Identification of Novel Plasmodium falciparum Hexokinase Inhibitors with Antiparasitic Activity

Davis, Mindy I.; Patrick, Stephen L.; Blanding, Walker M.; Dwivedi, Varun; Suryadi, Jimmy; Golden, Jennifer E.; Coussens, Nathan P.; Lee, Olivia W.; Shen, Min; Boxer, Matthew B.; Hall, Matthew D.; Sharlow, Elizabeth R.; Drew, Mark E.; Morris, James C.

doi:10.1128/aac.00914-16

Cited by 16 publications

(16 citation statements)

References 24 publications

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“…Screening Libraries. The libraries that were used for the screen included the Mechanism Interrogation Plate (MIPE) composed of 1912 compounds (Mathews Griner et al, 2014), the NCATS pharmaceutical collection comprising 2816 compounds (Huang et al, 2011), the NCATS Pharmacologically Active Chemical Toolbox (NPACT) (Davis et al, 2016) composed of 5099 compounds, and a kinase inhibitor library comprising 977 compounds, for a total of 10,804 compounds.…”

Section: Methodsmentioning

confidence: 99%

A High-Throughput Screen of a Library of Therapeutics Identifies Cytotoxic Substrates of P-glycoprotein

Lee

Brimacombe

et al. 2019

Mol Pharmacol

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The ATP-binding cassette transporter P-glycoprotein (P-gp) is known to limit both brain penetration and oral bioavailability of many chemotherapy drugs. Although US Food and Drug Administration guidelines require that potential interactions of investigational drugs with P-gp be explored, often this information does not enter the literature. In response, we developed a high-throughput screen to identify substrates of P-gp from a series of chemical libraries, testing a total of 10,804 compounds, most of which have known mechanisms of action. We used the CellTiter-Glo viability assay to test library compounds against parental KB-3-1 human cervical adenocarcinoma cells and the colchicine-selected subline KB-8-5-11 that overexpresses P-gp. KB-8-5-11 cells were also tested in the presence of a P-gp inhibitor (tariquidar) to assess reversibility of transporter-mediated resistance. Of the tested compounds, a total of 90 P-gp substrates were identified, including 55 newly identified compounds. Substrates were confirmed using an orthogonal killing assay against human embryonic kidney-293 cells overexpressing P-gp. We confirmed that AT7159 (cyclindependent kinase inhibitor), AT9283, (Janus kinase 2/3 inhibitor), ispinesib (kinesin spindle protein inhibitor), gedatolisib (PKI-587, phosphoinositide 3-kinase/mammalian target of rampamycin inhibitor), GSK-690693 (AKT inhibitor), and KW-2478 (heatshock protein 90 inhibitor) were substrates. In addition, we assessed direct ATPase stimulation. ABCG2 was also found to confer high levels of resistance to AT9283, GSK-690693, and gedatolisib, whereas ispinesib, AT7519, and KW-2478 were weaker substrates. Combinations of P-gp substrates and inhibitors were assessed to demonstrate on-target synergistic cell killing. These data identified compounds whose oral bioavailability or brain penetration may be affected by P-gp. SIGNIFICANCE STATEMENT The ATP-binding cassette transporter P-glycoprotein (P-gp) is known to be expressed at barrier sites, where it acts to limit oral bioavailability and brain penetration of substrates. In order to identify novel compounds that are transported by P-gp, we developed a high-throughput screen using the KB-3-1 cancer cell line and its colchicine-selected subline KB-8-5-11. We screened the Mechanism Interrogation Plate (MIPE) library, the National Center for Advancing Translational Science (NCATS) pharmaceutical collection (NPC), the NCATS Pharmacologically Active Chemical Toolbox (NPACT), and a kinase inhibitor library comprising 977 compounds, for a total of 10,804 compounds. Of the 10,804 compounds screened, a total of 90 substrates were identified of which 55 were novel. P-gp expression may adversely affect the oral bioavailability or brain penetration of these compounds.

