James Audia | Yale University (original) (raw)

Papers by James Audia

Cancer Research, May 29, 2023

Although the treatment landscape of advanced urothelial cancer (UC) has changed dynamically over ... more Although the treatment landscape of advanced urothelial cancer (UC) has changed dynamically over the last decade, metastatic UC still has amongst the worst 5-year recurrence rate of any cancer type, highlighting the need for new therapeutic options for these patients. Analogous to breast cancer, advanced UC is a heterogeneous disease, with luminal and basal subtypes. For luminal breast cancer, small molecule targeting of the lineage-defining transcription factor estrogen receptor (ER) is a demonstrated and effective therapeutic strategy. Two-thirds of advanced UC is classified as luminal, which is characterized by overexpression of the urothelial lineage-defining transcription factor PPARG. Thus, small molecule targeting of PPARG in luminal UC may serve as a therapeutic strategy analogous to ER in luminal breast cancer. While PPARG agonists have been well studied in the context of metabolic disease, PPARG inhibitors have received far less attention. Previously reported PPARG inhibitors display minimal phenotypic activity in pre-clinical UC models, calling into question the performance of these molecules, or alternatively, the role of PPARG as a survival lineage oncogene in UC. Leveraging reconstituted, multi-component PPARG biochemical systems, we first identified the limitations of previous tool compounds and then discovered a novel, covalent lead series with unique, context-dependent electrophilic properties. These efforts lead to the discovery of FX-909, a first-in-class covalent PPARG inverse agonist with powerful repressive conformational biasing activity. FX-909 is highly potent in cells (IC50=1 nM), demonstrates high specificity for PPARG (>2000-fold selective over PPARA/PPARD), and elicits robust in vitro growth inhibition in UC cell lines with activated PPARG signaling. Tumor regression was observed in UMUC9 (PPARG amplification) and HT1197 (RXRAS427F hotspot mutation) xenograft models of UC with oral dosing at 1 mg/kg. Predictable, on-target and reversible pharmacology was observed at FX-909 doses above 1 mg/kg, mimicking PPARG loss-of-function mutations with notable tissue remodeling in adipose tissue and the normal urothelium. These collective findings corroborate the role of PPARG as a key UC survival oncogene and suggest that FX-909 will be an effective therapy for patients with advanced UC harboring the luminal subtype. Citation Format: Robert Sims, Jennifer A. Mertz, Jonathan E. Wilson, James E. Audia, Kaylyn E. Williamson, Yong Li, Miljan Kuljanin, Will W. Motely, Byron DeLaBarre, Michaela Bowden, Jacob I. Stuckey. Discovery of FX-909, a first-in-class inverse agonist of the peroxisome proliferator-activated receptor gamma (PPARG) lineage transcription factor, to potentially treat patients with the luminal subtype of advanced urothelial cancer (UC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr ND08.

The Notch pathway is a highly conserved signaling system that plays an important role in developm... more The Notch pathway is a highly conserved signaling system that plays an important role in development and tissue homeostasis. While Notch mutations are well characterized and implicated in hematological malignancies such as T-cell acute lymphoblastic leukemia, in solid tumors ligand or receptor over-expression may lead to enhanced/sustained Notch function, triggering increased tumor cell proliferation/survival, angiogenesis and tumor formation. In order to delineate an oncogenic role of activated Notch in tumors of epithelial origin, we carried out a series of in-vitro and in-vivo studies. We demonstrated that the activated Notch1 receptor (a - secretase-dependent Notch1 E with the transmembrane domain and a-secretase-independent constitutively activated Notch1 intracellular fragment) can transform normal rat cells, RK3E. These transformed cells formed colonies in soft agar, confirming their anchorage-independent growth potential, and when implanted subcutaneously, formed tumors in athymic nude mice. Inhibition of Notch signaling through a small molecule inhibitor of -secretase, a key regulator of Notch processing, may provide an attractive targeted cancer therapeutic strategy. We have identified and characterized a novel small molecule that is an exquisitely potent inhibitor of Notch signaling in tumor cell lines and endothelial cells with an IC50 ranging from 0.005 nM to 20 nM. The Notch inhibitor meets all pharmacokinetic criteria in pre-clinical species. In a xenograft tumor model, the novel compound inhibited Notch cleavage in a dose-dependent manner at 6 hours after a single oral dose. This inhibition of Notch cleavage resulted in the induction of apoptosis (as measured by activated caspase-3 levels) that was statistically significant at 24 hours after a single oral dose of 3 mg/kg. Analysis of tumors from animals treated with the Notch inhibitor revealed inhibition of angiogenesis through formation of leaky vasculature which may also contribute to observed anti-tumor activity. Furthermore, Notch inhibition produced tumor regression in the Notch-dependent tumor models. Anti-tumor activity was also observed in several human xenograft tumors of epithelial origin. To mitigate mucoid gasteroentropathy due to Notch inhibition, PK/PD data were incorporated in devising dosing strategies that identified an optimal intermittent dosing schedule without negatively impacting efficacy. Furthermore, the mucoid gastroentropathy was also mitigated by the prophylactic administration of dexamethasone without negatively impacting Notch inhibitor mediated efficacy. In summary, we have characterized an orally bio-available small molecule Notch inhibitor that may provide therapeutic benefit to cancer patients. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):B188.

