Impact of polymer geometry on the interactions of protein-PEG conjugates

Zarzar, Jonathan; Shatz, Whitney; Peer, Nikhil; Taing, Rosalynn; McGarry, Brian E.; Greene, Daniel G.; Zarraga, Isidro E.

doi:10.1016/j.bpc.2017.10.003

Cited by 15 publications

(22 citation statements)

References 29 publications

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“…We have developed a novel, multimeric Fab + PEG soluble system, administrable by IVT injection, which has yielded an increase in vitreous half-life and favorable ocular tolerability in NHP in vivo studies. By demonstrating this relationship in a primate species, these results corroborate and broaden a previously reported hypothesis that diffusion is a significant driver of ocular elimination [26], now within the context of a preclinical model species with ocular anatomy and physiology more relevant to human. These findings support a novel platform for delivery of therapeutics to the back of the eye, with the potential for every other month dosing in humans, resulting in fewer IVT injections per year than monthly dosing of non-conjugated Fab.…”

Section: Discussionsupporting

confidence: 86%

“…Clearance from vitreous humor was consistent with first order kinetics for both AFD.v14 Fab and Fab + PEG. Similar to other Fabs [26] the non-conjugated Fab had a vitreal half-life of 2.7 days, whereas the 8X Fab + 8-arm PEG had an increased half-life of 4.2 ± 1.1 days (Table 5 and Fig 6B); dose normalized exposure was approximately two-fold higher for octamer (2120 ± 17 μ g/mL*day/mg Fab) vs. Fab (980 μ g/mL*day/mg). Vitreal exposure for octamer dosed at 1.3 or 4.3 mg/eye (1.1 or 3.9 mg Fab/eye) was dose proportional on the basis of AUC (2140 μ g/mL*day/mg Fab vs. 2110 μ g/mL*day/mg Fab); however, a higher dose-normalized C max (310 μ g/mL/mg vs. 250 μ g/mL/mg) and lower volume of distribution (3.1 mL vs. 3.4 mL) were observed for the 1.3 mg/eye dose group relative to the 4.3 mg/eye dose group.…”

Section: Resultsmentioning

confidence: 59%

“…Conjugation to PEG has been widely employed for systemic half-life extension and is generally considered safe for use in a wide variety of settings [26]. The recent availability of multimeric PEG structures has yielded new insights into the influence of PEG on the biophysical and biochemical properties of a Fab.…”

Section: Discussionmentioning

confidence: 99%

“…Since IVT drug delivery is limited by the maximal volume of injection (≤100 ul), a high concentration formulation is important for delivery of the maximum feasible dose. However, it has been shown that coupling a therapeutic Fab to PEG has the potential to increase viscosity by increasing the excluded volume of the Fab therapeutic at high concentrations [26]. Therefore, understanding visco-elastic properties of the top candidates at a range of formulation concentrations intended to enable high dose scenarios (>50 mg/mL) was of importance in their evaluation.…”

Section: Discussionmentioning

confidence: 99%

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Identification and characterization of an octameric PEG-protein conjugate system for intravitreal long-acting delivery to the back of the eye

Shatz¹,

Hass²,

Peer³

et al. 2019

PLoS ONE

Self Cite

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Innovative protein engineering and chemical conjugation technologies have yielded an impressive number of drug candidates in clinical development including >80 antibody drug conjugates, >60 bispecific antibodies, >35 Fc-fusion proteins and >10 immuno-cytokines. Despite these innovations, technological advances are needed to address unmet medical needs with new pharmacological mechanisms. Age-related eye diseases are among the most common causes of blindness and poor vision in the world. Many such diseases affect the back of the eye, where the inaccessibility of the site of action necessitates therapeutic delivery via intravitreal (IVT) injection. Treatments administered via this route typically have vitreal half-lives <10 days in humans, requiring frequent administration. Since IVT injection is burdensome to patients, there exists a strong need to develop therapeutics with prolonged residence time in the eye. We report here a strategy to increase retention of a therapeutic fragment antibody (Fab) in the eye, using an anti-complement factor D Fab previously optimized for ocular delivery. Polyethylene glycol structures, varying in length, geometry and degree of branching, were coupled to the Fab via maleimide-activated termini. A screening strategy was developed to allow for key determinants of ocular half-life to be measured in vitro. After compound selection, a scalable process was established to enable tolerability and pharmacokinetic studies in cynomolgus monkeys, demonstrating an increase in vitreal half-life with no associated adverse events. Further, we show that the technique for compound selection, analytical characterization, and scalable production is general for a range of antibody fragments. The application of the technology has broad impact in across many therapeutic areas with the first major advancement in the treatment of an important ocular disease.

