TY - JOUR
T1 - A Reactive Oxygen Species-Scavenging Stealth' Polymer, Poly(thioglycidyl glycerol), Outperforms Poly(ethylene glycol) in Protein Conjugates and Nanocarriers and Enhances Protein Stability to Environmental and Biological Stressors
AU - D'Arcy, Richard
AU - El Mohtadi, Farah
AU - Francini, Nora
AU - Dejulius, Carlisle R.
AU - Back, Hyunmoon
AU - Gennari, Arianna
AU - Geven, Mike
AU - Lopez-Cavestany, Maria
AU - Turhan, Zulfiye Yesim
AU - Yu, Fang
AU - Lee, Jong Bong
AU - King, Michael R.
AU - Kagan, Leonid
AU - Duvall, Craig L.
AU - Tirelli, Nicola
N1 - Funding Information:
Financial support is gratefully acknowledged (a) from the Engineering and Physical Sciences Research Council (EPSRC) for a Doctoral Prize fellowship to R.d.A. during his time at the University of Manchester; (b) from the King Saud University (Riyadh, Kingdom of Saudi Arabia), Office of the Vice Rector for Graduate Studies & Scientific Research, for the collaborative grant “Nanotechnology for drug delivery” that funded F.E.M.; (c) from the European Union’s Horizon 2020 research and innovation programme, grant agreement No. 824074 (GrowBot; a FET Pro-Active project) that funded M.G.; (d) from the Republic of Turkey’s Ministry of National Education for the PhD studentship to Z.Y.T.; and (e) from the Italian Association for Cancer Research (AIRC), Investigator grant No 24628, for general financial support.
Publisher Copyright:
©
PY - 2022/11/23
Y1 - 2022/11/23
N2 - This study addresses well-known shortcomings of poly(ethylene glycol) (PEG)-based conjugates. PEGylation is by far the most common method employed to overcome immunogenicity and suboptimal pharmacokinetics of, for example, therapeutic proteins but has significant drawbacks. First, PEG offers no protection from denaturation during lyophilization, storage, or oxidation (e.g., by biological oxidants, reactive oxygen species); second, PEG's inherent immunogenicity, leading to hypersensitivity and accelerated blood clearance (ABC), is a growing concern. We have here developed an active-stealth' polymer, poly(thioglycidyl glycerol)(PTGG), which in human plasma is less immunogenic than PEG (35% less complement activation) and features a reactive oxygen species-scavenging and anti-inflammatory action (50% less TNF-α in LPS-stimulated macrophages at only 0.1 mg/mL). PTGG was conjugated to proteins via a one-pot process; molar mass- and grafting density-matched PTGG-lysozyme conjugates were superior to their PEG analogues in terms of enzyme activity and stability against freeze-drying or oxidation; the latter is due to sacrificial oxidation of methionine-mimetic PTGG chains. Both in mice and rats, PTGG-ovalbumin displayed circulation half-lives up to twice as long as PEG-ovalbumin, but most importantly and differently from PEG without any associated ABC effect seen either in the time dependency of blood concentration, in the liver/splenic accumulation, or in antipolymer IgM/IgG titers. Furthermore, similar pharmacokinetic results were obtained with PTGGylated/PEGylated liposomal nanocarriers. PTGG's active-stealth' character therefore makes it a highly promising alternative to PEG for conjugation to biologics or nanocarriers.
AB - This study addresses well-known shortcomings of poly(ethylene glycol) (PEG)-based conjugates. PEGylation is by far the most common method employed to overcome immunogenicity and suboptimal pharmacokinetics of, for example, therapeutic proteins but has significant drawbacks. First, PEG offers no protection from denaturation during lyophilization, storage, or oxidation (e.g., by biological oxidants, reactive oxygen species); second, PEG's inherent immunogenicity, leading to hypersensitivity and accelerated blood clearance (ABC), is a growing concern. We have here developed an active-stealth' polymer, poly(thioglycidyl glycerol)(PTGG), which in human plasma is less immunogenic than PEG (35% less complement activation) and features a reactive oxygen species-scavenging and anti-inflammatory action (50% less TNF-α in LPS-stimulated macrophages at only 0.1 mg/mL). PTGG was conjugated to proteins via a one-pot process; molar mass- and grafting density-matched PTGG-lysozyme conjugates were superior to their PEG analogues in terms of enzyme activity and stability against freeze-drying or oxidation; the latter is due to sacrificial oxidation of methionine-mimetic PTGG chains. Both in mice and rats, PTGG-ovalbumin displayed circulation half-lives up to twice as long as PEG-ovalbumin, but most importantly and differently from PEG without any associated ABC effect seen either in the time dependency of blood concentration, in the liver/splenic accumulation, or in antipolymer IgM/IgG titers. Furthermore, similar pharmacokinetic results were obtained with PTGGylated/PEGylated liposomal nanocarriers. PTGG's active-stealth' character therefore makes it a highly promising alternative to PEG for conjugation to biologics or nanocarriers.
KW - Animals
KW - Glycerol
KW - Humans
KW - Mice
KW - Ovalbumin
KW - Polyethylene Glycols/metabolism
KW - Polymers/pharmacology
KW - Protein Stability
KW - Rats
KW - Reactive Oxygen Species
U2 - 10.1021/jacs.2c09232
DO - 10.1021/jacs.2c09232
M3 - Article
C2 - 36367536
AN - SCOPUS:85141956191
SN - 0002-7863
VL - 144
SP - 21304
EP - 21317
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 46
ER -