Bioorthogonal Probes for the Study of MDM2-p53 Inhibitors in Cells and Development of High-Content Screening Assays for Drug Discovery

Background: In recent years, there has been an improvement in the in vitro and in vivo methodology for the screening of anti-chagasic compounds. Millions of compounds can now have their activity evaluated (in large compound libraries) by means of high throughput in vitro screening assays. Objective: Current approaches to drug discovery for Chagas disease. Method: This review article examines the contribution of these methodological advances in medicinal chemistry in the last four years, focusing on Trypanosoma cruzi infection, obtained from the PubMed, Web of Science, and Scopus databases. Results: Here, we have shown that the promise is increasing each year for more lead compounds for the development of a new drug against Chagas disease. Conclusion: There is increased optimism among those working with the objective to find new drug candidates for optimal treatments against Chagas disease.

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Integrating High-Content Analysis into a Multiplexed Screening Approach to Identify and Characterize GPCR Agonists

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057114533146 ◽

2014 ◽

Vol 19 (7) ◽

pp. 1079-1089 ◽

Cited By ~ 6

Author(s):

Yingjie Zhu ◽

John Watson ◽

Mengjie Chen ◽

Ding Ren Shen ◽

Melissa Yarde ◽

...

Keyword(s):

Drug Discovery ◽

G Protein Coupled Receptors ◽

High Content Imaging ◽

Biased Agonism ◽

Receptor Oligomerization ◽

Sphingosine 1 Phosphate ◽

High Content Analysis ◽

Screening Assays ◽

Hit Identification ◽

G Protein Coupled

G protein–coupled receptors (GPCRs) are one of the most popular and proven target classes for therapeutic intervention. The increased appreciation for allosteric modulation, receptor oligomerization, and biased agonism has led to the development of new assay platforms that seek to capitalize on these aspects of GPCR biology. High-content screening is particularly well suited for GPCR drug discovery given the ability to image and quantify changes in multiple cellular parameters, to resolve subcellular structures, and to monitor events within a physiologically relevant environment. Focusing on the sphingosine-1-phosphate (S1P1) receptor, we evaluated the utility of high-content approaches in hit identification efforts by developing and applying assays to monitor β-arrestin translocation, GPCR internalization, and GPCR recycling kinetics. Using these approaches in combination with more traditional GPCR screening assays, we identified compounds whose unique pharmacological profiles would have gone unnoticed if using a single platform. In addition, we identified a compound that induces an atypical pattern of β-arrestin translocation and GPCR recycling kinetics. Our results highlight the value of high-content imaging in GPCR drug discovery efforts and emphasize the value of a multiassay approach to study pharmacological properties of compounds of interest.

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Development of a High-Content Screening Assay Panel to Accelerate Mechanism of Action Studies for Oncology Research

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057112450050 ◽

2012 ◽

Vol 17 (8) ◽

pp. 1005-1017 ◽

Cited By ~ 16

Author(s):

Danli L. Towne ◽

Emily E. Nicholl ◽

Kenneth M. Comess ◽

Scott C. Galasinski ◽

Philip J. Hajduk ◽

...

Keyword(s):

Drug Discovery ◽

Mechanism Of Action ◽

Large Scale ◽

Morphological Changes ◽

Single Cells ◽

High Content Screening ◽

Individual Compound ◽

Screening Assay ◽

Oncology Research ◽

Protein Functions

Efficient elucidation of the biological mechanism of action of novel compounds remains a major bottleneck in the drug discovery process. To address this need in the area of oncology, we report the development of a multiparametric high-content screening assay panel at the level of single cells to dramatically accelerate understanding the mechanism of action of cell growth–inhibiting compounds on a large scale. Our approach is based on measuring 10 established end points associated with mitochondrial apoptosis, cell cycle disruption, DNA damage, and cellular morphological changes in the same experiment, across three multiparametric assays. The data from all of the measurements taken together are expected to help increase our current understanding of target protein functions, constrain the list of possible targets for compounds identified using phenotypic screens, and identify off-target effects. We have also developed novel data visualization and phenotypic classification approaches for detailed interpretation of individual compound effects and navigation of large collections of multiparametric cellular responses. We expect this general approach to be valuable for drug discovery across multiple therapeutic areas.

