60 results on '"Audia JE"'
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2. Neurogenic dural protein extravasation induced by meta-chlorophenylpiperazine (mCPP) involves nitric oxide and 5-HT2B receptor activation.
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Johnson, KW, Nelson, DL, Dieckman, DK, Wainscott, DB, Lucaites, VL, Audia, JE, Owton, WM, and Phebus, LA
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- *
HEADACHE , *BLOOD proteins , *GUINEA pigs - Abstract
&figfu1; The compound m-chlorophenylpiperazine (mCPP), which is known to trigger migraine-like head pain in some subjects, was evaluated for its ability to induce dural plasma protein extravasation (PPE) in guinea pigs. Intravenous mCPP dose-dependently increased PPE. This effect was inhibited by non-selective 5-HT2 receptor antagonists (methysergide, LY53857, LY215840), by a peripherally restricted 5-HT2 receptor antagonist (xylamidine) and by a 5-HT2B selective receptor antagonist (LY202146). These data suggests that peripheral 5-HT2B receptors mediate mCPP-induced PPE. The nitric oxide synthase inhibitor L-NAME and 5-HT1 agonist sumatriptan also blocked mCPP-induced PPE, suggesting a role for nitric oxide (NO) and the trigeminal system, respectively. NO release has been linked to activation of the 5-HT2B receptor on the vascular endothelium. However, LY202146 did not block PPE induced by electrical stimulation of the trigeminal ganglion. These data are consistent with activation of peripheral 5-HT2B receptors initiating PPE and the theory that selective 5-HT2B antagonists might be effective prophylactic therapies for migraine. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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3. Collaboration and knowledge integration for successful brain therapeutics - lessons learned from the pandemic.
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Loza MI, Hmeljak J, Bountra C, Audia JE, Chowdhury S, Weiman S, Merchant K, and Blanco MJ
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- Humans, Translational Research, Biomedical, Brain, Pandemics, COVID-19
- Abstract
Brain diseases are a major cause of death and disability worldwide and contribute significantly to years of potential life lost. Although there have been considerable advances in biological mechanisms associated with brain disorders as well as drug discovery paradigms in recent years, these have not been sufficiently translated into effective treatments. This Special Article expands on Keystone Symposia's pre- and post-pandemic panel discussions on translational neuroscience research. In the article, we discuss how lessons learned from the COVID-19 pandemic can catalyze critical progress in translational research, with efficient collaboration bridging the gap between basic discovery and clinical application. To achieve this, we must place patients at the center of the research paradigm. Furthermore, we need commitment from all collaborators to jointly mitigate the risk associated with the research process. This will require support from investors, the public sector and pharmaceutical companies to translate disease mechanisms into world-class drugs. We also discuss the role of scientific publishing in supporting these models of open innovation. Open science journals can now function as hubs to accelerate progress from discovery to treatments, in neuroscience in particular, making this process less tortuous by bringing scientists together and enabling them to exchange data, tools and knowledge effectively. As stakeholders from a broad range of scientific professions, we feel an urgency to advance brain disease therapies and encourage readers to work together in tackling this challenge., Competing Interests: Competing interests M.-J.B., M.I.L. and K.M. were the organizers of the Keystone Symposia events discussed in this article. J.H. is Senior Editor at DMM but was not involved in any aspect of the editorial handling of the article. S.C. is Head of Research at The Michael J. Fox Foundation for Parkinson's Research and thus has extensive knowledge of this funder's priorities., (© 2022. Published by The Company of Biologists Ltd.)
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- 2022
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4. GNE-064: A Potent, Selective, and Orally Bioavailable Chemical Probe for the Bromodomains of SMARCA2 and SMARCA4 and the Fifth Bromodomain of PBRM1.
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Taylor AM, Bailey C, Belmont LD, Campbell R, Cantone N, Côté A, Crawford TD, Cummings R, DeMent K, Duplessis M, Flynn M, Good AC, Huang HR, Joshi S, Leblanc Y, Murray J, Nasveschuk CG, Neiss A, Poy F, Romero FA, Sandy P, Tang Y, Tsui V, Zawadzke L, Sims RJ 3rd, Audia JE, Bellon SF, Magnuson SR, Albrecht BK, and Cochran AG
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- DNA-Binding Proteins, Humans, Nuclear Proteins, Protein Domains, DNA Helicases, Transcription Factors
- Abstract
Bromodomains are acetyllysine recognition domains present in a variety of human proteins. Bromodomains also bind small molecules that compete with acetyllysine, and therefore bromodomains have been targets for drug discovery efforts. Highly potent and selective ligands with good cellular permeability have been proposed as chemical probes for use in exploring the functions of many of the bromodomain proteins. We report here the discovery of a class of such inhibitors targeting the family VIII bromodomains of SMARCA2 (BRM) and SMARCA4 (BRG1), and PBRM1 (polybromo-1) bromodomain 5. We propose one example from this series, GNE-064, as a chemical probe for the bromodomains SMARCA2, SMARCA4, and PBRM1(5) with the potential for in vivo use.
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- 2022
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5. Ilaprazole and other novel prazole-based compounds that bind Tsg101 inhibit viral budding of HSV-1/2 and HIV from cells.
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Leis J, Luan CH, Audia JE, Dunne SF, and Heath CM
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In many enveloped virus families, including HIV and HSV, a crucial, yet unexploited, step in the viral life cycle is releasing particles from the infected cell membranes. This release process is mediated by host ESCRT complex proteins, which are recruited by viral structural proteins and provides the mechanical means for membrane scission and subsequent viral budding. The prazole drug, tenatoprazole, was previously shown to bind to ESCRT complex member Tsg101 and to quantitatively block the release of infectious HIV-1 from cells in culture. In this report we show that tenatoprazole and a related prazole drug, ilaprazole, effectively block infectious Herpes Simplex Virus (HSV)-1/2 release from Vero cells in culture. By electron microscopy, we found that both prazole drugs block the transit of HSV particles through the cell nuclear membrane resulting in their accumulation in the nucleus. Ilaprazole also quantitatively blocks the release of HIV-1 from 293T cells with an EC
50 of 0.8-1.2 μM, which is much more potent than tenatoprazole. Our results indicate that prazole-based compounds may represent a class of drugs with potential to be broad-spectrum antiviral agents against multiple enveloped viruses, by interrupting cellular Tsg101 interaction with maturing virus, thus blocking the budding process that releases particles from the cell. Importance These results provide the basis for the development of drugs that target enveloped virus budding that can be used ultimately to control multiple virus infections in humans., (Copyright © 2021 Leis et al.)- Published
- 2021
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6. Identification and characterization of second-generation EZH2 inhibitors with extended residence times and improved biological activity.
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Stuckey JI, Cantone NR, Côté A, Arora S, Vivat V, Ramakrishnan A, Mertz JA, Khanna A, Brenneman J, Gehling VS, Moine L, Sims RJ 3rd, Audia JE, Trojer P, Levell JR, and Cummings RT
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- Allosteric Regulation drug effects, Animals, Drug Discovery, Enhancer of Zeste Homolog 2 Protein metabolism, Female, HeLa Cells, Humans, Mice, SCID, Small Molecule Libraries chemistry, Small Molecule Libraries pharmacology, Mice, Enhancer of Zeste Homolog 2 Protein antagonists & inhibitors, Enzyme Inhibitors chemistry, Enzyme Inhibitors pharmacology
- Abstract
The histone methyltransferase EZH2 has been the target of numerous small-molecule inhibitor discovery efforts over the last 10+ years. Emerging clinical data have provided early evidence for single agent activity with acceptable safety profiles for first-generation inhibitors. We have developed kinetic methodologies for studying EZH2-inhibitor-binding kinetics that have allowed us to identify a unique structural modification that results in significant increases in the drug-target residence times of all EZH2 inhibitor scaffolds we have studied. The unexpected residence time enhancement bestowed by this modification has enabled us to create a series of second-generation EZH2 inhibitors with sub-pM binding affinities. We provide both biophysical evidence validating this sub-pM potency and biological evidence demonstrating the utility and relevance of such high-affinity interactions with EZH2., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
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- 2021
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7. Early Drug-Discovery Efforts towards the Identification of EP300/CBP Histone Acetyltransferase (HAT) Inhibitors.
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Huhn AJ, Gardberg AS, Poy F, Brucelle F, Vivat V, Cantone N, Patel G, Patel C, Cummings R, Sims R, Levell J, Audia JE, Bommi-Reddy A, and Wilson JE
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- Crystallography, X-Ray, Dose-Response Relationship, Drug, Enzyme Inhibitors chemical synthesis, Enzyme Inhibitors chemistry, Humans, Models, Molecular, Molecular Structure, Structure-Activity Relationship, p300-CBP Transcription Factors metabolism, Drug Discovery, Enzyme Inhibitors pharmacology, p300-CBP Transcription Factors antagonists & inhibitors
- Abstract
EP300 and CBP (KAT3A/3B) are two highly homologous, multidomain, epigenetic coregulators that play central roles in transcription through the acetylation of lysine residues on histones and other proteins. Both enzymes have been implicated in human diseases, especially cancer. From a high-throughput screen of 191 000 compounds searching for EP300/CBP histone acetyltransferase (HAT) inhibitors, 18 compounds were characterized by a suite of biochemical enzymatic assays and biophysical methods, including X-ray crystallography and native mass spectrometry. This work resulted in the discovery of three distinct mechanistic classes of EP300/CBP HAT inhibitors, including two classes not previously described. The profiles of an example of each class of inhibitor are described in detail. A subsequent medicinal chemistry effort led to the development of a novel class of orally bioavailable AcCoA-competitive EP300/CBP HAT inhibitors with in vivo activity. We believe that this work will prove to be a useful guide for other groups interested in the development of HAT inhibitors., (© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)
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- 2020
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8. Design and Synthesis of Styrenylcyclopropylamine LSD1 Inhibitors.
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Gehling VS, McGrath JP, Duplessis M, Khanna A, Brucelle F, Vaswani RG, Côté A, Stuckey J, Watson V, Cummings RT, Balasubramanian S, Iyer P, Sawant P, Good AC, Albrecht BK, Harmange JC, Audia JE, Bellon SF, Trojer P, and Levell JR
- Abstract
Leveraging the catalytic machinery of LSD1 (KDM1A), a series of covalent styrenylcyclopropane LSD1 inhibitors were identified. These inhibitors represent a new class of mechanism-based inhibitors that target and covalently label the FAD cofactor of LSD1. The series was rapidly progressed to potent biochemical and cellular LSD1 inhibitors with good physical properties. This effort resulted in the identification of 34 , a highly potent (<4 nM biochemical, 2 nM cell, and 1 nM GI
50 ), and selective LSD1 inhibitor. In-depth kinetic profiling of 34 confirmed its covalent mechanism of action, validated the styrenylcyclopropane as an FAD-directed warhead, and demonstrated that the potency of this inhibitor is driven by improved non-covalent binding ( KI ). 34 demonstrated robust cell-killing activity in a panel of AML cell lines and robust antitumor activity in a Kasumi-1 xenograft model of AML when dosed orally at 1.5 mg/kg once daily., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)- Published
- 2020
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9. Design, Synthesis, and Pharmacological Evaluation of Second Generation EZH2 Inhibitors with Long Residence Time.
