3 research outputs found
Definitive Metabolite Identification Coupled with Automated Ligand Identification System (ALIS) Technology: A Novel Approach to Uncover Structure–Activity Relationships and Guide Drug Design in a Factor IXa Inhibitor Program
A potent
and selective Factor IXa (FIXa) inhibitor was subjected
to a series of liver microsomal incubations, which generated a number
of metabolites. Using automated ligand identification system-affinity
selection (ALIS-AS) methodology, metabolites in the incubation mixture
were prioritized by their binding affinities to the FIXa protein.
Microgram quantities of the metabolites of interest were then isolated
through microisolation analytical capabilities, and structurally characterized
using MicroCryoProbe heteronuclear 2D NMR techniques. The isolated
metabolites recovered from the NMR experiments were then submitted
directly to an <i>in vitro</i> FIXa enzymatic assay. The
order of the metabolites’ binding affinity to the Factor IXa
protein from the ALIS assay was completely consistent with the enzymatic
assay results. This work showcases an innovative and efficient approach
to uncover structure–activity relationships (SARs) and guide
drug design via microisolation–structural characterization
and ALIS capabilities
Definitive Metabolite Identification Coupled with Automated Ligand Identification System (ALIS) Technology: A Novel Approach to Uncover Structure–Activity Relationships and Guide Drug Design in a Factor IXa Inhibitor Program
A potent
and selective Factor IXa (FIXa) inhibitor was subjected
to a series of liver microsomal incubations, which generated a number
of metabolites. Using automated ligand identification system-affinity
selection (ALIS-AS) methodology, metabolites in the incubation mixture
were prioritized by their binding affinities to the FIXa protein.
Microgram quantities of the metabolites of interest were then isolated
through microisolation analytical capabilities, and structurally characterized
using MicroCryoProbe heteronuclear 2D NMR techniques. The isolated
metabolites recovered from the NMR experiments were then submitted
directly to an <i>in vitro</i> FIXa enzymatic assay. The
order of the metabolites’ binding affinity to the Factor IXa
protein from the ALIS assay was completely consistent with the enzymatic
assay results. This work showcases an innovative and efficient approach
to uncover structure–activity relationships (SARs) and guide
drug design via microisolation–structural characterization
and ALIS capabilities
Discovery of Orally Active Inhibitors of Brahma Homolog (BRM)/SMARCA2 ATPase Activity for the Treatment of Brahma Related Gene 1 (BRG1)/SMARCA4-Mutant Cancers
SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin subfamily A member 2 (SMARCA2), also known as Brahma homologue (BRM), is a Snf2-family DNA-dependent ATPase. BRM and its close homologue Brahma-related gene 1 (BRG1), also known as SMARCA4, are mutually exclusive ATPases of the large ATP-dependent SWI/SNF chromatin-remodeling complexes involved in transcriptional regulation of gene expression. No small molecules have been reported that modulate SWI/SNF chromatin-remodeling activity via inhibition of its ATPase activity, an important goal given the well-established dependence of BRG1-deficient cancers on BRM. Here, we describe allosteric dual BRM and BRG1 inhibitors that downregulate BRM-dependent gene expression and show antiproliferative activity in a BRG1-mutant-lung-tumor xenograft model upon oral administration. These compounds represent useful tools for understanding the functions of BRM in BRG1-loss-of-function settings and should enable probing the role of SWI/SNF functions more broadly in different cancer contexts and those of other diseases