The long-term goal of our program is to advance the knowledge of RNA-ligand interactions and to apply the principles uncovered to the design, synthesis and discovery of new antiretroviral agents. The guiding hypothesis is that small molecules that target viral-specific RNA sites and prevent the formation of key RNA-protein and RNA-RNA complexes are promising candidates for antiretroviral drug discovery.
The specific aims of this proposal are: (1) Develop rapid assays for the discovery of potent and specific viral RNA binders. To complement our Rev-RRE assay, we will develop a rapid assay for identifying high affinity and selective Tat-TAR inhibitors. We will also develop a fluorescence-based assay for the discovery of HIV DIS binders. (2) Design and synthesize RNA-selective ligands. We will conjugate """"""""RNA-friendly"""""""" scaffolds (e.g., functionalized beta-and gamma-peptides, conformationally-constrained amino- and guanidine-glycosides) to sequence- and structure-selective elements (e.g., nucleobase analogs, RNA-selective intercalators, cis-Pt) to provide new families of potent and selective viral RNA binders. (3) Determine affinity and selectivity to HIV RNA sites. We will assess the Rev-RRE and Tat-TAR inhibitory activity as well as DIS affinity of all binders using the novel assays. (4) Evaluate anti HIV activity. Most promising binders will be evaluated for: (a) their ability to interfere with viral replication in HIV-1 infected cells, (b) their toxicity, and (c) their potential site(s) of interference. The rapid emergence of resistant viral strains necessitates the development of novel strategies for anti-HIV drug design. Small molecules that target pivotal viral RNA sites, and can specifically prevent the formation of key regulatory RNA-protein and RNA-RNA complexes, are proposed as promising candidates for antiretroviral drug development. Our integrated approach brings together organic synthesis, chemical diversity, novel solution and solid-phase assays, systematic structure-activity relationship studies, as well as in vivo anti-HIV tests, toxicity evaluation and mechanistic studies. It will yield new families of effective and specific binders that target key HIV-1 RNA sites. Potent and specific effectors of essential regulatory events will significantly advance the development of highly active antiretroviral agents.
| Belousoff, Matthew J; Ung, Phuc; Forsyth, Craig M et al. (2009) New macrocyclic terbium(III) complex for use in RNA footprinting experiments. J Am Chem Soc 131:1106-14 |
| Staple, David W; Venditti, Vincenzo; Niccolai, Neri et al. (2008) Guanidinoneomycin B recognition of an HIV-1 RNA helix. Chembiochem 9:93-102 |
| Srivatsan, Seergazhi G; Tor, Yitzhak (2007) Fluorescent pyrimidine ribonucleotide: synthesis, enzymatic incorporation, and utilization. J Am Chem Soc 129:2044-53 |
| Tam, Victor K; Kwong, Denise; Tor, Yitzhak (2007) Fluorescent HIV-1 Dimerization Initiation Site: design, properties, and use for ligand discovery. J Am Chem Soc 129:3257-66 |
| Srivatsan, Seergazhi G; Tor, Yitzhak (2007) Using an emissive uridine analogue for assembling fluorescent HIV-1 TAR constructs. Tetrahedron 63:3601-3607 |
