Cell-based and virtual fragment screening for adrenergic α2C receptor agonists

Szollosi, Edit; Bobok, Amrita; Kiss, László; Vass, Márton; Kurkó, Dalma; Kolok, Sándor; Visegrády, András; Keserü, György M.

doi:10.1016/j.bmc.2015.01.013

Cited by 11 publications

(7 citation statements)

References 43 publications

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“…A structure-based VS and a functional cell-based screening were undertaken in order to identify adrenergic a 2C AR receptor agonists. 233 A homology model of the activated a 2C AR was built based on the human active-state b 2 AR crystal structure, and the best conformation for VS was chosen based on retrospective docking. A library of 3071 fragments was experimentally screened, and also docked to the model, exhibiting a hit rate of 6.7% and an EF of 12.…”

Section: Discovery Of New Ligands Through Virtual Screeningmentioning

confidence: 99%

Expanding the horizons of G protein-coupled receptor structure-based ligand discovery and optimization using homology models

Cavasotto

Palomba

2015

Chem. Commun.

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With >800 members in humans, the G protein-coupled receptor (GPCR) super-family is the target for more than 30% of the marketed drugs. The recent boom in GPCR crystallography has enabled the solution of ∼30 different GPCR structures, which boosted the identification and optimization of novel modulators and new chemical entities through structure-based strategies. However, the number of available structures represents a small part of the human GPCR druggable target space, and its complete coverage in the near future seems unlikely. Homology modelling represents a reliable tool to fill this gap, and hence to vastly expand the horizons of structure-based drug discovery and design. In this Feature Article, we show from a wealth of retrospective and prospective studies that in spite of the pitfalls of and standing challenges in homology modelling, structural models have been critical for the blossoming and success of GPCR structure-based lead discovery and optimization endeavours; in addition, they have also been instrumental in characterizing receptor-ligand interaction, guiding the design of site-directed mutagenesis and SAR studies, and assessing off-target effects. Considering though their current limitations, we also discuss the most pressing issues to develop more accurate homology modelling strategies, with a special focus on the integration of computational tools with biochemical, biophysical and QSAR data, highlighting methodological aspects and recent progress. The teachings of the three GPCR Dock community-wide assessments and the fresh developments in GPCR classes B, C and F are commented. This is a fast growing and highly promising field of research, and in the coming years, the use of high-quality models should enable the discovery of a growing number of potent, selective and efficient GPCR drug leads with high therapeutic potential through receptor structure-based strategies.

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Section: Discovery Of New Ligands Through Virtual Screeningmentioning

confidence: 99%

Expanding the horizons of G protein-coupled receptor structure-based ligand discovery and optimization using homology models

Cavasotto

Palomba

2015

Chem. Commun.

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“…Hence, developing a highly sensitive cell-based fragment assays presents a cornerstone with the potential to identify specific, bioactive small molecule ligands. Yet, only a handful of cell-based fragment assays have been reported, indicating the complexity of these screening approaches. − One reason might be the difficulty to find sensitive readout methods. Moreover, high compound concentrations, which are necessary in fragment-based applications, have a higher tendency to unspecifically interfere with cellular processes, complicating the search for specific small molecules that are active in living cells.…”

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confidence: 99%

CellFy: A Cell-Based Fragment Screen against C-Type Lectins

et al. 2018

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Fragment-based drug discovery is a powerful complement to conventional high-throughput screening, especially for difficult targets. Screening low-molecular-weight fragments usually requires highly sensitive biophysical methods, because of the generally low affinity of the identified ligands. Here, we developed a cell-based fragment screening assay (cellFy) that allows sensitive identification of fragment hits in a physiologically more relevant environment, in contrast to isolated target screenings in solution. For this, a fluorescently labeled multivalent reporter was employed, enabling direct measurement of displacement by low-molecular-weight fragments without requiring enzymatic reactions or receptor activation. We applied this technique to identify hits against two challenging targets of the C-type lectin receptor (CLR) family: Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Nonintegrin (DC-SIGN) and Langerin. Both receptors are involved in pathogen recognition and initiation of an immune response, which renders them attractive targets for immune modulation. Because of their shallow and hydrophilic primary binding site, hit identification for CLRs is challenging and druglike ligands for CLRs are sparse. Screening of a fragment library followed by hit validation identified several promising candidates for further fragment evolution for DC-SIGN. In addition, a multiplexed assay format was developed for simultaneous screening against multiple CLRs, allowing a selectivity counterscreening. Overall, this sensitive cell-based fragment screening assay provides a powerful tool for rapid identification of bioactive fragments, even for difficult targets.

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“…Weak affinity chromatography was developed as a tool to screen hits in FBDD ( Duong-Thi et al, 2011 ). Fragment-based screening was also carried out using cell-based assays ( Szõllõsi et al, 2015 ; Schulze et al, 2018 ). A study showed that fragments reacting with cysteine residues were able to identify proteins that formed interactions with these compounds ( Backus et al, 2016 ).…”

Section: Fragment Screening Methodsmentioning

confidence: 99%

Application of Fragment-Based Drug Discovery to Versatile Targets

2020

Front. Mol. Biosci.

125

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Fragment-based drug discovery (FBDD) is a powerful method to develop potent smallmolecule compounds starting from fragments binding weakly to targets. As FBDD exhibits several advantages over high-throughput screening campaigns, it becomes an attractive strategy in target-based drug discovery. Many potent compounds/inhibitors of diverse targets have been developed using this approach. Methods used in fragment screening and understanding fragment-binding modes are critical in FBDD. This review elucidates fragment libraries, methods utilized in fragment identification/confirmation, strategies applied in growing the identified fragments into drug-like lead compounds, and applications of FBDD to different targets. As FBDD can be readily carried out through different biophysical and computer-based methods, it will play more important roles in drug discovery.

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Cell-based and virtual fragment screening for adrenergic α2C receptor agonists

Cited by 11 publications

References 43 publications

Expanding the horizons of G protein-coupled receptor structure-based ligand discovery and optimization using homology models

Expanding the horizons of G protein-coupled receptor structure-based ligand discovery and optimization using homology models

CellFy: A Cell-Based Fragment Screen against C-Type Lectins

Application of Fragment-Based Drug Discovery to Versatile Targets

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