(194e) Formulation of Peptide and Protein Therapeutics into Nanoparticles for Prolonged Activity and Improved Delivery

Ristroph, K. D., Princeton University
Prud'homme, R. K., Princeton University
Rummaneethorn, P., Princeton University

AIChE 2018 annual conference abstracts

IP peptides and proteins

“Formulation of peptide and protein therapeutics into
nanoparticles for prolonged activity and improved delivery”

Kurt Ristroph, Paradorn Rummaneethorn, and Robert K.

Department of Chemical & Biological Engineering,
Princeton University, Princeton, New Jersey, United States

Biologics, the fastest-growing sector of the pharmaceutical
marketplace, are an attractive class of therapeutics because of their
impressive potency, high selectivity, and reduced off-target effects. But while
the effectiveness of these drugs outclasses many of their small-molecule
predecessors, administering biologics remains a challenge. Physiological
barriers such as chemical digestion (when taken orally), rapid blood clearance
(when injected), or thick pulmonary mucus (when inhaled) chemically or
physically prevent biologics from reaching their targets and working as
designed. To reduce the frequency of dosing, strategies of protecting these
proteins and peptides within delivery vehicles have arisen, but the majority of
these processes suffer from high losses and poor scalability. We here present a
scalable and continuous method of encapsulating water-soluble charged biologics
into polymeric nanoparticles. This is done by simultaneously reversibly
ionically modifying the biologics of interest with hydrophobic counterions and
controllably precipitating the newly-formed hydrophobic complex into
nanoparticles via the polymer-directed Flash NanoPrecipitation technique. This
combined technique, termed hydrophobic ion pairing Flash NanoPrecipitation
(HIP-FNP), is applicable to a wide variety of peptides and proteins, both
anionic and cationic. Importantly, the process is continuous, scalable, and
achieves encapsulation efficiencies greater than 95%. We herein demonstrate
encapsulation of two model proteins: the cationic enzyme lysozyme (MW 14,300 D)
and the anionic protein ovalbumin (MW 42,700 D). By altering the identity or
amount of hydrophobic counterion used, we can tune protein release rates, an
important consideration for prolonged delivery. Importantly, we also show that
the proteins’ activity has been retained throughout the processing steps. We
believe this technique offers a route forward for improving the delivery of
many biologic therapeutics and may improve patient comfort and compliance by
simplifying dosing regimens.

Schematic of hydrophobic ion pairing Flash NanoPrecipitation
(HIP-FNP) used to ionically complex a hydrophilic biologic molecule and encapsulate
it into a nanoparticle.

Release rates of lysozyme from NPs. Fraction of lysozyme
released over time when using various charge ratios of lysozyme to hydrophobic
counterion (here, dodecyl sulfate).