Publications by authors named "James L Ellis"

30 Publications

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Combining a High Dose of Metformin With the SIRT1 Activator, SRT1720, Reduces Life Span in Aged Mice Fed a High-Fat Diet.

J Gerontol A Biol Sci Med Sci 2020 10;75(11):2037-2041

Translational Gerontology Branch, National Institute on Aging, National Institutes of Health, Baltimore, Maryland.

SRT1720, a sirtuin1-activator, and metformin (MET), an antidiabetic drug, confer health and life-span benefits when administered individually. It is unclear whether combination of the two compounds could lead to additional benefits. Groups of 56-week-old C57BL/6J male mice were fed a high-fat diet (HFD) alone or supplemented with either SRT1720 (2 g/kg food), a high dose of MET (1% wt/wt food), or a combination of both. Animals were monitored for survival, body weight, food consumption, body composition, and rotarod performance. Mice treated with MET alone did not have improved longevity, and life span was dramatically reduced by combination of MET with SRT1720. Although all groups of animals were consuming similar amounts of food, mice on MET or MET + SRT1720 showed a sharp reduction in body weight. SRT1720 + MET mice also had lower percent body fat combined with better performance on the rotarod compared to controls. These data suggest that co-treatment of SRT1720 with MET is detrimental to survival at the doses used and, therefore, risk-benefits of combining life-span-extending drugs especially in older populations needs to be systematically evaluated.
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http://dx.doi.org/10.1093/gerona/glaa148DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7750506PMC
October 2020

Sirtuin activators and inhibitors: Promises, achievements, and challenges.

Pharmacol Ther 2018 08 22;188:140-154. Epub 2018 Mar 22.

Department of Biochemistry and Research Center for Bio-Macromolecules, University of Bayreuth, 95440 Bayreuth, Germany. Electronic address:

The NAD-dependent protein lysine deacylases of the Sirtuin family regulate various physiological functions, from energy metabolism to stress responses. The human Sirtuin isoforms, SIRT1-7, are considered attractive therapeutic targets for aging-related diseases, such as type 2 diabetes, inflammatory diseases and neurodegenerative disorders. We review the status of Sirtuin-targeted drug discovery and development. Potent and selective pharmacological Sirt1 activators and inhibitors are available, and initial clinical trials have been carried out. Several promising inhibitors and activators have also been described for other isoforms. Progress in understanding the mechanisms of Sirtuin modulation by such compounds provides a rational basis for further drug development.
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http://dx.doi.org/10.1016/j.pharmthera.2018.03.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6342514PMC
August 2018

Nicotinamide Improves Aspects of Healthspan, but Not Lifespan, in Mice.

Cell Metab 2018 03;27(3):667-676.e4

Experimental Gerontology Section, Translational Gerontology Branch, National Institute on Aging, NIH, Baltimore, MD 21224, USA. Electronic address:

The role in longevity and healthspan of nicotinamide (NAM), the physiological precursor of NAD, is elusive. Here, we report that chronic NAM supplementation improves healthspan measures in mice without extending lifespan. Untargeted metabolite profiling of the liver and metabolic flux analysis of liver-derived cells revealed NAM-mediated improvement in glucose homeostasis in mice on a high-fat diet (HFD) that was associated with reduced hepatic steatosis and inflammation concomitant with increased glycogen deposition and flux through the pentose phosphate and glycolytic pathways. Targeted NAD metabolome analysis in liver revealed depressed expression of NAM salvage in NAM-treated mice, an effect counteracted by higher expression of de novo NAD biosynthetic enzymes. Although neither hepatic NAD nor NADP was boosted by NAM, acetylation of some SIRT1 targets was enhanced by NAM supplementation in a diet- and NAM dose-dependent manner. Collectively, our results show health improvement in NAM-supplemented HFD-fed mice in the absence of survival effects.
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http://dx.doi.org/10.1016/j.cmet.2018.02.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5854409PMC
March 2018

PHARMACOLOGICAL SIRT1 ACTIVATION IMPROVES MORTALITY AND MARKEDLY ALTERS TRANSCRIPTIONAL PROFILES THAT ACCOMPANY EXPERIMENTAL SEPSIS.

Shock 2016 Apr;45(4):411-8

*Infectious Disease Division, Memorial Hospital of RI, Pawtucket †The Alpert Medical School of Brown University, Providence, Rhode Island ‡Sirtris, A GSK company, Cambridge, Massachusetts §GlaxoSmithKline, Collegeville, Pennsylvania ||GlaxoSmithKline, Research Triangle Park, North Carolina ¶Pathology Department, Memorial Hospital of RI, Pawtucket, Rhode Island.

