Journal of Biomolecular Screening

 

Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here to sign up for SAGE Journal Email Alerts today!

Sign In to gain access to subscriptions and/or personal tools.
This Article
Right arrow Abstract Freely available
Right arrow Free Full Text (Free PDF) Free
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via ISI Web of Science (1)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zhang, S.-P.
Right arrow Articles by Codd, E. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zhang, S.-P.
Right arrow Articles by Codd, E. E.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
This version was published on September 1, 2006
Journal of Biomolecular Screening, Vol. 11, No. 6, 672-677 (2006)
DOI: 10.1177/1087057106289210

High-Throughput Screening for N-Type Calcium Channel Blockers Using a Scintillation Proximity Assay

Sui-Po Zhang

Drug Discovery, Johnson & Johnson Pharmaceutical Research and Development, LLC, Spring House, Pennsylvania

Jack Kauffman

Drug Discovery, Johnson & Johnson Pharmaceutical Research and Development, LLC, Spring House, Pennsylvania

Susan K. Yagel

Drug Discovery, Johnson & Johnson Pharmaceutical Research and Development, LLC, Spring House, Pennsylvania

Ellen E. Codd

Drug Discovery, Johnson & Johnson Pharmaceutical Research and Development, LLC, Spring House, Pennsylvania

N-type calcium channels located on presynaptic nerve terminals regulate neurotransmitter release, including that from the spinal terminations of primary afferent nociceptors. Accordingly, N-type calcium channel blockers may have clinical utility as analgesic drugs. A selective N-type calcium channel inhibitor, ziconotide (Prialt), is a neuroactive peptide recently marketed as a novel nonopioid treatment for severe chronic pain. To develop a small-molecule N-type calcium channel blocker, the authors developed a 96-well plate high-throughput screening scintillation proximity assay (SPA) for N-type calcium channel blockers using [125I]-labeled {omega}-conotoxin GVIA as a channel-specific ligand. Assay reagents were handled using Caliper’s Allegro automation system, and bound ligands were detected using a PerkinElmer TopCount. Using this assay, more than 150,000 compounds were screened at 10 µM and approximately 340 compounds were identified as hits, exhibiting at least 40% inhibition of [125I]GVIA binding. This is the 1st demonstration of the use of [125I]-labeled peptides with SPA beads to provide a binding assay for the evaluation of ligand binding to calcium channels. This assay could be a useful tool for drug discovery.

