Potent and selective inhibition of SARS coronavirus replication by aurintricarboxylic acid

He, Runtao; Adonov, Anton; Traykova-Adonova, Maya; Cao, Jingxin; Cutts, Todd; Grudesky, Elsie; Deschambaul, Yvon; Berry, Jody D.; Drebot, Michael A.; Li, Xuguang

doi:10.1016/j.bbrc.2004.06.076

Cited by 77 publications

(92 citation statements)

References 22 publications

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“…In addition, ATA has also been reported to inhibit protein synthesis by blocking mRNA association with ribosomes [22], disrupt cellular signalling pathways by diverse mechanisms [23][24][25][26] and promote cell survival in response to variety of insults in various cell types [23,26,27]. The therapeutic potential of ATA is highlighted by its powerful antiviral activity against HIV [18,21], vesicular stomatitis virus [28], severe acute respiratory syndrome coronavirus (SARS-CoV) [29], Rauscher leukaemia virus [18,21] and vaccinia virus [30].…”

mentioning

confidence: 99%

Characterization of Aurintricarboxylic Acid as a Potent Hepatitis C Virus Replicase Inhibitor

Chen

Waffo

Basu

et al. 2009

Antivir Chem Chemother

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Background: Hepatitis C virus (HCV) NS5B is an essential component of the viral replication machinery and an important target for antiviral intervention. Aurintricarboxylic acid (ATA), a broad-spectrum antiviral agent, was evaluated and characterized for its anti-NS5B activity in vitro and in HCV replicon cells. Methods: Recombinant NS5B, HCV replicase and Huh-7 cells harbouring the subgenomic HCV replicon of genotype 1b were employed for biochemical and mechanistic investigations. Results: Analysis of ATA activity in vitro yielded equipotent inhibition of recombinant NS5B and HCV replicase in the submicromolar range (50% inhibition concentration [IC 50 ] approximately 150 nM). Biochemical and mechanistic studies revealed a bimodal mechanism of ATA inhibition with characteristics of pyrophosphate mimics and non-nucleoside inhibitors. Molecular modelling and competition displacement studies were consistent with these parameters, suggesting that ATA might bind to the benzothiadiazine allosteric pocket 3 of NS5B or at its catalytic centre. Kinetic studies revealed a mixed mode of ATA inhibition with respect to both RNA and UTP substrates. Under single-cycle assay conditions, ATA inhibited HCV NS5B initiation and elongation from pre-bound RNA, but with ≥fivefold decreased potency compared with continuous polymerization conditions. The IC 50 value of ATA for the native replicase complex was 145 nM. In HCV replicon cells, ATA treatment ablated HCV RNA replication (50% effective concentration =75 nM) with concomitant decrease in NS5B expression and no apparent cytotoxic effects. Conclusions: This study identified ATA as a potent anti-NS5B inhibitor and suggests that its unique mode of action might be exploited for structural refinement and development of novel anti-NS5B agents.Hepatitis C virus (HCV) is a significant human bloodborne pathogen affecting an estimated 3% of the world's population, of whom 80% progress to chronic infection [1,2]. Sequelae include fibrosis, cirrhosis and hepatocellular carcinoma, making HCV the leading cause of liver transplantation in the USA [2,3]. The currently approved anti-HCV therapy of pegylated interferon-α (PEG-IFN-α) in combination with ribavirin exhibit sustained virological response (SVR) rates of 40-50% for genotype 1 and ≤80% for genotypes 2 and 3, and are associated with severe side effects resulting in limited patient compliance [4]. Additional challenges are posed by the high cost of therapy and the emergence of HCV quasispecies during treatment [5]; therefore, it is paramount to develop new and improved anti-HCV therapeutic agents. A prime area of focus is the development of small molecule inhibitors targeting the essential viral enzymes.HCV is an enveloped, single-stranded RNA virus with approximately 9.6 kb genome of positive polarity that encodes three structural and seven non-structural (NS) proteins [6,7]. Among these, the NS5B RNAdependent RNA polymerase (RdRp), a crucial and unique component of the viral replication machinery with no functional equivalent in the ho...

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confidence: 99%

Characterization of Aurintricarboxylic Acid as a Potent Hepatitis C Virus Replicase Inhibitor

Chen

Waffo

Basu

et al. 2009

Antivir Chem Chemother

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“…Other HIV-1 protease inhibitors (nelfinavir, indinavir, amprenavir, or saquinavir) did not show any anti-HCoV-NL63 activity. We observed inhibition of HCoV-NL63 with aurintricarboxylic acid (32,36), an RNase and polymerase inhibitor (22,30), at a concentration of ϳ60 M, but we could not exclude the possibility that the effect was the result of an increased pH of the medium (Table 3). We measured no antiHCoV-NL63 activity in the following compounds: calpain inhibitors VI and III (4), glycyrrhizin (18), valinomycin (58), ȩscin (58), ribavirin (41), dipyridamole (3), actinomycin D (38), and pentoxifylline (8).…”

