Discovery of α-Aminoazaheterocycle-Methylglyoxal Adducts as a New Class of High-Affinity Inhibitors of Cystic Fibrosis Transmembrane Conductance Regulator Chloride Channels

Routaboul, C.; Norez, Caroline; Melin, Patricia; Molina, M.-C.; Boucherle, Benjamin; Bossard, Florian; Noël, Sabrina; Robert, Renaud; Gauthier, Chantal; Becq, Frédéric; Décout, Jean‐Luc

doi:10.1124/jpet.107.123307

Cited by 15 publications

(31 citation statements)

References 41 publications

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“…The cell viability of HeLa-F508del-CFTR cells exposed to different concentrations of compounds was evaluated using the mitochondrial-dependent reduction of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Sigma-Aldrich] colorimetric assay, as described elsewhere (Routaboul et al, 2007). Cells were incubated at 37°C with the test compound at the concentration and time of incubation previously defined.…”

Section: Methodsmentioning

confidence: 99%

Searching for Combinations of Small-Molecule Correctors to Restore F508del–Cystic Fibrosis Transmembrane Conductance Regulator Function and Processing

Boinot

Souchet

Ferru-Clément

et al. 2014

J Pharmacol Exp Ther

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The mutated protein F508del-cystic fibrosis transmembrane conductance regulator (CFTR) failed to traffic properly as a result of its retention in the endoplasmic reticulum and functions as a chloride (Cl 2 ) channel with abnormal gating and endocytosis. Small chemicals (called correctors) individually restore F508del-CFTR trafficking and Cl 2 transport function, but recent findings indicate that synergistic pharmacology should be considered to address CFTR defects more clearly. We studied the function and maturation of F508del-CFTR expressed in HeLa cells using a combination of five correctors [miglustat,, and suberoylamilide hydroxamic acid (SAHA)]. Using the whole-cell patch-clamp technique, the current density recorded in response to CFTR activators (forskolin 1 genistein) was significantly increased in the presence of the following combinations: VX-809 1 IsoLAB; VX-809 1 miglustat 1 SAHA; VX-809 1 miglustat 1 IsoLAB; VX-809 1 IsoLAB 1 SAHA; VX-809 1 miglustat 1 IsoLAB 1 SAHA. These combinations restored the activity of F508del-CFTR but with a differential effect on the appearance of mature c-band of F508del-CFTR proteins. Focusing on the VX-809 1 IsoLAB cocktail, we recorded a level of correction higher at 37°C versus room temperature, but without amelioration of the thermal instability of CFTR. The level of functional rescue with VX-809 1 IsoLAB after 4 hours of incubation was maximal and similar to that obtained in optimal conditions of use for each compound (i.e., 24 hours for VX-809 1 4 hours for IsoLAB).

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Section: Methodsmentioning

confidence: 99%

Searching for Combinations of Small-Molecule Correctors to Restore F508del–Cystic Fibrosis Transmembrane Conductance Regulator Function and Processing

Boinot

Souchet

Ferru-Clément

et al. 2014

J Pharmacol Exp Ther

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“…Similar experiments were performed with the minor isomer GPact-11b leading to an EC50 of 2.9¡0.9 mM (data not shown, n54). The effect of GPact-11a was fully inhibited by CFTR inh -172, GlyH-101 and GPinh-5a, three selective CFTR inhibitors [13,26,27] but was not affected by DIDS and calixarene, two non-CFTR inhibitors [28] (fig. 1e).…”

Section: Effect Of Gpact-11a On Wt-cftr Activitymentioning

confidence: 99%

“…GPinh-5a was synthesised as previously described [13]. Miglustat was from Toronto research chemicals (Toronto, ON, Canada).…”

Section: Other Chemicalsmentioning

confidence: 99%

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Identification of a novel water-soluble activator of wild-type and F508del CFTR: GPact-11a

