6 results on '"Maloney, David J."'
Search Results
2. A high-throughput screen to identify novel small molecule inhibitors of the Werner Syndrome Helicase-Nuclease (WRN).
- Author
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Sommers, Joshua A., Kulikowicz, Tomasz, Croteau, Deborah L., Dexheimer, Thomas, Dorjsuren, Dorjbal, Jadhav, Ajit, Maloney, David J., Simeonov, Anton, Bohr, Vilhelm A., and Jr.Brosh, Robert M.
- Subjects
SMALL molecules ,WERNER'S syndrome ,DNA helicases ,DNA replication ,CANCER treatment - Abstract
Werner syndrome (WS), an autosomal recessive genetic disorder, displays accelerated clinical symptoms of aging leading to a mean lifespan less than 50 years. The WS helicase-nuclease (WRN) is involved in many important pathways including DNA replication, recombination and repair. Replicating cells are dependent on helicase activity, leading to the pursuit of human helicases as potential therapeutic targets for cancer treatment. Small molecule inhibitors of DNA helicases can be used to induce synthetic lethality, which attempts to target helicase-dependent compensatory DNA repair pathways in tumor cells that are already genetically deficient in a specific pathway of DNA repair. Alternatively, helicase inhibitors may be useful as tools to study the specialized roles of helicases in replication and DNA repair. In this study, approximately 350,000 small molecules were screened based on their ability to inhibit duplex DNA unwinding by a catalytically active WRN helicase domain fragment in a high-throughput fluorometric assay to discover new non-covalent small molecule inhibitors of the WRN helicase. Select compounds were screened to exclude ones that inhibited DNA unwinding by other helicases in the screen, bound non-specifically to DNA, acted as irreversible inhibitors, or possessed unfavorable chemical properties. Several compounds were tested for their ability to impair proliferation of cultured tumor cells. We observed that two of the newly identified WRN helicase inhibitors inhibited proliferation of cancer cells in a lineage-dependent manner. These studies represent the first high-throughput screen for WRN helicase inhibitors and the results have implications for anti-cancer strategies targeting WRN in different cancer cells and genetic backgrounds. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
3. A High-Content Assay Enables the Automated Screening and Identification of Small Molecules with Specific ALDH1A1-Inhibitory Activity.
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Yasgar, Adam, Titus, Steven A., Wang, Yuhong, Danchik, Carina, Yang, Shyh-Ming, Vasiliou, Vasilis, Jadhav, Ajit, Maloney, David J., Simeonov, Anton, and Martinez, Natalia J.
- Subjects
ALDEHYDE dehydrogenase ,ENZYME inhibitors ,CANCER stem cells ,ALDEHYDES ,STRUCTURE-activity relationships - Abstract
Aldehyde dehydrogenase enzymes (ALDHs) have a broad spectrum of biological activities through the oxidation of both endogenous and exogenous aldehydes. Increased expression of ALDH1A1 has been identified in a wide-range of human cancer stem cells and is associated with cancer relapse and poor prognosis, raising the potential of ALDH1A1 as a therapeutic target. To facilitate quantitative high-throughput screening (qHTS) campaigns for the discovery, characterization and structure-activity-relationship (SAR) studies of small molecule ALDH1A1 inhibitors with cellular activity, we show herein the miniaturization to 1536-well format and automation of a high-content cell-based ALDEFLUOR assay. We demonstrate the utility of this assay by generating dose-response curves on a comprehensive set of prior art inhibitors as well as hundreds of ALDH1A1 inhibitors synthesized in house. Finally, we established a screening paradigm using a pair of cell lines with low and high ALDH1A1 expression, respectively, to uncover novel cell-active ALDH1A1-specific inhibitors from a collection of over 1,000 small molecules. [ABSTRACT FROM AUTHOR]
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- 2017
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4. A High-Throughput Screen Identifies 2,9-Diazaspiro[5.5]Undecanes as Inducers of the Endoplasmic Reticulum Stress Response with Cytotoxic Activity in 3D Glioma Cell Models.
- Author
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Martinez, Natalia J., Rai, Ganesha, Yasgar, Adam, Lea, Wendy A., Sun, Hongmao, Wang, Yuhong, Luci, Diane K., Yang, Shyh-Ming, Nishihara, Kana, Takeda, Shunichi, Sagor, Mohiuddin, Earnshaw, Irina, Okada, Tetsuya, Mori, Kazutoshi, Wilson, Kelli, Riggins, Gregory J., Xia, Menghang, Grimaldi, Maurizio, Jadhav, Ajit, and Maloney, David J.
