title: Engineering Highly Potent and Selective Microproteins against Nav1.7 Sodium Channel for Treatment of Pain.
abstract: The prominent role of voltage-gated sodium channel 1.7 (Nav1.7) in nociception was revealed by remarkable human clinical and genetic evidence. Development of potent and subtype-selective inhibitors of this ion channel is crucial for obtaining therapeutically useful analgesic compounds. Microproteins isolated from animal venoms have been identified as promising therapeutic leads for ion channels, because they naturally evolved to be potent ion channel blockers. Here, we report the engineering of highly potent and selective inhibitors of the Nav1.7 channel based on tarantula ceratotoxin-1 (CcoTx1). We utilized a combination of directed evolution, saturation mutagenesis, chemical modification, and rational drug design to obtain higher potency and selectivity to the Nav1.7 channel. The resulting microproteins are highly potent (IC50 to Nav1.7 of 2.5 nm) and selective. We achieved 80- and 20-fold selectivity over the closely related Nav1.2 and Nav1.6 channels, respectively, and the IC50 on skeletal (Nav1.4) and cardiac (Nav1.5) sodium channels is above 3000 nm The lead molecules have the potential for future clinical development as novel therapeutics in the treatment of pain.
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Anatoly
authors.initials: A
authors.lastname: Shcherbatko
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rossi
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Davide
authors.initials: D
authors.lastname: Foletti
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Guoyun
authors.initials: G
authors.lastname: Zhu
authors.affiliation: Amunix, Mountain View, California 94043.
authors.forename: Oren
authors.initials: O
authors.lastname: Bogin
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Galindo Casas
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Mathias
authors.initials: M
authors.lastname: Rickert
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Adela
authors.initials: A
authors.lastname: Hasa-Moreno
authors.affiliation: Amunix, Mountain View, California 94043.
authors.forename: Victor
authors.initials: V
authors.lastname: Bartsevich
authors.affiliation: Amunix, Mountain View, California 94043.
authors.forename: Andreas
authors.initials: A
authors.lastname: Crameri
authors.affiliation: Sutro Biopharma, South San Francisco, California 94080.
authors.forename: Alexander R
authors.initials: AR
authors.lastname: Steiner
authors.affiliation: Sutro Biopharma, South San Francisco, California 94080.
authors.forename: Robert
authors.initials: R
authors.lastname: Henningsen
authors.affiliation: Sutro Biopharma, South San Francisco, California 94080.
authors.forename: Avinash
authors.initials: A
authors.lastname: Gill
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Jaume
authors.initials: J
authors.lastname: Pons
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: David L
authors.initials: DL
authors.lastname: Shelton
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080.
authors.forename: Arvind
authors.initials: A
authors.lastname: Rajpal
authors.affiliation: Rinat Laboratories, Pfizer Inc., South San Francisco, California 94080,. Electronic address: pavel.strop@bms.com.
authors.forename: Pavel
authors.initials: P
authors.lastname: Strop
mesh_terms: D057809:HEK293 Cells; D006801:Humans; D062556:NAV1.7 Voltage-Gated Sodium Channel / Q000737:chemistry* / Q000187:drug effects; D059408:Pain Management / Q000379:methods*; D018408:Patch-Clamp Techniques; D010802:Phylogeny; D015202:Protein Engineering; D013111:Spider Venoms / Q000737:chemistry; D061567:Voltage-Gated Sodium Channel Blockers / Q000494:pharmacology*
chemical_list: D062556:NAV1.7 Voltage-Gated Sodium Channel; D013111:Spider Venoms; D061567:Voltage-Gated Sodium Channel Blockers
keywords: Nav1.7; ceratotoxin; directed evolution; engineering; pain; sodium channel; structure-function; toxin