ML221 is a potent apelin /APJ functional antagonist, inhibiting apelin-13-mediated activation of APJ, with IC50s of 0.70 μM in the cAMP assay, and 1.75 μM in the β-arrestin assay, and EC80 of 10 nM in both assays.

APJ:1.75 μM

ML221是一种强效的apelin/APJ功能拮抗剂，能够抑制apelin-13介导的APJ激活，其在cAMP实验中的IC50为0.70 μM，在β-arrestin实验中的IC50为1.75 μM，且在两种实验中EC80均为10 nM。ML221对于与之密切相关的血管紧张素II型1(AT1)受体的选择性>37倍（IC50，>79 μM），且在细胞中显示出来。ML221对一系列GPCR表现出有限的交叉反应性，除了κ-阿片类受体和苯二氮卓受体（<50/<70%I at 10 μM）。

ML221 is dissolved in DMSO. Cells (angiotensin II receptor-like 1 (AGTRL-1) cell line) are seeded at 1000 cell/well (1536 plate) in 4 μL and grown overnight (16-18 h) at 37°C, 5% CO2, 100% humidity, then 60 nL of either DMSO control or 2 mM stock test compounds (ML221, etc.) in DMSO are transferred to each well, followed by 2 μL of 30 nM Apelin-13 to negative control and test compound wells, and 2 μL of assay media (F12 nutrient mix HAMs supplemented with 10% hi-FBS, 1× penicillin/streptomycin) to positive control wells. This yields a final concentration of test compound (ML221, etc.) of 20 μM and 1% final DMSO. Assay is incubated for 90 min at room temperature, and then developed with 3 μL of detection reagent for 60 min and luminescence read on a ViewLux. They are for reference only.