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Khanna A, Côté A, Arora S, Moine L, Gehling VS, Brenneman J, Cantone N, Stuckey JI, Apte S, Ramakrishnan A, Bruderek K, Bradley WD, Audia JE, Cummings RT, Sims RJ 3rd, Trojer P, and Levell JR
- Abstract
Histone methyltransferase EZH2, which is the catalytic subunit of the PRC2 complex, catalyzes the methylation of histone H3K27-a transcriptionally repressive post-translational modification (PTM). EZH2 is commonly mutated in hematologic malignancies and frequently overexpressed in solid tumors, where its expression level often correlates with poor prognosis. First generation EZH2 inhibitors are beginning to show clinical benefit, and we believe that a second generation EZH2 inhibitor could further build upon this foundation to fully realize the therapeutic potential of EZH2 inhibition. During our medicinal chemistry campaign, we identified 4-thiomethyl pyridone as a key modification that led to significantly increased potency and prolonged residence time. Leveraging this finding, we optimized a series of EZH2 inhibitors, with enhanced antitumor activity and improved physiochemical properties, which have the potential to expand the clinical use of EZH2 inhibition., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)
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- 2020
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10. Mentoring Matters in Drug Discovery.
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Blanco MJ and Audia JE
- Abstract
Mentoring is an essential opportunity to be embraced by medicinal chemists seeking a gratifying career. In this Viewpoint, we highlight the importance of developing an intentional mindset about mentoring to achieve professional goals. The impact of mentoring is even more critical for underrepresented minorities in drug discovery and in particular for women aspiring to achieve leadership positions in the field., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)
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- 2020
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11. Preparation and biological evaluation of BACE1 inhibitors: Leveraging trans-cyclopropyl moieties as ligand efficient conformational constraints.
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Winneroski LL, Erickson JA, Green SJ, Lopez JE, Stout SL, Porter WJ, Timm DE, Audia JE, Barberis M, Beck JP, Boggs LN, Borders AR, Boyer RD, Brier RA, Hembre EJ, Hendle J, Garcia-Losada P, Minguez JM, Mathes BM, May PC, Monk SA, Rankovic Z, Shi Y, Watson BM, Yang Z, and Mergott DJ
- Subjects
- Alzheimer Disease drug therapy, Alzheimer Disease metabolism, Amyloid Precursor Protein Secretases metabolism, Aspartic Acid Endopeptidases metabolism, Crystallography, X-Ray, Cyclopropanes chemical synthesis, Cyclopropanes chemistry, Dose-Response Relationship, Drug, Humans, Ligands, Models, Molecular, Molecular Conformation, Protease Inhibitors chemical synthesis, Protease Inhibitors chemistry, Structure-Activity Relationship, Amyloid Precursor Protein Secretases antagonists & inhibitors, Aspartic Acid Endopeptidases antagonists & inhibitors, Cyclopropanes pharmacology, Protease Inhibitors pharmacology
- Abstract
Inhibition of BACE1 has become an important strategy in the quest for disease modifying agents to slow the progression of Alzheimer's disease. We previously reported the fragment-based discovery of LY2811376, the first BACE1 inhibitor reported to demonstrate robust reduction of human CSF Aβ in a Phase I clinical trial. We also reported on the discovery of LY2886721, a potent BACE1 inhibitor that reached phase 2 clinical trials. Herein we describe the preparation and structure activity relationships (SAR) of a series of BACE1 inhibitors utilizing trans-cyclopropyl moieties as conformational constraints. The design, details of the stereochemically complex organic synthesis, and biological activity of these BACE1 inhibitors is described., (Copyright © 2019 Elsevier Ltd. All rights reserved.)
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- 2020
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12. GNE-371, a Potent and Selective Chemical Probe for the Second Bromodomains of Human Transcription-Initiation-Factor TFIID Subunit 1 and Transcription-Initiation-Factor TFIID Subunit 1-like.
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Wang S, Tsui V, Crawford TD, Audia JE, Burdick DJ, Beresini MH, Côté A, Cummings R, Duplessis M, Flynn EM, Hewitt MC, Huang HR, Jayaram H, Jiang Y, Joshi S, Murray J, Nasveschuk CG, Pardo E, Poy F, Romero FA, Tang Y, Taylor AM, Wang J, Xu Z, Zawadzke LE, Zhu X, Albrecht BK, Magnuson SR, Bellon S, and Cochran AG
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- Humans, Models, Molecular, Protein Conformation, Protein Domains, Benzimidazoles metabolism, Drug Design, Molecular Probes metabolism, Transcription Factor TFIID chemistry, Transcription Factor TFIID metabolism
- Abstract
The biological functions of the dual bromodomains of human transcription-initiation-factor TFIID subunit 1 (TAF1(1,2)) remain unknown, although TAF1 has been identified as a potential target for oncology research. Here, we describe the discovery of a potent and selective in vitro tool compound for TAF1(2), starting from a previously reported lead. A cocrystal structure of lead compound 2 bound to TAF1(2) enabled structure-based design and structure-activity-relationship studies that ultimately led to our in vitro tool compound, 27 (GNE-371). Compound 27 binds TAF1(2) with an IC
50 of 10 nM while maintaining excellent selectivity over other bromodomain-family members. Compound 27 is also active in a cellular-TAF1(2) target-engagement assay (IC50 = 38 nM) and exhibits antiproliferative synergy with the BET inhibitor JQ1, suggesting engagement of endogenous TAF1 by 27 and further supporting the use of 27 in mechanistic and target-validation studies.- Published
- 2018
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13. GNE-886: A Potent and Selective Inhibitor of the Cat Eye Syndrome Chromosome Region Candidate 2 Bromodomain (CECR2).
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Crawford TD, Audia JE, Bellon S, Burdick DJ, Bommi-Reddy A, Côté A, Cummings RT, Duplessis M, Flynn EM, Hewitt M, Huang HR, Jayaram H, Jiang Y, Joshi S, Kiefer JR, Murray J, Nasveschuk CG, Neiss A, Pardo E, Romero FA, Sandy P, Sims RJ 3rd, Tang Y, Taylor AM, Tsui V, Wang J, Wang S, Wang Y, Xu Z, Zawadzke L, Zhu X, Albrecht BK, Magnuson SR, and Cochran AG
- Abstract
The biological function of bromodomains, epigenetic readers of acetylated lysine residues, remains largely unknown. Herein we report our efforts to discover a potent and selective inhibitor of the bromodomain of cat eye syndrome chromosome region candidate 2 (CECR2). Screening of our internal medicinal chemistry collection led to the identification of a pyrrolopyridone chemical lead, and subsequent structure-based drug design led to a potent and selective CECR2 bromodomain inhibitor (GNE-886) suitable for use as an in vitro tool compound.
- Published
- 2017
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14. US immigration order strikes against biotech.
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Levin JM, Holtzman SH, Maraganore J, Hastings PJ, Cohen R, Dahiyat B, Adams J, Adams C, Ahrens B, Albers J, Aspinall MG, Audia JE, Babler M, Barrett P, Barry Z, Bermingham N, Bloch S, Blum RI, Bolno PB, Bonney MW, Booth B, Bradbury DM, Brauer SK, Byers B, Cagnoni PJ, Cali BM, Ciechanover I, Clark C, Clayman MD, Cleland JL, Cobb P, Cooper R, Currie MG, Diekman J, Dobmeier EL, Doerfler D, Donley EL, Dunsire D, During M, Eckstein JW, Elenko E, Exter NA, Fleming JJ, Flesher GJ, Formela JF, Forrester R, Francois C, Franklin H, Freeman MW, Furst H, Gage LP, Galakatos N, Gallagher BM, Geraghty JA, Gill S, Goeddel DV, Goldsmith MA, Gowen M, Goyal V, Graney T, Grayzel D, Greene B, Grint P, Gutierrez-Ramos JC, Haney B, Ha-Ngoc T, Harris T, Hasnain F, Hata YS, Hecht P, Henshaw L, Heyman R, Hoppenot H, Horvitz HR, Hughes TE, Hutton WS, Isaacs ST, Jenkins A, Jonker J, Kaplan J, Karsen P, Keiper J, Kim J, Kindler J, King R, King V, Kjellson N, Koenig S, Koenig G, Kolchinsky P, Laikind P, Langer RB, Lee JJ, Leff JS, Leicher BA, Leschly N, Levin A, Levin M, Levine AJ, Levy A, Liu DR, Lodish HF, Lopatin U, Love TW, Macdonald G, Maderis GJ, Mahadevia A, Mahanthappa NK, Martin JF, Martin A, Martucci WE, McArthur JG, McCann CM, McCarthy SA, McDonough CG, Mendlein J, Miller L, Miralles D, Moch KI, More B, Myers AG, Narachi MA, Nashat A, Nelson W, Newell WJ, Olle B, Osborn JE, Owens JC, Pande A, Papadopoulos S, Parker HS, Parmar KM, Patterson MR, Paul SM, Perez R, Perry M, Pfeffer CG, Powell M, Pruzanski M, Purcell DJ, Rakhit A, Ramamoorthi K, Rastetter W, Rawcliffe AA, Reid LE, Renaud RC, Rhodes JP, Rieflin WJ, Robins C, Rocklage SM, Rosenblatt M, Rosin JG, Rutter WJ, Saha S, Samuels C, Sato VL, Scangos G, Scarlett JA, Schenkein D, Schreiber SL, Schwab A, Sekhri P, Shah R, Shenk T, Siegall CB, Simon NJ, Simonian N, Stein J, Su M, Szela MT, Taglietti M, Tandon N, Termeer H, Thornberry NA, Tolar M, Ulevitch R, Vaishnaw AK, VanLent A, Varsavsky M, Vlasuk GP, Vounatsos M, Waksal SG, Warma N, Watts RJ, Werber Y, Westphal C, Wierenga W, Williams DE, Williams LR, Xanthopoulos KG, Zohar D, and Zweifach SS
- Subjects
- Humans, Population Dynamics, Biotechnology legislation & jurisprudence, Emigration and Immigration legislation & jurisprudence, Public Policy legislation & jurisprudence
- Published
- 2017
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15. Identification of (R)-N-((4-Methoxy-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2-methyl-1-(1-(1-(2,2,2-trifluoroethyl)piperidin-4-yl)ethyl)-1H-indole-3-carboxamide (CPI-1205), a Potent and Selective Inhibitor of Histone Methyltransferase EZH2, Suitable for Phase I Clinical Trials for B-Cell Lymphomas.
