ABCMdb

PMID: 15722457

Pedemonte N, Sonawane ND, Taddei A, Hu J, Zegarra-Moran O, Suen YF, Robins LI, Dicus CW, Willenbring D, Nantz MH, Kurth MJ, Galietta LJ, Verkman AS
Phenylglycine and sulfonamide correctors of defective delta F508 and G551D cystic fibrosis transmembrane conductance regulator chloride-channel gating.
Mol Pharmacol. 2005 May;67(5):1797-807. Epub 2005 Feb 18., [PubMed]

Sentences

No. Mutations Sentence Comment

3 ABCC7 p.Gly551Asp The ⌬F508 mutation produces defects in channel gating and cellular processing, whereas the G551D mutation produces primarily a gating defect. Login to comment 5 ABCC7 p.Gly551Asp Secondary analysis and testing of Ͼ1000 structural analogs yielded two novel classes of correctors of defective ⌬F508-CFTR gating ("potentiators") with nanomolar potency that were active in human ⌬F508 and G551D cells. Login to comment 6 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp The most potent compound of the phenylglycine class, 2-[(2-1H-indol-3-yl-acetyl)-methylamino]-N-(4-isopro- pylphenyl)-2-phenylacetamide, reversibly activated ⌬F508-CFTR in the presence of forskolin with Ka ϳ 70 nM and also activated the CFTR gating mutants G551D and G1349D with Ka values of ϳ1100 and 40 nM, respectively. Login to comment 28 ABCC7 p.Gly551Asp The most common of the CFTR gating mutants is G551D, with a worldwide frequency of 3.1% among CF chromosomes (Hamosh et al., 1992), although people of Celtic descent have frequencies as high as 8% (Cashman et al., 1995). Login to comment 32 ABCC7 p.Gly551Asp Potentiators may also be useful as monotherapy for CF caused by gating mutants of CFTR such as G551D. Login to comment 35 ABCC7 p.Gly551Asp Flavones at high concentrations also are able to correct defective gating in G551D-CFTR (Illek et al., 1999; Zegarra-Moran et al., 2002). Login to comment 38 ABCC7 p.Gly551Asp However, activation required high concentrations of cAMP agonists, and the benzothiophenes did not activate CFTR gating mutants such as G551D. Login to comment 41 ABCC7 p.Gly551Asp These compounds were potent in ⌬F508-CFTR-transfected and natively expressing human cells, active in the presence of relatively low concentrations of cAMP agonists, and active against multiple CFTR gating mutants, including G551D. Login to comment 50 ABCC7 p.Gly551Asp Some measurements were done using stably transfected FRT cells expressing YFP-H148Q and wild-type or G551D-CFTR (Galietta et al., 2001b). Login to comment 188 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Gly1349Asp Activation of G551Dand G1349D-CFTR mutants. Login to comment 189 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Gly1349Asp A and B, stimulation of apical membrane Cl- current by genistein (top) and PG-01 (bottom) in G551Dand G1349D-CFTR-expressing FRT cells. Login to comment 191 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Gly1349Asp C and D, dose-responses for the PG-01 and genistein for activation of G551Dand G1349D-CFTR (S.E., n ϭ 4). Login to comment 217 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp Measurements were done in the "class III" mutants G551D and G1349D, which produce a severe gating defect without impairment in protein trafficking (Gregory et al., 1991). Login to comment 222 ABCC7 p.Gly551Asp B, G551D-CFTR cells. Login to comment 223 ABCC7 p.Asp1152His C, D1152H-CFTR cells. Login to comment 225 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp The G551D and G1349D mutant CFTRs produced little Cl- current after the addition of maximal forskolin (Fig. 6, A and B). Login to comment 226 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Gly1349Asp Genistein, a known activator of G551Dand G1349D-CFTR, increased Cl- cur- rent substantially, albeit at high micromolar concentrations (Fig. 6, A and B, top curves). Login to comment 227 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Gly1349Asp PG-01 produced large currents in both G551Dand G1349D-CFTR-expressing cells as shown in Fig. 6, A and B (bottom curves) and summarized in Fig. 6, C and D. Login to comment 229 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Gly1349Asp In contrast, PG-01, SF-01, and the benzothiophene ⌬F508act-02 did not increase Cl- currents in G551Dand G1349D-CFTR-expressing cells (data not shown). Login to comment 240 ABCC7 p.Gly551Asp Nasal epithelial cells from a subject with the G551D mutation (Zegarra-Moran et al., 2002) showed a large response to PG-01 after forskolin stimulation (Fig. 7B). Login to comment 241 ABCC7 p.Asp1152His Cells were also tested from a subject having D1152H and ⌬F508 CFTR mutations, with the former mutation affecting the second nucleotide-binding domain and causing a decrease in channel activity (Vankeerberghen et al., 1998). Login to comment 242 ABCC7 p.Asp1152His The D1152H/⌬F508 cells maintained at 37°C showed large CFTR currents in response to PG-01 (Fig. 7C). Login to comment 283 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp ABCC7 p.Asp1152His The phenyglycines corrected defective gating in a number of CF-causing CFTR mutants including ⌬F508, G551D, G1349D, and D1152H. Login to comment 284 ABCC7 p.Gly551Asp ABCC7 p.Gly1349Asp G551D and G1349D affect critical glycine residues in nucleotide binding domains 1 and 2 of CFTR, respectively (Hyde et al., 1990), producing a severe gating defect (Gregory et al., 1991; Logan et al., 1994; Derand et al., 2002; Zegarra-Moran et al., 2002). Login to comment 286 ABCC7 p.Gly551Asp The apparent Kd for PG-01 for G551D-CFTR activation was ϳ1 ␮M, approximately 100-fold better than that of genistein. Login to comment 287 ABCC7 p.Gly1349Asp The potency for activation G1349D-CFTR by PG-01 was even better at ϳ40 nM. Login to comment 294 ABCC7 p.Gly551Asp ABCC7 p.Asp1152His The best phenylglycine was also effective on cells cultured from subjects with CF having G551D and D1152H CFTR mutations, supporting the possible use of this class of compounds for monotherapy of CF caused by some mutations. Login to comment