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A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo
Abstract
Polypeptides composed entirely of d-amino acids and the achiral amino acid glycine (d-proteins) inherently have in vivo properties that are proposed to be near-optimal for a large molecule therapeutic agent. Specifically, d-proteins are resistant to degradation by proteases and are anticipated to be nonimmunogenic. Furthermore, d-proteins are manufactured chemically and can be engineered to have other desirable properties, such as improved stability, affinity, and pharmacokinetics. Thus, a well-designed d-protein therapeutic would likely have significant advantages over l-protein drugs. Toward the goal of developing d-protein therapeutics, we previously generated RFX001.D, a d-protein antagonist of natural vascular endothelial growth factor A (VEGF-A) that inhibited binding to its receptor. However, RFX001.D is unstable at physiological temperatures (Tm = 33 °C). Here, we describe RFX037.D, a variant of RFX001.D with extreme thermal stability (Tm > 95 °C), high affinity for VEGF-A (Kd = 6 nM), and improved receptor blocking. Comparison of the two enantiomeric forms of RFX037 revealed that the d-protein is more stable in mouse, monkey, and human plasma and has a longer half-life in vivo in mice. Significantly, RFX037.D was nonimmunogenic in mice, whereas the l-enantiomer generated a strong immune response. These results confirm the potential utility of synthetic d-proteins as alternativesmore »
- Authors:
-
- Banting and Best Department of Medical Research and Department of Molecular Genetics, the Donnelly Centre, University of Toronto, Toronto, Ontario, Canada M5S 3E1
- Department of Chemistry, Department of Biochemistry and Molecular Biology, Institute for Biophysical Dynamics, University of Chicago, Chicago, Illinois 60637, United States
- Pharmacokinetics &, Drug Metabolism, Amgen, Inc., Thousand Oaks, California 91320, United States
- Therapeutic Discovery, Amgen, Inc., Thousand Oaks, California 91320, United States
- Antibody Solutions, Sunnyvale, California 94089, United States
- Reflexion Pharmaceuticals, Inc., San Francisco, California 94104, United States
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1236725
- Alternate Identifier(s):
- OSTI ID: 1249238
- Grant/Contract Number:
- AC02-06CH11357
- Resource Type:
- Journal Article: Published Article
- Journal Name:
- ACS Chemical Biology
- Additional Journal Information:
- Journal Name: ACS Chemical Biology Journal Volume: 11 Journal Issue: 4; Journal ID: ISSN 1554-8929
- Publisher:
- American Chemical Society (ACS)
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; immunology; peptides and proteins; rodent models; plasma; stability
Citation Formats
Uppalapati, Maruti, Lee, Dong Jun, Mandal, Kalyaneswar, Li, Hongyan, Miranda, Les P., Lowitz, Joshua, Kenney, John, Adams, Jarrett J., Ault-Riché, Dana, Kent, Stephen B. H., and Sidhu, Sachdev S. A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo. United States: N. p., 2016.
Web. doi:10.1021/acschembio.5b01006.
Uppalapati, Maruti, Lee, Dong Jun, Mandal, Kalyaneswar, Li, Hongyan, Miranda, Les P., Lowitz, Joshua, Kenney, John, Adams, Jarrett J., Ault-Riché, Dana, Kent, Stephen B. H., & Sidhu, Sachdev S. A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo. United States. https://doi.org/10.1021/acschembio.5b01006
Uppalapati, Maruti, Lee, Dong Jun, Mandal, Kalyaneswar, Li, Hongyan, Miranda, Les P., Lowitz, Joshua, Kenney, John, Adams, Jarrett J., Ault-Riché, Dana, Kent, Stephen B. H., and Sidhu, Sachdev S. 2016.
"A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo". United States. https://doi.org/10.1021/acschembio.5b01006.
@article{osti_1236725,
title = {A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo},
author = {Uppalapati, Maruti and Lee, Dong Jun and Mandal, Kalyaneswar and Li, Hongyan and Miranda, Les P. and Lowitz, Joshua and Kenney, John and Adams, Jarrett J. and Ault-Riché, Dana and Kent, Stephen B. H. and Sidhu, Sachdev S.},
abstractNote = {Polypeptides composed entirely of d-amino acids and the achiral amino acid glycine (d-proteins) inherently have in vivo properties that are proposed to be near-optimal for a large molecule therapeutic agent. Specifically, d-proteins are resistant to degradation by proteases and are anticipated to be nonimmunogenic. Furthermore, d-proteins are manufactured chemically and can be engineered to have other desirable properties, such as improved stability, affinity, and pharmacokinetics. Thus, a well-designed d-protein therapeutic would likely have significant advantages over l-protein drugs. Toward the goal of developing d-protein therapeutics, we previously generated RFX001.D, a d-protein antagonist of natural vascular endothelial growth factor A (VEGF-A) that inhibited binding to its receptor. However, RFX001.D is unstable at physiological temperatures (Tm = 33 °C). Here, we describe RFX037.D, a variant of RFX001.D with extreme thermal stability (Tm > 95 °C), high affinity for VEGF-A (Kd = 6 nM), and improved receptor blocking. Comparison of the two enantiomeric forms of RFX037 revealed that the d-protein is more stable in mouse, monkey, and human plasma and has a longer half-life in vivo in mice. Significantly, RFX037.D was nonimmunogenic in mice, whereas the l-enantiomer generated a strong immune response. These results confirm the potential utility of synthetic d-proteins as alternatives to therapeutic antibodies.},
doi = {10.1021/acschembio.5b01006},
url = {https://www.osti.gov/biblio/1236725},
journal = {ACS Chemical Biology},
issn = {1554-8929},
number = 4,
volume = 11,
place = {United States},
year = {Wed Feb 03 00:00:00 EST 2016},
month = {Wed Feb 03 00:00:00 EST 2016}
}
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