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A novel multi-parametric high content screening assay in ciPTEC-OAT1 to predict drug-induced nephrotoxicity during drug discovery

Archives of Toxicology ◽

10.1007/s00204-018-2284-y ◽

2018 ◽

Vol 92 (10) ◽

pp. 3175-3190 ◽

Cited By ~ 19

Author(s):

Anna-Karin Sjögren ◽

Katarina Breitholtz ◽

Ernst Ahlberg ◽

Lucas Milton ◽

Malin Forsgard ◽

...

Keyword(s):

Drug Discovery ◽

High Content Screening ◽

Screening Assay ◽

Drug Induced

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The IUPHAR/BPS Guide to PHARMACOLOGY in 2020: extending immunopharmacology content and introducing the IUPHAR/MMV Guide to MALARIA PHARMACOLOGY

Nucleic Acids Research ◽

10.1093/nar/gkz951 ◽

2019 ◽

Cited By ~ 7

Author(s):

Jane F Armstrong ◽

Elena Faccenda ◽

Simon D Harding ◽

Adam J Pawson ◽

Christopher Southan ◽

...

Keyword(s):

Drug Discovery ◽

Open Access ◽

Molecular Interactions ◽

Malaria Parasite ◽

Antimalarial Activity ◽

Molecular Targets ◽

Interaction Data ◽

Data Types ◽

Global Challenge ◽

Screening Assays

Abstract The IUPHAR/BPS Guide to PHARMACOLOGY (www.guidetopharmacology.org) is an open-access, expert-curated database of molecular interactions between ligands and their targets. We describe significant updates made over the seven releases during the last two years. The database is notably enhanced through the continued linking of relevant pharmacology with key immunological data types as part of the IUPHAR Guide to IMMUNOPHARMACOLOGY (www.guidetoimmunopharmacology.org) and by a major new extension, the IUPHAR/MMV Guide to Malaria PHARMACOLOGY (www.guidetomalariapharmacology.org). The latter has been constructed in partnership with the Medicines for Malaria Venture, an organization dedicated to identifying, developing and delivering new antimalarial therapies that are both effective and affordable. This is in response to the global challenge of over 200 million cases of malaria and 400 000 deaths worldwide, with the majority in the WHO Africa Region. It provides new pharmacological content, including molecular targets in the malaria parasite, interaction data for ligands with antimalarial activity, and establishes curation of data from screening assays, used routinely in antimalarial drug discovery, against the whole organism. A dedicated portal has been developed to provide quick and focused access to these new data.

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Drug Discovery for Chagas Disease: Impact of Different Host Cell Lines on Assay Performance and Hit Compound Selection

Tropical Medicine and Infectious Disease ◽

10.3390/tropicalmed4020082 ◽

2019 ◽

Vol 4 (2) ◽

pp. 82 ◽

Cited By ~ 5

Author(s):

Caio Haddad Franco ◽

Laura Maria Alcântara ◽

Eric Chatelain ◽

Lucio Freitas-Junior ◽

Carolina Borsoi Moraes

Keyword(s):

Drug Discovery ◽

Host Cell ◽

Cell Lines ◽

Chagas Disease ◽

Host Cells ◽

Mammalian Cell Lines ◽

Starting Point ◽

Disease Impact ◽

Screening Assays ◽

Host Parasite

Cell-based screening has become the major compound interrogation strategy in Chagas disease drug discovery. Several different cell lines have been deployed as host cells in screening assays. However, host cell characteristics and host-parasite interactions may play an important role when assessing anti-T. cruzi compound activity, ultimately impacting on hit discovery. To verify this hypothesis, four distinct mammalian cell lines (U2OS, THP-1, Vero and L6) were used as T. cruzi host cells in High Content Screening assays. Rates of infection varied greatly between different host cells. Susceptibility to benznidazole also varied, depending on the host cell and parasite strain. A library of 1,280 compounds was screened against the four different cell lines infected with T. cruzi, resulting in the selection of a total of 82 distinct compounds as hits. From these, only two hits were common to all four cell lines assays (2.4%) and 51 were exclusively selected from a single assay (62.2%). Infected U2OS cells were the most sensitive assay, as 55 compounds in total were identified as hits; infected THP-1 yielded the lowest hit rates, with only 16 hit compounds. Of the selected hits, compound FPL64176 presented selective anti-T. cruzi activity and could serve as a starting point for the discovery of new anti-chagasic drugs.

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