Synthesis, SAR, Crystal Structure, and Biological Evaluation of Benzoquinoliziniums as Activators of Wild-Type and Mutant Cystic Fibrosis Transmembrane Conductance Regulator Channels

Marivingt-Mounir, Cécile; Norez, Caroline; Dérand, Renaud; Bulteau-Pignoux, Laurence; Dung, Nguyen‐Huy; Viossat, Β.; Morgant, Georges; Becq, Frédéric; Vierfond, Jean‐Michel; Mettey, Yvette

doi:10.1021/jm0308848

Cited by 50 publications

(74 citation statements)

References 39 publications

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“…5A), highlighting the important role of cAMP in the activation of the water channel. To confirm the key role of cAMP in the interaction between CFTR protein and water channels, we also used MPB-91, a benzo[c]quinolizinium compound activating the chloride channel activity of CFTR through a cAMP-independent mechanism (Becq et al, 1999;Marivingt-Mounir et al, 2004). Similarly to genistein, a transient application of MPB-91 (50-100 mM; 5 minutes) was unable to The bar chart shows the significant difference between different populations of CHO cells (*P,0.05, ***P,0.005, unpaired t-test) after application of forskolin (1 mM, 5 minutes).…”

Section: Resultsmentioning

confidence: 99%

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The human CFTR protein expressed in CHO cells activates an aquaporin 3 in a cAMP dependent pathway: study by Digital Holographic Microscopy

Jourdain

Becq

Lengacher

et al. 2013

Journal of Cell Science

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The transmembrane water movements during cellular processes and their relationship to ionic channel activity remain largely unknown. As an example, in epithelial cells it was proposed that the movement of water could be directly linked to cystic fibrosis transmembrane conductance regulator (CFTR) protein activity through a cAMP-stimulated aqueous pore, or be dependent on aquaporin. Here, we used digital holographic microscopy (DHM) an interferometric technique to quantify in situ the transmembrane water fluxes during the activity of the epithelial chloride channel, CFTR, measured by patch-clamp and iodide efflux techniques. We showed that the water transport measured by DHM is fully inhibited by the selective CFTR blocker CFTR inh172 and is absent in cells lacking CFTR. Of note, in cells expressing the mutated version of CFTR (F508del-CFTR), which mimics the most common genetic alteration encountered in cystic fibrosis, we also show that the water movement is profoundly altered but restored by pharmacological manipulation of F508del-CFTRdefective trafficking. Importantly, whereas activation of this endogenous water channel required a cAMP-dependent stimulation of CFTR, activation of CFTR or F508del-CFTR by two cAMP-independent CFTR activators, genistein and MPB91, failed to trigger water movements. Finally, using a specific smallinterfering RNA against the endogenous aquaporin AQP3, the water transport accompanying CFTR activity decreased. We conclude that water fluxes accompanying CFTR activity are linked to AQP3 but not to a cAMP-stimulated aqueous pore in the CFTR protein.

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Section: Resultsmentioning

confidence: 99%

“…The CFTR activator benzo[c]quinolizinium compound MPB-91 (5-butyl-10-chloro-6-hydroxybenzo[c]quinolizinium chloride) was synthesised in our laboratory as previously described (Marivingt-Mounir et al, 2004). Forskolin was purchased from LC Laboratories (PKC Pharmaceuticals, Woburn, MA).…”

Section: Iodide Efflux Assaysmentioning

confidence: 99%

The human CFTR protein expressed in CHO cells activates an aquaporin 3 in a cAMP dependent pathway: study by Digital Holographic Microscopy

Jourdain

Becq

Lengacher

et al. 2013

Journal of Cell Science

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“…MPB-07 and -91 are good specific CFTR activators, whereas MPB-80 is a very poor CFTR activator but a useful control [6,7,21]. Unlike MPB-80, MPB-07 and MPB-91 induced strong vasorelaxation in IPA rings whether endothelium-denuded or not (figs 5 and 6b and figs 5 and 6a, respectively).…”

Section: Role Of Cftr In Intrapulmonary Arterial Vasorelaxationmentioning

confidence: 98%

“…As determined in preliminary experiments, tissues were set at optimal length by equilibration against a passive load of 0. 3-300 mM, prepared as described previously [21]) by constructing a cumulative concentration-response curve. Some experiments were performed in endothelium-denuded rings.…”

Section: Isometric Tension Measurementsmentioning

confidence: 99%

Expression and function of cystic fibrosis transmembrane conductance regulator in rat intrapulmonary arteries