The sirtuin family consists of seven NAD+-dependent enzymes affecting a broad array of regulatory protein networks by primarily catalyzing the deacetylation of key lysine residues in regulatory proteins. The enzymatic activity of SIRT1 can be enhanced by small molecule activators known as SIRT1 activator compounds (STACs). We tested the therapeutic potential of the STAC SRT3025 in two preclinical models of severe infection, the murine cecal ligation and puncture (CLP) model to induce peritonitis and intratracheal installation of Streptococcus pneumoniae to induce severe bacterial pneumonia. SRT3025 provided significant survival benefits over vehicle control in both the peritonitis and pneumococcal pneumonia models when administered with appropriate antimicrobial agents. The survival benefit of SRT3025 in the CLP model was absent in SIRT1 knockout showing the SIRT1 dependency of SRT3025's effects. SRT3025 administration promoted bacterial clearance and significantly reduced inflammatory cytokines from the lungs of animals challenged with S. pneumoniae. SRT3025 treatment was also accompanied by striking changes in the transcription profiles in multiple inflammatory and metabolic pathways in liver, spleen, small bowel, and lung tissue. Remarkably, these organ-specific changes in the transcriptome analyses were similar following CLP or pneumococcal challenge despite different sets of pathogens at disparate sites of infection. Pharmacologic activation of SIRT1 modulates the innate host response and could represent a novel treatment strategy for severe infection.
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http://dx.doi.org/10.1097/SHK.0000000000000528DOI Listing
April 2016

Crystallographic structure of a small molecule SIRT1 activator-enzyme complex.

Nat Commun 2015 Jul 2;6:7645. Epub 2015 Jul 2.

Sirtris, a GlaxoSmithKline Company, 200 Technology Square, Suite 300, Cambridge, Massachusetts 02139, USA.

SIRT1, the founding member of the mammalian family of seven NAD(+)-dependent sirtuins, is composed of 747 amino acids forming a catalytic domain and extended N- and C-terminal regions. We report the design and characterization of an engineered human SIRT1 construct (mini-hSIRT1) containing the minimal structural elements required for lysine deacetylation and catalytic activation by small molecule sirtuin-activating compounds (STACs). Using this construct, we solved the crystal structure of a mini-hSIRT1-STAC complex, which revealed the STAC-binding site within the N-terminal domain of hSIRT1. Together with hydrogen-deuterium exchange mass spectrometry (HDX-MS) and site-directed mutagenesis using full-length hSIRT1, these data establish a specific STAC-binding site and identify key intermolecular interactions with hSIRT1. The determination of the interface governing the binding of STACs with human SIRT1 facilitates greater understanding of STAC activation of this enzyme, which holds significant promise as a therapeutic target for multiple human diseases.
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http://dx.doi.org/10.1038/ncomms8645DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4506539PMC
July 2015

Sirtuin 1 activator SRT2104 protects Huntington's disease mice.

Ann Clin Transl Neurol 2014 Dec 31;1(12):1047-52. Epub 2014 Oct 31.

Division of Neurobiology, Department of Psychiatry and Behavioral Sciences, School of Medicine, Johns Hopkins University Baltimore, Maryland, 21287 ; Department of Neuroscience, School of Medicine, Johns Hopkins University Baltimore, Maryland, 21287 ; Program in Cellular and Molecular Medicine, School of Medicine, Johns Hopkins University Baltimore, Maryland, 21287.

Sirtuin 1 is a nicotinamide adenine dinucleotide-dependent protein deacetylase which regulates longevity and improves metabolism. Activation of Sirtuin 1 confers beneficial effects in models of neurodegenerative diseases. We and others have provided convincing evidence that overexpression of Sirtuin 1 plays a neuroprotective role in mouse models of Huntington's disease. In this study, we report that SRT2104, a small molecule Sirtuin 1 activator, penetrated the blood-brain barrier, attenuated brain atrophy, improved motor function, and extended survival in a mouse model of Huntington's disease. These findings imply a novel therapeutic strategy for Huntington's disease by targeting Sirtuin 1.
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http://dx.doi.org/10.1002/acn3.135DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4284130PMC
December 2014

SRT2104 extends survival of male mice on a standard diet and preserves bone and muscle mass.

Aging Cell 2014 Oct 16;13(5):787-96. Epub 2014 Jun 16.

Translational Gerontology Branch, National Institute on Aging, National Institutes of Health, Baltimore, MD, 21224, USA.

Increased expression of SIRT1 extends the lifespan of lower organisms and delays the onset of age-related diseases in mammals. Here, we show that SRT2104, a synthetic small molecule activator of SIRT1, extends both mean and maximal lifespan of mice fed a standard diet. This is accompanied by improvements in health, including enhanced motor coordination, performance, bone mineral density, and insulin sensitivity associated with higher mitochondrial content and decreased inflammation. Short-term SRT2104 treatment preserves bone and muscle mass in an experimental model of atrophy. These results demonstrate it is possible to design a small molecule that can slow aging and delay multiple age-related diseases in mammals, supporting the therapeutic potential of SIRT1 activators in humans.
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http://dx.doi.org/10.1111/acel.12220DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4172519PMC
October 2014

The Sirt1 activator SRT3025 provides atheroprotection in Apoe-/- mice by reducing hepatic Pcsk9 secretion and enhancing Ldlr expression.

Eur Heart J 2015 Jan 6;36(1):51-9. Epub 2014 Mar 6.

