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EXECUTIVE SUMMARY
New Variants of Reverse Transcriptase Enzyme from HIV to be used in High-Throughput Protein Crystallography to Enhance Structure Based Drug Design
SUMMARY: Using a systematic protein engineering approach, Professor Eddy Arnold and colleagues in the Department of Chemistry and Chemical Biology at Rutgers University have developed new variants of the reverse transcriptase (RT) enzyme from HIV. These engineered proteins offer opportunities to carry out efficient and expeditious iterative co-crystallization experiments that will facilitate drug design and lead optimization processes for the discovery of novel drugs targeting HIV-1 RT (both polymerase and RNase H inhibitors) for the treatment of HIV infection. Siuta Consulting has been retained by Rutgers to identify partners for this technology. BACKGROUND: Effective treatment of HIV infections is constantly threatened by the emergence of drug-resistant viral strains. New drugs are needed for overcoming the effects of drug-resistance mutations. RT is a bifunctional enzyme with both polymerase and RNase H activities. More than half of the current HIV drugs target RT polymerization but not one targets RNase H. RT remains as an important target for new anti-AIDS drug development and several pharmaceutical and biotech companies are actively engaged in finding RT polymerase/RNase H inhibitors for the treatment of AIDS. TECHNOLOGY: With an aim to facilitate the understanding of the three-dimensional structure of the enzyme RT and its interactions with inhibitors, new variants of HIV-1 RT have been developed for use in protein crystallography. The new RT constructs to are expressed in large quantity and demonstrate both polymerase and RNase H activity. Some of these engineered forms of HIV-1 reverse transcriptase crystallize in highly desirable forms. The crystals diffract X-rays to high resolution (better than 2 Ĺ), thus enabling the determination of high precision structures of RT in enzyme-inhibitor complexes. These high resolution structures of RT are ideal for structure-based design of new RT inhibitors that target polymerase and/or RNase H activities. PUBLICATIONS: The following four publications and meeting presentation are available upon request: K. Das, J. D. Bauman, A. D. Clark, Jr., Y. V. Frenkel, P. J. Lewi, A. J. Shatkin, S. H. Hughes, and E. Arnold, High Resolution Structures of HIV-1 RT/TMC278 Complexes: Strategic Flexibility Explains Potency Against Resistance Mutations, Proc. Natl. Acad. Sci., 105, 1466-1471 (2008) Structures of HIV-1 Reverse Transcriptase Complexed with NNRTI TMC278: Conformational and Positional Adaptability Overcomes Resistance Mutations, Abstract 88, 14th Conference on Retroviruses and Opportunistic Infections (2007) K. Das, P. J. Lewi, S. H. Hughes and E. Arnold, Crystallography and the Design of Anti-AIDS Drugs: Conformational Flexibility and Positional Adaptability are Important in the Design of Non-Nucleoside HIV-1 Reverse Transcriptase Inhibitors, Prog. Biophys. Mol. Biol., 88, 209-231 (2005) P. A. J. Janssen, P. J. Lewi, E. Arnold, F. Daeyaert, M. de Jonge, J. Heeres, L. Koymans, M. Vinkers, J. Guillemont, E. Pasquier, M. Kukla, D. Ludovici, K. Andries, M.-P. de Béthune, R. Pauwels, K. Das, A. D. Clark, Jr., Y. V. Frenkel, S. H. Hughes, B. Medaer, F. De Knaep, H. Bohets, F. De Clerck, A. Lampo, P. Williams and P. Stoffels, In Search of a Novel Anti-HIV Drug: Multidisciplinary Coordination in the Discovery of 4-[[4-[[4-[(1E)-2-2cyanoethenyl]-2,6-dimethylphenyl]amino]-2-pyrimidinyl]amino]-benzonitrile (R278474, rilpivirine), J. Med. Chem., 48, 1901-1909 (2005) K. Das, A. D. Clark, Jr., P. J. Lewi, J. Heeres, M. R. de Jonge, L. M. H. Koymans, H. M. Vinkers, F. Daeyaert, D. W. Ludovici, M. J. Kukla, B. De Corte, R. W. Kavash, C. Y. Ho, H. Ye, M. A. Lichtenstein, K. Andries, R. Pauwels, M.-P. de Béthune, P. L. Boyer, P. Clark, S. H. Hughes, P. A. J. Janssen and E. Arnold, Roles of Conformational and Positional Adaptability in Structure-Based Design of TMC125-R165335 (etravirine) and Related Non-Nucleoside Reverse Transcriptase Inhibitors that are Highly Potent and Effective Against Wild-Type and Drug-Resistant HIV-1 Variants, J. Med. Chem. 47, 2550-2560 (2004) PATENT STATUS: A patent application covering this new technology has recently been filed which claims both composition of matter to the new variants of reverse transcriptase enzyme as well as use of these proteins in structural biology studies. LICENSE TERMS: This research tool is available for licensing. |