A Novel Method—Amenable for High-throughput Screening Purposes—to Quantify Antiviral Activity Against Viruses that Induce Limited CPE

High-throughput screening (HTS) previously identified benzimidazole 1 (JMN3-003) as a compound with broad antiviral activity against different influenza viruses and paramyxovirus strains. In pursuit of a lead compound from this series for development, we sought to increase both the potency and the aqueous solubility of 1. Lead optimization has achieved compounds with potent antiviral activity against a panel of myxovirus family members (EC50 values in the low nanomolar range) and much improved aqueous solubilities relative to that of 1. Additionally, we have devised a robust synthetic strategy for preparing 1 and congeners in an enantio-enriched fashion, which has allowed us to demonstrate that the (S)-enantiomers are generally 7- to 110-fold more potent than the corresponding (R)-isomers.

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DRABAL: novel method to mine large high-throughput screening assays using Bayesian active learning

Journal of Cheminformatics ◽

10.1186/s13321-016-0177-8 ◽

2016 ◽

Vol 8 (1) ◽

Cited By ~ 11

Author(s):

Othman Soufan ◽

Wail Ba-Alawi ◽

Moataz Afeef ◽

Magbubah Essack ◽

Panos Kalnis ◽

...

Keyword(s):

Active Learning ◽

High Throughput ◽

High Throughput Screening ◽

Novel Method ◽

Screening Assays

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Matrix-Based Activity Pattern Classification as a Novel Method for the Characterization of Enzyme Inhibitors Derived from High-Throughput Screening

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057116667255 ◽

2016 ◽

Vol 21 (10) ◽

pp. 1075-1089 ◽

Cited By ~ 2

Author(s):

Douglas S. Auld ◽

Marta Jimenez ◽

Kimberley Yue ◽

Scott Busby ◽

Yu-Chi Chen ◽

...

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Enzyme Inhibitors ◽

Classification Method ◽

Shift Method ◽

Lead Selection ◽

Small Matrix ◽

Novel Method ◽

Good Agreement

One of the central questions in the characterization of enzyme inhibitors is determining the mode of inhibition (MOI). Classically, this is done with a number of low-throughput methods in which inhibition models are fitted to the data. The ability to rapidly characterize the MOI for inhibitors arising from high-throughput screening in which hundreds to thousands of primary inhibitors may need to be characterized would greatly help in lead selection efforts. Here we describe a novel method for determining the MOI of a compound without the need for curve fitting of the enzyme inhibition data. We provide experimental data to demonstrate the utility of this new high-throughput MOI classification method based on nonparametric analysis of the activity derived from a small matrix of substrate and inhibitor concentrations (e.g., from a 4S × 4I matrix). Lists of inhibitors from four different enzyme assays are studied, and the results are compared with the previously described IC50-shift method for MOI classification. The MOI results from this method are in good agreement with the known MOI and compare favorably with those from the IC50-shift method. In addition, we discuss some advantages and limitations of the method and provide recommendations for utilization of this MOI classification method.

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Novel method for high-throughput phenotyping of sleep in mice

Physiological Genomics ◽

10.1152/physiolgenomics.00139.2006 ◽

2007 ◽

Vol 28 (2) ◽

pp. 232-238 ◽

Cited By ~ 125

Author(s):

Allan I. Pack ◽

Raymond J. Galante ◽

Greg Maislin ◽

Jacqueline Cater ◽

Dimitris Metaxas ◽

...

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Time Of Day ◽

Chemical Mutagenesis ◽

High Throughput Phenotyping ◽

Novel Method ◽

Initial Implantation ◽

Method Of Assessment ◽

Digital Video Analysis ◽

Electroencephalogram Eeg

Assessment of sleep in mice currently requires initial implantation of chronic electrodes for assessment of electroencephalogram (EEG) and electromyogram (EMG) followed by time to recover from surgery. Hence, it is not ideal for high-throughput screening. To address this deficiency, a method of assessment of sleep and wakefulness in mice has been developed based on assessment of activity/inactivity either by digital video analysis or by breaking infrared beams in the mouse cage. It is based on the algorithm that any episode of continuous inactivity of ≥40 s is predicted to be sleep. The method gives excellent agreement in C57BL/6J male mice with simultaneous assessment of sleep by EEG/EMG recording. The average agreement over 8,640 10-s epochs in 24 h is 92% ( n = 7 mice) with agreement in individual mice being 88–94%. Average EEG/EMG determined sleep per 2-h interval across the day was 59.4 min. The estimated mean difference (bias) per 2-h interval between inactivity-defined sleep and EEG/EMG-defined sleep was only 1.0 min (95% confidence interval for mean bias −0.06 to +2.6 min). The standard deviation of differences (precision) was 7.5 min per 2-h interval with 95% limits of agreement ranging from −13.7 to +15.7 min. Although bias significantly varied by time of day ( P = 0.0007), the magnitude of time-of-day differences was not large (average bias during lights on and lights off was +5.0 and −3.0 min per 2-h interval, respectively). This method has applications in chemical mutagenesis and for studies of molecular changes in brain with sleep/wakefulness.