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Section: Discussionsupporting

confidence: 86%

Section: Resultsmentioning

confidence: 59%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Identification and characterization of an octameric PEG-protein conjugate system for intravitreal long-acting delivery to the back of the eye

Shatz¹,

Hass²,

Peer³

et al. 2019

PLoS ONE

Self Cite

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“…The measured torque determined the shear stress, from which the viscosity was calculated, as previously described. 69 …”

Section: Methodsmentioning

confidence: 99%

Targeted IgMs agonize ocular targets with extended vitreal exposure

Chen¹,

Paluch²,

Zorn³

et al. 2020

mAbs

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Treatment of ocular disease is hindered by the presence of the blood-retinal barrier, which restricts access of systemic drugs to the eye. Intravitreal injections bypass this barrier, delivering high concentrations of drug to the targeted tissue. However, the recommended dosing interval for approved biologics is typically 6–12 weeks, and frequent travel to the physician’s office poses a substantial burden for elderly patients with poor vision. Real-world data suggest that many patients are under-treated. Here, we investigate IgMs as a novel platform for treating ocular disease. We show that IgMs are well-suited to ocular administration due to moderate viscosity, long ocular exposure, and rapid systemic clearance. The complement-dependent cytotoxicity of IgMs can be readily removed with a P436G mutation, reducing safety liabilities. Furthermore, dodecavalent binding of IgM hexamers can potently activate pathways implicated in the treatment of progressive blindness, including the Tie2 receptor tyrosine kinase signaling pathway for the treatment of diabetic macular edema, or the death receptor 4 tumor necrosis family receptor pathway for the treatment of wet age-related macular degeneration. Collectively, these data demonstrate the promise of IgMs as therapeutic agonists for treating progressive blindness.

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How to Characterize the Protein Structure and Polymer Conformation in Protein‐Polymer Conjugates – a Perspective

Mathieu-Gaedke

Böker

Glebe

2023

Macro Chemistry & Physics

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Proteins, having miscellaneous properties perfected by nature over millions of years, are attractive for many biomedical and industrial applications. The exploitation of proteins offers great opportunities to increase, among others, the selectivity, biocompatibility, and sustainability of artificial materials. However, the stability of proteins is limited and exposure to conditions like heat or organic solvents often leads to denaturation or aggregation. The modification with synthetic polymers is a powerful possibility to increase the stability of proteins. Throughout the last few years, various methods have been established to characterize obtained conjugates regarding the conjugation efficiency, protein stability, their self-assembly behavior, and beyond. Nevertheless, it is still very challenging to determine if the protein structure is unaltered in the biohybrid materials and how the polymer chains arrange around the protein surface. Here, an overview of different analysis techniques is presented, their applicability and feasibility are discussed, and current literature examples are provided. The focus of this Perspective lies on nuclear magnetic resonance spectroscopy, size exclusion chromatography coupled with multi-angle laser light scattering, analytical ultracentrifugation, and small-angle scattering techniques as these methods shed light into the structure of proteins in conjugates and the polymer conformation around proteins.

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Impact of polymer geometry on the interactions of protein-PEG conjugates

Cited by 15 publications

References 29 publications

Identification and characterization of an octameric PEG-protein conjugate system for intravitreal long-acting delivery to the back of the eye

Identification and characterization of an octameric PEG-protein conjugate system for intravitreal long-acting delivery to the back of the eye

Targeted IgMs agonize ocular targets with extended vitreal exposure

How to Characterize the Protein Structure and Polymer Conformation in Protein‐Polymer Conjugates – a Perspective

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