title: Linker length matters, fynomer-Fc fusion with an optimized linker displaying picomolar IL-17A inhibition potency.
abstract: Fynomers are small binding proteins derived from the human Fyn SH3 domain. Using phage display technology, Fynomers were generated inhibiting the activity of the proinflammatory cytokine interleukin-17A (IL-17A). One specific Fynomer called 2C1 inhibited human IL-17A in vitro with an IC50 value of 2.2 nm. Interestingly, when 2C1 was genetically fused to the Fc part of a human antibody via four different amino acid linkers to yield bivalent IL-17A binding proteins (each linker differed in length), the 2C1-Fc fusion protein with the longest linker displayed the most potent inhibitory activity. It blocked homodimeric IL-17A with an IC50 value of 21 pm, which corresponds to a hundredfold improved IC50 value as compared to the value obtained with monovalent Fynomer 2C1. In contrast, the 2C1-Fc fusion with the shortest linker showed only an ∼8-fold improved IC50 value of 260 pm. Furthermore, in a mouse model of acute inflammation, we have shown that the most potent 2C1-Fc fusion protein is able to efficiently inhibit IL-17A in vivo. With their suitable biophysical properties, Fynomer-Fc fusion proteins represent new drug candidates for the treatment of IL-17A mediated inflammatory conditions such as psoriasis, psoriatic arthritis, or rheumatoid arthritis.
authors.affiliation: From Covagen AG, Wagistrasse 25, CH-8952 Schlieren, Switzerland.
authors.forename: Michela
authors.initials: M
authors.lastname: Silacci
authors.affiliation: From Covagen AG, Wagistrasse 25, CH-8952 Schlieren, Switzerland.
authors.forename: Nadja
authors.initials: N
authors.lastname: Baenziger-Tobler
authors.affiliation: From Covagen AG, Wagistrasse 25, CH-8952 Schlieren, Switzerland.
authors.forename: Wibke
authors.initials: W
authors.lastname: Lembke
authors.affiliation: From Covagen AG, Wagistrasse 25, CH-8952 Schlieren, Switzerland.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Zha
authors.affiliation: From Covagen AG, Wagistrasse 25, CH-8952 Schlieren, Switzerland.
authors.forename: Sarah
authors.initials: S
authors.lastname: Batey
authors.affiliation: From Covagen AG, Wagistrasse 25, CH-8952 Schlieren, Switzerland.
authors.forename: Julian
authors.initials: J
authors.lastname: Bertschinger
authors.affiliation: From Covagen AG, Wagistrasse 25, CH-8952 Schlieren, Switzerland dragan.grabulovski@covagen.com.
authors.forename: Dragan
authors.initials: D
authors.lastname: Grabulovski
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D020128:Inhibitory Concentration 50; D020381:Interleukin-17 / Q000037:antagonists & inhibitors*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D051076:Proto-Oncogene Proteins c-fyn / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000493:pharmacokinetics / Q000494:pharmacology*; D018909:src Homology Domains
chemical_list: D007141:Immunoglobulin Fc Fragments; D020381:Interleukin-17; D011993:Recombinant Fusion Proteins; D051076:Proto-Oncogene Proteins c-fyn
keywords: Drug Discovery; Fynomer; Inflammation; Interleukin; Protein Engineering; SH3 Domains