Key Words: calcium channel • SPA • {omega}-conotoxin GVIA • HTS

References

  • Augustine GJ, Charlton MP, Smith SJ: Calcium action in synaptic transmitter release. Annu Rev Neurosci 1987;10: 633-693.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Maggi CA, Tramontana M, Cecconi R, Santicioli P: Neurochemical evidence for the involvement of N-type calcium channels in transmitter secretion from peripheral endings of sensory nerves in guinea pigs. Neurosci Lett 1990;114: 203-206.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Holz GG IV, Dunlap K, Kream RM: Characterization of the electrically evoked release of substance P from dorsal root ganglion neurons: methods and dihydropyridine sensitivity. J Neurosci 1988;8: 463-471.[Abstract]
  • Santicioli P, Del Bianco E, Tramontana M, Geppetti P, Maggi CA: Release of calcitonin gene-related peptide like-immunoreactivity induced by electrical field stimulation from rat spinal afferents is mediated by conotoxin-sensitive calcium channels. Neurosci Lett 1992;136: 161-164.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Altier C, Zamponi GW: Targeting Ca2+ channels to treat pain: T-type versus N-type. Trends Pharmacol Sci 2004;25: 465-470.[CrossRef][Medline] [Order article via Infotrieve]
  • Malmberg AB, Yaksh TL: Effect of continuous intrathecal infusion of omega-conopeptides, N-type calcium-channel blockers, on behavior and antinociception in the formalin and hot-plate tests in rats. Pain 1995;60: 83-90.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Xiao WH, Bennett GJ: Synthetic omega-conopeptides applied to the site of nerve injury suppress neuropathic pains in rats. J Pharmacol Exp Ther 1995; 274: 666-672.[Abstract/Free Full Text]
  • White DM, Cousins MJ: Effect of subcutaneous administration of calcium channel blockers on nerve injury-induced hyperalgesia. Brain Res 1998;801: 50-58.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Bowersox SS, Luther R: Pharmacotherapeutic potential of omega-conotoxin MVIIA (SNX-111), an N-type neuronal calcium channel blocker found in the venom of Conus magus. Toxicon 1651;36: 1651-1658.[CrossRef]
  • Terlau H, Olivera BM: Conus venoms: a rich source of novel ion channel-targeted peptides. Physiol Rev 2004;84: 41-68.[Abstract/Free Full Text]
  • Miljanich GP: Ziconotide: neuronal calcium channel blocker for treating severe chronic pain. Curr Med Chem 2004;11: 3029-3040.[ISI][Medline] [Order article via Infotrieve]
  • Tsien RW, Lipscombe D, Madison DV, Bley KR, Fox AP: Multiple types of neuronal calcium channels and their selective modulation. Trends Neurosci 1988;11: 431-438.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Olivera BM, Miljanich GP, Ramachandran J, Adams ME: Calcium channel diversity and neurotransmitter release: the omega-conotoxins and omega-agatoxins. Annu Rev Biochem 1994;63: 823-867.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Dunlap K, Luebke JI, Turner TJ: Exocytotic Ca2+ channels in mammalian central neurons. Trends Neurosci 1995;18: 89-98.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Olivera BM, Cruz LJ, de Santos V, LeCheminant GW, Griffin D, Zeikus R, McIntosh, et al: Neuronal calcium channel antagonists: discrimination between calcium channel subtypes using omega-conotoxin from Conus magus venom. Biochemistry 1987;26: 2086-2090.[CrossRef][Medline] [Order article via Infotrieve]
  • Lewis RJ, Nielsen KJ, Craik DJ, Loughnan ML, Adams DA, Sharpe IA, et al: Novel omega-conotoxins from Conus catus discriminate among neuronal calcium channel subtypes. J Biol Chem 2000;275: 35335-35344.[Abstract/Free Full Text]
  • Favreau P, Gilles N, Lamthanh H, Bournaud R, Shimahara T, Bouet F, et al: A new omega-conotoxin that targets N-type voltage-sensitive calcium channels with unusual specificity. Biochemistry 2001;40: 14567-14575.[CrossRef][Medline] [Order article via Infotrieve]
  • Dos Santos RG, Van Renterghem C, Martin-Moutot N, Mansuelle P, Cordeiro MN, Diniz CR, et al: Phoneutria nigriventer omega-phonetoxin IIA blocks the Cav2 family of calcium channels and interacts with omega-conotoxin-binding sites. J Biol Chem 2002;277: 13856-13862.[Abstract/Free Full Text]
  • Yan L, Adams ME: The spider toxin omega-Aga IIIA defines a high affinity site on neuronal high voltage-activated calcium channels. J Biol Chem 2000;275: 21309-21316.[Abstract/Free Full Text]