Cardiovascular Research, Institute of Physiology, University of Zurich and University Heart Center, Cardiology, University Hospital Zurich, Raemistrasse 100, 8091 Zurich, Switzerland Zurich Center of Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland

Aims: The deacetylase sirtuin 1 (Sirt1) exerts beneficial effects on lipid metabolism, but its roles in plasma LDL-cholesterol regulation and atherosclerosis are controversial. Thus, we applied the pharmacological Sirt1 activator SRT3025 in a mouse model of atherosclerosis and in hepatocyte culture.

Methods And Results: Apolipoprotein E-deficient (Apoe(-/-)) mice were fed a high-cholesterol diet (1.25% w/w) supplemented with SRT3025 (3.18 g kg(-1) diet) for 12 weeks. In vitro, the drug activated wild-type Sirt1 protein, but not the activation-resistant Sirt1 mutant; in vivo, it increased deacetylation of hepatic p65 and skeletal muscle Foxo1. SRT3025 treatment decreased plasma levels of LDL-cholesterol and total cholesterol and reduced atherosclerosis. Drug treatment did not change mRNA expression of hepatic LDL receptor (Ldlr) and proprotein convertase subtilisin/kexin type 9 (Pcsk9), but increased their protein expression indicating post-translational effects. Consistent with hepatocyte Ldlr and Pcsk9 accumulation, we found reduced plasma levels of Pcsk9 after pharmacological Sirt1 activation. In vitro administration of SRT3025 to cultured AML12 hepatocytes attenuated Pcsk9 secretion and its binding to Ldlr, thereby reducing Pcsk9-mediated Ldlr degradation and increasing Ldlr expression and LDL uptake. Co-administration of exogenous Pcsk9 with SRT3025 blunted these effects. Sirt1 activation with SRT3025 in Ldlr(-/-) mice reduced neither plasma Pcsk9, nor LDL-cholesterol levels, nor atherosclerosis.

Conclusion: We identify reduction in Pcsk9 secretion as a novel effect of Sirt1 activity and uncover Ldlr as a prerequisite for Sirt1-mediated atheroprotection in mice. Pharmacological activation of Sirt1 appears promising to be tested in patients for its effects on plasma Pcsk9, LDL-cholesterol, and atherosclerosis.
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http://dx.doi.org/10.1093/eurheartj/ehu095DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4286317PMC
January 2015

The SIRT1 activator SRT1720 extends lifespan and improves health of mice fed a standard diet.

Cell Rep 2014 Mar 27;6(5):836-43. Epub 2014 Feb 27.

Translational Gerontology Branch, National Institute on Aging, National Institutes of Health, 251 Bayview Boulevard, Baltimore, MD 21224, USA. Electronic address:

The prevention or delay of the onset of age-related diseases prolongs survival and improves quality of life while reducing the burden on the health care system. Activation of sirtuin 1 (SIRT1), an NAD(+)-dependent deacetylase, improves metabolism and confers protection against physiological and cognitive disturbances in old age. SRT1720 is a specific SIRT1 activator that has health and lifespan benefits in adult mice fed a high-fat diet. We found extension in lifespan, delayed onset of age-related metabolic diseases, and improved general health in mice fed a standard diet after SRT1720 supplementation. Inhibition of proinflammatory gene expression in both liver and muscle of SRT1720-treated animals was noted. SRT1720 lowered the phosphorylation of NF-κB pathway regulators in vitro only when SIRT1 was functionally present. Combined with our previous work, the current study further supports the beneficial effects of SRT1720 on health across the lifespan in mice.
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http://dx.doi.org/10.1016/j.celrep.2014.01.031DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4010117PMC
March 2014

Carboxamide SIRT1 inhibitors block DBC1 binding via an acetylation-independent mechanism.

Cell Cycle 2013 Jul;12(14):2233-40

Department of Genetics, Harvard Medical School, Boston, MA, USA.

SIRT1 is an NAD (+) -dependent deacetylase that counteracts multiple disease states associated with aging and may underlie some of the health benefits of calorie restriction. Understanding how SIRT1 is regulated in vivo could therefore lead to new strategies to treat age-related diseases. SIRT1 forms a stable complex with DBC1, an endogenous inhibitor. Little is known regarding the biochemical nature of SIRT1-DBC1 complex formation, how it is regulated and whether or not it is possible to block this interaction pharmacologically. In this study, we show that critical residues within the catalytic core of SIRT1 mediate binding to DBC1 via its N-terminal region, and that several carboxamide SIRT1 inhibitors, including EX-527, can completely block this interaction. We identify two acetylation sites on DBC1 that regulate its ability to bind SIRT1 and suppress its activity. Furthermore, we show that DBC1 itself is a substrate for SIRT1. Surprisingly, the effect of EX-527 on SIRT1-DBC1 binding is independent of DBC1 acetylation. Together, these data show that protein acetylation serves as an endogenous regulatory mechanism for SIRT1-DBC1 binding and illuminate a new path to developing small-molecule modulators of SIRT1.
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http://dx.doi.org/10.4161/cc.25268DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3755073PMC
July 2013

Identification of a SIRT1 mutation in a family with type 1 diabetes.