Journal of Virology, May 10, 2021

In many enveloped virus families, including human immunodeficiency virus (HIV) and herpes simplex... more In many enveloped virus families, including human immunodeficiency virus (HIV) and herpes simplex virus (HSV), a crucial, yet unexploited, step in the viral life cycle is the release of particles from the infected cell membranes. This release process is mediated by host ESCRT complex proteins, which are recruited by viral structural proteins and provide the mechanical means for membrane scission and subsequent viral budding. The prazole drug tenatoprazole was previously shown to bind to the 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 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 a 50% effective concentration (EC 50) of 0.8 to 1.2mM, which is much more potent than tenatoprazole. Our results indicate that prazole-based compounds may represent a class of drugs with the potential to be broad-spectrum antiviral agents against multiple enveloped viruses by interrupting the 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. KEYWORDS budding of viruses, drugs to block budding, pan-antiviral agents T he advent of antibiotics had a major impact on controlling bacterial infections in patients worldwide, with a single drug being used to treat multiple infections. Unfortunately, antivirals have not had the same success. There are many factors contributing to this shortcoming, foremost the fact that few mechanisms are shared by different viruses, which limits targets for a broad-spectrum antiviral. Consequently, approved antivirals generally act against individual rather than groups of viruses, limiting a single drug's potential. Enveloped viruses bud from the host cell membranes and use the acquired lipid layer as a protective coat that also contains the glycoproteins required for infection of other cells. Enveloped viruses do not encode the machinery needed for budding and must recruit host cell proteins to bud from cells. In human immunodeficiency virus (HIV), ESCRT proteins are recruited to virus budding complexes through an interaction between the L domain (PT\SAP motif) in virus structural proteins (1-5, 67) and the

Encyclopedia of Reagents for Organic Synthesis, Apr 15, 2001

Chemischer Informationsdienst, Oct 18, 1983

Der Trimethylsilylkupfer‐lithiumcyanid‐Komplex (II) lagert sich stereospezifisch an den Alkincarb... more Der Trimethylsilylkupfer‐lithiumcyanid‐Komplex (II) lagert sich stereospezifisch an den Alkincarbonsäureester (I) an; der so gebildete silylierte Carbonsäureester (III) wird mit Diisobutylaluminiumhydrid zum Allylalkohol (IV) reduziert.

ACS Medicinal Chemistry Letters, Mar 26, 2020

Histone methyltransferase EZH2, which is the catalytic subunit of the PRC2 complex, catalyzes the... more 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.

Chemischer Informationsdienst, Oct 14, 1986

Drugs of The Future, 1995

Journal of Biological Chemistry, 2021

The histone methyltransferase EZH2 has been the target of numerous small-molecule inhibitor disco... more 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.

