School of Pharmacy, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
School of Pharmacy, Xi'an Jiaotong University, Xi'an, Shaanxi, China.
J Pharm Biomed Anal. 2021 Mar 20;196:113911. doi: 10.1016/j.jpba.2021.113911. Epub 2021 Jan 19.
Local anesthetic has a wide application in clinical practice. However, angioedema, an adverse reaction caused by local anesthetics, has been reported to be related to histamine H receptor (HR). Hence, an effective and practical method for investigating the interaction characteristics between local anesthetics and HR is needed. In this work, the competition binding assay and the relative standard method based on HR-HEK293/cell membrane chromatography (CMC) were developed to analyze the equilibrium dissociation constant (K) values of local anesthetics with HR. The activity of drugs toward HR was evaluated by intracellular Ca imaging assay. Molecular docking was used to verify the interaction modes that occurred at the activate pocket of HR protein. Results showed that the local anesthetics can directly occupy histamine binding sites on HR, and the K values obtained from different CMC methods exhibited positive correlations with each other (p < 0.01). The K values of tetracaine, procaine, and lidocaine were much closer to that of histamine than bupivacaine and ropivacaine. This was not only in line with the Ca responses in activating HR, but also consistent with the same amino acid residues shared with histamine in the HR active site. In conclusion, this study provided new insight into the interactions between local anesthetics and HR. The HR-HEK293/CMC methods developed in this study could be used to evaluate the interaction characteristics of those compounds acting on HR.
局部麻醉剂在临床实践中有广泛的应用。然而，局部麻醉剂引起的血管性水肿不良反应已被报道与组胺 H 受体（HR）有关。因此，需要一种有效且实用的方法来研究局部麻醉剂与 HR 之间的相互作用特征。在这项工作中，建立了基于 HR-HEK293/细胞膜色谱（CMC）的竞争结合测定和相对标准方法，以分析局部麻醉剂与 HR 的平衡解离常数（K）值。通过细胞内 Ca 成像测定评估药物对 HR 的活性。分子对接用于验证发生在 HR 蛋白活性口袋的相互作用模式。结果表明，局部麻醉剂可以直接占据 HR 上的组胺结合位点，并且从不同 CMC 方法获得的 K 值彼此呈正相关（p<0.01）。与布比卡因和罗哌卡因相比，丁卡因、普鲁卡因和利多卡因的 K 值更接近组胺。这不仅与激活 HR 时的 Ca 反应一致，而且与 HR 活性位点中与组胺共享的相同氨基酸残基一致。总之，本研究为局部麻醉剂与 HR 之间的相互作用提供了新的见解。本研究中开发的 HR-HEK293/CMC 方法可用于评估作用于 HR 的化合物的相互作用特征。