Bertrand¹,

Boucherle²,

Billet³

et al. 2010

European Respiratory Journal

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One of the major therapeutic strategy in cystic fibrosis aims at developing modulators of cystic fibrosis transmembrane conductance regulator (CFTR) channels. We recently discovered methylglyoxal a-aminoazaheterocycle adducts, as a new family of CFTR inhibitors. In a structure-activity relationship study, we have now identified GPact-11a, a compound able not to inhibit but to activate CFTR.Here, we present the effect of GPact-11a on CFTR activity using in vitro (iodide efflux, fluorescence imaging and patch-clamp recordings), ex vivo (short-circuit current measurements) and in vivo (salivary secretion) experiments.We report that GPact-11a: 1) is an activator of CFTR in several airway epithelial cell lines; 2) activates rescued F508del-CFTR in nasal, tracheal, bronchial, pancreatic cell lines and in human CF ciliated epithelial cells, freshly dissociated from lung samples; 3) stimulates ex vivo the colonic chloride secretion and increases in vivo the salivary secretion in cftr +/+ but not cftr -/-mice; and 4) is selective for CFTR because its effect is inhibited by CFTR inh -172, GlyH-101, glibenclamide and GPinh-5a. To conclude, this work identifies a selective activator of wild-type and rescued F508del-CFTR. This nontoxic and water-soluble agent represents a good candidate, alone or in combination with a F508del-CFTR corrector, for the development of a CFTR modulator in cystic fibrosis.

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“…The glycine hydrazides block the CFTR pore at its external surface, which allowed the synthesis of nonabsorbable polyethylene glycol and lectin adducts that do not cross membranes and block intestinal fluid secre-tion in animals models of cholera (Sonawane et al, 2006(Sonawane et al, , 2007. Routaboul et al (2007) recently reported that ␣-aminoazaheterocyclic-methylglyoxal adducts inhibit CFTR chloride channel function completely and reversibly, with low picomolar IC 50 values, more than 10,000-fold better than the most potent known CFTR inhibitors. The picomolar-potency compounds were identified by screening a collection of seven ␣-aminoazaheterocyclic-methylglyoxal adducts.…”

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confidence: 99%

α-Aminoazaheterocyclic-Methylglyoxal Adducts Do Not Inhibit Cystic Fibrosis Transmembrane Conductance Regulator Chloride Channel Activity

Sonawane

Zegarra‐Moran

Namkung

et al. 2008

J Pharmacol Exp Ther

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Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel have potential applications in the therapy of secretory diarrheas and polycystic kidney disease. In a recent study, several highly polar ␣-aminoazaheterocyclic-methylglyoxal adducts were reported to reversibly inhibit CFTR chloride channel activity with IC 50 values in the low picomolar range (J Pharmacol Exp Ther 322:1023-1035, 2007), more than 10,000-fold better than that of thiazolidinone and glycine hydrazide CFTR inhibitors previously identified by highthroughput screening. In this study, we resynthesized and evaluated the ␣-aminoazaheterocyclic-methylglyoxal adducts reported to have high CFTR inhibition potency (compounds 5, 7, and 8). We verified that the reported synthesis procedures produced the target compounds in high yield. However, we found that these compounds did not inhibit CFTR chloride channel function in multiple cell lines at up to 100 M concentration, using three independent assays of CFTR function including short-circuit current analysis, whole-cell patch-clamp experiments, and yellow fluorescence protein-fluorescence quenching. As positive controls, approximately 100% of CFTR inhibition was found by thiazolidinone and glycine hydrazide CFTR inhibitors. Our data provide direct evidence against CFTR inhibition by ␣-aminoazaheterocyclic-methylglyoxal adducts.

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Discovery of α-Aminoazaheterocycle-Methylglyoxal Adducts as a New Class of High-Affinity Inhibitors of Cystic Fibrosis Transmembrane Conductance Regulator Chloride Channels

Cited by 15 publications

References 41 publications

Searching for Combinations of Small-Molecule Correctors to Restore F508del–Cystic Fibrosis Transmembrane Conductance Regulator Function and Processing

Searching for Combinations of Small-Molecule Correctors to Restore F508del–Cystic Fibrosis Transmembrane Conductance Regulator Function and Processing

Identification of a novel water-soluble activator of wild-type and F508del CFTR: GPact-11a

α-Aminoazaheterocyclic-Methylglyoxal Adducts Do Not Inhibit Cystic Fibrosis Transmembrane Conductance Regulator Chloride Channel Activity

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