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Section: Methodsmentioning

confidence: 99%

A High-Throughput Screen of a Library of Therapeutics Identifies Cytotoxic Substrates of P-glycoprotein

Lee

Brimacombe

et al. 2019

Mol Pharmacol

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“…Compounds were tested for their effects on viability using OvCa cell lines and a resazurin-based CellTiter Blue (Promega, Fitchburg, WI) 2D assay system with previously described procedures [ 53 ]. All OvCa cell lines were cultured in complete growth medium with 500 cells/22 μL being seeded into each well of a 384 well black/clear microtiter plate (VWR).…”

Section: Methodsmentioning

confidence: 99%

Targeting ovarian cancer and endothelium with an allosteric PTP4A3 phosphatase inhibitor

et al. 2017

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Overexpression of protein tyrosine phosphatase PTP4A oncoproteins is common in many human cancers and is associated with poor patient prognosis and survival. We observed elevated levels of PTP4A3 phosphatase in 79% of human ovarian tumor samples, with significant overexpression in tumor endothelium and pericytes. Furthermore, PTP4A phosphatases appear to regulate several key malignant processes, such as invasion, migration, and angiogenesis, suggesting a pivotal regulatory role in cancer and endothelial signaling pathways. While phosphatases are attractive therapeutic targets, they have been poorly investigated because of a lack of potent and selective chemical probes. In this study, we disclose that a potent, selective, reversible, and noncompetitive PTP4A inhibitor, JMS-053, markedly enhanced microvascular barrier function after exposure of endothelial cells to vascular endothelial growth factor or lipopolysaccharide. JMS-053 also blocked the concomitant increase in RhoA activation and loss of Rac1. In human ovarian cancer cells, JMS-053 impeded migration, disrupted spheroid growth, and decreased RhoA activity. Importantly, JMS-053 displayed anticancer activity in a murine xenograft model of drug resistant human ovarian cancer. These data demonstrate that PTP4A phosphatases can be targeted in both endothelial and ovarian cancer cells, and confirm that RhoA signaling cascades are regulated by the PTP4A family.

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“…Screening Libraries. The libraries that were used for the screen included the Mechanism Interrogation Plate (MIPE) comprised of 1,912 compounds (Mathews Griner et al, 2014), the NCATS pharmaceutical collection (NPC) comprised of 2816 compounds (Huang et al, 2011), the NCATS Pharmacologically Active Chemical Toolbox (NPACT) (Davis et al, 2016) comprised of 5,099 compounds, and a kinase inhibitor library comprised of 977 compounds, for a total of 10,804 compounds.…”

Section: Methodsmentioning

confidence: 99%

A High-Throughput Screen of a Library of Therapeutics Identifies Substrates of P-glycoprotein

Lee

Lee²,

Brimacombe³

et al. 2019

Preprint

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The ATP-binding cassette transporter P-glycoprotein (P-gp) is known to limit brain penetration of many chemotherapy drugs. Although Food and Drug Administration guidelines require that potential interactions of investigational drugs with P-gp be explored, often this information does not enter into the literature. As such, we developed a high-throughput screen (HTS) to identify substrates of P-gp from a series of chemical libraries, testing a total of 10,804 compounds, most of which have known mechanisms of action. We used the CellTiter-Glo viability assay to test library compounds against parental KB-3-1 human cervical adenocarcinoma cells and the colchicine-selected sub-line KB-8-5-11 that over-expresses P-gp. KB-8-5-11 cells were also tested in the presence of a P-gp inhibitor (tariquidar) to assess reversability of transporter-mediated resistance. Of the tested compounds, a total of 90 P-gp substrates were identified including 55 newly identified P-gp substrates. Substrates were confirmed using an orthogonal killing assay against HEK-293 cells transfected with P-gp. We confirmed that AT7159 (cyclin-dependent kinase inhibitor); AT9283, (Janus kinase 2/3 inhibitor); ispinesib (kinesin spindle protein inhibitor); gedatolisib (PKI-587, phosphoinositide 3-kinase/mammalian target of rampamycin inhibitor); GSK-690693 (AKT inhibitor); and KW-2478 (heat shock protein 90 inhibitor) were substrates, and direct ATPase stimulation was assessed. ABCG2 was also found to confer high levels of resistance to AT9283, GSK-690693 and gedatolisib, while ispinesib, AT7519 and KW-2478 were weaker substrates. Combinations of P-gp substrates and inhibitors were assessed to demonstrate on-target synergistic cell killing. This data will be of use in determining understanding how chemotherapeutic agents will cross the blood-brain barrier.

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Identification of Novel Plasmodium falciparum Hexokinase Inhibitors with Antiparasitic Activity

Cited by 16 publications

References 24 publications

A High-Throughput Screen of a Library of Therapeutics Identifies Cytotoxic Substrates of P-glycoprotein

A High-Throughput Screen of a Library of Therapeutics Identifies Cytotoxic Substrates of P-glycoprotein

Targeting ovarian cancer and endothelium with an allosteric PTP4A3 phosphatase inhibitor

A High-Throughput Screen of a Library of Therapeutics Identifies Substrates of P-glycoprotein

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