(Change of Scope) Modified Project Summary (Change of Scope) U01 A1082015-O1 """"""""Broad Spectrum Therapeutics Targeting UO1 AI082015-01 """"""""Broad Spectrum Therapeutics Targeting Resolvase Enzymes"""""""" For applications in biodefense, it is desirable for small molecule inhibitors to biodefense, target multiple category A-C agents because it is difficult and expensive to develop even one small molecule inhibitor. We propose to develop inhibitors of molecule inhibitor. We propose Holliday junction resolvase enzymes, which are found in several category A-C Initial studies focused on resolvase enzymes agents. Initial studies focused on resolvase enzymes found in poxviruses. These enzymes catalyze a required replication step in which concatemers of the viral genomic DNA are cleaved into unit length genomes for packaging. The cleaved into unit length genomes for packaging. enzyme is also important in pathogenic fungi such as Coccidioldes, the causative Coccidioides, Fever, where agent of Valley Fever, where it is involved in mitochondrial DNA replication. In involved in mitochondrial DNA replication. previous work we developed a high throughput assay and screened >133,000 Our best compound so far has an IC50 small molecules for inhibitory activity. Our best compound so far has an 1C50 against purified resolvase of -100 nM, 1C50 against virus of 3 uM. We have -100 nM, IC50 against virus of 3 uM. revised our Aims in accordance with the reviewers comments, and to maximize our progress over the projected two years (instead of five years) of funding. We projected two years (instead of five years) of funding. will emphasize new compound identification, iterative compound synthesis, and increasingly stringent assays to develop inhibitors of poxvirus resolvases that are active in animal models. Inhibitors active against poxviruses in vivo will also be models. Inhibitors active against poxviruses tested in pathogenic fungi in an effort to develop """"""""dual use"""""""" pharmacotherapy. pathogelllic Funding of the project will generate jobs for more than five people (5.45 FTEs summed over all the participating groups). and two pieces of equipment will be purchased (a fluorescence plate reader and a -80?C freezer) from American sources. advancing the goals of the ARRA. sources, thereby advancing the goals of the ARRA.
The United States of America is at risk of terrorist attacks using poxviruses or using pathogenic fungi. Both are also current medical problems. We propose to fungi. Both are medical problems. develop improved small molecule therapy targeting the Holliday junction resolving enzymes encoded by these pathogens as new treatments for infection.
| Li, Huiguang; Hwang, Young; Perry, Kay et al. (2016) Structure and Metal Binding Properties of a Poxvirus Resolvase. J Biol Chem 291:11094-104 |
| Culyba, Matthew; Hwang, Young; Attar, Sana et al. (2012) Bulged DNA substrates for identifying poxvirus resolvase inhibitors. Nucleic Acids Res 40:e124 |