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GLIOMAS ,CELL-mediated cytotoxicity ,ENDOPLASMIC reticulum ,PROTEIN folding ,MOLECULAR chaperones ,HOMEOSTASIS ,PHYSIOLOGY - Abstract
The endoplasmic reticulum (ER) is involved in Ca
2+ signaling and protein folding. ER Ca2+ depletion and accumulation of unfolded proteins activate the molecular chaperone GRP78 (glucose-regulated protein 78) which in turn triggers the ER stress response (ERSR) pathway aimed to restore ER homeostasis. Failure to adapt to stress, however, results in apoptosis. We and others have shown that malignant cells are more susceptible to ERSR-induced apoptosis than their normal counterparts, implicating the ERSR as a potential target for cancer therapeutics. Predicated on these findings, we developed an assay that uses a GRP78 biosensor to identify small molecule activators of ERSR in glioma cells. We performed a quantitative high-throughput screen (qHTS) against a collection of ~425,000 compounds and a comprehensive panel of orthogonal secondary assays was formulated for stringent compound validation. We identified novel activators of ERSR, including a compound with a 2,9-diazaspiro[5.5]undecane core, which depletes intracellular Ca2+ stores and induces apoptosis-mediated cell death in several cancer cell lines, including patient-derived and 3D cultures of glioma cells. This study demonstrates that our screening platform enables the identification and profiling of ERSR inducers with cytotoxic activity and advocates for characterization of these compound in in vivo models. [ABSTRACT FROM AUTHOR]- Published
- 2016
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5. The function of phosphatidylinositol 5-phosphate 4-kinase ɣ (PI5P4Kɣ) explored using a specific inhibitor that targets the PI5P-binding site.
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Clarke, Jonathan H., Giudici, Maria-Luisa, Burke, John E., Williams, Roger L., Maloney, David J., Marugan, Juan, and Irvine, Robin F.
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PHOSPHATIDYLINOSITOL 3-kinases ,GENE targeting ,ENZYMES ,CHEMICAL synthesis ,ATP-binding cassette transporters ,CELL physiology ,BINDING sites ,AMINO acid residues - Abstract
NIH-12848 (NCGC00012848-02), a putative phosphatidylinositol 5-phosphate 4-kinase ɣ (PI5P4Kɣ) inhibitor, was explored as a tool for investigating this enigmatic, low activity, lipid kinase. PI5P4K assays in vitro showed that NIH-12848 inhibited PI5P4Kɣ with an IC50 of approximately 1 µM but did not inhibit the ɑ and β PI5P4K isoforms at concentrations up to 100 µM. A lack of inhibition of PI5P4Kɣ ATPase activity suggested that NIH-12848 does not interact with the enzyme's ATP-binding site and direct exploration of binding using hydrogen-deuterium exchange (HDX)-MS (HDX-MS) revealed the putative PI5P-binding site of PI5P4Kɣ to be the likely region of interaction. This was confirmed by a series of mutation experiments which led to the identification of a single PI5P4Kɣ amino acid residue that can be mutated to its PI5P4Ks ɑ and β homologue to render PI5P4Kɣ resistant NIH-12848 inhibition. NIH-12848 (10 µM) was applied to cultured mouse principal kidney cortical collecting duct (mpkCCD) cells which, we show, express PI5P4Kɣ that increases when the cells grow to confluence and polarize. NIH-12848 inhibited the translocation of Na
+ /K+ -ATPase to the plasma membrane that occurs when mpkCCD cells grow to confluence and also prevented reversibly their forming of 'domes' on the culture dish. Both these NIH-12848-induced effects were mimicked by specific RNAi knockdown of PI5P4Kɣ, but not that of PI5P4Ks ɑ or β. Overall, the data reveal a probable contribution of PI5P4Kɣ to the development and maintenance of epithelial cell functional polarity and show that NIH-12848 is a potentially powerful tool for exploring the cell physiology of PI5P4Ks. [ABSTRACT FROM AUTHOR]- Published
- 2015
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6. Discovery of a Novel Dual Fungal CYP51/Human 5-Lipoxygenase Inhibitor: Implications for Anti-Fungal Therapy.
- Author
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Hoobler, Eric K., Rai, Ganesha, Warrilow, Andrew G. S., Perry, Steven C., Smyrniotis, Christopher J., Jadhav, Ajit, Simeonov, Anton, Parker, Josie E., Kelly, Diane E., Maloney, David J., Kelly, S. L., and Holman, Theodore R.
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ANTIFUNGAL agents ,LIPOXYGENASES ,DRUG development ,DRUG synergism ,PHENYLENEDIAMINES ,PHARMACEUTICAL chemistry ,CLINICAL drug trials ,MYCOSES ,COMMUNICABLE disease treatment - Abstract
We report the discovery of a novel dual inhibitor targeting fungal sterol 14α-demethylase (CYP51 or Erg11) and human 5-lipoxygenase (5-LOX) with improved potency against 5-LOX due to its reduction of the iron center by its phenylenediamine core. A series of potent 5-LOX inhibitors containing a phenylenediamine core, were synthesized that exhibit nanomolar potency and >30-fold selectivity against the LOX paralogs, platelet-type 12-human lipoxygenase, reticulocyte 15-human lipoxygenase type-1, and epithelial 15-human lipoxygenase type-2, and >100-fold selectivity against ovine cyclooxygenase-1 and human cyclooxygnease-2. The phenylenediamine core was then translated into the structure of ketoconazole, a highly effective anti-fungal medication for seborrheic dermatitis, to generate a novel compound, ketaminazole. Ketaminazole was found to be a potent dual inhibitor against human 5-LOX (IC
50 = 700 nM) and CYP51 (IC50 = 43 nM) in vitro. It was tested in whole blood and found to down-regulate LTB4 synthesis, displaying 45% inhibition at 10 µM. In addition, ketaminazole selectively inhibited yeast CYP51 relative to human CYP51 by 17-fold, which is greater selectivity than that of ketoconazole and could confer a therapeutic advantage. This novel dual anti-fungal/anti-inflammatory inhibitor could potentially have therapeutic uses against fungal infections that have an anti-inflammatory component. [ABSTRACT FROM AUTHOR]- Published
- 2013
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