scholarly journals Target identification for repurposed drugs active against SARS-CoV-2 via high-throughput inverse docking

2021 ◽  
Author(s):  
Sergio R. Ribone ◽  
S. Alexis Paz ◽  
Cameron F. Abrams ◽  
Marcos A. Villarreal
Keyword(s):  
High Throughput ◽  
Target Identification ◽  
Repurposed Drugs

scholarly journals Target identification for repurposed drugs active against SARS-CoV-2 via high-throughput inverse docking

2021 ◽  
Author(s):  
Marcos A. Villarreal ◽  
Sergio P. Ribone ◽  
S. Alexis Paz ◽  
Cameron F. Abrams
Keyword(s):  
High Throughput ◽  
Target Identification ◽  
Repurposed Drugs

scholarly journals Transcriptional-Based Screens for Pathway-Specific, High-Throughput Target Discovery in Endothelial Cells

2004 ◽  
Vol 9 (8) ◽  
pp. 704-711 ◽  
Author(s):  
Robert L. Yauch ◽  
Edward E. Kadel ◽  
Cory Nicholas ◽  
Selwyna Tetangco ◽  
Douglas O. Clary
Keyword(s):  
Endothelial Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Target Identification ◽  
Gene Products ◽  
Target Discovery ◽  
Drug Target Identification ◽  
A Cell ◽  
Cytokine Stimulation ◽  
High Throughput Assay

With the sequence of the human genome at hand, target discovery strategies are needed that can rapidly identify novel gene products involved in human disease pathways. In this article, the authors describe a cell-based, high-throughput assay that can identify gene products capable of modulating the vascular endothelial growth factor (VEGF) and tumor necrosis factor • (TNFa) signaling pathways in human endothelial cells. The assay uses real-time PCRtechnology tomeasure downstreamreporter mRNA transcripts induced upon cytokine stimulation in a 96-well plate format and has been adapted for use with recombinant adenoviruses. The authors specifically demonstratemodulation of cytokine-driven reporter transcripts using drug inhibitors and through adenoviral-mediated expression of known signaling intermediates of the respective pathways. In addition, they have used an arrayed library of 350 recombinant adenoviruses to screen for novel modulators of the VEGF and TNF• pathways. The high-throughput screening capacity and sensitivity of this system make it a useful tool for new drug target identification.

scholarly journals Electroporation Knows No Boundaries

2015 ◽  
Vol 20 (8) ◽  
pp. 932-942 ◽  
Author(s):  
Christin Luft ◽  
Robin Ketteler
Keyword(s):  
Cell Lines ◽  
High Throughput ◽  
High Throughput Screening ◽  
High Efficiency ◽  
Target Identification ◽  
Cell Types ◽  
Rnai Technology ◽  
Drug Target Identification ◽  
Interfering Rna ◽  
Calcium Phosphate Precipitation

The discovery of RNA interference (RNAi) has enabled several breakthrough discoveries in the area of functional genomics. The RNAi technology has emerged as one of the major tools for drug target identification and has been steadily improved to allow gene manipulation in cell lines, tissues, and whole organisms. One of the major hurdles for the use of RNAi in high-throughput screening has been delivery to cells and tissues. Some cell types are refractory to high-efficiency transfection with standard methods such as lipofection or calcium phosphate precipitation and require different means. Electroporation is a powerful and versatile method for delivery of RNA, DNA, peptides, and small molecules into cell lines and primary cells, as well as whole tissues and organisms. Of particular interest is the use of electroporation for delivery of small interfering RNA oligonucleotides and clustered regularly interspaced short palindromic repeats/Cas9 plasmid vectors in high-throughput screening and for therapeutic applications. Here, we will review the use of electroporation in high-throughput screening in cell lines and tissues.

Protein Preparation Automatic Protocol for High-Throughput Inverse Virtual Screening: Accelerating the Target Identification by Computational Methods

2019 ◽  
Vol 59 (11) ◽  
pp. 4678-4690 ◽  
Author(s):  
Simona De Vita ◽  
Gianluigi Lauro ◽  
Dafne Ruggiero ◽  
Stefania Terracciano ◽  
Raffaele Riccio ◽  
...  
Keyword(s):  
Virtual Screening ◽  
Computational Methods ◽  
High Throughput ◽  
Target Identification ◽  
Protein Preparation

Application of Combination High-Throughput Phenotypic Screening and Target Identification Methods for the Discovery of Natural Product-Based Combination Drugs

2017 ◽  
Vol 38 (2) ◽  
pp. 504-524 ◽  
Author(s):  
Monica Isgut ◽  
Mukkavilli Rao ◽  
Chunhua Yang ◽  
Vangala Subrahmanyam ◽  
Padmashree C. G. Rida ◽  
...  
Keyword(s):  
Natural Product ◽  
High Throughput ◽  
Target Identification ◽  
Phenotypic Screening ◽  
Identification Methods ◽  
Combination Drugs

scholarly journals High-throughput screening and evaluation of repurposed drugs targeting the SARS-CoV-2 main protease