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Vaswani RG, Gehling VS, Dakin LA, Cook AS, Nasveschuk CG, Duplessis M, Iyer P, Balasubramanian S, Zhao F, Good AC, Campbell R, Lee C, Cantone N, Cummings RT, Normant E, Bellon SF, Albrecht BK, Harmange JC, Trojer P, Audia JE, Zhang Y, Justin N, Chen S, Wilson JR, and Gamblin SJ
- Subjects
- Animals, Antineoplastic Agents chemical synthesis, Antineoplastic Agents chemistry, Cell Proliferation drug effects, Dogs, Dose-Response Relationship, Drug, Drug Screening Assays, Antitumor, Enzyme Inhibitors chemical synthesis, Enzyme Inhibitors chemistry, Histone Methyltransferases, Histone-Lysine N-Methyltransferase metabolism, Humans, Indoles chemical synthesis, Indoles chemistry, Models, Molecular, Molecular Structure, Neoplasms, Experimental drug therapy, Neoplasms, Experimental pathology, Piperidines chemical synthesis, Piperidines chemistry, Rats, Structure-Activity Relationship, Antineoplastic Agents pharmacology, Clinical Trials, Phase I as Topic, Enzyme Inhibitors pharmacology, Histone-Lysine N-Methyltransferase antagonists & inhibitors, Indoles pharmacology, Lymphoma, B-Cell drug therapy, Piperidines pharmacology
- Abstract
Polycomb repressive complex 2 (PRC2) has been shown to play a major role in transcriptional silencing in part by installing methylation marks on lysine 27 of histone 3. Dysregulation of PRC2 function correlates with certain malignancies and poor prognosis. EZH2 is the catalytic engine of the PRC2 complex and thus represents a key candidate oncology target for pharmacological intervention. Here we report the optimization of our indole-based EZH2 inhibitor series that led to the identification of CPI-1205, a highly potent (biochemical IC
50 = 0.002 μM, cellular EC50 = 0.032 μM) and selective inhibitor of EZH2. This compound demonstrates robust antitumor effects in a Karpas-422 xenograft model when dosed at 160 mg/kg BID and is currently in Phase I clinical trials. Additionally, we disclose the co-crystal structure of our inhibitor series bound to the human PRC2 complex.- Published
- 2016
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16. Identification of potent, selective KDM5 inhibitors.
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Gehling VS, Bellon SF, Harmange JC, LeBlanc Y, Poy F, Odate S, Buker S, Lan F, Arora S, Williamson KE, Sandy P, Cummings RT, Bailey CM, Bergeron L, Mao W, Gustafson A, Liu Y, VanderPorten E, Audia JE, Trojer P, and Albrecht BK
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- Animals, Binding Sites, Blotting, Western, Cell Line, Drug Discovery, Enzyme Activation drug effects, Enzyme Inhibitors chemistry, Enzyme Inhibitors isolation & purification, Humans, Inhibitory Concentration 50, Mice, Microsomes, Liver enzymology, Models, Molecular, Rats, Enzyme Inhibitors pharmacology, Retinoblastoma-Binding Protein 2 antagonists & inhibitors
- Abstract
This communication describes the identification and optimization of a series of pan-KDM5 inhibitors derived from compound 1, a hit initially identified against KDM4C. Compound 1 was optimized to afford compound 20, a 10nM inhibitor of KDM5A. Compound 20 is highly selective for the KDM5 enzymes versus other histone lysine demethylases and demonstrates activity in a cellular assay measuring the increase in global histone 3 lysine 4 tri-methylation (H3K4me3). In addition compound 20 has good ADME properties, excellent mouse PK, and is a suitable starting point for further optimization., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
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17. Regulatory T Cell Modulation by CBP/EP300 Bromodomain Inhibition.
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Ghosh S, Taylor A, Chin M, Huang HR, Conery AR, Mertz JA, Salmeron A, Dakle PJ, Mele D, Cote A, Jayaram H, Setser JW, Poy F, Hatzivassiliou G, DeAlmeida-Nagata D, Sandy P, Hatton C, Romero FA, Chiang E, Reimer T, Crawford T, Pardo E, Watson VG, Tsui V, Cochran AG, Zawadzke L, Harmange JC, Audia JE, Bryant BM, Cummings RT, Magnuson SR, Grogan JL, Bellon SF, Albrecht BK, Sims RJ 3rd, and Lora JM
- Subjects
- Acetylation drug effects, CREB-Binding Protein chemistry, CREB-Binding Protein metabolism, Cell Differentiation drug effects, Cell Line, Cells, Cultured, E1A-Associated p300 Protein chemistry, E1A-Associated p300 Protein metabolism, Forkhead Transcription Factors metabolism, Histones metabolism, Humans, Molecular Docking Simulation, Protein Structure, Tertiary drug effects, T-Lymphocytes, Regulatory cytology, T-Lymphocytes, Regulatory metabolism, Transcriptome drug effects, CREB-Binding Protein antagonists & inhibitors, E1A-Associated p300 Protein antagonists & inhibitors, Small Molecule Libraries chemistry, Small Molecule Libraries pharmacology, T-Lymphocytes, Regulatory drug effects
- Abstract
Covalent modification of histones is a fundamental mechanism of regulated gene expression in eukaryotes, and interpretation of histone modifications is an essential feature of epigenetic control. Bromodomains are specialized binding modules that interact with acetylated histones, linking chromatin recognition to gene transcription. Because of their ability to function in a domain-specific fashion, selective disruption of bromodomain:acetylated histone interactions with chemical probes serves as a powerful means for understanding biological processes regulated by these chromatin adaptors. Here we describe the discovery and characterization of potent and selective small molecule inhibitors for the bromodomains of CREBBP/EP300 that engage their target in cellular assays. We use these tools to demonstrate a critical role for CREBBP/EP300 bromodomains in regulatory T cell biology. Because regulatory T cell recruitment to tumors is a major mechanism of immune evasion by cancer cells, our data highlight the importance of CREBBP/EP300 bromodomain inhibition as a novel, small molecule-based approach for cancer immunotherapy., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2016
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18. Diving into the Water: Inducible Binding Conformations for BRD4, TAF1(2), BRD9, and CECR2 Bromodomains.
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Crawford TD, Tsui V, Flynn EM, Wang S, Taylor AM, Côté A, Audia JE, Beresini MH, Burdick DJ, Cummings R, Dakin LA, Duplessis M, Good AC, Hewitt MC, Huang HR, Jayaram H, Kiefer JR, Jiang Y, Murray J, Nasveschuk CG, Pardo E, Poy F, Romero FA, Tang Y, Wang J, Xu Z, Zawadzke LE, Zhu X, Albrecht BK, Magnuson SR, Bellon S, and Cochran AG
- Subjects
- Binding Sites drug effects, Cell Cycle Proteins, Dose-Response Relationship, Drug, Fluorescence Resonance Energy Transfer, Fluorometry, Histone Acetyltransferases metabolism, Humans, Ligands, Models, Molecular, Molecular Conformation, Nuclear Proteins metabolism, Pyridones chemical synthesis, Pyridones chemistry, Pyrroles chemical synthesis, Pyrroles chemistry, Structure-Activity Relationship, TATA-Binding Protein Associated Factors metabolism, Transcription Factor TFIID metabolism, Transcription Factors metabolism, Histone Acetyltransferases antagonists & inhibitors, Nuclear Proteins antagonists & inhibitors, Pyridones pharmacology, Pyrroles pharmacology, TATA-Binding Protein Associated Factors antagonists & inhibitors, Transcription Factor TFIID antagonists & inhibitors, Transcription Factors antagonists & inhibitors, Water chemistry
- Abstract
The biological role played by non-BET bromodomains remains poorly understood, and it is therefore imperative to identify potent and highly selective inhibitors to effectively explore the biology of individual bromodomain proteins. A ligand-efficient nonselective bromodomain inhibitor was identified from a 6-methyl pyrrolopyridone fragment. Small hydrophobic substituents replacing the N-methyl group were designed directing toward the conserved bromodomain water pocket, and two distinct binding conformations were then observed. The substituents either directly displaced and rearranged the conserved solvent network, as in BRD4(1) and TAF1(2), or induced a narrow hydrophobic channel adjacent to the lipophilic shelf, as in BRD9 and CECR2. The preference of distinct substituents for individual bromodomains provided selectivity handles useful for future lead optimization efforts for selective BRD9, CECR2, and TAF1(2) inhibitors.
- Published
- 2016
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19. Histone Modifications and Cancer.
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Audia JE and Campbell RM
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- Acetylation, Drug Resistance, Neoplasm genetics, Gene Expression Regulation, Neoplastic, Methylation, Neoplasms therapy, Chromatin Assembly and Disassembly, Epigenesis, Genetic, Histone Code, Histones metabolism, Neoplasms genetics
- Abstract
Histone posttranslational modifications represent a versatile set of epigenetic marks involved not only in dynamic cellular processes, such as transcription and DNA repair, but also in the stable maintenance of repressive chromatin. In this article, we review many of the key and newly identified histone modifications known to be deregulated in cancer and how this impacts function. The latter part of the article addresses the challenges and current status of the epigenetic drug development process as it applies to cancer therapeutics., (Copyright © 2016 Cold Spring Harbor Laboratory Press; all rights reserved.)
- Published
- 2016
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20. Fragment-Based Discovery of a Selective and Cell-Active Benzodiazepinone CBP/EP300 Bromodomain Inhibitor (CPI-637).
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Taylor AM, Côté A, Hewitt MC, Pastor R, Leblanc Y, Nasveschuk CG, Romero FA, Crawford TD, Cantone N, Jayaram H, Setser J, Murray J, Beresini MH, de Leon Boenig G, Chen Z, Conery AR, Cummings RT, Dakin LA, Flynn EM, Huang OW, Kaufman S, Keller PJ, Kiefer JR, Lai T, Li Y, Liao J, Liu W, Lu H, Pardo E, Tsui V, Wang J, Wang Y, Xu Z, Yan F, Yu D, Zawadzke L, Zhu X, Zhu X, Sims RJ 3rd, Cochran AG, Bellon S, Audia JE, Magnuson S, and Albrecht BK
- Abstract
CBP and EP300 are highly homologous, bromodomain-containing transcription coactivators involved in numerous cellular pathways relevant to oncology. As part of our effort to explore the potential therapeutic implications of selectively targeting bromodomains, we set out to identify a CBP/EP300 bromodomain inhibitor that was potent both in vitro and in cellular target engagement assays and was selective over the other members of the bromodomain family. Reported here is a series of cell-potent and selective probes of the CBP/EP300 bromodomains, derived from the fragment screening hit 4-methyl-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one.
