The filoviruses Ebola (EBOV) and Marburg (MARV) cause the most severe hemorrhagic fever known, have no approved vaccines or treatments, are highly contagious and infectious In an aerosolized form, and can be used as weapons in acts of bioterrorism and biological warfare. The viruses have the RNA-dependent polymerase, which mediates both the transcription and replication of the genome. The EBOV polymerase complex consists of three subunits common for all Mononegavirales: nucleoprotein (N), phosphoprotein (VP35), and the large subunit of polymerase (L). In addition, the polymerase complex includes a filovirus-specific VP30, which is critical for the transcriptional, but not replication activity of EBOV polymerase, and can only support transcription In a dephosphorylated form. Our preliminary studies show that protein phosphatase 1 (PP1) dephosphorylates VP30 in cultured cells. Furthermore, PP1-targeted small molecule compound 1E7-03 that was developed through rounds of computational modeling followed by screening induced phosphorylation of VP30 and inhibited transcription of EBOV genes and replication of the virus. The 1E7-03 compound targeted a non-catalytic site of PP1 and had no toxicity or produced significant changes in cellular proteome. The compound inhibited the transcription but not replication of the viral genome, and completely suppressed replication of EBOV. Our findings suggest that targeting PP1 is a feasible approach for development of a drug against filoviruses. Furthermore, a remarkable similarity in the phosphorylation sites at the N-termini of the VP30 proteins of all species of EBOV and MARV suggests that this PP1 targeting approach can be used for developing a pan-filovirus drug. The proposed study is aimed at the detailed testing and optimization of the 1E7-03 compound as a drug against EBOV and MARV. The following Specific Aims will be pursued: 1) Continue optimization of the molecules for the inhibition of EBOV and MARV replication by using a medicinal chemistry approach; 2) Characterize the ability of the most potent small molecules to inhibit replication of EBOV and MARV in vitro and in vivo; 3) Non-clinical toxicology and PK research and development. The approach is innovative, because the developed compound targets PP1 without affecting Its general enzymatic activity, and this approach has never previously been used for any virus, with the exception of the Pi's recent experiments with EBOV and HIV-1. The proposed research is significant because It may open a new therapeutic avenue against multiple filovirus species.

Public Health Relevance

The proposed research is relevant to public health because It Is aimed at development of a drug which will target the interaction of a viral protein with a host enzyme and which is likely to be active against all species of filoviruses. The proposed research Is relevant to the NIH mission that pertains to a reduced vulnerability of the population to biothreat agents. Including hemorrhagic fever viruses.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI109664-06
Application #
9649156
Study Section
Special Emphasis Panel (ZAI1)
Program Officer
Maric, Maja
Project Start
Project End
2020-02-29
Budget Start
2018-03-01
Budget End
2019-02-28
Support Year
6
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Georgia State University
Department
Type
DUNS #
837322494
City
Atlanta
State
GA
Country
United States
Zip Code
30302
Dashti, Hesam; Wedell, Jonathan R; Westler, William M et al. (2018) Applications of Parametrized NMR Spin Systems of Small Molecules. Anal Chem 90:10646-10649
Fink, Susan L; Vojtech, Lucia; Wagoner, Jessica et al. (2018) The Antiviral Drug Arbidol Inhibits Zika Virus. Sci Rep 8:8989
Tigabu, Bersabeh; Ramanathan, Palaniappan; Ivanov, Andrey et al. (2018) PHOSPHORYLATED VP30 OF MARBURG VIRUS IS A REPRESSOR OF TRANSCRIPTION. J Virol :
Su, Zhaoming; Wu, Chao; Shi, Liuqing et al. (2018) Electron Cryo-microscopy Structure of Ebola Virus Nucleoprotein Reveals a Mechanism for Nucleocapsid-like Assembly. Cell 172:966-978.e12
Luthra, Priya; Liang, Jue; Pietzsch, Colette A et al. (2018) A high throughput screen identifies benzoquinoline compounds as inhibitors of Ebola virus replication. Antiviral Res 150:193-201
Knoverek, Catherine R; Amarasinghe, Gaya K; Bowman, Gregory R (2018) Advanced Methods for Accessing Protein Shape-Shifting Present New Therapeutic Opportunities. Trends Biochem Sci :
Ivanov, Andrey; Lin, Xionghao; Ammosova, Tatiana et al. (2018) HIV-1 Tat phosphorylation on Ser-16 residue modulates HIV-1 transcription. Retrovirology 15:39
Chanthamontri, C Ken; Jordan, David; Wang, Wenjie et al. (2018) Ebola Viral Protein 35 N-terminus is a Parallel Tetramer. Biochemistry :
Lin, Xionghao; Kumari, Namita; DeMarino, Catherine et al. (2017) Inhibition of HIV-1 infection in humanized mice and metabolic stability of protein phosphatase-1-targeting small molecule 1E7-03. Oncotarget 8:76749-76769
Postler, Thomas S; Clawson, Anna N; Amarasinghe, Gaya K et al. (2017) Possibility and Challenges of Conversion of Current Virus Species Names to Linnaean Binomials. Syst Biol 66:463-473

Showing the most recent 10 out of 47 publications