Publications by authors named "Stephen Altmann"

8 Publications

  • Page 1 of 1

Pharmacologic Characterization of AMG8379, a Potent and Selective Small Molecule Sulfonamide Antagonist of the Voltage-Gated Sodium Channel Na1.7.

J Pharmacol Exp Ther 2017 07 4;362(1):146-160. Epub 2017 May 4.

Department of Neuroscience (T.J.K., R.Y., S.A, C.P.I., M.J., D.J., J.H.L., S.G.L., J.Li., D.L., J.Lu., D.M., D.O., K.T., J.W., V.Y., D.X.D.Z., R.T.F., B.D.M.), Department of Medicinal Chemistry (M.M.W.), and Department of Pharmacokinetics and Drug Metabolism (X.B., V.B., J.R.), Amgen Inc., Cambridge, Massachusetts and Thousand Oaks, California

Potent and selective antagonists of the voltage-gated sodium channel Na1.7 represent a promising avenue for the development of new chronic pain therapies. We generated a small molecule atropisomer quinolone sulfonamide antagonist AMG8379 and a less active enantiomer AMG8380. Here we show that AMG8379 potently blocks human Na1.7 channels with an IC of 8.5 nM and endogenous tetrodotoxin (TTX)-sensitive sodium channels in dorsal root ganglion (DRG) neurons with an IC of 3.1 nM in whole-cell patch clamp electrophysiology assays using a voltage protocol that interrogates channels in a partially inactivated state. AMG8379 was 100- to 1000-fold selective over other Na family members, including Na1.4 expressed in muscle and Na1.5 expressed in the heart, as well as TTX-resistant Na channels in DRG neurons. Using an ex vivo mouse skin-nerve preparation, AMG8379 blocked mechanically induced action potential firing in C-fibers in both a time-dependent and dose-dependent manner. AMG8379 similarly reduced the frequency of thermally induced C-fiber spiking, whereas AMG8380 affected neither mechanical nor thermal responses. In vivo target engagement of AMG8379 in mice was evaluated in multiple Na1.7-dependent behavioral endpoints. AMG8379 dose-dependently inhibited intradermal histamine-induced scratching and intraplantar capsaicin-induced licking, and reversed UVB radiation skin burn-induced thermal hyperalgesia; notably, behavioral effects were not observed with AMG8380 at similar plasma exposure levels. AMG8379 is a potent and selective Na1.7 inhibitor that blocks sodium current in heterologous cells as well as DRG neurons, inhibits action potential firing in peripheral nerve fibers, and exhibits pharmacodynamic effects in translatable models of both itch and pain.
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http://dx.doi.org/10.1124/jpet.116.239590DOI Listing
July 2017

Application of a Parallel Synthetic Strategy in the Discovery of Biaryl Acyl Sulfonamides as Efficient and Selective NaV1.7 Inhibitors.

J Med Chem 2016 09 29;59(17):7818-39. Epub 2016 Aug 29.

Department of Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.

The majority of potent and selective hNaV1.7 inhibitors possess common pharmacophoric features that include a heteroaryl sulfonamide headgroup and a lipophilic aromatic tail group. Recently, reports of similar aromatic tail groups in combination with an acyl sulfonamide headgroup have emerged, with the acyl sulfonamide bestowing levels of selectivity over hNaV1.5 comparable to the heteroaryl sulfonamide. Beginning with commercially available carboxylic acids that met selected pharmacophoric requirements in the lipophilic tail, a parallel synthetic approach was applied to rapidly generate the derived acyl sulfonamides. A biaryl acyl sulfonamide hit from this library was elaborated, optimizing for potency and selectivity with attention to physicochemical properties. The resulting novel leads are potent, ligand and lipophilic efficient, and selective over hNaV1.5. Representative lead 36 demonstrates selectivity over other human NaV isoforms and good pharmacokinetics in rodents. The biaryl acyl sulfonamides reported herein may also offer ADME advantages over known heteroaryl sulfonamide inhibitors.
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http://dx.doi.org/10.1021/acs.jmedchem.6b00425DOI Listing
September 2016

Sirtuin 2 inhibitors rescue alpha-synuclein-mediated toxicity in models of Parkinson's disease.

