novel NNRTIs. These NNRTIs have excellent potency on WT and drug resistant strains of HIV, optimal pharmacological properties, synergy with clinically relevant HIV drugs, and efficacy in AIDS hu-mouse models including long acting and extended- release formulations. Comprehensive studies are described to further develop these compounds as preclinical candidates as combination therapy and PrEP. We discovered that some of these NNRTIs have potent activity working by a completely different mechanism involving Gag-Pol dimerization, premature HIV protease activation, and CARD8-mediated inflammasome cell-killing of HIV-1 infected cells via pyroptosis. Studies are described to develop this new class of Gag-Pol dimerizer NNRTIs as a complementary strategy to attack HIV reservoirs.