Kinetic binding and activation profiles of endogenous tachykinins targeting the NK1 receptor

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• 作者：I. Nederpelt ; D. Bleeker ; B. Tuijt ; A.P. IJzerman ; L.H. Heitman ; ^{l.h.heitman@lacdr.leidenuniv.nl" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Binding kinetics ; NK1 receptor ; Label-free ; Association rate ; GPCR ; Tachykinin
• 刊名：Biochemical Pharmacology
• 出版年：2016
• 出版时间：15 October 2016
• 年：2016
• 卷：118
• 期：Complete
• 页码：88-95
• 全文大小：1480 K

文摘

Ligand–receptor binding kinetics (i.e. association and dissociation rates) are emerging as important parameters for drug efficacy in vivo. Awareness of the kinetic behavior of endogenous ligands is pivotal, as drugs often have to compete with those. The binding kinetics of neurokinin 1 (NK1) receptor antagonists have been widely investigated while binding kinetics of endogenous tachykinins have hardly been reported, if at all. Therefore, the aim of this research was to investigate the binding kinetics of endogenous tachykinins and derivatives thereof and their role in the activation of the NK1 receptor.

We determined the binding kinetics of seven tachykinins targeting the NK1 receptor. Dissociation rate constants (k_off) ranged from 0.026 ± 0.0029 min⁻¹ (Sar⁹,Met(O₂)¹¹-SP) to 0.21 ± 0.015 min⁻¹ (septide). Association rate constants (k_on) were more diverse: substance P (SP) associated the fastest with a k_on value of 0.24 ± 0.046 nM⁻¹ min⁻¹ while neurokinin A (NKA) had the slowest association rate constant of 0.001 ± 0.0002 nM⁻¹ min⁻¹. Kinetic binding parameters were highly correlated with potency and maximal response values determined in label-free impedance-based experiments on U-251 MG cells.

Our research demonstrates large variations in binding kinetics of tachykinins which correlate to receptor activation. These findings provide new insights into the ligand–receptor interactions of tachykinins and underline the importance of measuring binding kinetics of both drug candidates and competing endogenous ligands.