Cell Metab 2013 Mar;17(3):448-455

Functional Genomic Center Zurich, Federal Institute of Technology University of Zurich, 8057 Zurich, Switzerland.

Type 1 diabetes is caused by autoimmune-mediated β cell destruction leading to insulin deficiency. The histone deacetylase SIRT1 plays an essential role in modulating several age-related diseases. Here we describe a family carrying a mutation in the SIRT1 gene, in which all five affected members developed an autoimmune disorder: four developed type 1 diabetes, and one developed ulcerative colitis. Initially, a 26-year-old man was diagnosed with the typical features of type 1 diabetes, including lean body mass, autoantibodies, T cell reactivity to β cell antigens, and a rapid dependence on insulin. Direct and exome sequencing identified the presence of a T-to-C exchange in exon 1 of SIRT1, corresponding to a leucine-to-proline mutation at residue 107. Expression of SIRT1-L107P in insulin-producing cells resulted in overproduction of nitric oxide, cytokines, and chemokines. These observations identify a role for SIRT1 in human autoimmunity and unveil a monogenic form of type 1 diabetes.
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http://dx.doi.org/10.1016/j.cmet.2013.02.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3746172PMC
March 2013

Evidence for a common mechanism of SIRT1 regulation by allosteric activators.

Science 2013 Mar;339(6124):1216-9

Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.

A molecule that treats multiple age-related diseases would have a major impact on global health and economics. The SIRT1 deacetylase has drawn attention in this regard as a target for drug design. Yet controversy exists around the mechanism of sirtuin-activating compounds (STACs). We found that specific hydrophobic motifs found in SIRT1 substrates such as PGC-1α and FOXO3a facilitate SIRT1 activation by STACs. A single amino acid in SIRT1, Glu(230), located in a structured N-terminal domain, was critical for activation by all previously reported STAC scaffolds and a new class of chemically distinct activators. In primary cells reconstituted with activation-defective SIRT1, the metabolic effects of STACs were blocked. Thus, SIRT1 can be directly activated through an allosteric mechanism common to chemically diverse STACs.
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http://dx.doi.org/10.1126/science.1231097DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3799917PMC
March 2013

Pharmacological modulation of circadian rhythms by synthetic activators of the deacetylase SIRT1.

Proc Natl Acad Sci U S A 2013 Feb 22;110(9):3333-8. Epub 2013 Jan 22.

Center for Epigenetics and Metabolism, Department of Biological Chemistry, University of California, Irvine, CA 92697, USA.

Circadian rhythms govern a wide variety of physiological and metabolic functions in many organisms, from prokaryotes to humans. We previously reported that silent information regulator 1 (SIRT1), a NAD(+)-dependent deacetylase, contributes to circadian control. In addition, SIRT1 activity is regulated in a cyclic manner in virtue of the circadian oscillation of the coenzyme NAD(+). Here we used specific SIRT1 activator compounds both in vitro and in vivo. We tested a variety of compounds to show that the activation of SIRT1 alters CLOCK:BMAL1-driven transcription in different systems. Activation of SIRT1 induces repression of circadian gene expression and decreases H3 K9/K14 acetylation at corresponding promoters in a time-specific manner. Specific activation of SIRT1 was demonstrated in vivo using liver-specific SIRT1-deficient mice, where the effect of SIRT1 activator compounds was shown to be dependent on SIRT1. Our findings demonstrate that SIRT1 can fine-tune circadian rhythm and pave the way to the development of pharmacological strategies to address a broad range of therapeutic indications.
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http://dx.doi.org/10.1073/pnas.1214266110DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3587185PMC
February 2013

SIRT1 activators suppress inflammatory responses through promotion of p65 deacetylation and inhibition of NF-κB activity.

PLoS One 2012 28;7(9):e46364. Epub 2012 Sep 28.

Sirtris, a GSK Company, Cambridge, Massachusetts, United States of America.

Chronic inflammation is a major contributing factor in the pathogenesis of many age-associated diseases. One central protein that regulates inflammation is NF-κB, the activity of which is modulated by post-translational modifications as well as by association with co-activator and co-repressor proteins. SIRT1, an NAD(+)-dependent protein deacetylase, has been shown to suppress NF-κB signaling through deacetylation of the p65 subunit of NF-κB resulting in the reduction of the inflammatory responses mediated by this transcription factor. The role of SIRT1 in the regulation of NF-κB provides the necessary validation for the development of pharmacological strategies for activating SIRT1 as an approach for the development of a new class of anti-inflammatory therapeutics. We report herein the development of a quantitative assay to assess compound effects on acetylated p65 protein in the cell. We demonstrate that small molecule activators of SIRT1 (STACs) enhance deacetylation of cellular p65 protein, which results in the suppression of TNFα-induced NF-κB transcriptional activation and reduction of LPS-stimulated TNFα secretion in a SIRT1-dependent manner. In an acute mouse model of LPS-induced inflammation, the STAC SRTCX1003 decreased the production of the proinflammatory cytokines TNFα and IL-12. Our studies indicate that increasing SIRT1-mediated NF-κB deacetylation using small molecule activating compounds is a novel approach to the development of a new class of therapeutic anti-inflammatory agents.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0046364PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3460821PMC
February 2013

SRT1720 improves survival and healthspan of obese mice.

