Safety screening in early drug discovery: An optimized assay panel

Bendels, Stefanie; Bissantz, Caterina; Fasching, Bernhard; Gerebtzoff, Grégori; Guba, Wolfgang; Kansy, Manfred; Migeon, Jacques; Mohr, Susanne; Peters, Jens‐Uwe; Tillier, Fabien; Wyler, R.; Lerner, C.; Krämer, Christian; Richter, Hans G.; Roberts, Sonia

doi:10.1016/j.vascn.2019.106609

Cited by 52 publications

(37 citation statements)

References 28 publications

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“…identify potential off-targets, compound 3b was screened against a customized panel of 50 representative proteins. 50 In this assay, 3b exhibited a very clean profile showing only a weak interaction with prostaglandin F receptor, which was considered not relevant due to the high test concentration of 10 µM (see SI Table S2).…”

Section: Resultsmentioning

confidence: 99%

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

et al. 2020

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Pharmacological modulation of cannabinoid type 2 receptor (CB2R) holds promise for the treatment of numerous conditions, including inflammatory diseases, autoimmune disorders, pain, and cancer. Despite the significance of this receptor, researchers lack reliable tools to address questions concerning the expression and complex mechanism of CB2R signaling, especially in cell-type and tissue-dependent context. Herein, we report for the first time a versatile ligand platform for the modular design of a collection of highly specific CB2R fluorescent probes, used successfully across applications, species and cell types. These include flow cytometry of endogenously expressing cells, real-time confocal microscopy of mouse splenocytes and human macrophages, as well as FRET-based kinetic and equilibrium binding assays. High CB2R specificity was demonstrated by competition experiments in living cells expressing CB2R at native levels. The probes were effectively applied to FACS analysis of microglial cells derived from a mouse model relevant to Alzheimer's disease and to the detection of CB2R in human breast cancer cells.

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Section: Resultsmentioning

confidence: 99%

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

et al. 2020

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“…For in vitro profiling, EOS789 was evaluated at 10 mmol/l in a total of 136 in vitro pharmacological assays in BioPrint panels (Eurofins Panlabs, Inc. St. Charles, MO). 21 The only targets that EOS789 inhibited >50% were the sigma and L type Ca 2þ channels (data not shown). The in vitro transporter uptake assays also indicated that EOS789 is a multi-, but selective, inhibitor against sodium-dependent phosphate transporters (Table 1 and Supplementary Table S1).…”

Section: Discussionmentioning

confidence: 96%

EOS789, a novel pan-phosphate transporter inhibitor, is effective for the treatment of chronic kidney disease–mineral bone disorder

Tsuboi¹,

Ohtomo²,

Ichida³

et al. 2020

Kidney International

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Chronic kidney disease is characterized as impaired renal function along with the imbalance and dysregulation of mineral metabolism; recognized as chronic kidney diseasemineral and bone disorder. Hyperphosphatemia, characterized by altered phosphate homeostasis along with elevated fibroblast growth factor-23 and intact parathyroid hormone, is such an alteration of mineral metabolism. We discovered a novel inhibitor, EOS789, that interacts with several sodium-dependent phosphate transporters (NaPi-IIb, PiT-1, and PiT-2) known to contribute to intestinal phosphate absorption. This inhibitor dose-dependently increased the fecal phosphorus excretion rate and inversely decreased the urinary phosphorus excretion rate in normal rats, suggesting inhibition of intestinal phosphorus absorption. In rats with adenine-induced hyperphosphatemia, EOS789 markedly decreased the serum phosphate, fibroblast growth factor-23, and intact parathyroid hormone below values found in normal control rats. Notably, this pan-phosphate transporter inhibitor exhibited a more potent effect on serum phosphate than a NaPi-IIb-selective inhibitor in rats with hyperphosphatemia indicating that PiT-1 and PiT-2 play important roles in intestinal phosphate absorption. Moreover, in a long-term study, EOS789 sustained the suppression of serum phosphorus in parallel with fibroblast growth factor-23 and intact parathyroid hormone and ameliorated ectopic calcification of the thoracic aorta. Additionally, EOS789 treatment also ameliorated kidney deterioration in rats with progressive kidney injury, probably due to the strict phosphate control. Thus, EOS789 has potent efficacy against hyperphosphatemia and its complications and could provide a significant benefit to patients who are ineffectively treated with phosphate binders.

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“…To identify potential off-targets, compound 3b was screened against a customized panel of 50 representative proteins, including classical cannabinoid targets PPARγ and 5-HT 1A . 53 In this assay, 3b exhibited very clean profile, showing only a weak interaction with prostaglandin F receptor, which was considered not relevant due to the high test concentration of 10 μM (see SI Table S2).…”

Section: ■ Results and Discussionmentioning

confidence: 98%

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

Sarott¹,

Westphal²,

Pfaff³

et al. 2020

Preprint

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Pharmacological modulation of cannabinoid type 2 receptor (CB2R) holds promise for the treatment of numerous conditions, including inflammatory diseases, autoimmune disorders, pain, and cancer. Despite the significance of this receptor, researchers lack reliable tools to address questions concerning the expression and complex mechanism of CB2R signaling, especially in cell-type and tissue-dependent context. Herein, we report for the first time a versatile ligand platform for the modular design of a collection of highly specific CB2R fluorescent probes, used successfully across applications, species and cell types. These include flow cytometry of endogenously expressing cells, real-time confocal microscopy of mouse splenocytes and human macrophages, as well as FRET-based kinetic and equilibrium binding assays. High CB2R specificity was demonstrated by competition experiments in living cells expressing CB2R at native levels. The probes were effectively applied to FACS analysis of microglial cells derived from a mouse model relevant to Alzheimer’s disease and to the detection of CB2R in human breast cancer cells.

show abstract

Safety screening in early drug discovery: An optimized assay panel

Cited by 52 publications

References 28 publications

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

EOS789, a novel pan-phosphate transporter inhibitor, is effective for the treatment of chronic kidney disease–mineral bone disorder

Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells

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