Section: Of 17 M [12])mentioning

confidence: 94%

Inhibition of HCoV-NL63 infection at early stages of the replication cycle

Hoek¹,

Bosch²,

Berkhout³

et al. 2006

Journal of Clinical Virology

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Human coronavirus NL63 (HCoV-NL63), a recently discovered member of the Coronaviridae family, has spread worldwide and is associated with acute respiratory illness in young children and elderly and immunocompromised persons. Further analysis of HCoV-NL63 pathogenicity seems warranted, in particular because the virus uses the same cellular receptor as severe acute respiratory syndrome-associated coronavirus. As there is currently no HCoV-NL63-specific and effective vaccine or drug therapy available, we evaluated several existing antiviral drugs and new synthetic compounds as inhibitors of HCoV-NL63, targeting multiple stages of the replication cycle. Of the 28 compounds that we tested, 6 potently inhibited HCoV-NL63 at early steps of the replication cycle. Intravenous immunoglobulins, heptad repeat 2 peptide, small interfering RNA1 (siRNA1), siRNA2, ␤-D-N 4 -hydroxycytidine, and 6-azauridine showed 50% inhibitory concentrations of 125 g/ml, 2 M, 5 nM, 3 nM, 400 nM, and 32 nM, respectively, and low 50% cytotoxicity concentrations (>10 mg/ml, >40 M, >200 nM, >200 nM, >100 M, and 80 M, respectively). These agents may be investigated further for the treatment of coronavirus infections.Coronaviruses are enveloped viruses with a positive, singlestranded RNA genome of approximately 27 to 32 kb. The 5Ј two-thirds of their genome encodes a polyprotein that contains all proteins necessary for RNA replication. The 3Ј one-third encodes several structural proteins, such as spike (S), envelope (E), membrane (M), and nucleocapsid (N), that, among other functions, participate in viral budding and are incorporated into the virus particle. Nonstructural accessory protein genes are also present in the 3Ј part of the genome, at a position and arrangement that is characteristic for each of the different coronavirus groups.Coronavirus infection starts with the recognition of a specific receptor on the host cell surface by an S protein, followed by virus internalization, which occurs either immediately by direct fusion with the plasma membrane or after endocytosis. Fusion of the viral membrane with the cellular membrane triggers the release of the viral RNA genome into the host cell cytoplasm. Viral RNA is copied by the viral replicase in membrane-associated replication centers (14). During the replication process, copies of the full-length genomic RNA and a nested set of subgenomic mRNAs are generated. These subgenomic mRNAs are functional templates for the translation of the structural proteins encoded in the 3Ј one-third of the genome. Fulllength viral RNA is encapsidated and released from the host cell as an infectious virus particle.Human coronavirus NL63 (HCoV-NL63), a recently discovered (45, 55) member of the Coronaviridae family, has spread worldwide, is observed most frequently in the winter season, and is associated with acute respiratory illness and croup in young children, elderly people, and immunocompromised patients (2,6,15,25,29,40,(53)(54)(55)(56). A recent report suggested that HCoV-NL63 is the causative agent of Kaw...

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“…At higher concentrations (up to 100 μM) ATA has been reported to have several effects on different cell types. For example, ATA has been described as an anti-viral [40][41][42][43][44][45] and anti-apoptotic agent [51][52][53]. It also inhibits several types of nucleases (DNAse I, RNAse A, SI nuclease, exonuclease) [46][47][48].…”

Section: Pmca4 Mutmentioning

confidence: 99%

Development and characterization of a novel fluorescent indicator protein PMCA4-GCaMP2 in cardiomyocytes

Mohamed

Abouleisa

Baudoin

et al. 2013

Journal of Molecular and Cellular Cardiology

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Isoform 4 of the plasma membrane calcium/calmodulin dependent ATPase (PMCA4) has recently emerged as an important regulator of several key pathophysiological processes in the heart, such as contractility and hypertrophy. However, direct monitoring of PMCA4 activity and assessment of calcium dynamics in its vicinity in cardiomyocytes are difficult due to the lack of molecular tools. In this study, we developed novel calcium fluorescent indicators by fusing the GCaMP2 calcium sensor to the N-terminus of PMCA4 to generate the PMCA4-GCaMP2 fusion molecule. We also identified a novel specific inhibitor of PMCA4, which might be useful for studying the role of this molecule in cardiomyocytes and other cell types. Using an adenoviral system we successfully expressed PMCA4-GCaMP2 in both neonatal and adult rat cardiomyocytes. This fusion molecule was correctly targeted to the plasma membrane and co-localised with caveolin-3. It could monitor signal oscillations in electrically stimulated cardiomyocytes. The PMCA4-GCaMP2 generated a higher signal amplitude and faster signal decay rate compared to a mutant inactive PMCA4 mut GCaMP2 fusion protein, in electrically stimulated neonatal and adult rat cardiomyocytes. A small molecule library screen enabled us to identify a novel selective inhibitor for PMCA4, which we found to reduce signal amplitude of PMCA4-GCaMP2 and prolong the time of signal decay (Tau) to a level comparable with the signal generated by PMCA4 mut GCaMP2. In addition, PMCA4-GCaMP2 but not the mutant form produced an enhanced signal in response to β-adrenergic stimulation. Together, the PMCA4-GCaMP2 and PMCA4 mut GCaMP2 demonstrate calcium dynamics in the vicinity of the pump under active or inactive conditions, respectively. In summary, the PMCA4-GCaMP2 together with the novel specific inhibitor provides new means with which to monitor calcium dynamics in the vicinity of a calcium transporter in cardiomyocytes and may become a useful tool to further study the biological functions of PMCA4. In addition, similar approaches could be useful for studying the activity of other calcium transporters during excitation-contraction coupling in the heart.

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Potent and selective inhibition of SARS coronavirus replication by aurintricarboxylic acid

Cited by 77 publications

References 22 publications

Characterization of Aurintricarboxylic Acid as a Potent Hepatitis C Virus Replicase Inhibitor

Characterization of Aurintricarboxylic Acid as a Potent Hepatitis C Virus Replicase Inhibitor

Inhibition of HCoV-NL63 infection at early stages of the replication cycle

Development and characterization of a novel fluorescent indicator protein PMCA4-GCaMP2 in cardiomyocytes

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