Sci Rep 2011 18;1:70. Epub 2011 Aug 18.

Laboratory of Experimental Gerontology, National Institute on Aging, National Institutes of Health, 251 Bayview Boulevard, Baltimore, MD 21224, USA.

Sirt1 is an NAD(+)-dependent deacetylase that extends lifespan in lower organisms and improves metabolism and delays the onset of age-related diseases in mammals. Here we show that SRT1720, a synthetic compound that was identified for its ability to activate Sirt1 in vitro, extends both mean and maximum lifespan of adult mice fed a high-fat diet. This lifespan extension is accompanied by health benefits including reduced liver steatosis, increased insulin sensitivity, enhanced locomotor activity and normalization of gene expression profiles and markers of inflammation and apoptosis, all in the absence of any observable toxicity. Using a conditional SIRT1 knockout mouse and specific gene knockdowns we show SRT1720 affects mitochondrial respiration in a Sirt1- and PGC-1α-dependent manner. These findings indicate that SRT1720 has long-term benefits and demonstrate for the first time the feasibility of designing novel molecules that are safe and effective in promoting longevity and preventing multiple age-related diseases in mammals.
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http://dx.doi.org/10.1038/srep00070DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3216557PMC
August 2013

SIRT1 modulation as a novel approach to the treatment of diseases of aging.

J Med Chem 2011 Jan 16;54(2):417-32. Epub 2010 Nov 16.

Sirtris, A GSK Company, 200 Technology Square, Cambridge, Massachusetts 02139, United States.

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http://dx.doi.org/10.1021/jm100861pDOI Listing
January 2011

Comparative gene expression profiling in three primary human cell lines after treatment with a novel inhibitor of Rho kinase or atorvastatin.

Blood Coagul Fibrinolysis 2008 Oct;19(7):709-18

Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, 4301 West Markham, Little Rock, AR 72205, USA.

Inhibitors of Rho kinase (ROCK) are a relatively new class of drugs with potential benefits in oncology, neurology, and fibrotic and cardiovascular diseases. ROCK inhibitors modulate many cellular functions, some of which are similar to the pleiotropic effects of statins, suggesting additive or synergistic properties. Studies to date have used compounds that inhibit both isoforms of ROCK, ROCK1 and ROCK2. This study was designed to compare gene expression profiles of atorvastatin with the newly developed ROCK2 inhibitor SLx-2119 in primary cultures of normal human endothelial cells, smooth muscle cells, and fibroblasts. Cells were treated with each compound for 24 h, after which total RNA was isolated and genome-wide gene-expression profiles were obtained with Illumina arrays. Because of the known effect of statins on the actin cytoskeleton and on connective tissue growth factor, a prominent growth factor involved in tissue fibrosis, the effects of SLx-2119 and atorvastatin on the actin cytoskeleton and connective tissue growth factor mRNA were also examined in cultures of smooth muscle cells with a fibrotic phenotype, isolated from biopsies of human intestine with radiation-induced fibrosis. Although SLx-2119 and atorvastatin affected expression of genes belonging to the same biological processes, individual genes were mostly different, consistent with synergistic or additive properties. Both SLx-2119 and atorvastatin reduced connective tissue growth factor mRNA and remodeled the actin cytoskeleton in fibrosis-derived smooth muscle cells, suggesting that both compounds have antifibrotic properties. These results form the basis for further studies to assess the possible therapeutic benefit of combined treatments.
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http://dx.doi.org/10.1097/MBC.0b013e32830b2891DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2713681PMC
October 2008

Structure-based design, synthesis, and biological evaluation of indomethacin derivatives as cyclooxygenase-2 inhibiting nitric oxide donors.

J Med Chem 2007 Dec 10;50(25):6367-82. Epub 2007 Nov 10.

NitroMed, Inc., 125 Spring Street, Lexington, Massachusetts 02421, USA.

Indomethacin, a nonselective cyclooxygenase (COX) inhibitor, was modified in three distinct regions in an attempt both to increase cyclooxygenase-2 (COX-2) selectivity and to enhance drug safety by covalent attachment of an organic nitrate moiety as a nitric oxide donor. A human whole-blood COX assay shows the modifications on the 3-acetic acid part of the indomethacin yielding an amide-nitrate derivative 32 and a sulfonamide-nitrate derivative 61 conferred COX-2 selectivity. Along with their respective des-nitrate analogs, for example, 31 and 62, the nitrates 32 and 61 were effective antiinflammatory agents in the rat air-pouch model. After oral dosing, though, only 32 increased nitrate and nitrite levels in rat plasma, indicating that its nitrate tether served as a nitric oxide donor in vivo. In a rat gastric injury model, examples 31 and 32 both show a 98% reduction in gastric lesion score compared to that of indomethacin. In addition, the nitrated derivative 32 inducing 85% fewer gastric lesions when coadministered with aspirin as compared to the combination of aspirin and valdecoxib.
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http://dx.doi.org/10.1021/jm0611861DOI Listing
December 2007

Rat model for evaluation of therapeutics on peripheral vascular resistance.