Science 2007 Jul 21;317(5837):516-9. Epub 2007 Jun 21.

Alzheimer's Research Unit, MGH, Harvard Medical School, CNY 114, 16th Street, Charlestown, MA 02129, USA.

The sirtuins are members of the histone deacetylase family of proteins that participate in a variety of cellular functions and play a role in aging. We identified a potent inhibitor of sirtuin 2 (SIRT2) and found that inhibition of SIRT2 rescued alpha-synuclein toxicity and modified inclusion morphology in a cellular model of Parkinson's disease. Genetic inhibition of SIRT2 via small interfering RNA similarly rescued alpha-synuclein toxicity. Furthermore, the inhibitors protected against dopaminergic cell death both in vitro and in a Drosophila model of Parkinson's disease. The results suggest a link between neurodegeneration and aging.
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http://dx.doi.org/10.1126/science.1143780DOI Listing
July 2007

Discovery of a novel small-molecule targeting selective clearance of mutant huntingtin fragments.

J Biomol Screen 2007 Apr 22;12(3):351-60. Epub 2007 Mar 22.

Massachusetts Institute of Technology, Cambridge, MA, USA.

CAG-triplet repeat extension, translated into polyglutamines within the coding frame of otherwise unrelated gene products, causes 9 incurable neurodegenerative disorders, including Huntington's disease. Although an expansion in the CAG repeat length is the autosomal dominant mutation that causes the fully penetrant neurological phenotypes, the repeat length is inversely correlated with the age of onset. The precise molecular mechanism(s) of neurodegeneration remains elusive, but compelling evidence implicates the protein or its proteolytic fragments as the cause for the gain of novel pathological function(s). The authors sought to identify small molecules that target the selective clearance of polypeptides containing pathological polyglutamine extension. In a high-throughput chemical screen, they identified compounds that facilitate the clearance of a small huntingtin fragment with extended polyglutamines fused to green fluorescent protein reporter. Identified hits were validated in dose-response and toxicity tests. Compounds have been further tested in an assay for clearance of a larger huntingtin fragment, containing either pathological or normal polyglutamine repeats. In this assay, the authors identified compounds selectively targeting the clearance of mutant but not normal huntingtin fragments. These compounds were subjected to a functional assay, which yielded a lead compound that rescues cells from induced mutant polyglutamine toxicity.
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http://dx.doi.org/10.1177/1087057107299428DOI Listing
April 2007

Discovery of bioactive small-molecule inhibitor of poly adp-ribose polymerase: implications for energy-deficient cells.

Chem Biol 2006 Jul;13(7):765-70

Massachusetts General Institute for Neurodegenerative Disease and Harvard Medical School, Department of Neurology, Massachusetts General Hospital, Charlestown, 02129, USA.

Poly (ADP-ribose) polymerase (PARP1) is a nuclear protein that, when overactivated by oxidative stress-induced DNA damage, ADP ribosylates target proteins leading to dramatic cellular ATP depletion. We have discovered a biologically active small-molecule inhibitor of PARP1. The discovered compound inhibited PARP1 enzymatic activity in vitro and prevented ATP loss and cell death in a surrogate model of oxidative stress in vivo. We also investigated a new use for PARP1 inhibitors in energy-deficient cells by using Huntington's disease as a model. Our results showed that insult with the oxidant hydrogen peroxide depleted cellular ATP in mutant cells below the threshold of viability. The protective role of PARP1 inhibitors against oxidative stress has been shown in this model system.
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http://dx.doi.org/10.1016/j.chembiol.2006.05.012DOI Listing
July 2006

Pharmacological promotion of inclusion formation: a therapeutic approach for Huntington's and Parkinson's diseases.

Proc Natl Acad Sci U S A 2006 Mar 6;103(11):4246-51. Epub 2006 Mar 6.

Center for Cancer Research, Massachusetts Institute of Technology, Room E18-505, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.