- Published
- 2016
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21. Identification of a Benzoisoxazoloazepine Inhibitor (CPI-0610) of the Bromodomain and Extra-Terminal (BET) Family as a Candidate for Human Clinical Trials.
- Author
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Albrecht BK, Gehling VS, Hewitt MC, Vaswani RG, Côté A, Leblanc Y, Nasveschuk CG, Bellon S, Bergeron L, Campbell R, Cantone N, Cooper MR, Cummings RT, Jayaram H, Joshi S, Mertz JA, Neiss A, Normant E, O'Meara M, Pardo E, Poy F, Sandy P, Supko J, Sims RJ 3rd, Harmange JC, Taylor AM, and Audia JE
- Subjects
- Animals, Antineoplastic Agents pharmacokinetics, Antineoplastic Agents pharmacology, Azepines pharmacokinetics, Azepines pharmacology, Cell Cycle Proteins, Cell Line, Tumor, Clinical Trials as Topic, Dogs, Genes, myc drug effects, Humans, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute metabolism, Mice, Nuclear Proteins chemistry, Nuclear Proteins metabolism, Proto-Oncogene Proteins c-myc genetics, Rats, Transcription Factors chemistry, Transcription Factors metabolism, Xenograft Model Antitumor Assays, Antineoplastic Agents chemistry, Antineoplastic Agents therapeutic use, Azepines chemistry, Azepines therapeutic use, Gene Expression Regulation, Neoplastic drug effects, Leukemia, Myeloid, Acute drug therapy, Nuclear Proteins antagonists & inhibitors, Transcription Factors antagonists & inhibitors
- Abstract
In recent years, inhibition of the interaction between the bromodomain and extra-terminal domain (BET) family of chromatin adaptors and acetyl-lysine residues on chromatin has emerged as a promising approach to regulate the expression of important disease-relevant genes, including MYC, BCL-2, and NF-κB. Here we describe the identification and characterization of a potent and selective benzoisoxazoloazepine BET bromodomain inhibitor that attenuates BET-dependent gene expression in vivo, demonstrates antitumor efficacy in an MV-4-11 mouse xenograft model, and is currently undergoing human clinical trials for hematological malignancies (CPI-0610).
- Published
- 2016
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22. Corrigendum: The promise and peril of chemical probes.
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Arrowsmith CH, Audia JE, Austin C, Baell J, Bennett J, Blagg J, Bountra C, Brennan PE, Brown PJ, Bunnage ME, Buser-Doepner C, Campbell RM, Carter AJ, Cohen P, Copeland RA, Cravatt B, Dahlin JL, Dhanak D, Edwards AM, Frederiksen M, Frye SV, Gray N, Grimshaw CE, Hepworth D, Howe T, Huber KV, Jin J, Knapp S, Kotz JD, Kruger RG, Lowe D, Mader MM, Marsden B, Mueller-Fahrnow A, Müller S, O'Hagan RC, Overington JP, Owen DR, Rosenberg SH, Ross R, Roth B, Schapira M, Schreiber SL, Shoichet B, Sundström M, Superti-Furga G, Taunton J, Toledo-Sherman L, Walpole C, Walters MA, Willson TM, Workman P, Young RN, and Zuercher WJ
- Published
- 2015
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23. Discovery, design, and synthesis of indole-based EZH2 inhibitors.
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Gehling VS, Vaswani RG, Nasveschuk CG, Duplessis M, Iyer P, Balasubramanian S, Zhao F, Good AC, Campbell R, Lee C, Dakin LA, Cook AS, Gagnon A, Harmange JC, Audia JE, Cummings RT, Normant E, Trojer P, and Albrecht BK
- Subjects
- Animals, Antineoplastic Agents chemical synthesis, Cell Line, Tumor, Chemistry Techniques, Synthetic, Drug Design, Drug Discovery, Drug Screening Assays, Antitumor, Drug Stability, Enhancer of Zeste Homolog 2 Protein, HeLa Cells drug effects, Humans, Inhibitory Concentration 50, Mice, Molecular Targeted Therapy methods, Small Molecule Libraries chemistry, Small Molecule Libraries pharmacology, Structure-Activity Relationship, Xenograft Model Antitumor Assays, Antineoplastic Agents chemistry, Antineoplastic Agents pharmacology, Indoles chemistry, Polycomb Repressive Complex 2 antagonists & inhibitors
- Abstract
The discovery and optimization of a series of small molecule EZH2 inhibitors is described. Starting from dimethylpyridone HTS hit (2), a series of indole-based EZH2 inhibitors were identified. Biochemical potency and microsomal stability were optimized during these studies and afforded compound 22. This compound demonstrates nanomolar levels of biochemical potency (IC50=0.002 μM), cellular potency (EC50=0.080 μM), and afforded tumor regression when dosed (200 mpk SC BID) in an EZH2 dependent tumor xenograft model., (Copyright © 2015. Published by Elsevier Ltd.)
- Published
- 2015
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24. The promise and peril of chemical probes.
- Author
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Arrowsmith CH, Audia JE, Austin C, Baell J, Bennett J, Blagg J, Bountra C, Brennan PE, Brown PJ, Bunnage ME, Buser-Doepner C, Campbell RM, Carter AJ, Cohen P, Copeland RA, Cravatt B, Dahlin JL, Dhanak D, Edwards AM, Frederiksen M, Frye SV, Gray N, Grimshaw CE, Hepworth D, Howe T, Huber KV, Jin J, Knapp S, Kotz JD, Kruger RG, Lowe D, Mader MM, Marsden B, Mueller-Fahrnow A, Müller S, O'Hagan RC, Overington JP, Owen DR, Rosenberg SH, Roth B, Ross R, Schapira M, Schreiber SL, Shoichet B, Sundström M, Superti-Furga G, Taunton J, Toledo-Sherman L, Walpole C, Walters MA, Willson TM, Workman P, Young RN, and Zuercher WJ
- Subjects
- Biomedical Research instrumentation, Humans, Intellectual Property, Internet, Molecular Probes pharmacology, Molecular Weight, Sensitivity and Specificity, Small Molecule Libraries pharmacology, Biomedical Research methods, Information Dissemination ethics, Molecular Probes chemistry, Small Molecule Libraries chemistry
- Published
- 2015
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25. Preparation and biological evaluation of conformationally constrained BACE1 inhibitors.
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Winneroski LL, Schiffler MA, Erickson JA, May PC, Monk SA, Timm DE, Audia JE, Beck JP, Boggs LN, Borders AR, Boyer RD, Brier RA, Hudziak KJ, Klimkowski VJ, Garcia Losada P, Mathes BM, Stout SL, Watson BM, and Mergott DJ
- Subjects
- Amyloid Precursor Protein Secretases chemistry, Amyloid Precursor Protein Secretases isolation & purification, Animals, Aspartic Acid Endopeptidases chemistry, Aspartic Acid Endopeptidases isolation & purification, Brain Chemistry, Bridged Bicyclo Compounds chemistry, Crystallography, X-Ray, Drug Design, Humans, Mice, Molecular Conformation, Molecular Docking Simulation, Protease Inhibitors chemistry, Stereoisomerism, Thiazines chemistry, Amyloid Precursor Protein Secretases antagonists & inhibitors, Aspartic Acid Endopeptidases antagonists & inhibitors, Bridged Bicyclo Compounds chemical synthesis, Protease Inhibitors chemical synthesis, Thiazines chemical synthesis
- Abstract
The BACE1 enzyme is a key target for Alzheimer's disease. During our BACE1 research efforts, fragment screening revealed that bicyclic thiazine 3 had low millimolar activity against BACE1. Analysis of the co-crystal structure of 3 suggested that potency could be increased through extension toward the S3 pocket and through conformational constraint of the thiazine core. Pursuit of S3-binding groups produced low micromolar inhibitor 6, which informed the S3-design for constrained analogs 7 and 8, themselves prepared via independent, multi-step synthetic routes. Biological characterization of BACE inhibitors 6-8 is described., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2015
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26. Development of methyl isoxazoleazepines as inhibitors of BET.
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Hewitt MC, Leblanc Y, Gehling VS, Vaswani RG, Côté A, Nasveschuk CG, Taylor AM, Harmange JC, Audia JE, Pardo E, Cummings R, Joshi S, Sandy P, Mertz JA, Sims RJ 3rd, Bergeron L, Bryant BM, Bellon S, Poy F, Jayaram H, Tang Y, and Albrecht BK
- Subjects
- Azepines chemical synthesis, Azepines chemistry, Cell Cycle Proteins, Dose-Response Relationship, Drug, Humans, Models, Molecular, Molecular Structure, Oxazoles chemical synthesis, Oxazoles chemistry, Structure-Activity Relationship, Azepines pharmacology, Drug Design, Nuclear Proteins antagonists & inhibitors, Oxazoles pharmacology, Protein Serine-Threonine Kinases antagonists & inhibitors, RNA-Binding Proteins antagonists & inhibitors, Transcription Factors antagonists & inhibitors
- Abstract
In this report we detail the evolution of our previously reported thiophene isoxazole BET inhibitor chemotype exemplified by CPI-3 to a novel bromodomain selective chemotype (the methyl isoxazoleazepine chemotype) exemplified by carboxamide 23. The methyl isoxazoleazepine chemotype provides potent inhibition of the bromodomains of the BET family, excellent in vivo PK across species, low unbound clearance, and target engagement in a MYC PK-PD model., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2015
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27. Discovery of Benzotriazolo[4,3-d][1,4]diazepines as Orally Active Inhibitors of BET Bromodomains.
- Author
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Taylor AM, Vaswani RG, Gehling VS, Hewitt MC, Leblanc Y, Audia JE, Bellon S, Cummings RT, Côté A, Harmange JC, Jayaram H, Joshi S, Lora JM, Mertz JA, Neiss A, Pardo E, Nasveschuk CG, Poy F, Sandy P, Setser JW, Sims RJ 3rd, Tang Y, and Albrecht BK
- Abstract
Inhibition of the bromodomains of the BET family, of which BRD4 is a member, has been shown to decrease myc and interleukin (IL) 6 in vivo, markers that are of therapeutic relevance to cancer and inflammatory disease, respectively. Herein we report substituted benzo[b]isoxazolo[4,5-d]azepines and benzotriazolo[4,3-d][1,4]diazepines as fragment-derived novel inhibitors of the bromodomain of BRD4. Compounds from these series were potent and selective in cells, and subsequent optimization of microsomal stability yielded representatives that demonstrated dose- and time-dependent reduction of plasma IL-6 in mice.