Curr Protoc Pharmacol 2006 Dec;Chapter 5:Unit5.44

Surface Logix, Inc., Brighton, Massachusetts, USA.

Described in this protocol is a rat preparation that serves as a sensitive, reliable in vivo assay of peripheral vascular resistance. Experiments are conducted on conscious animals 5 days following the surgical implantation of an arterial pressure catheter in the carotid artery and a transit time flow probe on the abdominal aorta. The transit time flow probe measures volume blood flow per unit time, making it possible to calculate a true vascular resistance, a reliable indicator of microvascular tone. After allowing appropriate recovery time between experiments, it is possible to use the animal preparation multiple times. This increases study efficiency by reducing the number of animals required and makes it possible to use paired statistical analysis since multiple treatment interventions, including control conditions, can be evaluated in each animal.
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http://dx.doi.org/10.1002/0471141755.ph0544s35DOI Listing
December 2006

Modulation of citric acid-induced cough following lipopolysaccharide-mediated neutrophilia in the guinea pig.

Pulm Pharmacol Ther 2007 14;20(1):90-7. Epub 2006 Feb 14.

UCB Research Inc., UCB Pharma, 840 Memorial Dr., Cambridge, MA 02139, USA.

This investigation examined a possible correlation between lipopolysaccharide (LPS)-induced pulmonary neutrophilia and cough. Conscious male guinea pigs were acutely exposed to aerosolized LPS and thereafter at various times challenged with citric acid aerosol (CA; 250mM) to induce cough followed by bronchoalveolar lavage (BAL) to quantitate inflammatory cell accumulation. LPS caused a hyporesponsive cough at 24h post-LPS with neutrophilia apparent from 2h post-LPS. By 96h post-LPS both cough and neutrophilia had returned towards normal. Dexamethasone (DEX, 2mgkg(-1)/day for 3 days prior) did not affect the cough hyporesponsiveness at 24h; however it attenuated LPS-induced BAL fluid neutrophilia. Since LPS can stimulate inducible nitric oxide synthase (iNOS) we hypothesized that the cough hyporesponsiveness may involve nitric oxide. To investigate this we treated animals with an aerosolized iNOS inhibitor 1400W (1mM) immediately prior to LPS. 1400W had no significant effect on either cough hyporesponsiveness or BAL fluid neutrophilia at 24h post-LPS. Despite differing effects on neutrophilia, these findings clearly indicate that neither DEX nor iNOS inhibition had any direct effect on LPS-induced cough hyporesponsiveness. The mechanism underlying the LPS-induced cough hyporesponsiveness does not appear to be directly linked to LPS-induced neutrophilic inflammation.
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http://dx.doi.org/10.1016/j.pupt.2005.12.009DOI Listing
May 2007

Synthesis and anti-inflammatory activity of a series of N-substituted naproxen glycolamides: nitric oxide-donor naproxen prodrugs.

Bioorg Med Chem 2006 Apr 13;14(8):2589-99. Epub 2005 Dec 13.

NitroMed, Inc., 125 Spring St., Lexington, MA 02421, USA.

A series of glycolamide naproxen prodrugs containing a nitrate group as a nitric oxide (NO) donor moiety has been synthesized. These compounds were evaluated for their anti-inflammatory activity, naproxen release, and gastric tolerance. Compounds 4a, 4b, 5a, 5b, 7b, and 7c exhibited anti-inflammatory activity equivalent to that of the parent NSAID, naproxen-Na, in the rat carrageenan paw edema model. At equimolar doses relative to naproxen-Na, the NO-donor glycolamide derivatives 4a, 4b, 5a, 5b, 7b, and 7c were gastro-sparing in the rat. Naproxen formation from these NO-donor glycolamides varied among the structures examined, with the N-substituent on the amide group having a particular influence, and demonstrated their prodrug nature. Compound 7b was selected for exemplary demonstration that the glycolamide nitrates can be bioactivated to release NO. These data open the possibility that naproxen glycolamide nitrates may represent a safer alternative to naproxen as anti-inflammatory medicines.
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http://dx.doi.org/10.1016/j.bmc.2005.11.040DOI Listing
April 2006

NMI-1182, a gastro-protective cyclo-oxygenase-inhibiting nitric oxide donor.

Inflammopharmacology 2005 ;12(5-6):521-34

NitroMed Inc., 125 Spring Street, Lexington, MA 02421-0781, USA.

