Synthesis and biological evaluation of [11C]SB366791: A new PET-radioligand for in vivo imaging of the TRPV1 receptor

Veghel, Daisy van; Cleynhens, Jan; Pearce, Larry V.; Blumberg, Peter M.; Laere, Koen Van; Verbruggen, Alfons; Bormans, Guy

doi:10.1016/j.nucmedbio.2012.08.011

Cited by 12 publications

(32 citation statements)

References 35 publications

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“…Tet-On induced CHO-hTRPV1 cells were cultured in maintaining medium (F12 supplemented with 10% TET-free FBS (Atlanta Biologicals, GA), 25 mM HEPES, 10 µg/mL blasticidin and 250 µg/mL geneticin (all from Invitrogen Life Sciences; Grand Island, NY)). TRPV1 protein was induced with induction medium (F12 supplemented with 10% FBS, 25 mM HEPES, and 1 µg/mL tetracycline) as described below for ligand binding and Ca 2+ uptake measurements [34]. …”

Section: Methodsmentioning

confidence: 99%

“…Assay conditions were as previously described [34]. CHO-hTRPV1 cells were plated in 24-well plates, reaching 40 to 60% confluence in maintaining medium after 24 h. The cells were washed once with Dulbecco’s phosphate-buffered saline (DPBS; Invitrogen) to remove antibiotics, and fresh medium with tetracycline (inducing medium) was added to induce TRPV1 expression.…”

Section: Methodsmentioning

confidence: 99%

“…Experiments were done approximately 24 h after induction. The cells were at least 90% confluent at the time of the assays [34]. …”

Section: Methodsmentioning

confidence: 99%

“…For Ca 2+ uptake assays [34] testing agonists, the inducing medium was aspirated and replaced by DMEM supplemented with bovine serum albumin (BSA, 0.25 mg/mL), Ca 2+ (37 kBq/mL) and 100 µL of increasing concentrations of the non-radioactive ligand for a total volume of 400 µL/well. The cells were incubated for 5 min in a water bath at 37°C.…”

Section: Methodsmentioning

confidence: 99%

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Multi-Functional Diarylurea Small Molecule Inhibitors of TRPV1 with Therapeutic Potential for Neuroinflammation

Feng

Pearce

Zhang

et al. 2016

AAPS J

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Transient receptor potential vanilloid type 1 (TRPV1), a heat-sensitive calcium channel protein, contributes to inflammation as well as to acute and persistent pain. Since TRPV1 occupies a central position in pathways of neuronal inflammatory signaling, it represents a highly attractive potential therapeutic target for neuroinflammation. In the present work, we have in silico identified a series of diarylurea analogues for hTRPV1, of which 11 compounds showed activity in the nanomolar to micromolar range as validated by in vitro biological assays. Then, we utilized molecular docking to explore the detailed interactions between TRPV1 and the compounds to understand the contributions of the different substituent groups. Tyr511, Leu518, Leu547, Thr550, Asn551, Arg557, and Leu670 were important for the recognition of the small molecules by TRPV1. A hydrophobic group in R2 or a polar/hydrophilic group in R1 contributed significantly to the activities of the antagonists at TRPV1. In addition, the subtle different binding pose of meta-chloro in place of para-fluoro in the R2 group converted antagonism into partial agonism, as was predicted by our short-term molecular dynamics (MD) simulation and validated by bioassay. Importantly, compound 15, one of our best TRPV1 inhibitors, also showed potential binding affinity (1.39 µM) at cannabinoid receptor 2 (CB2), which is another attractive target for immune-inflammation diseases. Furthermore, compound 1 and its diarylurea analogues were predicted to target the C-X-C chemokine receptor 2 (CXCR2), although bioassay validation of CXCR2 with these compounds still needs to be performed. This prediction from the modeling is of interest, since CXCR2 is also a potential therapeutic target for chronic inflammatory diseases. Our findings provide novel strategies to develop a small molecule inhibitor to simultaneously target two or more inflammation-related proteins for the treatment of a wide range of inflammatory disorders including neuroinflammation and neurodegenerative diseases with potential synergistic effect.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Experiments were done approximately 24 h after induction. The cells were at least 90% confluent at the time of the assays [34]. …”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Multi-Functional Diarylurea Small Molecule Inhibitors of TRPV1 with Therapeutic Potential for Neuroinflammation

