title: Discovery and Characterization of a Potent Interleukin-6 Binding Peptide with Neutralizing Activity In Vivo.
abstract: Interleukin-6 (IL-6) is an important member of the cytokine superfamily, exerting pleiotropic actions on many physiological processes. Over-production of IL-6 is a hallmark of immune-mediated inflammatory diseases such as Castleman's Disease (CD) and rheumatoid arthritis (RA). Antagonism of the interleukin IL-6/IL-6 receptor (IL-6R)/gp130 signaling complex continues to show promise as a therapeutic target. Monoclonal antibodies (mAbs) directed against components of this complex have been approved as therapeutics for both CD and RA. To potentially provide an additional modality to antagonize IL-6 induced pathophysiology, a peptide-based antagonist approach was undertaken. Using a combination of molecular design, phage-display, and medicinal chemistry, disulfide-rich peptides (DRPs) directed against IL-6 were developed with low nanomolar potency in inhibiting IL-6-induced pSTAT3 in U937 monocytic cells. Targeted PEGylation of IL-6 binding peptides resulted in molecules that retained their potency against IL-6 and had a prolongation of their pharmacokinetic (PK) profiles in rodents and monkeys. One such peptide, PN-2921, contained a 40 kDa polyethylene glycol (PEG) moiety and inhibited IL-6-induced pSTAT3 in U937 cells with sub-nM potency and possessed 23, 36, and 59 h PK half-life values in mice, rats, and cynomolgus monkeys, respectively. Parenteral administration of PN-2921 to mice and cynomolgus monkeys potently inhibited IL-6-induced biomarker responses, with significant reductions in the acute inflammatory phase proteins, serum amyloid A (SAA) and C-reactive protein (CRP). This potent, PEGylated IL-6 binding peptide offers a new approach to antagonize IL-6-induced signaling and associated pathophysiology.
authors.affiliation: Discovery Biology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Sheila
authors.initials: S
authors.lastname: Ranganath
authors.affiliation: Chemistry, Protagonist Therapeutics, Milpitas, CA, United States of America.
authors.forename: Ashok
authors.initials: A
authors.lastname: Bhandari
authors.affiliation: Discovery Pharmacology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Nicole
authors.initials: N
authors.lastname: Avitahl-Curtis
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.
authors.forename: Jaimee
authors.initials: J
authors.lastname: McMahon
authors.affiliation: DMPK, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Derek
authors.initials: D
authors.lastname: Wachtel
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.
authors.forename: Jenny
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Pharmaceutical Development, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Christopher
authors.initials: C
authors.lastname: Leitheiser
authors.affiliation: Discovery Biology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Sylvie G
authors.initials: SG
authors.lastname: Bernier
authors.affiliation: Discovery Biology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Guang
authors.initials: G
authors.lastname: Liu
authors.affiliation: Protagonist Pty Ltd, Therapeutics Pty Ltd, Brisbane, Australia.
authors.forename: Tran T
authors.initials: TT
authors.lastname: Tran
authors.affiliation: Chemistry, Protagonist Therapeutics, Milpitas, CA, United States of America.
authors.forename: Herodion
authors.initials: H
authors.lastname: Celino
authors.affiliation: Discovery Pharmacology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Jenny
authors.initials: J
authors.lastname: Tobin
authors.affiliation: Chemistry, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Joon
authors.initials: J
authors.lastname: Jung
authors.affiliation: Pharmaceutical Development, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.
authors.forename: Katie E
authors.initials: KE
authors.lastname: Glen
authors.affiliation: Discovery Pharmacology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Chris
authors.initials: C
authors.lastname: Graul
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.
authors.forename: Aliesha
authors.initials: A
authors.lastname: Griffin
authors.affiliation: Pharmaceutical Development, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Wayne C
authors.initials: WC
authors.lastname: Schairer
authors.affiliation: DMPK, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Carolyn
authors.initials: C
authors.lastname: Higgins
authors.affiliation: Discovery Toxicology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Tammi L
authors.initials: TL
authors.lastname: Reza
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.
authors.forename: Eva
authors.initials: E
authors.lastname: Mowe
authors.affiliation: Discovery Pharmacology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Sam
authors.initials: S
authors.lastname: Rivers
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.
authors.forename: Sonya
authors.initials: S
authors.lastname: Scott
authors.affiliation: Discovery Biology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Alex
authors.initials: A
authors.lastname: Monreal
authors.affiliation: Discovery Pharmacology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Courtney
authors.initials: C
authors.lastname: Shea
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.|Protagonist Pty Ltd, Therapeutics Pty Ltd, Brisbane, Australia.
authors.forename: Greg
authors.initials: G
authors.lastname: Bourne
authors.affiliation: Pharmaceutical Development, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Casey
authors.initials: C
authors.lastname: Coons
authors.affiliation: Discovery Toxicology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Adaline
authors.initials: A
authors.lastname: Smith
authors.affiliation: Discovery Pharmacology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Kim
authors.initials: K
authors.lastname: Tang
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.
authors.forename: Ramya A
authors.initials: RA
authors.lastname: Mandyam
authors.affiliation: Discovery Pharmacology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Jaime
authors.initials: J
authors.lastname: Masferrer
authors.affiliation: Biology, Protagonist Therapeutics, Milpitas, CA, United States of America.
authors.forename: David
authors.initials: D
authors.lastname: Liu
authors.affiliation: Chemistry, Protagonist Therapeutics, Milpitas, CA, United States of America.
authors.forename: Dinesh V
authors.initials: DV
authors.lastname: Patel
authors.affiliation: Pharmaceutical Development, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Angelika
authors.initials: A
authors.lastname: Fretzen
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.|Protagonist Pty Ltd, Therapeutics Pty Ltd, Brisbane, Australia.
authors.forename: Craig A
authors.initials: CA
authors.lastname: Murphy
authors.affiliation: Discovery Biology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: G Todd
authors.initials: GT
authors.lastname: Milne
authors.affiliation: Institute for Molecular Biosciences, The University of Queensland, Brisbane, Australia.|Protagonist Pty Ltd, Therapeutics Pty Ltd, Brisbane, Australia.
authors.forename: Mark L
authors.initials: ML
authors.lastname: Smythe
authors.affiliation: Discovery Biology, Ironwood Pharmaceuticals, Cambridge, MA, United States of America.
authors.forename: Kenneth E
authors.initials: KE
authors.lastname: Carlson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015195:Drug Design; D006207:Half-Life; D006801:Humans; D006825:Hybridomas; D015850:Interleukin-6 / Q000737:chemistry / Q000378:metabolism; D008252:Macaca fascicularis; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008813:Mice, Inbred ICR; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011487:Protein Conformation; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D019947:Receptors, Interleukin-6 / Q000737:chemistry; D011994:Recombinant Proteins / Q000494:pharmacology; D050796:STAT3 Transcription Factor / Q000378:metabolism; D013329:Structure-Activity Relationship; D020298:U937 Cells; D000093242:Interleukin-6 Inhibitors
chemical_list: D015850:Interleukin-6; D019151:Peptide Library; D010455:Peptides; D019947:Receptors, Interleukin-6; D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor; D000093242:Interleukin-6 Inhibitors; C508600:IL6 protein, human; C000608555:PN-2921; C494086:STAT3 protein, human