Misfolded proteins accumulate in many neurodegenerative diseases, including huntingtin in Huntington's disease and alpha-synuclein in Parkinson's disease. The disease-causing proteins can take various conformations and are prone to aggregate and form larger cytoplasmic or nuclear inclusions. One approach to the development of therapeutic intervention for these diseases has been to identify chemical compounds that reduce the size or number of inclusions. We have, however, identified a compound that promotes inclusion formation in cellular models of both Huntington's disease and Parkinson's disease. Of particular interest, this compound prevents huntingtin-mediated proteasome dysfunction and reduces alpha-synuclein-mediated toxicity. These results demonstrate that compounds that increase inclusion formation may actually lessen cellular pathology in both Huntington's and Parkinson's diseases, suggesting a therapeutic approach for neurodegenerative diseases caused by protein misfolding.
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http://dx.doi.org/10.1073/pnas.0511256103DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1449678PMC
March 2006

Cloning and characterization of an Mx gene and its corresponding promoter from the zebrafish, Danio rerio.

Dev Comp Immunol 2004 Apr;28(4):295-306

Department of Biochemistry, Microbiology, and Molecular Biology, University of Maine, Orono, ME 04469, USA.

Type I interferons (IFNs) represent a crucial component of the innate immune response to viruses. An important downstream effector of IFN is the Mx gene, which is activated solely through this pathway. Mx proteins are characterized by a tripartite GTP-binding domain, dynamin family signature, and leucine zipper motif. Mx genes are transcribed upon activation of an interferon-stimulated response element (ISRE) located in the Mx promoter region. In this article, we describe the cloning and analysis of an Mx gene and its corresponding promoter from the zebrafish (Danio rerio). The deduced amino acid sequence of zebrafish Mx contains the conserved GTP-binding domain, dynamin family signature, and leucine zipper motif common to Mx proteins, and shows a 50% identity to human MxA and 69% identity both to rainbow trout and to Atlantic salmon. Zebrafish liver cells produced high levels of Mx mRNA in response to induction by the known IFN-inducer polyinosinic-polycytidylic acid (Poly[I:C]). The zebrafish Mx promoter contains two ISREs homologous to those found in the promoter regions of many IFN-inducible genes, and was able to drive transcription of a luciferase reporter gene when induced by either purified zebrafish IFN or Poly[I:C].
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http://dx.doi.org/10.1016/j.dci.2003.09.001DOI Listing
April 2004

Molecular and functional analysis of an interferon gene from the zebrafish, Danio rerio.

J Virol 2003 Feb;77(3):1992-2002

Department of Biochemistry, Microbiology, and Molecular Biology, University of Maine, Orono, Maine 04469, USA.

The interferon (IFN) family consisting of alpha IFN (IFN-alpha), IFN-beta, IFN-omega, IFN-delta, IFN-kappa, and IFN-tau is a large group of cytokines involved in the innate immune response against various microorganisms. Genes for IFN have been cloned from a variety of mammalian and avian species; however, IFN genes from lower-order vertebrates have not been forthcoming. Here, we report the cloning and characterization of the IFN gene from the zebrafish, Danio rerio. Zebrafish IFN (zfIFN) is 185 amino acids in length, with the first 22 amino acids representing a putative signal peptide. Treatment with the known IFN inducer polyinosinic acid-polycytidylic acid (poly[I]-poly[C]) resulted in an increase in zfIFN mRNA transcripts. zfIFN was also able to activate the IFN-inducible Mx promoter when cotransfected with a plasmid containing the zebrafish Mx promoter upstream of a luciferase reporter gene. To demonstrate antiviral activity, zebrafish cells were transfected with zfIFN and challenged with a fish rhabdovirus. A 36% reduction in plaque number was seen in zfIFN-transfected cells, compared to cells transfected with a control vector. Phylogenetic analysis has shown zfIFN to be approximately equally divergent from avian and mammalian IFN, consistent with its origin from an IFN present in the most recent common ancestor of these divergent lineages. A putative IFN from puffer, Fugu rubripes, was also found when zfIFN was used to search the fugu genome database, demonstrating that zfIFN can be used to find additional fish IFN genes. These results demonstrate that zebrafish can be used as an effective model for studying innate immunity and immune response to infectious disease.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC140984PMC
http://dx.doi.org/10.1128/jvi.77.3.1992-2002.2003DOI Listing
February 2003