The Journal of Neuroscience, Oct 1, 2003

Soluble amyloid-␤ (A␤) peptide converts to structures with high ␤-sheet content in Alzheimer's di... more Soluble amyloid-␤ (A␤) peptide converts to structures with high ␤-sheet content in Alzheimer's disease (AD). Soluble A␤ is released by neurons into the brain interstitial fluid (ISF), in which it can convert into toxic aggregates. Because assessment of ISF A␤ levels may provide unique insights into A␤ metabolism and AD, an in vivo microdialysis technique was developed to measure it. Our A␤ 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 A␤ in hippocampus and cortex correlated with levels of APP in those tissues. After the onset of A␤ deposition, significant changes in the ISF A␤ 40 /A␤ 42 ratio developed without changes in A␤ 1-x. These changes differed from changes seen in tissue lysates from the same animals. By rapidly inhibiting A␤ production, we found that ISF A␤ half-life was short (ϳ2 hr) in young mice but was twofold longer in mice with A␤ deposits. This increase in half-life, without an increase in steady-state levels, suggests that inhibition of A␤ synthesis reveals a portion of the insoluble A␤ pool that is in dynamic equilibrium with ISF A␤. This now measurable in vivo pool is a likely target for new diagnostic and therapeutic strategies.

Cold Spring Harbor Perspectives in Biology, Apr 1, 2016

Histone posttranslational modifications represent a versatile set of epigenetic marks involved no... more 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.

Elsevier eBooks, 1991

Publisher Summary Research in the past decade has led to tremendous advances in the understanding... more Publisher Summary Research in the past decade has led to tremendous advances in the understanding of both the central and peripheral physiological effects of serotonin (5HT). This chapter discusses recent advances in drug discovery efforts toward serotonin modulators for cardiovascular (CV) and gastrointestinal (GI) disorders. The Serotonin Club Nomenclature Committee has recognized three major classes of serotonin receptors: 5HT1 receptors, which have a high affinity for serotonin itself; 5HT2 receptors, which have a low affinity for serotonin; and 5HT3 receptors, which have moderate affinity. The 5HT1 family is further subdivided into 5HT1A, 5HT1B, 5HT1C, and 5HT1D subtypes, each of which differs in the physiological roles, in distribution, and in effector pathways. To date, the 5HT1A, 5HT1C, 5HT1D, and 5HT2 receptors have been cloned and characterized at the molecular level, showing marked differences in protein sequences. Of the high affinity 5HT1 subtypes of serotonin receptors, the 5HT1A and 5HT1D subtypes have been most widely studied with respect to the CV/GI systems. In addition to the consensus subtypes, other serotonin binding sites have been proposed and their relevance to CV and GI disorders continues to unfold. The structural diversity, permitted by each receptor-binding site, allows the modification of the chemical and pharmacological properties of the compounds to address issues such as the central nervous system (CNS) penetration, bioavailability, and metabolic half-life. This chapter discusses the importance of 5HT in cardiovascular and gastrointestinal pathophysiology.

Molecular pharmacology, 1993

The rat 5-hydroxytryptamine (5-HT)2F (serotonin2F) receptor is a newly cloned member of the 5-HT2... more The rat 5-hydroxytryptamine (5-HT)2F (serotonin2F) receptor is a newly cloned member of the 5-HT2/1C receptor family. The pharmacology of the 5-HT2F receptor was explored using a variety of structurally different compounds in a radioligand binding assay. In addition, the 5-HT2F receptor was shown to stimulate production of inositol 1,4,5-trisphosphate in the transformed cells. Based on the affinities of the compounds tested, their known affinities for certain of the other 5-HT receptors, and the fact that activation of the cloned 5-HT2F receptor stimulates inositol 1,4,5-trisphosphate production, the 5-HT2F receptor was determined to be a novel receptor and a member of the 5-HT2/1C receptor family. In addition, several agonists and partial agonists were evaluated for contractile activity in the rat stomach fundus, and these activities were correlated with their binding affinities at the 5-HT2F receptor. A highly significant correlation was found, providing additional evidence that i...

Encyclopedia of Reagents for Organic Synthesis, 2006

Encyclopedia of Reagents for Organic Synthesis, 2001

Disease Models & Mechanisms, Dec 1, 2022

Brain diseases are a major cause of death and disability worldwide and contribute significantly t... more 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.

