Deciphering the shape selective conformational equilibrium of E- and Z-locked azobenzene–tetraethylammonium ion in regulating photo-switchable K+-ion channel blocking

Abstract

The search for photo-switchable optopharmacological agents that can block ion channels has been a prevalent area owing to its prime advantages of reversibility and specificity over the traditional blockers. However, the quest for a higher blocking ability shown by a less stable photo-isomer to perfectly suit the requirement of the optopharmacological agents is still ongoing. To date, only a marginal improvement in terms of blocking ability is observed by the less stable E-isomer of para-substituted locked azobenzene with TEA (LAB–TEA) for the K⁺-ion channel. Thus, rationalization of the limitation for achieving high activity by the E-isomer is rather essential to aid the improvement of the efficiency of photoswitchable blocker drugs. Herein, we report a molecular-level analysis on the mechanism of blocking by E- and Z-LAB–TEA with the bacterial KcsA K⁺-ion channel using Molecular Dynamics (MD) simulation and Quantum Mechanical (QM) calculations. The positively charged TEA fragment engages in stronger electrostatic interactions, while the neutral LAB fragment engages in weaker dispersive interactions. The binding free energy calculated by Molecular Mechanics Poisson–Boltzmann Surface Area (MMPBSA) for E-LAB–TEA (−22.3 kcal mol⁻¹) shows less thermodynamic preference for binding with K⁺-ion channels than Z-LAB–TEA (−21.6 kcal mol⁻¹) corroborating the experimental observation. The correlation between the structure and the binding ability of E- and Z-isomers of LAB–TEA indicates that the channel gate is narrow and acts as a bottleneck for the entry of the binder molecule inside the large cavity. Upon irradiation, the Z-isomer converts into a less stable but long and planar E-isomer (ΔE of photoisomerism = 7.0 kcal mol⁻¹, at SA2-CASPT2(6,4)/6-31+G(d)//CASSCF(6,4)/6-31+G(d)), which is structurally more suitable to fit into the narrow channel gate rather than the curved and non-planar Z-LAB–TEA. Thus, a reduction in the ionic current is observed owing to the preferential entry and subsequent blocking by E-LAB–TEA. Discontinuing the irradiation leads to conversion to the Z-isomer, the curved nature of which hinders its spontaneous release outside the cavity, thereby contributing only a small increase in the ionic current.