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to treat inflammation and to provide pain relief but suffer from a major liability concerning their propensity to cause gastric damage. As nitric oxide (NO) is known to be gastro-protective we have synthesized a NO-donating prodrug of naproxen named NMI-1182. We evaluated two cyclo-oxygenase (COX)-inhibiting nitric oxide donors (CINODs), NMI-1182 and AZD3582, for their ability to be gastro-protective compared to naproxen and for their anti-inflammatory activity. NMI-1182 and AZD3582 were found to produce similar inhibition of COX activity to that produced by naproxen. Both NMI-1182 and AZD3582 produced significantly less gastric lesions after oral administration than naproxen. All three compounds effectively inhibited paw swelling in the rat carrageenan paw edema model. In the carrageenan air pouch model all three compounds significantly reduced PGE2 levels in the pouch exudate but only NMI-1182 and naproxen inhibited leukocyte influx. These data demonstrate that NMI-1182 has comparable anti-inflammatory activity to naproxen but with a much reduced likelihood to cause gastric damage.
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http://dx.doi.org/10.1163/156856005774382661DOI Listing
May 2007

A comparison of the cyclooxygenase inhibitor-NO donors (CINOD), NMI-1182 and AZD3582, using in vitro biochemical and pharmacological methods.

Biochem Pharmacol 2005 Nov;70(9):1343-51

Departments of Biology, 125 Spring St., Lexington, MA 02421, USA.

Cyclooxygenase (COX, EC 1.14.99.1) inhibitor-nitric oxide (NO) donor (CINOD) hybrid compounds represent an attractive alternative to NSAID and coxib therapy. This report compares two CINODs, NMI-1182 (naproxen-glyceryl dinitrate) and AZD3582 (naproxen-n-butyl nitrate), for their ability to inhibit COX-1 and -2, deliver bioavailable nitric oxide, and release naproxen, using in vitro biochemical and pharmacological methods. In human whole blood, both CINODs showed inhibition, comparable to naproxen, of both COX isozymes and slowly released naproxen. Both CINODs donated bioavailable NO, as detected by cGMP induction in the pig kidney transformed cell line, LLC-PK1, but NMI-1182 was more potent by 30-100 times than AZD3582, GTN, GDN, and ISDN and considerably faster in inducing cGMP synthesis than AZD3582. The nitrate groups of GTN, NMI-1182, and AZD3582 appeared to be bioactivated via a common pathway, since each compound desensitized LLC-PK1 cells to subsequent challenge with the other compounds. Similar cGMP induction also occurred in normal, untransformed cells (human renal proximal tubule epithelial cells and hepatocytes from man, rat, and monkey); again, NMI-1182 was superior to AZD3582. NMI-1182 was also the more metabolically labile compound, releasing more absolute nitrate and nitrite (total NO(x)) in human stomach (in which NO is salutary) and liver S9 homogenates. Naproxen was also more rapidly freed from NMI-1182 than AZD3582 in human stomach, although liver S9 hydrolyzed both CINODs with similar rates. These in vitro tests revealed that NMI-1182 may be a better CINOD than AZD3582 because of its superior NO donating and naproxen liberating properties.
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http://dx.doi.org/10.1016/j.bcp.2005.08.004DOI Listing
November 2005

Cetirizine and loratadine-based antihistamines with 5-lipoxygenase inhibitory activity.

Bioorg Med Chem Lett 2004 Nov;14(22):5591-4

UCB Research, 840 Memorial Drive, Cambridge, MA 02139, USA.

A series of compounds possessing both H(1) histamine receptor antagonist and 5-lipoxygenase (5-LO) inhibitory activities was synthesized. The H(1)-binding scaffolds of cetirizine, efletirizine, and loratadine were linked to a lipophilic N-hydroxyurea, the 5-LO inhibiting moiety of zileuton. Both activities were observed in vivo, as was increased CYP3A4 inhibition compared to their respective single-function drugs. Selected analogs in the series were shown to be orally active in guinea pig models.
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http://dx.doi.org/10.1016/j.bmcl.2004.08.060DOI Listing
November 2004

Synthesis and selective cyclooxygenase-2 inhibitory activity of a series of novel, nitric oxide donor-containing pyrazoles.

J Med Chem 2004 Apr;47(9):2180-93

NitroMed Inc., 12 Oak Park Drive, Bedford, Massachusetts 01730, USA.

The synthesis of a series of novel pyrazoles containing a nitrate (ONO(2)) moiety as a nitric oxide (NO)-donor functionality is reported. Their COX-1 and COX-2 inhibitory activities in human whole blood are profiled. Our data demonstrate that pyrazole ring substituents play an important role in COX-2 selective inhibition, such that a cycloalkyl pyrazole (6b) was found to be a potent and selective COX-2 inhibitor. Other modifications at the 3 position of the central pyrazole ring (17b, 23b, 26b-I) enhanced COX-2 inhibitory potency. Among the pyrazoles synthesized, the oxime (23b) was identified as the most potent COX-2 selective inhibitor. Accordingly, 23b was profiled pharmacologically in the rat after oral administration and shown to possess potent antiinflammatory activity in the carrageenan-induced air-pouch model and less gastric toxicity than a standard COX-2 inhibitor when administered with background aspirin treatment. We suggest that the enhanced gastric tolerance of an NO-donor COX-2 selective inhibitor has the potential to augment the clinical profile of this drug class.
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http://dx.doi.org/10.1021/jm030276sDOI Listing
April 2004

5-lipoxygenase inhibitors with histamine H(1) receptor antagonist activity.

Bioorg Med Chem Lett 2004 May;14(9):2265-8

UCB Research, 840 Memorial Dr, Cambridge MA 02139, USA.

