In vitro activity: AMI-1 suppresses the transcriptional coactivator activity of PRMT1 and PRMT4 and it inhibits HIV-1 RT polymerase (IC50 = 5.0μM). PRMT1 methylates histone H4, and is essential for other subsequent histone modifications. AMI-1 is the most active nonpeptidic inhibitor reported to be selective against PRMT1. AMI-1 is a selective PRMT inhibitor with a bisanionic structure that is related to compounds known to generate pleiotropic interactions with many proteins, should be further optimized before exploring additional binding pockets. It is cell permeable and does not compete for the AdoMet (S-adenosyl-L-methionine, SAM) binding site. AMI-1 inhibits protein arginine N-methyltransferases (PRMTs), including human PRMT1 (IC50 = 8.8μM) and yeast-Hmt1p (IC50 = 3.0μM), by blocking peptide-substrate binding. AMI-1 is also a potent scavenger of NADPH-oxidase-derived superoxide and can modulate nuclear receptor-regulated transcription from estrogen and androgen response elements.

Kinase Assay: AMI-1 is a potent, cell-permeable and specific inhibitor of Histone Methyltransferase (HMT) with IC50 of 3.0 μM and 8.8 μM for yeast Hmt1p and human PRMT1, respectively. It is cell permeable and does not compete for the AdoMet (S-adenosyl-L-methionine, SAM) binding site. AMI-1 inhibits protein arginine N-methyltransferases (PRMTs), including human PRMT1 (IC50 = 8.8μM) and yeast-Hmt1p (IC50 = 3.0μM), by blocking peptide-substrate binding.

Cell Assay: In HeLa cells, AMI-1 inhibits methylation levels of GFP-Npl3 fusion and endogenous PRMT1-like activity. AMI-1 also inhibits nuclear receptor-mediated transactivation of a luciferase reporter in MCF7 cells. In addition, AMI-1 inhibits HIV-1 RT polymerase activity with IC50 of 5 μM and inhibits DNA binding to HIV-1 RT with Kd of 2 μM. In INS-1 cells, AMI-1 improves INS-1 cell function and mediates translocations of FOXO1 and PDX-1 intracellularly by regulating FOXO1 phosphorylation and methylation