ML290 is a biased allosteric agonist at the relaxin receptor RXFP1.
ML290 is a biased allosteric agonist at the relaxin receptor RXFP1.
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DOI：
10.1038/s41598-017-02916-5
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发表时间：
2017-06-07
影响因子：
4.6
通讯作者：
Summers RJ
中科院分区：
文献类型：
--
作者：
Kocan M;Sarwar M;Ang SY;Xiao J;Marugan JJ;Hossain MA;Wang C;Hutchinson DS;Samuel CS;Agoulnik AI;Bathgate RAD;Summers RJ
松弛素受体RXFP1的激活与急性心力衰竭患者生存率的提高有关。ML290是一种结构简单、半衰期长、稳定性高的小分子RXFP1激动剂。在这里，我们证明了ML290在表达RXFP1的人细胞中是一种偏向性激动剂，对人心脏成纤维细胞（HCFs）的纤维化标志物具有长期有益作用。ML290不直接与正位松弛素结合竞争，也不影响结合动力学，但确实增加了与RXFP1的结合。在HEK-RXFP1细胞中，ML290刺激cAMP积累和p38MAPK磷酸化，但不刺激cGMP积累或ERK1/2磷酸化，尽管事先添加ML290增加了p-ERK1/2对松弛素的反应。在内源性表达RXFP1的人原代血管内皮细胞和平滑肌细胞中，ML290增加cAMP和cGMP的积累，但不增加p-ERK1/2。在hcf中，ML290增加了cGMP积累，但不影响p-ERK1/2和慢性激活的MMP-2表达，并抑制TGF-β1诱导的Smad2和Smad3磷酸化。在血管细胞中，ML290对cGMP积累和p-p38MAPK的作用是cAMP积累的10倍。ML290引起RXFP1与Gαs和GαoB强耦合，而与Gαi3弱耦合。ML290在RXFP1上表现出信号偏倚，具有血管扩张剂和抗纤维化特性的信号谱。
Activation of the relaxin receptor RXFP1 has been associated with improved survival in acute heart failure. ML290 is a small molecule RXFP1 agonist with simple structure, long half-life and high stability. Here we demonstrate that ML290 is a biased agonist in human cells expressing RXFP1 with long-term beneficial actions on markers of fibrosis in human cardiac fibroblasts (HCFs). ML290 did not directly compete with orthosteric relaxin binding and did not affect binding kinetics, but did increase binding to RXFP1. In HEK-RXFP1 cells, ML290 stimulated cAMP accumulation and p38MAPK phosphorylation but not cGMP accumulation or ERK1/2 phosphorylation although prior addition of ML290 increased p-ERK1/2 responses to relaxin. In human primary vascular endothelial and smooth muscle cells that endogenously express RXFP1, ML290 increased both cAMP and cGMP accumulation but not p-ERK1/2. In HCFs, ML290 increased cGMP accumulation but did not affect p-ERK1/2 and given chronically activated MMP-2 expression and inhibited TGF-β1-induced Smad2 and Smad3 phosphorylation. In vascular cells, ML290 was 10x more potent for cGMP accumulation and p-p38MAPK than for cAMP accumulation. ML290 caused strong coupling of RXFP1 to Gαs and GαoB but weak coupling to Gαi3. ML290 exhibited signalling bias at RXFP1 possessing a signalling profile indicative of vasodilator and anti-fibrotic properties.