Optimization of 3‑Cyano-7-cyclopropylamino-pyrazolo[1,5‑<i>a</i>]pyrimidines toward the Development of an In Vivo Chemical Probe for CSNK2A

3-Cyano-7-cyclopropylamino-pyrazolo[1,5-a]pyrimidines, including the chemical probe SGC-CK2-1, are potent and selective inhibitors of CSNK2A in cells but have limited utility in animal models due to their poor pharmacokinetic properties. While developing analogues with reduced intrinsic clearance and the potential for sustained exposure in mice, we discovered that phase II conjugation by GST enzymes was a major metabolic transformation in hepatocytes. A protocol for codosing with ethacrynic acid, a covalent reversible GST inhibitor, was developed to improve the exposure of analogue 2h in mice. A double codosing protocol, using a combination of ethacrynic acid and irreversible P450 inhibitor 1-aminobenzotriazole, increased the blood level of 2h by 40-fold at a 5 h time point

Small Molecule Agonists of the Orphan Nuclear Receptors Steroidogenic Factor-1 (SF-1, NR5A1) and Liver Receptor Homologue-1 (LRH-1, NR5A2)

The crystal structure of LRH-1 ligand binding domain bound to our previously reported agonist 3-(<i>E</i>-oct-4-en-4-yl)-1-phenylamino-2-phenyl-<i>cis</i>-bicyclo[3.3.0]oct-2-ene <b>5</b> is described. Two new classes of agonists in which the bridgehead anilino group from our first series was replaced with an alkoxy or 1-ethenyl group were designed, synthesized, and tested for activity in a peptide recruitment assay. Both new classes gave very active compounds, particularly against SF-1. Structure−activity studies led to excellent dual-LRH-1/SF-1 agonists (e.g., RJW100) as well as compounds selective for LRH-1 (RJW101) and SF-1 (RJW102 and RJW103). The series based on 1-ethenyl substitution was acid stable, overcoming a significant drawback of our original bridgehead anilino-substituted series. Initial studies on the regulation of gene expression in human cell lines showed excellent, reproducible activity at endogenous target genes

DS_773045 – Supplemental material for Application of Integrated Drug Screening/Kinome Analysis to Identify Inhibitors of Gemcitabine-Resistant Pancreatic Cancer Cell Growth

<p>Supplemental material, DS_773045 for Application of Integrated Drug Screening/Kinome Analysis to Identify Inhibitors of Gemcitabine-Resistant Pancreatic Cancer Cell Growth by Linas J. Krulikas, Ian M. McDonald, Benjamin Lee, Denis O. Okumu, Michael P. East, Thomas S. K. Gilbert, Laura E. Herring, Brian T. Golitz, Carrow I. Wells, Allison D. Axtman, William J. Zuercher, Timothy M. Willson, Dmitri Kireev, Jen Jen Yeh, Gary L. Johnson, Antonio T. Baines, and Lee M. Graves in SLAS Discovery</p

Optimized Chemical Probes for REV-ERBα

REV-ERBα has emerged as an important target for regulation of circadian rhythm and its associated physiology. Herein, we report on the optimization of a series of REV-ERBα agonists based on GSK4112 (<b>1</b>) for potency, selectivity, and bioavailability. Potent REV-ERBα agonists <b>4</b>, <b>10</b>, <b>16</b>, and <b>23</b> are detailed for their ability to suppress BMAL and IL-6 expression from human cells while also demonstrating excellent selectivity over LXRα. Amine <b>4</b> demonstrated in vivo bioavailability after either iv or oral dosing

Representation of the kinome coverage of the virtual set of 457 narrow spectrum inhibitors.

<p>Red background shows the human protein kinases ranked by number of citations. Blue bars show the protein kinases for which an assay is available at one of 10 commercial vendors. Black bars identify protein kinases that are covered by the virtual set of inhibitors (subset of PKIS, PKIS2, and literature) described in the text.</p

Schematic: Utilization of the KCGS.

<p>(<b>a</b>) Disease relevant phenotypic screen highlights active compounds (<b>b</b>) Because all the targets are annotated, active molecules points to potential targets of interest. (<b>c</b>) Follow up experiments can be used to help confirm importance of these highlighted targets (<b>d</b>) Body of evidence implicates kinases that can be inhibited to impact disease.</p

The screenable protein kinome.

<p>Using assays from 10 vendors a total of 436 unique non-mutant human protein kinases can be readily screened. (<b>a</b>) Subfamily representation of the protein kinases that were not available through the 10 vendors. Nearly half of the unscreenable human kinome is composed of pseudokinases (<b>b</b>) Histogram illustrating how many vendors can be used to screen across the kinome. For example, there are 43 kinases that all 10 vendors have screens for (“all” bar). There are another 109 kinases screened by 9 out of 10 vendors (“9/10” bar). There are 38 kinases that only 1 out of the 10 vendors has assays for (“1/10” bar). <i>Note</i>: <i>Our definition of the “screenable kinome” is those kinases for which there is a commercial assay that can be accessed</i>. <i>It is likely that assays could indeed be configured for many of the kinases not currently on this list</i>.</p