SBIR-STTR Award

This project aims to develop and test a rapid and sensitive screen for potential anti-nucleocapsid agents. The nucleocapsid (NC) and NC-containing precursor proteins from HIV-1 are targeted for drug discovery. NC is responsible for recognizing and packaging genomic RNA into new virus particles. In addition, the protein participates in other important viral processes, so anti-NC drugs could interfere at several stages of the life cycle of the virus. The screening procedure is a novel application of fluorescence technology, that promises very high sensitivity and selectivity for anti-NC agents. It is capable of being easily scaled for high-throughput screening. The method can also provide an accurate determination of the equilibrium dissociation constants for NC-competitor complexes. In addition, the procedures for developing the fluorescent probes can be simply extended to additional targets in the HIV-1 system, and other pathogens.

Thesaurus Terms:
antiviral agent, drug design /synthesis /production, fluorescent dye /probe, high throughput technology, nucleocapsid, technology /technique development RNA, intermolecular interaction, virus RNA

This project aims at creating and using the tools to screen large libraries of chemical compounds to find and evaluate drug candidates for three targets in HIV-1. These are (1) the RNAnucleocapsid (NC) protein complex, (2) the TAR RNA-Tat protein-CyclinT1 protein complex, and (3) the Vif protein. No inhibitors of these targets are available in current AIDS therapies, in phase I of this project we succeeded in creating a molecular switch that changes its state in the presence of NC, and switches back in the presence of an inhibitor that mimics the viral RNA packaging signal. The inhibitor causes an increase in fluorescence within seconds that is easily monitored in high throughput screens (HTS) of candidate drugs. It is easy and unambiguous to verify that the complex occurs with a 1:1 ratio of protein and switch components (or not), and to determine the affinity constant, K/d. We propose limited prototyping and to create HTS- and Kdkits that others can use to discover and refine drug candidates to interrupt NC, Tat, and Vif function in HIV-I. In the future, similar assays will target all fifteen HIV proteins with new drugs.

Thesaurus Terms:
antiviral agent, drug design /synthesis /production, drug screening /evaluation, fluorescent dye /probe, high throughput technology, nucleocapsid AIDS therapy, chemical registry /resource, human immunodeficiency virus 1, protein purification, stoichiometry, virus protein