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Yan Li ◽  
Jinyong Zhang ◽  
Zilei Duan ◽  
Ning Wang ◽  
Xiangcheng Sun ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Main Protease ◽  
Repurposed Drugs

scholarly journals Drug Repurposing: A Paradigm Shift in Drug Discovery

2020 ◽  
Vol 5 (04) ◽  
pp. 60-68
Author(s):  
Saravanan Jayaram ◽  
Emdormi Rymbai ◽  
Deepa Sugumar ◽  
Divakar Selvaraj
Keyword(s):  
Drug Discovery ◽  
Success Rate ◽  
Chemical Structure ◽  
Target Identification ◽  
Drug Repurposing ◽  
New Drugs ◽  
Attractive Option ◽  
Traditional Drug ◽  
Repurposed Drugs

The traditional methods of drug discovery and drug development are a tedious, complex, and costly process. Target identification, target validation; lead identification; and lead optimization are a lengthy and unreliable process that further complicates the discovery of new drugs. A study of more than 15 years reports that the success rate in the discovery of new drugs in the fields of ophthalmology, cardiovascular, infectious disease, and oncology to be 32.6%, 25.5%, 25.2% and 3.4%, respectively. A tedious and costly process coupled with a very low success rate makes the traditional drug discovery a less attractive option. Therefore, an alternative to traditional drug discovery is drug repurposing, a process in which already existing drugs are repurposed for conditions other than which were originally intended. Typical examples of repurposed drugs are thalidomide, sildenafil, memantine, mirtazapine, mifepristone, etc. In recent times, several databases have been developed to hasten drug repurposing based on the side effect profile, the similarity of chemical structure, and target site. This work reviews the pivotal role of drug repurposing in drug discovery and the databases currently available for drug repurposing.

scholarly journals Identification and Characterization of Influenza Virus Entry Inhibitors through Dual Myxovirus High-Throughput Screening

2016 ◽  
Vol 90 (16) ◽  
pp. 7368-7387 ◽  
Author(s):  
Marco Weisshaar ◽  
Robert Cox ◽  
Zachary Morehouse ◽  
Shiva Kumar Kyasa ◽  
Dan Yan ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Small Molecule ◽  
High Throughput ◽  
In Silico ◽  
High Throughput Screening ◽  
Target Identification ◽  
Viral Escape ◽  
Reporter Strain ◽  
Secondary Resistance ◽  
Developmental Potential

ABSTRACTInfluenza A virus (IAV) infections cause major morbidity and mortality, generating an urgent need for novel antiviral therapeutics. We recently established a dual myxovirus high-throughput screening protocol that combines a fully replication-competent IAV-WSN strain and a respiratory syncytial virus reporter strain for the simultaneous identification of IAV-specific, paramyxovirus-specific, and broad-spectrum inhibitors. In the present study, this protocol was applied to a screening campaign to assess a diverse chemical library with over 142,000 entries. Focusing on IAV-specific hits, we obtained a hit rate of 0.03% after cytotoxicity testing and counterscreening. Three chemically distinct hit classes with nanomolar potency and favorable cytotoxicity profiles were selected. Time-of-addition, minigenome, and viral entry studies demonstrated that these classes block hemagglutinin (HA)-mediated membrane fusion. Antiviral activity extends to an isolate from the 2009 pandemic and, in one case, another group 1 subtype. Target identification through biolayer interferometry confirmed binding of all hit compounds to HA. Resistance profiling revealed two distinct escape mechanisms: primary resistance, associated with reduced compound binding, and secondary resistance, associated with unaltered binding. Secondary resistance was mediated, unusually, through two different pairs of cooperative mutations, each combining a mutation eliminating the membrane-proximal stalk N-glycan with a membrane-distal change in HA1 or HA2. Chemical synthesis of an analog library combined within silicodocking extracted a docking pose for the hit classes. Chemical interrogation spotlights IAV HA as a major druggable target for small-molecule inhibition. Our study identifies novel chemical scaffolds with high developmental potential, outlines diverse routes of IAV escape from entry inhibition, and establishes a path toward structure-aided lead development.IMPORTANCEThis study is one of the first to apply a fully replication-competent third-generation IAV reporter strain to a large-scale high-throughput screen (HTS) drug discovery campaign, allowing multicycle infection and screening in physiologically relevant human respiratory cells. A large number of potential druggable targets was thus chemically interrogated, but mechanistic characterization, positive target identification, and resistance profiling demonstrated that three chemically promising and structurally distinct hit classes selected for further analysis all block HA-mediated membrane fusion. Viral escape from inhibition could be achieved through primary and secondary resistance mechanisms.In silicodocking predicted compound binding to a microdomain located at the membrane-distal site of the prefusion HA stalk that was also previously suggested as a target site for chemically unrelated HA inhibitors. This study identifies an unexpected chemodominance of the HA stalk microdomain for small-molecule inhibitors in IAV inhibitor screening campaigns and highlights a novel mechanism of cooperative resistance to IAV entry blockers.

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