Potency data for probe JNJ-9350 and control JNJ-4545


In vitro potency


Target name Target information Species Assay description JNJ-9350 JNJ-4545 Publication of assay conditions
SMOX full length human SMOX with N-terminal his tag Homo sapiens HyPerBlu SMOX enzymatic assay: In this assay spermine oxidase catalyzes the formation of spermidine, 3 -aminopropanal and hydrogen peroxide using oxygen and spermine as substrates. The addition of Lumigen HyperBlu reagent quenches the reaction and directly reacts with hydrogen peroxide to generate a chemiluminescence signal measured on a BMG PHERAStar FS/FSX multilabel plate reader. IC50 = 10 nM IC50 > 31 µM Slide 6, 14-23 in JNJ-9350 Inhibitor for SMOX.pdf
SMOX Expression in E.coli; human SMOX, Isoform 1 Homo sapiens Horseradish Peroxidase (HRP) Fluorescence Assay: Reactions were monitored continuously for 1hr on the BMG PHERAStar plate reader using an FI optic module (ex: 540 nm, em: 590 nm). Ki = 9.9 nM Slide 24-26 in JNJ-9350 Inhibitor for SMOX.pdf
PAOX full length human PAOX with N-terminal his tag Homo sapiens HyPerBlu POAX enzymatic assay: In this assay Polyamine Oxidase catalyzes the formation of spermidine, 3-acetamidopropanal and hydrogen peroxide using oxygen and N1-acetylspermine as substrates. The addition of 25 µM chlorhexidine compound mixed with the Lumigen HyPerBlu reagent is added to quench the reaction and directly react with hydrogen peroxide, respectively, to generate a chemiluminescence signal measured on the BMG PHERAStar plate reader. IC50 = 790 nM IC50 = 2.1 µM Slide 27-37 in JNJ-9350 Inhibitor for SMOX.pdf
KDM1A Homo sapiens HyPerBlu KDM1A enzymatic assay IC50 > 60 µM

In cell potency


Target name Target information Species Assay description JNJ-9350 JNJ-4545 Publication of assay conditions
SMOX A549 cells (adenocarcinomic human alveolar basal epithelial cells that express relatively high levels of SMOX); full length, endogenous human SMOX Homo sapiens SMOX CETSA assay: Intact SMOX is detected using alphaLISA. The optimal melting temperature (54°C) of hSMOX for CETSA was used. IC50 = 1.2 µM (n=4) Slide 10,11, 38-43 in JNJ-9350 Inhibitor for SMOX.pdf