Development of a Novel Screening Strategy Designed to Discover a New Class of HIV Drugs
Authors
Cheng, NancyLee, Sook-Kyung
Donover, P. Scott
Reichman, Mel
Schiffer, Celia A.
Hull-Ryde, Emily A.
Swanstrom, Ronald I.
Janzen, William P.
UMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyDocument Type
Journal ArticlePublication Date
2013-12-04Keywords
BiochemistryBiochemistry, Biophysics, and Structural Biology
Laboratory and Basic Science Research
Medicinal Chemistry and Pharmaceutics
Medicinal-Pharmaceutical Chemistry
Molecular Biology
Pharmacology
Therapeutics
Metadata
Show full item recordAbstract
Current antiretroviral treatments target multiple pathways important for human immunodeficiency virus (HIV) multiplication, including viral entry, synthesis and integration of the DNA provirus, and the processing of viral polyprotein precursors. However, HIV is becoming increasingly resistant to these "combination therapies." Recent findings show that inhibition of HIV Gag protein cleavage into its two structural proteins, matrix (MA) and capsid (CA), has a devastating effect on viral production, revealing a potential new target class for HIV treatment. Unlike the widely used HIV protease inhibitors, this new class of inhibitor would target the substrate, not the protease enzyme itself. This approach offers a distinct advantage in that inhibitors of MA/CA would only need to affect a subset of the Gag molecules to disable viral replication. To discover MA/CA-specific inhibitors, we constructed a modified MA/CA fusion peptide (MA/CADelta) that contains the HIV protease (PR) cleavage site as well as a tetracysteine motif for fluorescent labeling. The HIV PR cleavage of MA/CADelta can then be monitored via fluorescence polarization (FP). We have adapted this FP assay for high-throughput screening and validated it according to industry standards using a 384-well plate format. We have currently tested 24,000 compounds in this assay and here detail the screening methodology and the results of this screening campaign.Source
J Lab Autom. 2013 Dec 4;19(3):297-303. Link to article on publisher's siteDOI
10.1177/2211068213513453Permanent Link to this Item
http://hdl.handle.net/20.500.14038/26071PubMed ID
24305957Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1177/2211068213513453