Feng

Pearce

Zhang

et al. 2016

AAPS J

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“…TRPV1 protein was induced with induction medium (F12 supplemented with 10% FBS, 25 mM HEPES, and 1 μ g/mL tetracycline) as described below for ligand binding and Ca uptake measurements. 45 …”

Section: Binding Competition Assay and Ca2+ Uptake Assaymentioning

confidence: 99%

Structural Insight into Tetrameric hTRPV1 from Homology Modeling, Molecular Docking, Molecular Dynamics Simulation, Virtual Screening, and Bioassay Validations

Feng¹,

Pearce

Xu³

et al. 2015

J. Chem. Inf. Model.

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The transient receptor potential vanilloid type 1 (TRPV1) is a heat-activated cation channel protein, which contributes to inflammation, acute and persistent pain. Antagonists of human TRPV1 (hTRPV1) represent a novel therapeutic approach for the treatment of pain. Developing various antagonists of hTRPV1, however, has been hindered by the unavailability of a 3D structure of hTRPV1. Recently, the 3D structures of rat TRPV1 (rTRPV1) in the presence and absence of ligand have been reported as determined by cryo-EM. rTRPV1 shares 85.7% sequence identity with hTRPV1. In the present work, we constructed and reported the 3D homology tetramer model of hTRPV1 based on the cryo-EM structures of rTRPV1. Molecular dynamics (MD) simulations, energy minimizations, and prescreen were applied to select and validate the best model of hTRPV1. The predicted binding pocket of hTRPV1 consists of two adjacent monomers subunits, which were congruent with the experimental rTRPV1 data and the cyro-EM structures of rTRPV1. The detailed interactions between hTRPV1 and its antagonists or agonists were characterized by molecular docking, which helped us to identify the important residues. Conformational changes of hTRPV1 upon antagonist/agonist binding were also explored by MD simulation. The different movements of compounds led to the different conformational changes of monomers in hTRPV1, indicating that TRPV1 works in a concerted way, resembling some other channel proteins such as aquaporins. We observed that the selective filter was open when hTRPV1 bound with an agonist during MD simulation. For the lower gate of hTRPV1, we observed large similarities between hTRPV1 bound with antagonist and with agonist. A five-point pharmacophore model based on several antagonists was established, and the structural model was used to screen in silico for new antagonists for hTRPV1. By using the 3D TRPV1 structural model above, the pilot in silico screening has begun to yield promising hits with activity as hTRPV1 antagonists, several of which showed substantial potency.

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PET Imaging of the Endocannabinoid System

Terry

Raymont

Horti

2020

PET and SPECT of Neurobiological Systems

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Synthesis and biological evaluation of [11C]SB366791: A new PET-radioligand for in vivo imaging of the TRPV1 receptor

Abstract: [¹¹C] SB366791 has favourable biodistribution characteristics in mice. However the obtained low binding affinity for TRPV1 may not be sufficient to use the current compound as PET tracer.

Cited by 12 publications

References 35 publications

Multi-Functional Diarylurea Small Molecule Inhibitors of TRPV1 with Therapeutic Potential for Neuroinflammation

Multi-Functional Diarylurea Small Molecule Inhibitors of TRPV1 with Therapeutic Potential for Neuroinflammation

Structural Insight into Tetrameric hTRPV1 from Homology Modeling, Molecular Docking, Molecular Dynamics Simulation, Virtual Screening, and Bioassay Validations

PET Imaging of the Endocannabinoid System

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