DMX-5804 is a potent, selective MAP4K4 inhibitor, with an IC50 of 3 nM.DMX-5804 enhances cardiomyocyte survival, and reduces ischemia-reperfusion injury in mice.

MAP4K4:3 nM

DMX-5804 能挽救细胞存活，提升线粒体功能。

DMX-5804通过超过50%的比例减轻了小鼠的缺血再灌注损伤。

Mitochondrial function in hiPSC-CMs was determined using a Seahorse XFe24 Analyzer.?vCor.4U cells (60,000/well) were transferred to 0.1% gelatin-coated XF24 plates and maintained for 5 d. On day 6, DMX-5804 was added 45 min prior to challenge with menadione for 2 h. The medium was replaced 1 hr before the assay, using bicarbonate-free Seahorse assay medium (8.3 g L-1 DMEM Base , 10 mM glucose, 2 mM L-alanyl-L-glutamine dipeptide, 1 mM sodium pyruvate , pH 7.4).?Cells were maintained at 37 C without supplemental CO2 starting 1 hr before the assay.?For each state measured, three assay cycles were performed (4 min mixing, 2 min wait, and 2 min measurement periods, with readings every 15 s). The basal oxygen consumption rate (OCR) and extracellular acidification rate were determined, followed by sequential injection of 1 μM oligomycin A to inhibit ATP synthase, 0.5 μM carbonyl cyanide-4-phenylhydrazone to uncouple oxidative phosphorylation, and 1 μM antimycin A/rotenone to inhibit mitochondrial complex III and I. For each condition, 12 wells were tested, comprising 4 independent experiments.

n vivo pharmacokinetic profiling was performed in female CD-1 mice, using 3 animals per time point.?First, 30% w/v Kleptose as excipient was dissolved in water and vortexed gently for several min. Next, 30 mg of DMX-5084?was dissolved into 0.6 mL of DMSO, for a concentration of 50 mg ml-1, and 4.5 mL of the Kleptose solution was added to 0.5 mL of the test compound solution.?A precipitate forms, which re-dissolves over 2-5 min, leaving a clear or slightly hazy solution with a final concentration of 5 mg/ml.?The dosing solution is used as soon as practicable, vortexing immediately prior to use.?This amount was sufficient formulation for 25 doses of 200 μL (20 g mouse).?Compounds were administered orally at 50 mg/kg, with terminal blood (plasma) sampling at 10 min, 30 min, 1 h, 2.5 h, 5 h, 10 h, and 20 h[1].