A Novel Fc-FGF21 With Improved Resistance to Proteolysis, Increased Affinity Toward β-Klotho, and Enhanced Efficacy in Mice and Cynomolgus Monkeys.
Fibroblast growth factor (FGF) 21 is a natural hormone that modulates glucose, lipid, and energy metabolism. Previously, we engineered an Fc fusion FGF21 variant with two mutations, Fc-FGF21(RG), to extend the half-life and reduce aggregation and in vivo degradation of FGF21. We now describe a new variant developed to reduce the extreme C-terminal degradation and improve the binding affinity to β-Klotho. We demonstrate, by introducing one additional mutation located at the C terminus of FGF21 (A180E), that the new molecule, Fc-FGF21(RGE), has gained many improved attributes. Compared with Fc-FGF21(RG), Fc-FGF21(RGE) has similar in vitro potency, preserves β-Klotho dependency, and maintains FGF receptor selectivity and cross-species reactivity. In vivo, Fc-FGF21(RGE) showed reduced susceptibility to extreme C-terminal degradation and increased plasma levels of the bioactive intact molecule. The circulating half-life of intact Fc-FGF21(RGE) increased twofold compared with that of Fc-FGF21(RG) in mice and cynomolgus monkeys. Additionally, Fc-FGF21(RGE) exhibited threefold to fivefold enhanced binding affinity to coreceptor β-Klotho across mouse, cynomolgus monkey, and human species. In obese and diabetic mouse and cynomolgus monkey models, Fc-FGF21(RGE) demonstrated greater efficacies to Fc-FGF21(RG), resulting in larger and more sustained improvements in multiple metabolic parameters. No increased immunogenicity was observed with Fc-FGF21(RGE). The superior biophysical, pharmacokinetic, and pharmacodynamic properties, as well as the positive metabolic effects across species, suggest that further clinical development of Fc-FGF21(RGE) as a metabolic therapy for diabetic and/or obese patients may be warranted.
Department of Cardiometabolic Disorders, Amgen Inc., Thousand Oaks, California 91320.
Shanaka
S
Stanislaus
Department of Biologics, Amgen Inc., Thousand Oaks, California 91320.
Randy
R
Hecht
Department of Cardiometabolic Disorders, Amgen Inc., Thousand Oaks, California 91320.
Junming
J
Yie
Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., Thousand Oaks, California 91320.
Todd
T
Hager
Department of Pharmacokinetics and Drug Metabolism, Amgen Inc., Thousand Oaks, California 91320.
Michael
M
Hall
Department of Biologics, Amgen Inc., Thousand Oaks, California 91320.
Chris
C
Spahr
Department of Biologics, Amgen Inc., Thousand Oaks, California 91320.
Wei
W
Wang
Department of Cardiometabolic Disorders, Amgen Inc., Thousand Oaks, California 91320.|Amgen Inc., South San Francisco, California 94080.
Jennifer
J
Weiszmann
Department of Cardiometabolic Disorders, Amgen Inc., Thousand Oaks, California 91320.|Amgen Inc., South San Francisco, California 94080.
Yang
Y
Li
Department of Cardiometabolic Disorders, Amgen Inc., Thousand Oaks, California 91320.
Liying
L
Deng
Department of Biologics, Amgen Inc., Thousand Oaks, California 91320.
Dwight
D
Winters
Department of Biologics, Amgen Inc., Thousand Oaks, California 91320.
Stephen
S
Smith
Department of Medical Science, Amgen Inc., Thousand Oaks, California 91320.
Lei
L
Zhou
Department of Biologics, Amgen Inc., Thousand Oaks, California 91320.
Yuesheng
Y
Li
Department of Cardiometabolic Disorders, Amgen Inc., Thousand Oaks, California 91320.
Murielle M
MM
Véniant
Department of Cardiometabolic Disorders, Amgen Inc., Thousand Oaks, California 91320.
Jing
J
Xu
D041721:3T3-L1 Cells; D000818:Animals; D019440:Anti-Obesity Agents / Q000138:chemical synthesis / Q000378:metabolism / Q000627:therapeutic use*; D003921:Diabetes Mellitus, Experimental / Q000188:drug therapy* / Q000378:metabolism; D004195:Disease Models, Animal; D004355:Drug Stability; D005346:Fibroblast Growth Factors / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D057809:HEK293 Cells; D006207:Half-Life; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D000090265:Klotho Proteins; D008252:Macaca fascicularis; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D009765:Obesity / Q000188:drug therapy* / Q000378:metabolism; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D016896:Treatment Outcome
D019440:Anti-Obesity Agents; D007141:Immunoglobulin Fc Fragments; C538995:KLB protein, human; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C414620:fibroblast growth factor 21; D005346:Fibroblast Growth Factors; D000090265:Klotho Proteins