title: Evidence for a multivalent interaction of symmetrical, N-linked, lidocaine dimers with voltage-gated Na+ channels.
abstract: The interaction of symmetrical lidocaine dimers with voltage-gated Na+ channels (VGSCs) was examined using a FLIPR membrane potential assay and voltage-clamp. The dimers, in which the tertiary amines of the lidocaine moieties are linked by an alkylene chain (two to six methylene units), inhibited VGSC activator-evoked depolarization of cells heterologously-expressing rat (r) Na(v)1.2a, human (h) Na(v)1.5, and rNa(v)1.8, with potencies 10- to 100-fold higher than lidocaine (compound 1). The rank order of potency (C4 (compound 4) > C3 (compound 3) > or = C2 (compound 2) = C5 (compound 5) = C6 (compound 6) >> compound 1) was similar at each VGSC. Compound 4 exhibited strong use-dependent inhibition of hNa(v)1.5 with pIC50 values < 4.5 and 6.0 for tonic and phasic block, respectively. Coincubation with local anesthetics but not tetrodotoxin attenuated compound 4-mediated inhibition of hNa(v)1.5. These data suggest that the compound 4 binding site(s) is identical, or allosterically coupled, to the local anesthetic receptor. The dissociation rate of the dimers from hNa(v)1.5 was dependent upon the linker length, with a rank order of compound 1 > compound 5 = compound 6 > compound 2 >> compound 3. The observation that both the potency and dissociation rate of the dimers was dependent upon linker length is consistent with a multivalent interaction at VGSCs. hNa(v)1.5 VGSCs did not recover from inhibition by compound 4. However, "chase" with free local anesthetic site inhibitors increased the rate of dissociation of compound 4. Together, these data support the hypothesis that compound 4 simultaneously occupies two binding sites on VGSCs, both of which can be bound by known local anesthetic site inhibitors.
authors.affiliation: Theravance Inc., 901 Gateway Blvd., South San Francisco, CA 94080, USA. jsmith@theravance.com
authors.forename: J A M
authors.initials: JA
authors.lastname: Smith
authors.forename: S M
authors.initials: SM
authors.lastname: Amagasu
authors.forename: J
authors.initials: J
authors.lastname: Hembrador
authors.forename: S
authors.initials: S
authors.lastname: Axt
authors.forename: R
authors.initials: R
authors.lastname: Chang
authors.forename: T
authors.initials: T
authors.lastname: Church
authors.forename: C
authors.initials: C
authors.lastname: Gee
authors.forename: J R
authors.initials: JR
authors.lastname: Jacobsen
authors.forename: T
authors.initials: T
authors.lastname: Jenkins
authors.forename: E
authors.initials: E
authors.lastname: Kaufman
authors.forename: N
authors.initials: N
authors.lastname: Mai
authors.forename: R G
authors.initials: RG
authors.lastname: Vickery
mesh_terms: D000779:Anesthetics, Local / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D001681:Biological Assay; D002462:Cell Membrane / Q000187:drug effects*; D002478:Cells, Cultured; D019281:Dimerization; D005727:Ganglia, Spinal / Q000187:drug effects; D006801:Humans; D008012:Lidocaine / Q000737:chemistry / Q000494:pharmacology*; D008564:Membrane Potentials; D051381:Rats; D026941:Sodium Channel Blockers / Q000737:chemistry / Q000494:pharmacology*; D015222:Sodium Channels / Q000187:drug effects*; D013779:Tetrodotoxin / Q000494:pharmacology
chemical_list: D000779:Anesthetics, Local; D026941:Sodium Channel Blockers; D015222:Sodium Channels; D013779:Tetrodotoxin; D008012:Lidocaine