Inhibitors of Nucleotidyltransferase Superfamily Enzymes Suppress Herpes Simplex Virus Replication

Tavis, John E.; Wang, Hong; Tollefson, Ann E.; Bai, Ying; Korom, Maria; Cheng, Xiaohong; Cao, Feng; Davis, Katie L.; Wold, William S.M.; Morrison, Lynda A.

doi:10.1128/aac.03875-14

Cited by 42 publications

(39 citation statements)

References 73 publications

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“…10 On the basis of our interest in developing inhibitors directed toward a broader class of viral nucleotidyltransferases, 26 the 21 novel β -thujaplicinol derivatives of Figure 4, substituted on their heptatriene ring while the metal binding pharmacophore is preserved, were synthesized. For comparative purposes, we initially monitored pUL15C inhibition by a subset of these compounds via agarose gel electrophoresis, the standard method of choice.…”

Section: Resultsmentioning

confidence: 99%

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Characterization of the C-Terminal Nuclease Domain of Herpes Simplex Virus pUL15 as a Target of Nucleotidyltransferase Inhibitors

et al. 2016

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The natural product α-hydroxytropolones manicol and β-thujaplicinol inhibit replication of herpes simplex viruses 1 and 2 (HSV-1 and HSV-2, respectively) at nontoxic concentrations. Because these were originally developed as divalent metal-sequestering inhibitors of the ribonuclease H activity of HIV-1 reverse transcriptase, α-hydroxytropolones likely target related HSV proteins of the nucleotidyltransferase (NTase) superfamily, which share an “RNase H-like” fold. One potential candidate is pUL15, a component of the viral terminase molecular motor complex, whose C-terminal nuclease domain, pUL15C, has recently been crystallized. Crystallography also provided a working model for DNA occupancy of the nuclease active site, suggesting potential protein–nucleic acid contacts over a region of ∼14 bp. In this work, we extend crystallographic analysis by examining pUL15C-mediated hydrolysis of short, closely related DNA duplexes. In addition to defining a minimal substrate length, this strategy facilitated construction of a dual-probe fluorescence assay for rapid kinetic analysis of wild-type and mutant nucleases. On the basis of its proposed role in binding the phosphate backbone, studies with pUL15C variant Lys700Ala showed that this mutation affected neither binding of duplex DNA nor binding of small molecule to the active site but caused a 17-fold reduction in the turnover rate (kcat), possibly by slowing conversion of the enzyme–substrate complex to the enzyme–product complex and/or inhibiting dissociation from the hydrolysis product. Finally, with a view of pUL15-associated nuclease activity as an antiviral target, the dual-probe fluorescence assay, in combination with differential scanning fluorimetry, was used to demonstrate inhibition by several classes of small molecules that target divalent metal at the active site.

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Section: Resultsmentioning

confidence: 99%

“…10 Although the viral target was not established, α -hydroxytropolones were previously shown to potently inhibit ribonuclease H (RNase H) activity of HIV-1 reverse transcriptase (RT) by sequestering the catalytically critical divalent metal at the active site. 11–13 Thus, a structurally related HSV enzyme seems to be a plausible HSV target.…”

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confidence: 99%

Characterization of the C-Terminal Nuclease Domain of Herpes Simplex Virus pUL15 as a Target of Nucleotidyltransferase Inhibitors

et al. 2016

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“…αHTs were previously documented to inhibit HSV-1 and −2 replication in cell culture and postulated targets included viral proteins with an RNase H-like domain (Ireland et al, 2016; Tavis et al, 2014). In HSV-1, these targets include DNA polymerase (pU L 30), single-stranded DNA binding protein (ICP8, pU L 29), alkaline nuclease (pU L 12) and the nuclease domain of the viral terminase (pU L 15C) (Boehmer and Lehman, 1997; Bryant et al, 2012; Liu et al, 2006; Schumacher et al, 2012; Selvarajan Sigamani et al, 2013; Yan et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…In HSV-1, these enzymes include the single stranded DNA binding protein (ICP8), alkaline nuclease (pU L 12), DNA polymerase, and the nuclease of the viral terminase (pU L 15C) (Boehmer and Lehman, 1997; Bryant et al, 2012; Liu et al, 2006; Schumacher et al, 2012; Selvarajan Sigamani et al, 2013; Yan et al, 2014). Given the enzymes’ similar structure, inhibitors of the HIV RNase and integrase were recently screened for activity against HSV-1 and HSV-2 (Tavis et al, 2014). Tropolones, most notably natural and synthetic α-hydroxytropolones (αHTs), were found to be potent anti-HSV inhibitors (Ireland et al, 2016; Tavis et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