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High-throughput screening of organic reactions in microdroplets using desorption electrospray ionization mass spectrometry (DESI-MS): hardware and software implementation

Analytical Methods ◽

10.1039/d0ay00072h ◽

2020 ◽

Vol 12 (28) ◽

pp. 3654-3669

Author(s):

Tiago Jose P. Sobreira ◽

Larisa Avramova ◽

Botond Szilagyi ◽

David L. Logsdon ◽

Bradley P. Loren ◽

...

Keyword(s):

Mass Spectrometry ◽

Electrospray Ionization ◽

High Throughput ◽

Electrospray Ionization Mass Spectrometry ◽

High Throughput Screening ◽

Desorption Electrospray Ionization ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Desi Ms ◽

Novel Method

Implementation of a novel method for high-throughput screening of reactions in microdroplets. The reaction and analysis steps are performed simultaneously using desorption electrospray ionization mass spectrometry (DESI-MS) at a rate of up to 1 reaction mixture per second.

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A novel method for mining highly imbalanced high-throughput screening data in PubChem

Bioinformatics ◽

10.1093/bioinformatics/btp589 ◽

2009 ◽

Vol 25 (24) ◽

pp. 3310-3316 ◽

Cited By ~ 45

Author(s):

Qingliang Li ◽

Yanli Wang ◽

Stephen H. Bryant

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Novel Method

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Discovery of Novel Benzoquinazolinones and Thiazoloimidazoles, Inhibitors of Influenza H5N1 and H1N1 Viruses, from a Cell-Based High-Throughput Screen

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057110384613 ◽

2010 ◽

Vol 16 (1) ◽

pp. 73-81 ◽

Cited By ~ 17

Author(s):

Joseph A. Maddry ◽

Xi Chen ◽

Colleen B. Jonsson ◽

Subramaniam Ananthan ◽

Judith Hobrath ◽

...

Keyword(s):

Antiviral Activity ◽

High Throughput ◽

High Throughput Screening ◽

Influenza A ◽

Active Compounds ◽

Influenza Activity ◽

A Cell ◽

Molecular Libraries ◽

Viral Cytopathic Effect

A highly reproducible and robust cell-based high-throughput screening (HTS) assay was adapted for screening of small molecules for antiviral activity against influenza virus strain A/Vietnam/1203/2004 (H5N1). The NIH Molecular Libraries Small Molecule Repository (MLSMR) Molecular Libraries Screening Centers Network (MLSCN) 100,000-compound library was screened at 50 µM. The “hit” rate (>25% inhibition of the viral cytopathic effect) from the single-dose screen was 0.32%. The hits were evaluated for their antiviral activity, cell toxicity, and selectivity in dose-response experiments. The screen yielded 5 active compounds (SI value >3). One compound showed an SI50 value of greater than 3, 3 compounds had SI values ranging from greater than 14 to 34, and the most active compound displayed an SI value of 94. The active compounds represent 2 different classes of molecules, benzoquinazolinones and thiazoloimidazoles, which have not been previously identified as having antiviral/anti-influenza activity. These molecules were also effective against influenza A/California/04/2009 virus (H1N1) and other H1N1 and H5N1 virus strains in vitro but not H3N2 strains. Real-time qRT-PCR results reveal that these chemotypes significantly reduced M1 RNA levels as compared to the no-drug influenza-infected Madin Darby canine kidney cells.

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Inducible APOBEC3G-Vif Double Stable Cell Line as a High-Throughput Screening Platform To Identify Antiviral Compounds

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00775-09 ◽

2009 ◽

Vol 54 (1) ◽

pp. 78-87 ◽

Cited By ~ 3

Author(s):

Boris Nowotny ◽

Thomas Schneider ◽

Gabriele Pradel ◽

Tanja Schirmeister ◽

Axel Rethwilm ◽

...

Keyword(s):

Antiviral Activity ◽

Cell Line ◽

High Throughput ◽

High Throughput Screening ◽

Yellow Fluorescent Protein ◽

26S Proteasome ◽

Stable Cell Line ◽

Antiviral Compound ◽

Compound Libraries ◽

Antiviral Compounds

ABSTRACT Inhibition of the interaction of the human cytidine-deaminase APOBEC3G (A3G) with the human immunodeficiency virus (HIV) type 1-specific viral infectivity factor (Vif) represents a novel therapeutic approach in which a cellular factor with potent antiviral activity (A3G) plays a key role. In HIV-infected cells, the interaction of Vif with A3G leads to the subsequent degradation of A3G by the 26S proteasome via the ubiquitin pathway and to the loss of antiviral activity. To establish a stable and convenient cellular testing platform for the high-throughput screening of potential antiviral compound libraries, we engineered a double transgenic cell line constitutively expressing an enhanced yellow fluorescent protein expressor (EYFP-A3G) fusion as well as a Tet-Off controllable Vif protein. With this cell line, we were able to measure precisely the Vif-induced degradation of A3G in the presence of potential antiviral compounds in an easy-to-handle, robust, and practical high-throughput multiwell plate format with an excellent screening window coefficient (Z factor) of 0.67.

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