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Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319

Fibroblast growth factor 21 is a novel hormonal regulator with the potential to treat a broad variety of metabolic abnormalities, such as type 2 diabetes, obesity, hepatic steatosis, and cardiovascular disease. Human recombinant wild type FGF21 (FGF21) has been shown to ameliorate metabolic disorder...

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Autores principales: Kharitonenkov, Alexei, Beals, John M., Micanovic, Radmila, Strifler, Beth A., Rathnachalam, Radhakrishnan, Wroblewski, Victor J., Li, Shun, Koester, Anja, Ford, Amy M., Coskun, Tamer, Dunbar, James D., Cheng, Christine C., Frye, Christopher C., Bumol, Thomas F., Moller, David E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3594191/
https://www.ncbi.nlm.nih.gov/pubmed/23536797
http://dx.doi.org/10.1371/journal.pone.0058575
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author Kharitonenkov, Alexei
Beals, John M.
Micanovic, Radmila
Strifler, Beth A.
Rathnachalam, Radhakrishnan
Wroblewski, Victor J.
Li, Shun
Koester, Anja
Ford, Amy M.
Coskun, Tamer
Dunbar, James D.
Cheng, Christine C.
Frye, Christopher C.
Bumol, Thomas F.
Moller, David E.
author_facet Kharitonenkov, Alexei
Beals, John M.
Micanovic, Radmila
Strifler, Beth A.
Rathnachalam, Radhakrishnan
Wroblewski, Victor J.
Li, Shun
Koester, Anja
Ford, Amy M.
Coskun, Tamer
Dunbar, James D.
Cheng, Christine C.
Frye, Christopher C.
Bumol, Thomas F.
Moller, David E.
author_sort Kharitonenkov, Alexei
collection PubMed
description Fibroblast growth factor 21 is a novel hormonal regulator with the potential to treat a broad variety of metabolic abnormalities, such as type 2 diabetes, obesity, hepatic steatosis, and cardiovascular disease. Human recombinant wild type FGF21 (FGF21) has been shown to ameliorate metabolic disorders in rodents and non-human primates. However, development of FGF21 as a drug is challenging and requires re-engineering of its amino acid sequence to improve protein expression and formulation stability. Here we report the design and characterization of a novel FGF21 variant, LY2405319. To enable the development of a potential drug product with a once-daily dosing profile, in a preserved, multi-use formulation, an additional disulfide bond was introduced in FGF21 through Leu118Cys and Ala134Cys mutations. FGF21 was further optimized by deleting the four N-terminal amino acids, His-Pro-Ile-Pro (HPIP), which was subject to proteolytic cleavage. In addition, to eliminate an O-linked glycosylation site in yeast a Ser167Ala mutation was introduced, thus allowing large-scale, homogenous protein production in Pichia pastoris. Altogether re-engineering of FGF21 led to significant improvements in its biopharmaceutical properties. The impact of these changes was assessed in a panel of in vitro and in vivo assays, which confirmed that biological properties of LY2405319 were essentially identical to FGF21. Specifically, subcutaneous administration of LY2405319 in ob/ob and diet-induced obese (DIO) mice over 7–14 days resulted in a 25–50% lowering of plasma glucose coupled with a 10–30% reduction in body weight. Thus, LY2405319 exhibited all the biopharmaceutical and biological properties required for initiation of a clinical program designed to test the hypothesis that administration of exogenous FGF21 would result in effects on disease-related metabolic parameters in humans.
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spelling pubmed-35941912013-03-27 Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319 Kharitonenkov, Alexei Beals, John M. Micanovic, Radmila Strifler, Beth A. Rathnachalam, Radhakrishnan Wroblewski, Victor J. Li, Shun Koester, Anja Ford, Amy M. Coskun, Tamer Dunbar, James D. Cheng, Christine C. Frye, Christopher C. Bumol, Thomas F. Moller, David E. PLoS One Research Article Fibroblast growth factor 21 is a novel hormonal regulator with the potential to treat a broad variety of metabolic abnormalities, such as type 2 diabetes, obesity, hepatic steatosis, and cardiovascular disease. Human recombinant wild type FGF21 (FGF21) has been shown to ameliorate metabolic disorders in rodents and non-human primates. However, development of FGF21 as a drug is challenging and requires re-engineering of its amino acid sequence to improve protein expression and formulation stability. Here we report the design and characterization of a novel FGF21 variant, LY2405319. To enable the development of a potential drug product with a once-daily dosing profile, in a preserved, multi-use formulation, an additional disulfide bond was introduced in FGF21 through Leu118Cys and Ala134Cys mutations. FGF21 was further optimized by deleting the four N-terminal amino acids, His-Pro-Ile-Pro (HPIP), which was subject to proteolytic cleavage. In addition, to eliminate an O-linked glycosylation site in yeast a Ser167Ala mutation was introduced, thus allowing large-scale, homogenous protein production in Pichia pastoris. Altogether re-engineering of FGF21 led to significant improvements in its biopharmaceutical properties. The impact of these changes was assessed in a panel of in vitro and in vivo assays, which confirmed that biological properties of LY2405319 were essentially identical to FGF21. Specifically, subcutaneous administration of LY2405319 in ob/ob and diet-induced obese (DIO) mice over 7–14 days resulted in a 25–50% lowering of plasma glucose coupled with a 10–30% reduction in body weight. Thus, LY2405319 exhibited all the biopharmaceutical and biological properties required for initiation of a clinical program designed to test the hypothesis that administration of exogenous FGF21 would result in effects on disease-related metabolic parameters in humans. Public Library of Science 2013-03-11 /pmc/articles/PMC3594191/ /pubmed/23536797 http://dx.doi.org/10.1371/journal.pone.0058575 Text en © 2013 Kharitonenkov et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Kharitonenkov, Alexei
Beals, John M.
Micanovic, Radmila
Strifler, Beth A.
Rathnachalam, Radhakrishnan
Wroblewski, Victor J.
Li, Shun
Koester, Anja
Ford, Amy M.
Coskun, Tamer
Dunbar, James D.
Cheng, Christine C.
Frye, Christopher C.
Bumol, Thomas F.
Moller, David E.
Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319
title Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319
title_full Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319
title_fullStr Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319
title_full_unstemmed Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319
title_short Rational Design of a Fibroblast Growth Factor 21-Based Clinical Candidate, LY2405319
title_sort rational design of a fibroblast growth factor 21-based clinical candidate, ly2405319
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3594191/
https://www.ncbi.nlm.nih.gov/pubmed/23536797
http://dx.doi.org/10.1371/journal.pone.0058575
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