Bioluminescent assays for ADMET

Cali, James J.; Niles, Andrew L.; Valley, Michael P.; O’Brien, Martha A.; Riss, Terry; Shultz, John

doi:10.1517/17425255.4.1.103

Cited by 54 publications

(29 citation statements)

References 76 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The difference between low and high polarization can be used to detect NR ligand binding. One disadvantage of FP assays is their high level of background, which translates into a lower signal-to-noise ratio and decreased sensitivity [62].…”

Section: Ligand Binding Assays Scintillation Proximity Assays (Spa) Amentioning

confidence: 99%

Current In Vitro High Throughput Screening Approaches to Assess Nuclear Receptor Activation

Raucy¹,

Lasker²

2010

CDM

View full text Add to dashboard Cite

The screening of new drug candidates for nuclear receptor activation can identify agents with the potential to produce drug-drug interactions or elicit adverse drug effects. The nuclear receptors of interest are those that control the expression of drug metabolizing enzymes and drug transporters, and include the constitutive androstane receptor (CAR, NR1I3), the pregnane X receptor (PXR, NR1I2) and the aryl hydrocarbon receptor (AhR). This review will focus on the methods currently used to assess activation of these receptors. Assessment of nuclear receptor activation can be accomplished using direct or indirect approaches. Indirect methods quantify specific gene products that result from nuclear receptor activation while direct approaches measure either the binding of ligands to the receptors or the transcriptional events produced by ligand binding. Assays that directly quantify nuclear receptor activation are growing in popularity and, importantly, are amenable to high throughput screening (HTS). Several ligand binding assays are currently being utilized, including radioligand competition binding, where compounds compete with radiolabelled ligand for binding to PXR or CAR, such as the scintillation proximity binding assay that measures the reaction of ligands with receptor-coated beads. A fluorescence resonance energy transfer assay has also been developed, where the fluorescent signal is generated via the ligand-dependent interaction between the fluorescently-labeled ligand binding domain of a nuclear receptor and co-activator proteins. Other in vitro activation assays include transient- and stably-transfected cell lines incorporating an expression vector for PXR, CAR or AhR plus a reporter gene vector containing response elements. The methods focused on in this review will be limited to the more direct in vitro approaches that are amenable to high throughput screening.

show abstract

Section: Ligand Binding Assays Scintillation Proximity Assays (Spa) Amentioning

confidence: 99%

Current In Vitro High Throughput Screening Approaches to Assess Nuclear Receptor Activation

Raucy¹,

Lasker²

2010

CDM

View full text Add to dashboard Cite

show abstract

“…The activity was measured by the luciferase assay (Cali et al 2008). The instrument response is correlated to the chemical concentrations of components of luciferase pathway reactions, so the light intensity can be used to associate an observable parameter with a molecular process.…”

Section: Resultsmentioning

confidence: 99%

Design, conformational studies and analysis of structure–function relationships of PTH (1–11) analogues: the essential role of Val in position 2

et al. 2011

View full text Add to dashboard Cite

The N-terminal 1-34 segment of parathyroid hormone (PTH) is fully active in vitro and in vivo and it elicits all the biological responses characteristic of the native intact PTH. Recent studies reported potent helical analogues of the PTH (1-11) with helicity-enhancing substitutions. This work describes the synthesis, biological activity, and conformational studies of analogues obtained from the most active non-natural PTH (1-11) peptide H-Aib-Val-Aib-Glu-Ile-Gln-Leu-Nle-His-Gln-Har-NH2; specifically, the replacement of Val in position 2 with D-Val, L-(αMe)-Val and N-isopropyl-Gly was studied. The synthesized analogues were characterized functionally by in-cell assays and their structures were determined by CD and NMR spectroscopy. To clarify the relationship between the structure and activity, the structural data were used to generate a pharmacophoric model, obtained overlapping all the analogues. This model underlines the fundamental functional role of the side chain of Val2 and, at the same time, reveals that the introduction of conformationally constrained Cα-tetrasubstituted α-amino acids in the peptides increases their helical content, but does not necessarily ensure significant biological activity.

show abstract

“…BLI modalities acquire macroscopic information in vivo through two basic strategies: (1) genetically encoded reporters (luciferase assays) for studying the regulation of a gene of interest and (2) assays of injected luciferin derivative for studying the functional activity of membrane transporters (103).…”

Section: Blimentioning

confidence: 99%

Molecular Imaging of Membrane Transporters’ Activity in Cancer: a Picture is Worth a Thousand Tubes

Mann

Semenenko

Meir

et al. 2015

AAPS J

View full text Add to dashboard Cite

Abstract. Molecular imaging allows the non-invasive assessment of membrane transporter expression and function in living subjects. Such technologies have the potential to become diagnostic and prognostic tools, allowing detection, localization, and prediction of response of tumors and their metastases to therapy. Beyond tumors, imaging can also help understand the role of transporters in adverse drug effects and drug clearance. Here, we review molecular imaging technologies that monitor transporter-mediated processes. We emphasize emerging probe substrates and potential clinical applications of imaging the function of membrane transporters in cancer.

show abstract

Bioluminescent assays for ADMET

Cited by 54 publications

References 76 publications

Current In Vitro High Throughput Screening Approaches to Assess Nuclear Receptor Activation

Current In Vitro High Throughput Screening Approaches to Assess Nuclear Receptor Activation

Design, conformational studies and analysis of structure–function relationships of PTH (1–11) analogues: the essential role of Val in position 2

Molecular Imaging of Membrane Transporters’ Activity in Cancer: a Picture is Worth a Thousand Tubes

Contact Info

Product

Resources

About