Small-molecule Ca_Vα₁⋅Ca_Vβ antagonist suppresses neuronal voltage-gated calcium-channel trafficking.

Abstract

Extracellular calcium flow through neuronal voltage-gated Ca_V2.2 calcium channels converts action potential-encoded information to the release of pronociceptive neurotransmitters in the dorsal horn of the spinal cord, culminating in excitation of the postsynaptic central nociceptive neurons. The Ca_V2.2 channel is composed of a pore-forming α₁ subunit (Ca_Vα₁) that is engaged in protein-protein interactions with auxiliary α₂/δ and β subunits. The high-affinity Ca_V2.2α₁⋅Ca_Vβ₃ protein-protein interaction is essential for proper trafficking of Ca_V2.2 channels to the plasma membrane. Here, structure-based computational screening led to small molecules that disrupt the Ca_V2.2α₁⋅Ca_Vβ₃ protein-protein interaction. The binding mode of these compounds reveals that three substituents closely mimic the side chains of hot-spot residues located on the α-helix of Ca_V2.2α₁ Site-directed mutagenesis confirmed the critical nature of a salt-bridge interaction between the compounds and Ca_Vβ₃ Arg-307. In cells, compounds decreased trafficking of Ca_V2.2 channels to the plasma membrane and modulated the functions of the channel. In a rodent neuropathic pain model, the compounds suppressed pain responses. Small-molecule α-helical mimetics targeting ion channel protein-protein interactions may represent a strategy for developing nonopioid analgesia and for treatment of other neurological disorders associated with calcium-channel trafficking.