- Published
- 2015
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28. The potent BACE1 inhibitor LY2886721 elicits robust central Aβ pharmacodynamic responses in mice, dogs, and humans.
- Author
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May PC, Willis BA, Lowe SL, Dean RA, Monk SA, Cocke PJ, Audia JE, Boggs LN, Borders AR, Brier RA, Calligaro DO, Day TA, Ereshefsky L, Erickson JA, Gevorkyan H, Gonzales CR, James DE, Jhee SS, Komjathy SF, Li L, Lindstrom TD, Mathes BM, Martényi F, Sheehan SM, Stout SL, Timm DE, Vaught GM, Watson BM, Winneroski LL, Yang Z, and Mergott DJ
- Subjects
- Amyloid beta-Peptides blood, Amyloid beta-Peptides cerebrospinal fluid, Animals, Disease Models, Animal, Dogs, Heterocyclic Compounds, 2-Ring pharmacokinetics, Heterocyclic Compounds, 2-Ring therapeutic use, Humans, Mice, Picolinic Acids pharmacokinetics, Picolinic Acids therapeutic use, Protease Inhibitors pharmacokinetics, Protease Inhibitors therapeutic use, Alzheimer Disease drug therapy, Amyloid Precursor Protein Secretases antagonists & inhibitors, Amyloid beta-Peptides metabolism, Aspartic Acid Endopeptidases antagonists & inhibitors, Heterocyclic Compounds, 2-Ring pharmacology, Picolinic Acids pharmacology, Protease Inhibitors pharmacology
- Abstract
BACE1 is a key protease controlling the formation of amyloid β, a peptide hypothesized to play a significant role in the pathogenesis of Alzheimer's disease (AD). Therefore, the development of potent and selective inhibitors of BACE1 has been a focus of many drug discovery efforts in academia and industry. Herein, we report the nonclinical and early clinical development of LY2886721, a BACE1 active site inhibitor that reached phase 2 clinical trials in AD. LY2886721 has high selectivity against key off-target proteases, which efficiently translates in vitro activity into robust in vivo amyloid β lowering in nonclinical animal models. Similar potent and persistent amyloid β lowering was observed in plasma and lumbar CSF when single and multiple doses of LY2886721 were administered to healthy human subjects. Collectively, these data add support for BACE1 inhibition as an effective means of amyloid lowering and as an attractive target for potential disease modification therapy in AD., (Copyright © 2015 the authors 0270-6474/15/351199-12$15.00/0.)
- Published
- 2015
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29. EZH2 inhibitor efficacy in non-Hodgkin's lymphoma does not require suppression of H3K27 monomethylation.
- Author
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Bradley WD, Arora S, Busby J, Balasubramanian S, Gehling VS, Nasveschuk CG, Vaswani RG, Yuan CC, Hatton C, Zhao F, Williamson KE, Iyer P, Méndez J, Campbell R, Cantone N, Garapaty-Rao S, Audia JE, Cook AS, Dakin LA, Albrecht BK, Harmange JC, Daniels DL, Cummings RT, Bryant BM, Normant E, and Trojer P
- Subjects
- Amino Acid Sequence, Animals, Cell Line, Tumor, Cell Proliferation drug effects, Disease Models, Animal, Enzyme Inhibitors chemistry, Enzyme Inhibitors therapeutic use, Histones chemistry, Humans, Kinetics, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Large B-Cell, Diffuse pathology, Lymphoma, Non-Hodgkin metabolism, Lymphoma, Non-Hodgkin pathology, Methylation, Mice, Mice, Nude, Mutation, Peptides analysis, Polycomb Repressive Complex 2 genetics, Polycomb Repressive Complex 2 metabolism, Small Molecule Libraries chemistry, Small Molecule Libraries therapeutic use, Transplantation, Heterologous, Apoptosis drug effects, Enzyme Inhibitors toxicity, Histones metabolism, Polycomb Repressive Complex 2 antagonists & inhibitors, Small Molecule Libraries toxicity
- Abstract
The histone lysine methyltransferase (MT) Enhancer of Zeste Homolog 2 (EZH2) is considered an oncogenic driver in a subset of germinal center B-cell-like diffuse large B cell lymphoma (GCB-DLBCL) and follicular lymphoma due to the presence of recurrent, monoallelic mutations in the EZH2 catalytic domain. These genomic data suggest that targeting the EZH2 MT activity is a valid therapeutic strategy for the treatment of lymphoma patients with EZH2 mutations. Here we report the identification of highly potent and selective EZH2 small molecule inhibitors, their validation by a cellular thermal shift assay, application across a large cell panel representing various non-Hodgkin's lymphoma (NHL) subtypes, and their efficacy in EZH2mutant-containing GCB-DLBCL xenograft models. Surprisingly, our EZH2 inhibitors selectively affect the turnover of trimethylated, but not monomethylated histone H3 lysine 27 at pharmacologically relevant doses. Importantly, we find that these inhibitors are broadly efficacious also in NHL models with wild-type EZH2.
- Published
- 2014
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30. A practical synthesis of indoles via a Pd-catalyzed C-N ring formation.
- Author
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Vaswani RG, Albrecht BK, Audia JE, Côté A, Dakin LA, Duplessis M, Gehling VS, Harmange JC, Hewitt MC, Leblanc Y, Nasveschuk CG, and Taylor AM
- Subjects
- Amines chemistry, Catalysis, Cyclization, Indoles chemistry, Molecular Structure, Indoles chemical synthesis, Palladium chemistry
- Abstract
A method for the synthesis of N-functionalized C2-/C3-substituted indoles via Pd-catalyzed C-N bond coupling of halo-aryl enamines is described. The general strategy utilizes a variety of amines and β-keto esters which are elaborated into halo-aryl enamines as latent precursors to indoles. The preferred conditions comprising the RuPhos precatalyst and RuPhos in the presence of NaOMe in 1,4-dioxane tolerate a variety of substituents and are scalable for the construction of indoles in multigram quantities.
- Published
- 2014
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31. Discovery and Optimization of Tetramethylpiperidinyl Benzamides as Inhibitors of EZH2.
- Author
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Nasveschuk CG, Gagnon A, Garapaty-Rao S, Balasubramanian S, Campbell R, Lee C, Zhao F, Bergeron L, Cummings R, Trojer P, Audia JE, Albrecht BK, and Harmange JC
- Abstract
The identification and development of a novel series of small molecule Enhancer of Zeste Homologue 2 (EZH2) inhibitors is described. A concise and modular synthesis enabled the rapid development of structure-activity relationships, which led to the identification of 44 as a potent, SAM-competitive inhibitor of EZH2 that dose-dependently decreased global H3K27me3 in KARPAS-422 lymphoma cells.
- Published
- 2014
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32. Identification of EZH2 and EZH1 small molecule inhibitors with selective impact on diffuse large B cell lymphoma cell growth.
- Author
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Garapaty-Rao S, Nasveschuk C, Gagnon A, Chan EY, Sandy P, Busby J, Balasubramanian S, Campbell R, Zhao F, Bergeron L, Audia JE, Albrecht BK, Harmange JC, Cummings R, and Trojer P
- Subjects
- Antineoplastic Agents chemistry, Cell Proliferation drug effects, Dose-Response Relationship, Drug, Drug Screening Assays, Antitumor, Enhancer of Zeste Homolog 2 Protein, HeLa Cells, Humans, Lymphoma, Large B-Cell, Diffuse metabolism, Molecular Structure, Polycomb Repressive Complex 2 metabolism, Small Molecule Libraries chemistry, Structure-Activity Relationship, Tumor Cells, Cultured, Antineoplastic Agents pharmacology, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Large B-Cell, Diffuse pathology, Polycomb Repressive Complex 2 antagonists & inhibitors, Small Molecule Libraries pharmacology
- Abstract
The histone methyltransferase enhancer of Zeste homolog 2 (EZH2) is a candidate oncogene due to its prevalent overexpression in malignant diseases, including late stage prostate and breast cancers. The dependency of cancer cells on EZH2 activity is also predicated by recurrent missense mutations residing in the catalytic domain of EZH2 that have been identified in subtypes of diffuse large B cell lymphoma, follicular lymphoma and melanoma. Herein, we report the identification of a highly selective small molecule inhibitor series of EZH2 and EZH1. These compounds inhibit wild-type and mutant versions of EZH2 with nanomolar potency, suppress global histone H3-lysine 27 methylation, affect gene expression, and cause selective proliferation defects. These compounds represent a structurally distinct EZH2 inhibitor chemotype for the exploration of the role of Polycomb Repressive Complex 2-mediated H3K27 methylation in various biological contexts., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
- Published
- 2013
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33. Discovery, Design, and Optimization of Isoxazole Azepine BET Inhibitors.
- Author
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Gehling VS, Hewitt MC, Vaswani RG, Leblanc Y, Côté A, Nasveschuk CG, Taylor AM, Harmange JC, Audia JE, Pardo E, Joshi S, Sandy P, Mertz JA, Sims RJ 3rd, Bergeron L, Bryant BM, Bellon S, Poy F, Jayaram H, Sankaranarayanan R, Yellapantula S, Bangalore Srinivasamurthy N, Birudukota S, and Albrecht BK
- Abstract
The identification of a novel series of small molecule BET inhibitors is described. Using crystallographic binding modes of an amino-isoxazole fragment and known BET inhibitors, a structure-based drug design effort lead to a novel isoxazole azepine scaffold. This scaffold showed good potency in biochemical and cellular assays and oral activity in an in vivo model of BET inhibition.
- Published
- 2013
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34. Robust central reduction of amyloid-β in humans with an orally available, non-peptidic β-secretase inhibitor.