A series of novel compounds with both 5-lipoxygenase (5-LO) inhibitory and histamine H(1) receptor antagonist activity were designed for the treatment of asthma. These dual-function compounds were made by connecting 5-LO and H(1) pharmacophores,N-hydroxyureas and benzhydryl piperazines, respectively. A range of in vitro activities was observed, with the furan analog 10 demonstrating both activities in an animal model. The activities observed were compared to single-function drugs.
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http://dx.doi.org/10.1016/j.bmcl.2004.02.005DOI Listing
May 2004

Antitussive activity of sigma-1 receptor agonists in the guinea-pig.

Br J Pharmacol 2004 Jan 22;141(2):233-40. Epub 2003 Dec 22.

UCB Research Inc., UCB Pharma, 840 Memorial Drive, Cambridge, MA 02139, USA.

1. Current antitussive medications have limited efficacy and often contain the opiate-like agent dextromethorphan (DEX). The mechanism whereby DEX inhibits cough is ill defined. DEX displays affinity at both NMDA and sigma receptors, suggesting that the antitussive activity may involve central or peripheral activity at either of these receptors. This study examined and compared the antitussive activity of DEX and various putative sigma receptor agonists in the guinea-pig citric-acid cough model. 2. Intraperitoneal (i.p.) administration of DEX (30 mg kg(-1)) and the sigma-1 agonists SKF-10,047 (1-5 mg kg(-1)), Pre-084 (5 mg kg(-1)), and carbetapentane (1-5 mg kg(-1)) inhibited citric-acid-induced cough in guinea-pigs. Intraperitoneal administration of a sigma-1 antagonist, BD 1047 (1-5 mg kg(-1)), reversed the inhibition of cough elicited by SKF-10,047. In addition, two structurally dissimilar sigma agonists SKF-10,047 (1 mg ml(-1)) and Pre-084 (1 mg ml(-1)) inhibited cough when administered by aerosol. 3. Aerosolized BD 1047 (1 mg ml(-1), 30 min) prevented the antitussive action of SKF-10,047 (5 mg kg(-1)) or DEX (30 mg kg(-1)) given by i.p. administration and, likewise, i.p. administration of BD 1047 (5 mg kg(-1)) prevented the antitussive action of SKF-10,047 given by aerosol (1 mg ml(-1)). 4. These results therefore support the argument that antitussive effects of DEX may be mediated via sigma receptors, since both systemic and aerosol administration of sigma-1 receptor agonists inhibit citric-acid-induced cough in guinea-pigs. While significant systemic exposure is possible with aerosol administration, the very low doses administered (estimated <0.3 mg kg(-1)) suggest that there may be a peripheral component to the antitussive effect.
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http://dx.doi.org/10.1038/sj.bjp.0705605DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1574192PMC
January 2004

Effect of olvanil on the afferent and efferent function of capsaicin-sensitive C-fibers in guinea pig airways.

Eur J Pharmacol 2003 Jun;471(3):205-11

UCB Research Inc., 840 Memorial Drive, Cambridge, MA 02139, USA.

The aim of the present study was to examine the ability of the nonpungent vanilloid VR1 receptor agonist, olvanil, to activate the afferent and efferent function of capsaicin-sensitive C-fibers in guinea pig airways. We found that while capsaicin (10 nM-10 microM) and resiniferatoxin (0.1 nM-1.0 microM) evoked a robust contraction of the guinea pig trachea in vitro, olvanil (10 nM-10 microM) was a weak spasmogen. In addition, pretreatment with olvanil caused only a minor reduction of subsequent responses to capsaicin or resiniferatoxin. Using single fiber recording from guinea pig airway C-fibers, we found that olvanil (10 microM) did not evoke action potential discharge although these fibers responded vigorously to capsaicin after prolonged treatment with olvanil (10 microM). These findings are indicative of significant differences in the relative sensitivity of vanilloid VR1 receptor-transfected cells and the peripheral terminals of airway C-fibers to pungent and nonpungent vanilloid VR1 receptor agonists.
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http://dx.doi.org/10.1016/s0014-2999(03)01833-8DOI Listing
June 2003

The study of airway primary afferent neuron excitability.

Curr Opin Pharmacol 2002 Jun;2(3):216-9

UCB Research Inc., 840 Memorial Drive, Cambridge, Massachusetts 02139, USA.

The activation of airway afferent neurons initiates a variety of reflexes including cough and bronchoconstriction. Like somatic afferent neurons involved in inflammation-induced hyperalgesia, the excitability of airway afferent neurons is not fixed but, rather, can be increased by the action of a variety of mediators produced during inflammation. A variety of techniques have been applied to study the pharmacological modulation of the excitability of afferent neurons. Although airway afferent neurons have not been studied to the same extent as fibers involved in hyperalgesia, similar pathways may control their excitability. The ability to study the pharmacological modulation of airway afferent neuron excitability is crucial to our understanding of airway afferent neurophysiology and may also provide insight into novel therapeutic targets for various inflammatory lung diseases.
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http://dx.doi.org/10.1016/s1471-4892(02)00147-9DOI Listing
June 2002