title: Single Residue Substitutions That Confer Voltage-Gated Sodium Ion Channel Subtype Selectivity in the NaV1.7 Inhibitory Peptide GpTx-1.
abstract: There is interest in the identification and optimization of new molecular entities selectively targeting ion channels of therapeutic relevance. Peptide toxins represent a rich source of pharmacology for ion channels, and we recently reported GpTx-1 analogs that inhibit NaV1.7, a voltage-gated sodium ion channel that is a compelling target for improved treatment of pain. Here we utilize multi-attribute positional scan (MAPS) analoging, combining high-throughput synthesis and electrophysiology, to interrogate the interaction of GpTx-1 with NaV1.7 and related NaV subtypes. After one round of MAPS analoging, we found novel substitutions at multiple residue positions not previously identified, specifically glutamic acid at positions 10 or 11 or lysine at position 18, that produce peptides with single digit nanomolar potency on NaV1.7 and 500-fold selectivity against off-target sodium channels. Docking studies with a NaV1.7 homology model and peptide NMR structure generated a model consistent with the key potency and selectivity modifications mapped in this work.
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Justin K
authors.initials: JK
authors.lastname: Murray
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Jason
authors.initials: J
authors.lastname: Long
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Anruo
authors.initials: A
authors.lastname: Zou
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Joseph
authors.initials: J
authors.lastname: Ligutti
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Kristin L
authors.initials: KL
authors.lastname: Andrews
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Leszek
authors.initials: L
authors.lastname: Poppe
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Kaustav
authors.initials: K
authors.lastname: Biswas
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Bryan D
authors.initials: BD
authors.lastname: Moyer
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Stefan I
authors.initials: SI
authors.lastname: McDonough
authors.affiliation: Therapeutic Discovery and ‡Neuroscience, Amgen Inc. , One Amgen Center Drive, Thousand Oaks, California 91320, United States.|Therapeutic Discovery and ∥Neuroscience, Amgen Inc. , 360 Binney Street, Cambridge, Massachusetts 02142, United States.
authors.forename: Les P
authors.initials: LP
authors.lastname: Miranda
mesh_terms: D000595:Amino Acid Sequence; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D062556:NAV1.7 Voltage-Gated Sodium Channel / Q000187:drug effects*; D010455:Peptides / Q000494:pharmacology*; D026941:Sodium Channel Blockers / Q000737:chemistry* / Q000494:pharmacology*; D013111:Spider Venoms / Q000494:pharmacology*; D013379:Substrate Specificity
chemical_list: C000609915:GpTx-1 protein, Grammostola porteri; D062556:NAV1.7 Voltage-Gated Sodium Channel; D010455:Peptides; D026941:Sodium Channel Blockers; D013111:Spider Venoms