Robert¹,

Savineau²,

Norez³

et al. 2007

Eur Respir J

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The cystic fibrosis transmembrane conductance regulator (CFTR) gene encodes a cyclic adenosine monophosphate (cAMP)-dependent chloride channel located mainly at the apical membrane of epithelial cells. In myocytes of pulmonary arteries, numerous chloride channels have been identified and described, but not the CFTR. Thus the presence and function of the CFTR was investigated in rat intrapulmonary arteries.CFTR expression, localisation and function were analysed in cultured smooth muscle cells using Reverse transcriptase (RT)-PCR and immunoprecipitation followed by protein kinase A phosphorylation, immunolocalisation and an iodide efflux assay, respectively. The role of the CFTR in pulmonary vasoreactivity was determined in arterial rings using an organ bath system.RT-PCR and immunoprecipitation analyses, as well as the immunolocalisation study, revealed the expression of CFTR gene transcripts and protein. The iodide efflux assay showed the existence of functional cAMP-, calcium-and volume-dependent chloride channels. Furthermore, the following effects were found: 1) inhibition of forskolin/genistein-activated iodide efflux by glibenclamide, diphenylamine-2-carboxylic acid and CFTR-specific inhibitor (CFTRinh)-172; 2) activation of iodide efflux by the benzoquinolizinium derivative CFTR activators MPB-07 and MPB-91; and 3) inhibition of MPB-dependent efflux by CFTRinh-172. Finally, CFTR activators induced concentration-dependent vasorelaxation in rings preconstricted with phenylephrine, in the presence or absence of endothelium.The present results are the first to reveal functional cyclic adenosine monophosphate-regulated cystic fibrosis transmembrane conductance regulator contributing to endothelium-independent vasorelaxation in rat intrapulmonary arterial myocytes.

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“…Cells were cultured in multiwell plates in order to perform parallel experiments and comparison analysis using a robot-based system (MultiProbe II , PerkinElmer Life Sciences, Courtaboeuf, France) adapted to the iodide efflux method [20,21]. The cells were washed with an efflux buffer containing (in mM) the following: 136.9 NaCl, 5.4 KCl, 0.3 KH 2 PO 4 , 0.3 NaH 2 PO 4 , 4.2 NaHCO 3 , 2 CaCl 2 , 0.5 MgCl 2 , 0.4 MgSO 4 , 5.6 glucose and 10 HEPES, pH 7.4.…”

Section: Radiotracer Efflux Experimentsmentioning

confidence: 99%

Pharmacological profile of inhibition of the chloride channels activated by extracellular acid in cultured rat Sertoli cells

et al. 2006

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-Sertoli cells from mammalian testis are key cells involved in the development and maintenance of stem cell spermatogonia as well as in the secretion of a Cl − and K + -rich fluid into the lumen of seminiferous tubules. The pharmacology and contribution of Cl − channels to the physiology of Sertoli cells were investigated using whole-cell patch clamp and iodide efflux experiments applied to cultured rat Sertoli cells. We characterized an outwardly rectifying Cl − current stimulated by various acid species including the physiologically relevant lactic acid. Using the iodide efflux technique, the pharmacological properties of this Cl − current, noted ICl acid , revealed Ca 2+ -independent inhibition by DIDS (IC 50 = 27 µM), glibenclamide (IC 50 = 31 µM) and DPC (IC 50 = 86 µM). ICl acid was neither affected by calix[4]arene nor by 9-AC. The order of potency for inhibition of ICl acid is DIDS ≈ glibenclamide > DPC >> calix[4]arene, 9-AC. For comparison, the inhibitory profile of the swelling-and ATP-activated Cl − currents in Sertoli cells is DPC = DIDS >> glibenclamide = 9-AC for ICl swell and DPC = 9-AC = DIDS >> glibenclamide for ICl ATP . This description provides new insights into the physiology and pharmacology of the endogenous Cl − channels expressed and potentially involved in fluid secretion in Sertoli cells.Cl − channels / pharmacology / proton-activation / ATP / cell volume / glibenclamide

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Synthesis, SAR, Crystal Structure, and Biological Evaluation of Benzoquinoliziniums as Activators of Wild-Type and Mutant Cystic Fibrosis Transmembrane Conductance Regulator Channels

Cited by 50 publications

References 39 publications

The human CFTR protein expressed in CHO cells activates an aquaporin 3 in a cAMP dependent pathway: study by Digital Holographic Microscopy

The human CFTR protein expressed in CHO cells activates an aquaporin 3 in a cAMP dependent pathway: study by Digital Holographic Microscopy

Expression and function of cystic fibrosis transmembrane conductance regulator in rat intrapulmonary arteries

Pharmacological profile of inhibition of the chloride channels activated by extracellular acid in cultured rat Sertoli cells

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