Encyclopedia of Reagents for Organic Synthesis, Apr 15, 2001

Encyclopedia of Reagents for Organic Synthesis, Apr 15, 2001

Chemischer Informationsdienst, Jul 16, 1985

Journal of Organic Chemistry, 1994

Cancer Research, May 29, 2023

Although the treatment landscape of advanced urothelial cancer (UC) has changed dynamically over ... more Although the treatment landscape of advanced urothelial cancer (UC) has changed dynamically over the last decade, metastatic UC still has amongst the worst 5-year recurrence rate of any cancer type, highlighting the need for new therapeutic options for these patients. Analogous to breast cancer, advanced UC is a heterogeneous disease, with luminal and basal subtypes. For luminal breast cancer, small molecule targeting of the lineage-defining transcription factor estrogen receptor (ER) is a demonstrated and effective therapeutic strategy. Two-thirds of advanced UC is classified as luminal, which is characterized by overexpression of the urothelial lineage-defining transcription factor PPARG. Thus, small molecule targeting of PPARG in luminal UC may serve as a therapeutic strategy analogous to ER in luminal breast cancer. While PPARG agonists have been well studied in the context of metabolic disease, PPARG inhibitors have received far less attention. Previously reported PPARG inhibitors display minimal phenotypic activity in pre-clinical UC models, calling into question the performance of these molecules, or alternatively, the role of PPARG as a survival lineage oncogene in UC. Leveraging reconstituted, multi-component PPARG biochemical systems, we first identified the limitations of previous tool compounds and then discovered a novel, covalent lead series with unique, context-dependent electrophilic properties. These efforts lead to the discovery of FX-909, a first-in-class covalent PPARG inverse agonist with powerful repressive conformational biasing activity. FX-909 is highly potent in cells (IC50=1 nM), demonstrates high specificity for PPARG (>2000-fold selective over PPARA/PPARD), and elicits robust in vitro growth inhibition in UC cell lines with activated PPARG signaling. Tumor regression was observed in UMUC9 (PPARG amplification) and HT1197 (RXRAS427F hotspot mutation) xenograft models of UC with oral dosing at 1 mg/kg. Predictable, on-target and reversible pharmacology was observed at FX-909 doses above 1 mg/kg, mimicking PPARG loss-of-function mutations with notable tissue remodeling in adipose tissue and the normal urothelium. These collective findings corroborate the role of PPARG as a key UC survival oncogene and suggest that FX-909 will be an effective therapy for patients with advanced UC harboring the luminal subtype. Citation Format: Robert Sims, Jennifer A. Mertz, Jonathan E. Wilson, James E. Audia, Kaylyn E. Williamson, Yong Li, Miljan Kuljanin, Will W. Motely, Byron DeLaBarre, Michaela Bowden, Jacob I. Stuckey. Discovery of FX-909, a first-in-class inverse agonist of the peroxisome proliferator-activated receptor gamma (PPARG) lineage transcription factor, to potentially treat patients with the luminal subtype of advanced urothelial cancer (UC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr ND08.

The Notch pathway is a highly conserved signaling system that plays an important role in developm... more The Notch pathway is a highly conserved signaling system that plays an important role in development and tissue homeostasis. While Notch mutations are well characterized and implicated in hematological malignancies such as T-cell acute lymphoblastic leukemia, in solid tumors ligand or receptor over-expression may lead to enhanced/sustained Notch function, triggering increased tumor cell proliferation/survival, angiogenesis and tumor formation. In order to delineate an oncogenic role of activated Notch in tumors of epithelial origin, we carried out a series of in-vitro and in-vivo studies. We demonstrated that the activated Notch1 receptor (a - secretase-dependent Notch1 E with the transmembrane domain and a-secretase-independent constitutively activated Notch1 intracellular fragment) can transform normal rat cells, RK3E. These transformed cells formed colonies in soft agar, confirming their anchorage-independent growth potential, and when implanted subcutaneously, formed tumors in athymic nude mice. Inhibition of Notch signaling through a small molecule inhibitor of -secretase, a key regulator of Notch processing, may provide an attractive targeted cancer therapeutic strategy. We have identified and characterized a novel small molecule that is an exquisitely potent inhibitor of Notch signaling in tumor cell lines and endothelial cells with an IC50 ranging from 0.005 nM to 20 nM. The Notch inhibitor meets all pharmacokinetic criteria in pre-clinical species. In a xenograft tumor model, the novel compound inhibited Notch cleavage in a dose-dependent manner at 6 hours after a single oral dose. This inhibition of Notch cleavage resulted in the induction of apoptosis (as measured by activated caspase-3 levels) that was statistically significant at 24 hours after a single oral dose of 3 mg/kg. Analysis of tumors from animals treated with the Notch inhibitor revealed inhibition of angiogenesis through formation of leaky vasculature which may also contribute to observed anti-tumor activity. Furthermore, Notch inhibition produced tumor regression in the Notch-dependent tumor models. Anti-tumor activity was also observed in several human xenograft tumors of epithelial origin. To mitigate mucoid gasteroentropathy due to Notch inhibition, PK/PD data were incorporated in devising dosing strategies that identified an optimal intermittent dosing schedule without negatively impacting efficacy. Furthermore, the mucoid gastroentropathy was also mitigated by the prophylactic administration of dexamethasone without negatively impacting Notch inhibitor mediated efficacy. In summary, we have characterized an orally bio-available small molecule Notch inhibitor that may provide therapeutic benefit to cancer patients. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):B188.

