Highly active antiretroviral therapy (HAART) drugs primarily target three of the four HIV enzymes: reverse transcriptase (RT) DNA polymerase, protease, and integrase. Although HAART is very effective in suppressing viral load in HIV-infected patients, prolonged treatment inevitably leads to the emergence of drug-resistant viral strains. Hence, it is essential to develop agents that act on novel HIV targets. The fourth HIV enzyme, RT- associated ribonuclease H (RNH) is one such target. RNH degrades the viral RNA genome during reverse transcription and is essential for HIV replication. It is the only enzymatic activity of HIV that has yet to be addressed by antiretroviral drugs. Such drugs will likely be active against all current drug-resistant viral strains. Our goal is to develop potent RNH inhibitors (RNHIs) with nanomolar efficacy in cell-based replication assays and for this we will use a multidisciplinary approach based on our extensive expertise in structural biology, computational biology, medicinal chemistry, enzymology, biochemistry and virology. We will pursue improvement of current leads and achievement of nM potencies of antiviral activities through a structure-based design process involving iterative cycles of structure determination and computational analysis of RNH-RNHI complexes, medicinal chemistry, and biochemical and virological characterization of newly synthesized inhibitors. To this end we propose three specific aims:
Specific Aim 1. Structure-Activity Relationships (SAR) and Chemical Synthesis. We will prepare a database of the validated RNHI screening hits and novel scaffolds that we have developed. We will perform complete SAR for two different classes of RNHIs based on the database analysis and the structural information gained from crystallographic and molecular docking studies in aim 2.
Specific Aim 2. Crystallographic and computational analysis of RT-RNHI interaction. Structure-based design is a main focus of this application. We will use crystallographic tools that are already established in our lab and that routinely result in high resolution structures of RT and/or RNH in complex with inhibitors (resolutions up to 1.5 ?). The structural information will be used to guide the design of new inhibitors.
Specific Aim 3. Biochemical and virologic profiling of RNHIs. We will use biochemical and virological assays to assess selected validated screening hits and new RNHIs to be prepared in Aim 1. This information will be integrated in the iterative SAR-mediated design of new inhibitors. Our multidisciplinary approach will lead to new inhibitors of HIV that will be effective against both wild- type and drug-resistant viral strains.

Public Health Relevance

This project will discover compounds that target an essential biological activity of the HIV virus, that currently approved drugs do not address. These studies should lead to the development of future therapeutics that complement existing drugs and should also strengthen our efforts to block drug- resistant strains of HIV.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI100890-03
Application #
8680130
Study Section
Special Emphasis Panel (ZRG1-AARR-E (04))
Program Officer
Turk, Steven R
Project Start
2012-07-01
Project End
2017-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
3
Fiscal Year
2014
Total Cost
$748,607
Indirect Cost
$103,140
Name
University of Missouri-Columbia
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
153890272
City
Columbia
State
MO
Country
United States
Zip Code
65211
Puray-Chavez, Maritza; Tedbury, Philip R; Huber, Andrew D et al. (2017) Multiplex single-cell visualization of nucleic acids and protein during HIV infection. Nat Commun 8:1882
Vernekar, Sanjeev Kumar V; Tang, Jing; Wu, Bulan et al. (2017) Double-Winged 3-Hydroxypyrimidine-2,4-diones: Potent and Selective Inhibition against HIV-1 RNase H with Significant Antiviral Activity. J Med Chem 60:5045-5056
Tang, Jing; Kirby, Karen A; Huber, Andrew D et al. (2017) 6-Cyclohexylmethyl-3-hydroxypyrimidine-2,4-dione as an inhibitor scaffold of HIV reverase transcriptase: Impacts of the 3-OH on inhibiting RNase H and polymerase. Eur J Med Chem 128:168-179
Huber, Andrew D; Michailidis, Eleftherios; Tang, Jing et al. (2017) 3-Hydroxypyrimidine-2,4-Diones as Novel Hepatitis B Virus Antivirals Targeting the Viral Ribonuclease H. Antimicrob Agents Chemother 61:
Kankanala, Jayakanth; Kirby, Karen A; Huber, Andrew D et al. (2017) Design, synthesis and biological evaluations of N-Hydroxy thienopyrimidine-2,4-diones as inhibitors of HIV reverse transcriptase-associated RNase H. Eur J Med Chem 141:149-161
Tang, Jing; Vernekar, Sanjeev Kumar V; Chen, Yue-Lei et al. (2017) Synthesis, biological evaluation and molecular modeling of 2-Hydroxyisoquinoline-1,3-dione analogues as inhibitors of HIV reverse transcriptase associated ribonuclease H and polymerase. Eur J Med Chem 133:85-96
Kirby, Karen A; Myshakina, Nataliya A; Christen, Martin T et al. (2017) A 2-Hydroxyisoquinoline-1,3-Dione Active-Site RNase H Inhibitor Binds in Multiple Modes to HIV-1 Reverse Transcriptase. Antimicrob Agents Chemother 61:
Wu, Bulan; Tang, Jing; Wilson, Daniel J et al. (2016) 3-Hydroxypyrimidine-2,4-dione-5-N-benzylcarboxamides Potently Inhibit HIV-1 Integrase and RNase H. J Med Chem 59:6136-48
Kankanala, Jayakanth; Kirby, Karen A; Liu, Feng et al. (2016) Design, Synthesis, and Biological Evaluations of Hydroxypyridonecarboxylic Acids as Inhibitors of HIV Reverse Transcriptase Associated RNase H. J Med Chem 59:5051-62
Tang, Jing; Liu, Feng; Nagy, Eva et al. (2016) 3-Hydroxypyrimidine-2,4-diones as Selective Active Site Inhibitors of HIV Reverse Transcriptase-Associated RNase H: Design, Synthesis, and Biochemical Evaluations. J Med Chem 59:2648-59

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