title: High affinity interaction of mibefradil with voltage-gated calcium and sodium channels.
abstract: Mibefradil is a novel Ca(2+) antagonist which blocks both high-voltage activated and low voltage-activated Ca(2+) channels. Although L-type Ca(2+) channel block was demonstrated in functional experiments its molecular interaction with the channel has not yet been studied. We therefore investigated the binding of [(3)H]-mibefradil and a series of mibefradil analogues to L-type Ca(2+) channels in different tissues. [(3)H]-Mibefradil labelled a single class of high affinity sites on skeletal muscle L-type Ca(2+) channels (K(D) of 2.5+/-0.4 nM, B(max)=56.4+/-2.3 pmol mg(-1) of protein). Mibefradil (and a series of analogues) partially inhibited (+)-[(3)H]-isradipine binding to skeletal muscle membranes but stimulated binding to brain L-type Ca(2+) channels and alpha1C-subunits expressed in tsA201 cells indicating a tissue-specific, non-competitive interaction between the dihydropyridine and mibefradil binding domain. [(3)H]-Mibefradil also labelled a heterogenous population of high affinity sites in rabbit brain which was inhibited by a series of nonspecific Ca(2+) and Na(+)-channel blockers. Mibefradil and its analogue RO40-6040 had high affinity for neuronal voltage-gated Na(+)-channels as confirmed in binding (apparent K(i) values of 17 and 1.0 nM, respectively) and functional experiments (40% use-dependent inhibition of Na(+)-channel current by 1 microM mibefradil in GH3 cells). Our data demonstrate that mibefradil binds to voltage-gated L-type Ca(2+) channels with very high affinity and is also a potent blocker of voltage-gated neuronal Na(+)-channels. More lipophilic mibefradil analogues may possess neuroprotective properties like other nonselective Ca(2+)-/Na(+)-channel blockers.
authors.affiliation: Institut für Biochemische Pharmakologie, Peter-Mayrstr. 1, A-6020 Innsbruck, Austria.
authors.forename: P
authors.initials: P
authors.lastname: Eller
authors.forename: S
authors.initials: S
authors.lastname: Berjukov
authors.forename: S
authors.initials: S
authors.lastname: Wanner
authors.forename: I
authors.initials: I
authors.lastname: Huber
authors.forename: S
authors.initials: S
authors.lastname: Hering
authors.forename: H G
authors.initials: HG
authors.lastname: Knaus
authors.forename: G
authors.initials: G
authors.lastname: Toth
authors.forename: S D
authors.initials: SD
authors.lastname: Kimball
authors.forename: J
authors.initials: J
authors.lastname: Striessnig
mesh_terms: D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000187:drug effects*; D004558:Electric Stimulation; D004594:Electrophysiology; D006168:Guinea Pigs; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects*; D020748:Mibefradil / Q000494:pharmacology*; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011817:Rabbits; D011869:Radioligand Assay; D026941:Sodium Channel Blockers
chemical_list: D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D026941:Sodium Channel Blockers; D020748:Mibefradil