Journal of Virology, May 10, 2021

In many enveloped virus families, including human immunodeficiency virus (HIV) and herpes simplex... more In many enveloped virus families, including human immunodeficiency virus (HIV) and herpes simplex virus (HSV), a crucial, yet unexploited, step in the viral life cycle is the release of particles from the infected cell membranes. This release process is mediated by host ESCRT complex proteins, which are recruited by viral structural proteins and provide the mechanical means for membrane scission and subsequent viral budding. The prazole drug tenatoprazole was previously shown to bind to the 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 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 a 50% effective concentration (EC 50) of 0.8 to 1.2mM, which is much more potent than tenatoprazole. Our results indicate that prazole-based compounds may represent a class of drugs with the potential to be broad-spectrum antiviral agents against multiple enveloped viruses by interrupting the 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. KEYWORDS budding of viruses, drugs to block budding, pan-antiviral agents T he advent of antibiotics had a major impact on controlling bacterial infections in patients worldwide, with a single drug being used to treat multiple infections. Unfortunately, antivirals have not had the same success. There are many factors contributing to this shortcoming, foremost the fact that few mechanisms are shared by different viruses, which limits targets for a broad-spectrum antiviral. Consequently, approved antivirals generally act against individual rather than groups of viruses, limiting a single drug's potential. Enveloped viruses bud from the host cell membranes and use the acquired lipid layer as a protective coat that also contains the glycoproteins required for infection of other cells. Enveloped viruses do not encode the machinery needed for budding and must recruit host cell proteins to bud from cells. In human immunodeficiency virus (HIV), ESCRT proteins are recruited to virus budding complexes through an interaction between the L domain (PT\SAP motif) in virus structural proteins (1-5, 67) and the

Encyclopedia of Reagents for Organic Synthesis, Apr 15, 2001

Chemischer Informationsdienst, Oct 18, 1983

Der Trimethylsilylkupfer‐lithiumcyanid‐Komplex (II) lagert sich stereospezifisch an den Alkincarb... more Der Trimethylsilylkupfer‐lithiumcyanid‐Komplex (II) lagert sich stereospezifisch an den Alkincarbonsäureester (I) an; der so gebildete silylierte Carbonsäureester (III) wird mit Diisobutylaluminiumhydrid zum Allylalkohol (IV) reduziert.

ACS Medicinal Chemistry Letters, Mar 26, 2020

Histone methyltransferase EZH2, which is the catalytic subunit of the PRC2 complex, catalyzes the... more 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.

Chemischer Informationsdienst, Oct 14, 1986

Drugs of The Future, 1995

Journal of Biological Chemistry, 2021

The histone methyltransferase EZH2 has been the target of numerous small-molecule inhibitor disco... more 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.