- Author
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May PC, Dean RA, Lowe SL, Martenyi F, Sheehan SM, Boggs LN, Monk SA, Mathes BM, Mergott DJ, Watson BM, Stout SL, Timm DE, Smith Labell E, Gonzales CR, Nakano M, Jhee SS, Yen M, Ereshefsky L, Lindstrom TD, Calligaro DO, Cocke PJ, Greg Hall D, Friedrich S, Citron M, and Audia JE
- Subjects
- Adult, Alzheimer Disease drug therapy, Amyloid Precursor Protein Secretases analysis, Amyloid beta-Peptides cerebrospinal fluid, Amyloid beta-Protein Precursor cerebrospinal fluid, Amyloid beta-Protein Precursor genetics, Analysis of Variance, Animals, Aspartic Acid Endopeptidases analysis, Cells, Cultured, Cerebral Cortex cytology, Crystallography methods, Disease Models, Animal, Dogs, Dose-Response Relationship, Drug, Embryo, Mammalian, Enzyme Inhibitors blood, Enzyme-Linked Immunosorbent Assay methods, Female, Humans, Male, Mice, Mice, Transgenic, Middle Aged, Models, Chemical, Mutation genetics, Peptide Fragments cerebrospinal fluid, Pyrimidines pharmacology, Pyrimidines therapeutic use, Rats, Rats, Sprague-Dawley, Thiazines pharmacology, Thiazines therapeutic use, Time Factors, Young Adult, Alzheimer Disease cerebrospinal fluid, Amyloid Precursor Protein Secretases antagonists & inhibitors, Amyloid beta-Peptides metabolism, Aspartic Acid Endopeptidases antagonists & inhibitors, Enzyme Inhibitors pharmacology, Neurons drug effects
- Abstract
According to the amyloid cascade hypothesis, cerebral deposition of amyloid-β peptide (Aβ) is critical for Alzheimer's disease (AD) pathogenesis. Aβ generation is initiated when β-secretase (BACE1) cleaves the amyloid precursor protein. For more than a decade, BACE1 has been a prime target for designing drugs to prevent or treat AD. However, development of such agents has turned out to be extremely challenging, with major hurdles in cell penetration, oral bioavailability/metabolic clearance, and brain access. Using a fragment-based chemistry strategy, we have generated LY2811376 [(S)-4-(2,4-difluoro-5-pyrimidin-5-yl-phenyl)-4-methyl-5,6-dihydro-4H-[1,3]thiazin-2-ylamine], the first orally available non-peptidic BACE1 inhibitor that produces profound Aβ-lowering effects in animals. The biomarker changes obtained in preclinical animal models translate into man at doses of LY2811376 that were safe and well tolerated in healthy volunteers. Prominent and long-lasting Aβ reductions in lumbar CSF were measured after oral dosing of 30 or 90 mg of LY2811376. This represents the first translation of BACE1-driven biomarker changes in CNS from preclinical animal models to man. Because of toxicology findings identified in longer-term preclinical studies, this compound is no longer progressing in clinical development. However, BACE1 remains a viable target because the adverse effects reported here were recapitulated in LY2811376-treated BACE1 KO mice and thus are unrelated to BACE1 inhibition. The magnitude and duration of central Aβ reduction obtainable with BACE1 inhibition positions this protease as a tractable small-molecule target through which to test the amyloid hypothesis in man.
- Published
- 2011
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35. [3H]LY334370, a novel radioligand for the 5-HT1F receptor. II. Autoradiographic localization in rat, guinea pig, monkey and human brain.
- Author
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Lucaites VL, Krushinski JH, Schaus JM, Audia JE, and Nelson DL
- Subjects
- Animals, Autoradiography, Brain drug effects, Guinea Pigs, Humans, Ligands, Macaca mulatta, Male, Rats, Rats, Sprague-Dawley, Receptors, Serotonin drug effects, Species Specificity, Sumatriptan pharmacology, Tritium, Receptor, Serotonin, 5-HT1F, Benzamides pharmacology, Brain metabolism, Indoles pharmacology, Receptors, Serotonin analysis, Serotonin Receptor Agonists pharmacology
- Abstract
LY334370 is a high affinity, selective agonist at the 5-HT(1F) receptor. On this basis, the tritiated compound was examined for its utility in autoradiography to localize the 5-HT(1F) receptor in rat and guinea pig brain regions. Specific 5-HT(1F) receptor binding in rat brain was found in layers 4-5 of all cortical regions examined, as well as olfactory bulb and tubercle, nucleus accumbens, caudate putamen, parafascicular nucleus of the thalamus, medial mammillary nucleus, the CA3 region of the hippocampus, subiculum, and several amygdaloid nuclei. In guinea pig brain, the [(3)H]LY334370 binding sites were found at highest density in claustrum, but also in a layer of the cortex, caudate putamen, nucleus accumbens, thalamus, and medial mammillary nucleus. Some species differences in the distribution of the 5-HT(1F) receptor were noted. Side by side comparison of rat brain autoradiography with [(3)H]LY334370 and [(3)H]sumatriptan showed labeling in the same brain regions. Preliminary binding studies in rhesus monkey and human brain sections showed [(3)H]LY334370 binding in cortical layers 4-5, subiculum (in the monkey), and the granule cell layer of the cerebellum. These findings suggest a discrete localization of the 5-HT(1F) receptor in the rat, guinea pig, monkey and human brain, and confirms the utility of [(3)H]LY334370 as a potential tool to explore further the localization and possible functions of the 5-HT(1F) receptor.
- Published
- 2005
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36. [3H]LY334370, a novel radioligand for the 5-HT1F receptor. I. In vitro characterization of binding properties.
- Author
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Wainscott DB, Krushinski JH Jr, Audia JE, Schaus JM, Zgombick JM, Lucaites VL, and Nelson DL
- Subjects
- Animals, Brain drug effects, Brain metabolism, Cells, Cultured, Guanosine 5'-O-(3-Thiotriphosphate) pharmacology, Guinea Pigs, Humans, In Vitro Techniques, Ligands, Radioligand Assay, Rats, Receptors, Serotonin drug effects, Receptors, Serotonin genetics, Transfection, Tritium, Receptor, Serotonin, 5-HT1F, Benzamides pharmacology, Indoles pharmacology, Receptors, Serotonin analysis, Serotonin Receptor Agonists pharmacology
- Abstract
[(3)H]LY334370 was developed as a radioligand to study the characteristics of this compound's interaction with the 5-HT(1F) receptor. Monovalent or divalent cations did not enhance the binding of [(3)H]LY334370 to the cloned human 5-HT(1F) receptor. In the presence of MgCl(2), the time to reach equilibrium was approximately 2 h, while in its absence equilibrium was reached in less than 1 h. [(3)H]LY334370 had high affinity for the cloned human 5-HT(1F) receptor (K(d)=0.446 nM) and the 5-HT(1F) receptor in rat brain (K(d)=0.388 nM). The expression density of 5-HT(1F) receptors, as determined by binding to homogenates of cortical regions from rat, was low (B(max)=79.1 fmol/mg protein). There was a statistically significant correlation between the apparent pK(i) for inhibition of [(3)H]LY334370 binding and the pEC(50) for stimulation of [(35)S]GTPgammaS binding to homogenates of cells expressing the cloned human 5-HT(1F) receptor. In addition, there was a statistically significant correlation between the apparent pK(i) for inhibition of [(3)H]LY334370 binding to the cloned human 5-HT(1F) receptor and the pID(50) for inhibition of trigeminal nerve stimulated dural plasma protein extravasation in the guinea pig. The conclusion from these studies is that [(3)H]LY334370 is a high affinity radioligand which can be used for the study of the 5-HT(1F) receptor in rat brain or in cells transformed with the human 5-HT(1F) receptor.
- Published
- 2005
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37. In vivo assessment of brain interstitial fluid with microdialysis reveals plaque-associated changes in amyloid-beta metabolism and half-life.
- Author
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Cirrito JR, May PC, O'Dell MA, Taylor JW, Parsadanian M, Cramer JW, Audia JE, Nissen JS, Bales KR, Paul SM, DeMattos RB, and Holtzman DM
- Subjects
- Age Factors, Alzheimer Disease pathology, Amyloid Precursor Protein Secretases, Amyloid beta-Peptides cerebrospinal fluid, Amyloid beta-Protein Precursor metabolism, Animals, Aspartic Acid Endopeptidases, Brain pathology, Cerebral Cortex metabolism, Cerebrospinal Fluid chemistry, Disease Progression, Endopeptidases drug effects, Enzyme Inhibitors pharmacology, Enzyme-Linked Immunosorbent Assay, Extracellular Space chemistry, Hippocampus metabolism, Humans, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microdialysis methods, Plaque, Amyloid pathology, Alzheimer Disease metabolism, Amyloid beta-Peptides metabolism, Brain metabolism, Extracellular Space metabolism, Plaque, Amyloid metabolism
- Abstract
Soluble amyloid-beta (Abeta) peptide converts to structures with high beta-sheet content in Alzheimer's disease (AD). Soluble Abeta is released by neurons into the brain interstitial fluid (ISF), in which it can convert into toxic aggregates. Because assessment of ISF Abeta levels may provide unique insights into Abeta metabolism and AD, an in vivo microdialysis technique was developed to measure it. Our Abeta microdialysis technique was validated ex vivo with human CSF and then in vivo in awake, freely moving mice. Using human amyloid precursor protein (APP) transgenic mice, we found that, before the onset of AD-like pathology, ISF Abeta in hippocampus and cortex correlated with levels of APP in those tissues. After the onset of Abeta deposition, significant changes in the ISF Abeta40/Abeta42 ratio developed without changes in Abeta1-x. These changes differed from changes seen in tissue lysates from the same animals. By rapidly inhibiting Abeta production, we found that ISF Abeta half-life was short ( approximately 2 hr) in young mice but was twofold longer in mice with Abeta deposits. This increase in half-life, without an increase in steady-state levels, suggests that inhibition of Abeta synthesis reveals a portion of the insoluble Abeta pool that is in dynamic equilibrium with ISF Abeta. This now measurable in vivo pool is a likely target for new diagnostic and therapeutic strategies.
- Published
- 2003
38. A series of C-terminal amino alcohol dipeptide A beta inhibitors.
- Author
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Garofalo AW, Wone DW, Phuc A, Audia JE, Bales CA, Dovey HF, Dressen DB, Folmer B, Goldbach EG, Guinn AC, Latimer LH, Mabry TE, Nissen JS, Pleiss MA, Sohn S, Thorsett ED, Tung JS, and Wu J
- Subjects
- Amyloid Precursor Protein Secretases, Amyloid beta-Peptides genetics, Aspartic Acid Endopeptidases, Cell Line, Cell Survival drug effects, Endopeptidases metabolism, Humans, Mutation, Structure-Activity Relationship, Transfection, Amino Alcohols chemical synthesis, Amino Alcohols pharmacology, Amyloid beta-Peptides antagonists & inhibitors, Dipeptides chemical synthesis, Dipeptides pharmacology
- Abstract
Potent, small molecule A beta inhibitors have been prepared that incorporate an alanine core bracketed by an N-terminal arylacetyl group and various C-terminal amino alcohols. The compounds exhibit stereospecific inhibition as demonstrated in an in vitro assay.
