Verapamil block of T-type calcium channels

Mol Pharmacol. 2011 Mar;79(3):411-9. doi: 10.1124/mol.110.069492. Epub 2010 Dec 13.

Abstract

Verapamil is a prototypical phenylalkylamine (PAA), and it was the first calcium channel blocker to be used clinically. It tonically blocks L-type channels in the inner pore with micromolar affinity, and its affinity increases at depolarized membrane potentials. In T-type calcium channels, verapamil blocks with micromolar affinity and has modestly increased affinity at depolarized potentials. We found that a related PAA, 4-desmethoxyverapamil (D888), is comparable with verapamil both in affinity and in state-dependence. Permanently charged verapamil was more effective intracellularly than neutral verapamil. Charged PAAs were able to access their binding site from both inside and outside the cell. Furthermore, membrane-impermeant [2-(trimethylammonium)ethyl]methanethiosulfonate was able to access the inner pore from outside of the cell. We examined a homology model of the T-type calcium channel to look for possible routes of drug entry. Mutation of L1825W produced a channel that was blocked significantly more slowly by charged verapamil from the outside, with an increase in apparent affinity when the drug was applied from the inside. Data suggest that T-type channels have a back pathway through which charged drugs can access the inner pore of the channel without passing through the plasma membrane.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Calcium Channel Blockers / pharmacology*
  • Calcium Channels, T-Type / chemistry
  • Calcium Channels, T-Type / drug effects*
  • Dose-Response Relationship, Drug
  • Extracellular Space / drug effects
  • HEK293 Cells
  • Humans
  • Membrane Potentials / drug effects
  • Mutagenesis, Site-Directed
  • Protein Structure, Tertiary
  • Verapamil / analogs & derivatives
  • Verapamil / pharmacology*

Substances

  • Calcium Channel Blockers
  • Calcium Channels, T-Type
  • 4-desmethoxyverapamil
  • Verapamil