  • Hillyard DR, Monje VD, Mintz IM, Bean BP, Nadasdi L, Ramachandran J, et al: A new Conus peptide ligand for mammalian presynaptic Ca2+ channels. Neuron 1992; 9: 69-77.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Wheeler DB, Randall A, Tsien RW: Roles of N-type and Q-type Ca2+ channels in supporting hippocampal synaptic transmission. Science 1994;264: 107-111.[Abstract/Free Full Text]
  • Sasaki T, Kobayashi K, Kohno T, Sato K: Combinatorial synthesis of omega- conotoxin MVIIC analogues and their binding with N- and P/Q-type calcium channels. FEBS Lett 2000;466: 125-129.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Chen JQ, Zhang YQ, Dai J, Luo ZM, Liang SP: Antinociceptive effects of intrathecally administered huwentoxin-I, a selective N-type calcium channel blocker, in the formalin test in conscious rats. Toxicon 2005;45: 15-20.[Medline] [Order article via Infotrieve]
  • Cruz LJ, Olivera BM: Calcium channel antagonists: omega-conotoxin defines a new high affinity site. J Biol Chem 1986;261: 6230-6233.[Abstract/Free Full Text]
  • Jan CR, Titeler M, Schneider AS: Identification of omega-conotoxin binding sites on adrenal medullary membranes: possibility of multiple calcium channels in chromaffin cells. J Neurochem 1990;54: 355-358.[ISI][Medline] [Order article via Infotrieve]
  • Ichida S, Abe J, Zhang YA, Sugihara K, Imoto K, Wada T, et al: Characteristics of the inhibitory effect of calmodulin on specific [125I]omega-conotoxin GVIA binding to crude membranes from chick brain. Neurochem Res 2000;25: 1629-1635.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Stoehr SJ, Dooley DJ: Characteristics of [125I]omega-conotoxin MVIIA binding to rat neocortical membranes. Neurosci Lett 1993;161: 113-116.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Kristipati R, Nadasdi L, Tarczy-Hornoch K, Lau K, Miljanich GP, Ramachandran J, et al: Characterization of the binding of omega-conopeptides to different classes of non-L-type neuronal calcium channels. Mol Cell Neurosci 1994;5: 219-228.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Zhang SP, Wang HY, Lovenberg TW, Codd EE: Functional studies of bradykinin receptors in Chinese hamster ovary cells stably expressing the human B2 bradykinin receptor. Int Immunopharmacol 2001;1: 955-965.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Sato K, Raymond C, Martin-Moutot N, Sasaki T, Omori A, Ohtake A, et al: Binding of chimeric analogs of omega-conotoxin MVIIA and MVIIC to the N- and P/Q-type calcium channels. FEBS Lett 1997;414: 480-484.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Mintz IM, Venema VJ, Swiderek KM, Lee TD, Bean BP, Adams ME: P-type calcium channels blocked by the spider toxin omega-Aga-IVA. Nature 1992;355: 827-829.[CrossRef][Medline] [Order article via Infotrieve]
  • Randall A, Tsien RW: Pharmacological dissection of multiple types of Ca2+ channel currents in rat cerebellar granule neurons. J Neurosci 1995; 15: 2995-3012.[Abstract]
  • Bourinet E, Soong TW, Sutton K, Slaymaker S, Mathews E, Monteil A, et al: Splicing of alpha 1A subunit gene generates phenotypic variants of P- and Q-type calcium channels. Nat Neurosci 1999;2: 407-415.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Zhang J-H, Chung TDY, Oldenburg KR: A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 1999;4: 67-73.[Abstract]
  • Sher E, Pandiella A, Clementi F: Omega-conotoxin binding and effects on calcium channel function in human neuroblastoma and rat pheochromocytoma cell lines. FEBS Lett 1988;235: 178-182.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Schramm M, Towart R: Modulation of calcium channel function by drugs. Life Sci 1985;37: 1843-1860.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Liem LK, Simard JM, Song Y, Tewari K: The patch clamp technique. Neurosurgery 1995;36: 382-392.[ISI][Medline] [Order article via Infotrieve]
  • Jurkat-Rott K, Lehmann-Horn F: The patch clamp technique in ion channel research. Curr Pharm Biotechnol 2004;5: 387-395.[CrossRef][ISI][Medline] [Order article via Infotrieve]
  • Sullivan E, Tucker EM, Dale IL: Measurement of [Ca2+] using the fluoro-metric imaging plate reader (FLIPR). Methods Mol Biol 1999;114: 125-133.[Medline] [Order article via Infotrieve]

Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?



This Article
Right arrow Abstract Freely available
Right arrow Free Full Text (Free PDF) Free
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via ISI Web of Science (1)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Zhang, S.-P.
Right arrow Articles by Codd, E. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zhang, S.-P.
Right arrow Articles by Codd, E. E.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?