“…Given the enzymes’ similar structure, inhibitors of the HIV RNase and integrase were recently screened for activity against HSV-1 and HSV-2 (Tavis et al, 2014). Tropolones, most notably natural and synthetic α-hydroxytropolones (αHTs), were found to be potent anti-HSV inhibitors (Ireland et al, 2016; Tavis et al, 2014). While tropolones are well known metalloenzyme inhibitors due to the high negative charge character on both the carbonyl and adjacent oxygen at physiological pH, αHTs provide an additional contiguous oxygen that makes them particularly good inhibitors of dinuclear metalloenzymes (Bentley, 2008; Piettre et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Broad anti-herpesviral activity of α-hydroxytropolones

Dehghanpir

Birkenheuer

Yang

et al. 2018

Veterinary Microbiology

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Herpesviruses are ubiquitous in animals and cause economic losses concomitant with many diseases. Most of the domestic animal herpesviruses are within the subfamily Alphaherpesvirinae, which includes human herpes simplex virus 1 (HSV-1). Suppression of HSV-1 replication has been reported with α-hydroxytropolones (αHTs), aromatic ring compounds that have broad bioactivity due to potent chelating activity. It is postulated that αHTs inhibit enzymes within the nucleotidyltransferase superfamily (NTS). These enzymes require divalent cations for nucleic acid cleavage activity. Potential targets include the nuclease component of the herpesvirus terminase (pUL15C), a highly conserved NTS-like enzyme that cleaves viral DNA into genomic lengths prior to packaging into capsids. Inhibition of pUL15C activity in biochemical assays by various αHTs previously revealed a spectrum of potencies. Interestingly, the most potent anti-pUL15C αHT inhibited HSV-1 replication to a limited extent in cell culture. The aim of this study was to evaluate three different αHT molecules with varying biochemical anti-pUL15C activity for a capacity to inhibit replication of veterinary herpesviruses (BoHV-1, EHV-1, and FHV-1) and HSV-1. Given the known discordant potencies between anti-pUL15C and HSV-1 replication inhibition, a second objective was to elucidate the mechanism of action of these compounds. The results show that αHTs broadly inhibit herpesviruses, with similar inhibitory effect against HSV-1, BoHV-1, EHV-1, and FHV-1. Based on immunoblotting, Southern blotting, and real-time qPCR, the compounds were found to specifically inhibit viral DNA replication. Thus, αHTs represent a new class of broadly active anti-herpesviral compounds with potential veterinary applications.

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Synthesis of Polyoxygenated Tropolones and their Antiviral Activity against Hepatitis B Virus and Herpes Simplex Virus‐1

Schiavone

Kapkayeva

et al. 2022

Chemistry A European J

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Polyoxygenated tropolones possess a broad range of biological activity, and as a result are promising lead structures or fragments for drug development. However, structure–function studies and subsequent optimization have been challenging, in part due to the limited number of readily available tropolones and the obstacles to their synthesis. Oxidopyrylium [5+2] cycloaddition can effectively generate a diverse array of seven‐membered ring carbocycles, and as a result can provide a highly general strategy for tropolone synthesis. Here, we describe the use of 3‐hydroxy‐4‐pyrone‐based oxidopyrylium cycloaddition chemistry in the synthesis of functionalized 3,7‐dimethoxytropolones, 3,7‐dihydroxytropolones, and isomeric 3‐hydroxy‐7‐methoxytropolones through complementary benzyl alcohol‐incorporating procedures. The antiviral activity of these molecules against herpes simplex virus‐1 and hepatitis B virus is also described, highlighting the value of this approach and providing new structure–function insights relevant to their antiviral activity.

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Inhibitors of Nucleotidyltransferase Superfamily Enzymes Suppress Herpes Simplex Virus Replication

Cited by 42 publications

References 73 publications

Characterization of the C-Terminal Nuclease Domain of Herpes Simplex Virus pUL15 as a Target of Nucleotidyltransferase Inhibitors

Characterization of the C-Terminal Nuclease Domain of Herpes Simplex Virus pUL15 as a Target of Nucleotidyltransferase Inhibitors

Broad anti-herpesviral activity of α-hydroxytropolones

Synthesis of Polyoxygenated Tropolones and their Antiviral Activity against Hepatitis B Virus and Herpes Simplex Virus‐1

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