- Published
- 2002
- Full Text
- View/download PDF
39. Comparison of the effects of serotonin selective, norepinephrine selective, and dual serotonin and norepinephrine reuptake inhibitors on lower urinary tract function in cats.
- Author
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Katofiasc MA, Nissen J, Audia JE, and Thor KB
- Subjects
- Acetic Acid pharmacology, Administration, Intravesical, Animals, Duloxetine Hydrochloride, Electromyography, Female, Fluoxetine analogs & derivatives, Fluoxetine pharmacology, Humans, Methiothepin pharmacology, Serotonin Antagonists pharmacology, Urethra physiology, Urinary Bladder physiology, Venlafaxine Hydrochloride, Adrenergic Uptake Inhibitors pharmacology, Cats physiology, Cyclohexanols pharmacology, Selective Serotonin Reuptake Inhibitors pharmacology, Thiophenes pharmacology, Urethra drug effects, Urinary Bladder drug effects
- Abstract
Previous studies showed that the dual serotonin (5-hydroxytryptamine, 5-HT) and norepinephrine (NE) reuptake inhibitor, duloxetine, increases bladder capacity and urethral sphincter electromyographic (EMG) activity in a cat model of acetic acid-induced bladder irritation. The present study aimed to determine the relative importance of 5-HT versus NE reuptake inhibition for mediating these effects by examining drugs that are selective for either the 5-HT or NE system or both. Similar to duloxetine, venlafaxine (0.1 to 10 mg/kg), also a dual serotonin and norepinephrine reuptake inhibitor, produced marked increases in bladder capacity and EMG activity that were reversed by methiothepin (0.3 mg/kg). S-norfluoxetine (0.01 to 10 mg/kg), a serotonin selective reuptake inhibitor, produced small but significant increases in bladder capacity and EMG activity at doses of 3 and 10 mg/kg. Thionisoxetine (0.01 to 3.0 mg/kg), a NE selective reuptake inhibitor, produced no effects on bladder capacity or sphincter EMG activity. Surprisingly, co-administration of thionisoxetine and s-norfluoxetine up to doses of 1 mg/kg of each compound produced no effect on lower urinary tract function. These doses were the maximum that could be administered in combination due to drug-induced emergence of skeletal muscle activity in chloralose-anesthetized animals. These results indicate that there are unexplained pharmacological differences between the effects of single compounds that exhibit dual NE and 5-HT reuptake inhibition and a combination of compounds that exhibit selective NE and 5-HT reuptake inhibition on lower urinary tract function.
- Published
- 2002
- Full Text
- View/download PDF
40. N-[3-(2-Dimethylaminoethyl)-2-methyl-1H- indol-5-yl]-4-fluorobenzamide: a potent, selective, and orally active 5-HT(1F) receptor agonist potentially useful for migraine therapy.
- Author
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Xu YC, Johnson KW, Phebus LA, Cohen M, Nelson DL, Schenck K, Walker CD, Fritz JE, Kaldor SW, LeTourneau ME, Murff RE, Zgombick JM, Calligaro DO, Audia JE, and Schaus JM
- Subjects
- Animals, Benzamides chemistry, Benzamides metabolism, Benzamides pharmacology, Cell Line, Dura Mater drug effects, Guinea Pigs, Humans, In Vitro Techniques, Indoles chemistry, Indoles metabolism, Indoles pharmacology, Inflammation, Muscle Contraction drug effects, Muscle, Smooth, Vascular drug effects, Muscle, Smooth, Vascular physiology, Rabbits, Rats, Receptors, Neurotransmitter metabolism, Receptors, Serotonin metabolism, Saphenous Vein drug effects, Saphenous Vein physiology, Serotonin Receptor Agonists chemistry, Serotonin Receptor Agonists metabolism, Serotonin Receptor Agonists pharmacology, Structure-Activity Relationship, Receptor, Serotonin, 5-HT1F, Benzamides chemical synthesis, Indoles chemical synthesis, Migraine Disorders drug therapy, Receptors, Serotonin drug effects, Serotonin Receptor Agonists chemical synthesis
- Abstract
Recent studies have demonstrated that selective 5-HT(1F) receptor agonists inhibit neurogenic dural inflammation, a model of migraine headache, indicating that these compounds may be effective therapies for the treatment of migraine pain. This communication describes the synthesis and discovery of a novel compound, N-[3-(2-(dimethylamino)ethyl)-2-methyl-1H-indol-5-yl]-4-fluorobenzamide (4), which possesses high binding affinity and selectivity at the 5-HT(1F) receptor relative to more than 40 other serotonergic and nonserotonergic receptors examined.
- Published
- 2001
- Full Text
- View/download PDF
41. Functional gamma-secretase inhibitors reduce beta-amyloid peptide levels in brain.
- Author
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Dovey HF, John V, Anderson JP, Chen LZ, de Saint Andrieu P, Fang LY, Freedman SB, Folmer B, Goldbach E, Holsztynska EJ, Hu KL, Johnson-Wood KL, Kennedy SL, Kholodenko D, Knops JE, Latimer LH, Lee M, Liao Z, Lieberburg IM, Motter RN, Mutter LC, Nietz J, Quinn KP, Sacchi KL, Seubert PA, Shopp GM, Thorsett ED, Tung JS, Wu J, Yang S, Yin CT, Schenk DB, May PC, Altstiel LD, Bender MH, Boggs LN, Britton TC, Clemens JC, Czilli DL, Dieckman-McGinty DK, Droste JJ, Fuson KS, Gitter BD, Hyslop PA, Johnstone EM, Li WY, Little SP, Mabry TE, Miller FD, and Audia JE
- Subjects
- Administration, Oral, Alzheimer Disease drug therapy, Alzheimer Disease genetics, Amyloid Precursor Protein Secretases, Amyloid beta-Protein Precursor genetics, Amyloid beta-Protein Precursor metabolism, Animals, Aspartic Acid Endopeptidases, Brain cytology, Brain drug effects, Cells, Cultured, Disease Models, Animal, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, Endopeptidases drug effects, Enzyme Inhibitors administration & dosage, Female, Humans, Injections, Subcutaneous, Kidney cytology, Kidney drug effects, Kidney metabolism, Male, Mice, Mice, Transgenic, Neurons cytology, Neurons drug effects, Neurons metabolism, Peptide Fragments metabolism, Alzheimer Disease metabolism, Amyloid beta-Peptides metabolism, Brain metabolism, Dipeptides administration & dosage, Endopeptidases metabolism
- Abstract
Converging lines of evidence implicate the beta-amyloid peptide (Ass) as causative in Alzheimer's disease. We describe a novel class of compounds that reduce A beta production by functionally inhibiting gamma-secretase, the activity responsible for the carboxy-terminal cleavage required for A beta production. These molecules are active in both 293 HEK cells and neuronal cultures, and exert their effect upon A beta production without affecting protein secretion, most notably in the secreted forms of the amyloid precursor protein (APP). Oral administration of one of these compounds, N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester, to mice transgenic for human APP(V717F) reduces brain levels of Ass in a dose-dependent manner within 3 h. These studies represent the first demonstration of a reduction of brain A beta in vivo. Development of such novel functional gamma-secretase inhibitors will enable a clinical examination of the A beta hypothesis that Ass peptide drives the neuropathology observed in Alzheimer's disease.
- Published
- 2001
- Full Text
- View/download PDF
42. Kinetic analysis of LY320236: competitive inhibitor of type I and non-competitive inhibitor of type II human steroid 5alpha-reductase.
- Author
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McNulty AM, Audia JE, Bemis KG, Goode RL, Rocco VP, and Neubauer BL
- Subjects
- 3-Oxo-5-alpha-Steroid 4-Dehydrogenase classification, 3-Oxo-5-alpha-Steroid 4-Dehydrogenase metabolism, Androstadienes chemistry, Androstadienes metabolism, Androstadienes pharmacology, Benzoquinones chemistry, Binding, Competitive, Computer Simulation, Enzyme Inhibitors chemistry, Enzyme Inhibitors pharmacology, Humans, Inhibitory Concentration 50, Isoenzymes antagonists & inhibitors, Isoenzymes classification, Isoenzymes metabolism, Kinetics, Male, Prostate enzymology, Scalp enzymology, Thermodynamics, 5-alpha Reductase Inhibitors, Benzoquinones metabolism, Benzoquinones pharmacology, Enzyme Inhibitors metabolism
- Abstract
Type I and type II steroid 5alpha-reductases (5alpha-R) catalyze the conversion of testosterone (T) to dihydrotestosterone (DHT). LY320236 is a benzoquinolinone (BQ) that inhibits 5alpha-R activity in human scalp skin (Ki(typeI)=28.7+/-1.87 nM) and prostatic homogenates (Ki(typeII)=10.6+/-4.5 nM). Lineweaver-Burk, Dixon, and non-linear analysis methods were used to evaluate the kinetics of 5alpha-R inhibition by LY320236. Non-linear modeling of experimental data evaluated V(max) in the presence or absence of LY320236. Experimental data modeled to the following equation 1v=+ fixing the In0c value equal to 1.0 or 0 are consistent with non-competitive or competitive inhibition, respectively. LY320236 is a competitive inhibitor of type I 5alpha-R (In0c=0, Ki=3.39+/-0.38, RMSE = 1.300) and a non-competitive inhibitor of type II 5alpha-R (In0c=1, Ki=29. 7+/-3.4, RMSE = 0.0592). These data are in agreement with linear transformation of the data using Lineweaver-Burk and Dixon analyses. These enzyme kinetic data support the contention that the BQ LY320236 is a potent dual inhibitor with differing modes of activity against the two known human 5alpha-reductase isozymes. LY320236 represents a class of non-steroidal 5alpha-R inhibitors with potential therapeutic utility in treating a variety of androgen dependent disorders.
- Published
- 2000
- Full Text
- View/download PDF
43. N-Methyl-5-tert-butyltryptamine: A novel, highly potent 5-HT1D receptor agonist.
- Author
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Xu YC, Schaus JM, Walker C, Krushinski J, Adham N, Zgombick JM, Liang SX, Kohlman DT, and Audia JE
- Subjects
- Cell Line, Humans, Magnetic Resonance Spectroscopy, Mass Spectrometry, Radioligand Assay, Receptor, Serotonin, 5-HT1D, Serotonin Receptor Agonists chemical synthesis, Serotonin Receptor Agonists chemistry, Structure-Activity Relationship, Tryptamines chemical synthesis, Tryptamines chemistry, Receptors, Serotonin drug effects, Serotonin Receptor Agonists pharmacology, Tryptamines pharmacology
- Abstract
It has been observed that reported 5-HT1D receptor agonists have at least one heteroatom (N, O, or S) on the 5-substituent of the indole. This has led to the hypothesis that a 5-substituent capable of participating in hydrogen bonding is critical for conveying high affinity. This article describes the synthesis and biological evaluation of a new series of 5-alkyltryptamine analogues, which does not have a heteroatom in the 5-substituent group. In contrast to the hypothesis, 5-alkyltryptamines all exhibit high binding affinities for the human 5-HT1D receptor. The size of the lipophilic alkyl group at the 5-position of the indole has significant impact on the 5-HT1D binding affinity. Compounds with a tert-butyl group at the 5-position such as 9d, 10, and 11 were identified. These analogues display high binding affinity (Ki < 1 nM) and moderate receptor selectivity in comparison with known antimigraine agents such as sumatriptan, naratriptan, rizatriptan, and VML-251.