The Journal of Neuroscience, Oct 1, 2003

Soluble amyloid-␤ (A␤) peptide converts to structures with high ␤-sheet content in Alzheimer's di... more Soluble amyloid-␤ (A␤) peptide converts to structures with high ␤-sheet content in Alzheimer's disease (AD). Soluble A␤ is released by neurons into the brain interstitial fluid (ISF), in which it can convert into toxic aggregates. Because assessment of ISF A␤ levels may provide unique insights into A␤ metabolism and AD, an in vivo microdialysis technique was developed to measure it. Our A␤ 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 A␤ in hippocampus and cortex correlated with levels of APP in those tissues. After the onset of A␤ deposition, significant changes in the ISF A␤ 40 /A␤ 42 ratio developed without changes in A␤ 1-x. These changes differed from changes seen in tissue lysates from the same animals. By rapidly inhibiting A␤ production, we found that ISF A␤ half-life was short (ϳ2 hr) in young mice but was twofold longer in mice with A␤ deposits. This increase in half-life, without an increase in steady-state levels, suggests that inhibition of A␤ synthesis reveals a portion of the insoluble A␤ pool that is in dynamic equilibrium with ISF A␤. This now measurable in vivo pool is a likely target for new diagnostic and therapeutic strategies.

Cold Spring Harbor Perspectives in Biology, Apr 1, 2016

Histone posttranslational modifications represent a versatile set of epigenetic marks involved no... more 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.

Elsevier eBooks, 1991

Publisher Summary Research in the past decade has led to tremendous advances in the understanding... more Publisher Summary Research in the past decade has led to tremendous advances in the understanding of both the central and peripheral physiological effects of serotonin (5HT). This chapter discusses recent advances in drug discovery efforts toward serotonin modulators for cardiovascular (CV) and gastrointestinal (GI) disorders. The Serotonin Club Nomenclature Committee has recognized three major classes of serotonin receptors: 5HT1 receptors, which have a high affinity for serotonin itself; 5HT2 receptors, which have a low affinity for serotonin; and 5HT3 receptors, which have moderate affinity. The 5HT1 family is further subdivided into 5HT1A, 5HT1B, 5HT1C, and 5HT1D subtypes, each of which differs in the physiological roles, in distribution, and in effector pathways. To date, the 5HT1A, 5HT1C, 5HT1D, and 5HT2 receptors have been cloned and characterized at the molecular level, showing marked differences in protein sequences. Of the high affinity 5HT1 subtypes of serotonin receptors, the 5HT1A and 5HT1D subtypes have been most widely studied with respect to the CV/GI systems. In addition to the consensus subtypes, other serotonin binding sites have been proposed and their relevance to CV and GI disorders continues to unfold. The structural diversity, permitted by each receptor-binding site, allows the modification of the chemical and pharmacological properties of the compounds to address issues such as the central nervous system (CNS) penetration, bioavailability, and metabolic half-life. This chapter discusses the importance of 5HT in cardiovascular and gastrointestinal pathophysiology.

Molecular pharmacology, 1993

The rat 5-hydroxytryptamine (5-HT)2F (serotonin2F) receptor is a newly cloned member of the 5-HT2... more The rat 5-hydroxytryptamine (5-HT)2F (serotonin2F) receptor is a newly cloned member of the 5-HT2/1C receptor family. The pharmacology of the 5-HT2F receptor was explored using a variety of structurally different compounds in a radioligand binding assay. In addition, the 5-HT2F receptor was shown to stimulate production of inositol 1,4,5-trisphosphate in the transformed cells. Based on the affinities of the compounds tested, their known affinities for certain of the other 5-HT receptors, and the fact that activation of the cloned 5-HT2F receptor stimulates inositol 1,4,5-trisphosphate production, the 5-HT2F receptor was determined to be a novel receptor and a member of the 5-HT2/1C receptor family. In addition, several agonists and partial agonists were evaluated for contractile activity in the rat stomach fundus, and these activities were correlated with their binding affinities at the 5-HT2F receptor. A highly significant correlation was found, providing additional evidence that i...

Encyclopedia of Reagents for Organic Synthesis, 2006

Encyclopedia of Reagents for Organic Synthesis, 2001

Disease Models & Mechanisms, Dec 1, 2022

Brain diseases are a major cause of death and disability worldwide and contribute significantly t... more 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.

Encyclopedia of Reagents for Organic Synthesis, Apr 15, 2001

Encyclopedia of Reagents for Organic Synthesis, Apr 15, 2001

Chemischer Informationsdienst, Jul 16, 1985

Journal of Organic Chemistry, 1994