Positive allosteric modulation of the cannabinoid CB1 receptor potentiates endocannabinoid signalling and changes ERK1/2 phosphorylation kinetics

IF 6.8 2区医学 Q1 PHARMACOLOGY & PHARMACY

British Journal of Pharmacology Pub Date : 2024-06-03 DOI:10.1111/bph.16433

Hayley M. Green, Jamie J. Manning, Ian R. Greig, Ruth A. Ross, David B. Finlay, Michelle Glass

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Abstract

Background and Purpose

Activation of CB₁ by exogenous agonists causes adverse effects in vivo. Positive allosteric modulation may offer improved therapeutic potential and a reduced on-target adverse effect profile compared with orthosteric agonists, due to reduced desensitisation/tolerance, but this has not been directly tested. This study investigated the ability of PAMs/ago-PAMs to induce receptor regulation pathways, including desensitisation and receptor internalisation.

Experimental Approach

Bioluminescence resonance energy transfer (BRET) assays in HEK293 cells were performed to investigate G protein dissociation, ERK1/2 phosphorylation and β-arrestin 2 translocation, while immunocytochemistry was performed to measure internalisation of CB₁ in response to the PAMs ZCZ011, GAT229 and ABD1236 alone and in combination with the orthosteric agonists AEA, 2-AG, and AMB-FUBINACA.

Key Results

ZCZ011, GAT229 and ABD1236 were allosteric agonists in all pathways tested. The ago-PAM ZCZ011 induced a biphasic ERK1/2 phosphorylation time course compared to transient activation by orthosteric agonists. In combination with 2-AG but not AEA or AMB-FUBINACA, ZCZ011 and ABD1236 caused the transient peak of ERK1/2 phosphorylation to become sustained. All PAMs increased the potency and efficacy of AEA-induced signalling in all pathways tested; however, no notable potentiation of 2-AG or AMB-FUBINACA was observed.

Conclusion and Implications

Ago-PAMs can potentiate endocannabinoid CB₁ agonism by AEA to a larger extent compared with 2-AG. However, all compounds were found to be allosteric agonists and induce activation of CB₁ in the absence of endocannabinoid, including β-arrestin 2 recruitment and internalisation. Thus, the spatiotemporal signalling of endogenous cannabinoids will not be retained in vivo.

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大麻素 CB1 受体的正异构调节可增强内源性大麻素信号并改变 ERK1/2 磷酸化动力学。

背景和目的：外源性激动剂激活 CB1 会在体内产生不良反应。与正异位激动剂相比，正异位调节可能会因脱敏/耐受性降低而提高治疗潜力并减少靶向不良反应，但这一点尚未得到直接测试。本研究调查了 PAMs/ago-PAMs 诱导受体调节途径的能力，包括脱敏和受体内化：实验方法：在 HEK293 细胞中进行生物发光共振能量转移（BRET）测定，研究 G 蛋白解离、ERK1/2 磷酸化和 β-restin 2 转位；同时进行免疫细胞化学分析，测量 CB1 在 PAMs ZCZ011、GAT229 和 ABD1236 单独使用或与正交激动剂 AEA、2-AG 和 AMB-FUBINACA 结合使用时的内化情况：ZCZ011、GAT229和ABD1236是所有测试途径中的异位激动剂。与正表型激动剂的瞬时激活相比，前表型激动剂 ZCZ011 可诱导双相 ERK1/2 磷酸化时间过程。与 2-AG 而非 AEA 或 AMB-FUBINACA 结合使用时，ZCZ011 和 ABD1236 会使 ERK1/2 磷酸化的瞬时峰值变得持续。在所有测试途径中，所有 PAMs 都提高了 AEA 诱导信号的效力和功效；但是，没有观察到 2-AG 或 AMB-FUBINACA 有明显的增效作用：与 2-AG 相比，Ago-PAMs 能在更大程度上增强 AEA 对内源性大麻素 CB1 的激动作用。然而，研究发现所有化合物都是异位激动剂，在没有内源性大麻素的情况下也能诱导 CB1 的活化，包括 β-restin 2 的招募和内化。因此，内源性大麻素的时空信号不会在体内保留。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

British Journal of Pharmacology 医学-药学

CiteScore

15.40

自引率

12.30%

发文量

270

审稿时长

2.0 months

期刊介绍： The British Journal of Pharmacology (BJP) is a biomedical science journal offering comprehensive international coverage of experimental and translational pharmacology. It publishes original research, authoritative reviews, mini reviews, systematic reviews, meta-analyses, databases, letters to the Editor, and commentaries. Review articles, databases, systematic reviews, and meta-analyses are typically commissioned, but unsolicited contributions are also considered, either as standalone papers or part of themed issues. In addition to basic science research, BJP features translational pharmacology research, including proof-of-concept and early mechanistic studies in humans. While it generally does not publish first-in-man phase I studies or phase IIb, III, or IV studies, exceptions may be made under certain circumstances, particularly if results are combined with preclinical studies.

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