- Published
- 1999
- Full Text
- View/download PDF
44. Synthesis and 5 alpha-reductase inhibitory activity of 8-substituted benzo[f]quinolinones derived from palladium mediated coupling reactions.
- Author
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Smith EC, McQuaid LA, Goode RL, McNulty AM, Neubauer BL, Rocco VP, and Audia JE
- Subjects
- Cholestenone 5 alpha-Reductase, Enzyme Inhibitors chemistry, Humans, Male, Prostate drug effects, Prostate enzymology, Quinolones chemistry, Scalp drug effects, Scalp enzymology, Structure-Activity Relationship, Enzyme Inhibitors chemical synthesis, Enzyme Inhibitors pharmacology, Oxidoreductases antagonists & inhibitors, Palladium chemistry, Quinolones chemical synthesis, Quinolones pharmacology
- Abstract
Benzoquinolinones have been shown to be potent, selective inhibitors of the Type I 5 alpha-reductase enzyme, which is responsible for the production of dihydrotestosterone from testosterone localized in the scalp. In an effort to identify compounds that demonstrate inhibition of both 5 alpha-reductase isozymes, we have employed 8-bromobenzoquinolinone as an advanced intermediate for participation in a variety of palladium mediated carbon-carbon bond forming reactions. By varying the 8-substituent it is possible to alter the selectivity profile of the series.
- Published
- 1998
- Full Text
- View/download PDF
45. 5-HT1F receptor agonists inhibit neurogenic dural inflammation in guinea pigs.
- Author
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Johnson KW, Schaus JM, Durkin MM, Audia JE, Kaldor SW, Flaugh ME, Adham N, Zgombick JM, Cohen ML, Branchek TA, and Phebus LA
- Subjects
- Animals, Disease Models, Animal, Guinea Pigs, In Situ Hybridization, Inflammation drug therapy, Male, Piperidines pharmacology, RNA, Messenger metabolism, Rabbits, Tryptamines, Receptor, Serotonin, 5-HT1F, Benzamides pharmacology, Carbazoles pharmacology, Indoles pharmacology, Pyrazoles pharmacology, Receptors, Serotonin drug effects, Serotonin Receptor Agonists pharmacology, Trigeminal Ganglion drug effects
- Abstract
The serotonin (5-HT) receptor subtype mediating inhibition of neurogenic dural inflammation in guinea pigs was investigated using a series of serotonin agonists with differing affinities for the 5-HT1B, 5-HT1D and 5-HT1F receptors. When agonist potencies for inhibiting neurogenic inflammation were compared with affinities for these receptor subtypes, a significant positive correlation was seen only with the 5-HT1F receptor. The potency of agonists in inhibiting adenylate cyclase in cells transfected with human 5-HT1F receptor was also highly correlated with their potency in the animal model of migraine. In situ hybridization demonstrated 5-HT1F receptor mRNA in guinea pig trigeminal ganglion neurons. These data suggest that the 5-HT1F receptor is a rational target for migraine therapeutics.
- Published
- 1997
- Full Text
- View/download PDF
46. "Pictet-Spengler-like" Synthesis of Tetrahydro-beta-carbolines under Hydrolytic Conditions. Direct Use of Azalactones as Phenylacetaldehyde Equivalents.
- Author
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Audia JE, Droste JJ, Nissen JS, Murdoch GL, and Evrard DA
- Published
- 1996
- Full Text
- View/download PDF
47. Amyloid beta-mediated oxidative and metabolic stress in rat cortical neurons: no direct evidence for a role for H2O2 generation.
- Author
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Zhang Z, Rydel RE, Drzewiecki GJ, Fuson K, Wright S, Wogulis M, Audia JE, May PC, and Hyslop PA
- Subjects
- Adenosine Triphosphate metabolism, Analysis of Variance, Animals, Benzothiazoles, Catalase antagonists & inhibitors, Cell Survival, Cells, Cultured, Coloring Agents, Fetus, Fluoresceins, Free Radical Scavengers, Glyceraldehyde-3-Phosphate Dehydrogenases metabolism, Glycolysis drug effects, Glyoxylates pharmacology, L-Lactate Dehydrogenase, Neurons drug effects, Neurotoxins pharmacology, Rats, Reactive Oxygen Species metabolism, Thiazoles, Amyloid beta-Peptides pharmacology, Cerebral Cortex metabolism, Glucose metabolism, Hydrogen Peroxide metabolism, Neurons metabolism, Oxazines, Oxidative Stress, Peptide Fragments pharmacology, Xanthenes
- Abstract
H2O2 and free radical-mediated oxidative stresses have been implicated in mediating amyloid beta (1-40) [A beta (1-40)] neurotoxicity to cultured neurons. In this study, we confirm that addition of the H2O2-scavenging enzyme catalase protects neurons in culture against A beta-mediated toxicity; however, it does so by a mechanism that does not involve its ability to scavenge H2O2. A beta-mediated elevation in intracellular H2O2 production is suppressed by addition of a potent H2O2 scavenger without any significant neuroprotection. Three intracellular biochemical markers of H2O2-mediated oxidative stress were unchanged by A beta treatment: (a) glyceraldehyde-3-phosphate dehydrogenase activity, (b) hexose monophosphate shunt activity, and (c) glucose oxidation via the tricarboxylic acid cycle. lonspray mass spectra of A beta in the incubation medium indicated that A beta itself is an unlikely source of reactive oxygen species. In this study we demonstrate that intracellular ATP concentration is compromised during the first 24-h exposure of neurons to A beta. Our results challenge a pivotal role for H2O2 generation in mediating A beta toxicity, and we suggest that impairment of energy homeostasis may be a more significant early factor in the neurodegenerative process.
- Published
- 1996
- Full Text
- View/download PDF
48. Potent, selective tetrahydro-beta-carboline antagonists of the serotonin 2B (5HT2B) contractile receptor in the rat stomach fundus.
- Author
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Audia JE, Evrard DA, Murdoch GR, Droste JJ, Nissen JS, Schenck KW, Fludzinski P, Lucaites VL, Nelson DL, and Cohen ML
- Subjects
- Animals, Carbolines chemical synthesis, Cell Line, Cricetinae, Gastric Fundus, In Vitro Techniques, Male, Mesocricetus, Mice, Molecular Structure, Muscle Contraction drug effects, Rats, Rats, Wistar, Receptor, Serotonin, 5-HT2A, Receptor, Serotonin, 5-HT2B, Receptor, Serotonin, 5-HT2C, Receptors, Serotonin genetics, Receptors, Serotonin metabolism, Serotonin Antagonists chemical synthesis, Structure-Activity Relationship, Yohimbine chemistry, Carbolines pharmacology, Muscle, Smooth drug effects, Receptors, Serotonin drug effects, Serotonin Antagonists pharmacology
- Abstract
A series of potent, selective 5HT2B receptor antagonists has been identified based upon yohimbine, with SAR studies resulting in a 1000-fold increase in 5HT2B receptor affinity relative to the starting structure (-log KBS > 10.0 have been obtained). These high-affinity tetrahydro-beta-carboline antagonists are able to discriminate among the 5HT2 family of serotonin receptors, with members of the series showing selectivities of more than 100-fold versus both the 5HT2A and 5HT2C receptors based upon radioligand binding and functional assays. As the first compounds reported with such selectivity and enhanced receptor affinity, these tetrahydro-beta-carboline antagonists are useful tools for elucidating the role of serotonin acting at the 5HT2B receptor in normal and disease physiology.
- Published
- 1996
- Full Text
- View/download PDF
49. Resolution of delta-Lactams Provides Access to Nonracemic Benzoquinolinones: The Synthesis of LY300502 and LY300503.
- Author
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Astleford BA, Audia JE, Deeter J, Heath PC, Janisse SK, Kress TJ, Wepsiec JP, and Weigel LO
- Published
- 1996
- Full Text
- View/download PDF
50. LY191704 inhibits type I steroid 5 alpha-reductase in human scalp.
- Author
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Neubauer BL, Gray HM, Hanke CW, Hirsch KS, Hsiao KC, Jones CD, Kumar MV, Lawhorn DE, Lindzey J, McQuaid L, Tindall DJ, Toomey RE, Yao RC, and Audia JE
- Subjects
- Animals, Binding, Competitive, Dihydrotestosterone metabolism, Humans, Male, Mice, Osmolar Concentration, Quinolones metabolism, 5-alpha Reductase Inhibitors, Isoenzymes antagonists & inhibitors, Quinolones pharmacology, Scalp enzymology
- Abstract
Conversion of testosterone to dihydrotestosterone (DHT) has been demonstrated to be catalyzed by two isoforms of steroid 5 alpha-reductase, designated types I and II. Although several classes of steroid-based inhibitors of the type II isoform have been identified, these agents have not demonstrated highly selective pharmacological activity against human type I 5 alpha-reductase. LY191704 is representative of a series of nonsteroidal agents that have potent [apparent inhibitory constant (Ki) = 11.3 nM] inhibitory activity in human scalp skin homogenates (pH 7.5), a source of type I 5 alpha-reductase. [3H]-DHT production in the presence and absence of LY191704 is consistent with a noncompetitive mode of inhibition. In human prostatic homogenates (pH 5.5), a source of type II 5 alpha-reductase, LY191704 is virtually inactive as an inhibitor [concentration of inhibitor producing 50% inhibition of enzymatic activity (IC50) > 1,000 nM] of [3H]-DHT formation. LY191704 does not inhibit the type I or type II isoforms of rat 5 alpha-reductase, nor does the compound compete for binding to the murine androgen receptor expressed in SF9 cells using a baculo virus expression system. The benzoquinolinones, as exemplified by LY191704, possess exquisite pharmacological selectivity and provide a tool to understand the role of human type I 5 alpha-reductase in normal and pathophysiological states. These agents may also find clinical utility in treating androgen-dependent dermatological conditions.
- Published
- 1996
- Full Text
- View/download PDF
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