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Polymeric Nanocarrier Formulations of Biologics Using Inverse Flash NanoPrecipitation

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Abstract

The encapsulation of water-soluble therapeutics and biologics into nanocarriers to produce novel therapeutics has been envisioned for decades, but clinical translation has been hampered by complex synthesis strategies. The methods that have been developed are often limited by poor encapsulation efficiency/loading or complex processing to achieve therapeutic loadings high enough to be medically relevant. To address this unmet need, we introduce a solubility-driven self-assembly process to form polymeric nanocarriers comprising a biologic in a hydrophilic core, encapsulated by a poly(lactic acid) shell, and stabilized by a poly(ethylene glycol) brush. Called “inverse Flash NanoPrecipitation (iFNP),” the technique achieves biologic loadings (wt% of total formulation) that are 5–15× higher than typical values (9–27% versus < 2%). In contrast to liposomes and polymersomes, we sequentially assemble the polymer layers to form the final nanocarrier. Installation of the poly(lactic acid) shell before water exposure sequesters the biologic in the core and results in the improved loadings that are achieved. We demonstrate the broad applicability of the process and illustrate its implementation by formulating over a dozen different oligosaccharides, antibiotics, peptides, proteins, and RNA into nanocarriers with narrow size distributions, at high loadings, and with high reproducibility. Lysozyme and horseradish peroxidase are shown to retain 99% activity after processing. These results demonstrate the potential for commercial implementation of this technology, enabling the translation of novel treatments in immunology, oncology, or enzyme therapies.

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Acknowledgments

The authors wish to thank Dr. Simon McManus, Brian Wilson, and Kurt Ristroph for the helpful discussions. Kurt Ristroph provided MATLAB code used in the analysis of SEC data. Circular dichroism spectra were collected in the Princeton Dept. of Chemistry Biophysics Core Facility. Funding was provided by Optimeos Life Sciences, in which RFP and RKP have a financial interest, by the Helen Shipley Hunt Fund, and by the Industrial Innovation Fund of Princeton University. CEM was partially funded by a PhRMA Foundation Pre-Doctoral Fellowship in Pharmaceutics. The authors acknowledge the use of Princeton’s Imaging and Analysis Center, which is partially supported by the Princeton Center for Complex Materials, a National Science Foundation (NSF)-MRSEC program (DMR-1420541). The authors thank Brian Wilson for his assistance in staining particles for TEM imaging. We also acknowledge the support of Howard Bowman of Evonik Corporation for supplying the PLA-b-PEG polymers.

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Author notes

  1. Ava N. Hejazi

    Present address: Department of Chemistry, University of California, Berkeley, Berkeley, California, USA

  2. Akiva G. R. Gordon

    Present address: Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

  3. Alexandra L. Thompson

    Present address: Cardiothoracic Surgery, Children’s Hospital Los Angeles, Los Angeles, California, USA

  4. Chester E. Markwalter and Robert F. Pagels contributed equally to this work.

Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Princeton University, A301 Engineering Quad, Olden St., Princeton, New Jersey, 08544, USA

    Chester E. Markwalter, Robert F. Pagels, Ava N. Hejazi, Akiva G. R. Gordon, Alexandra L. Thompson & Robert K. Prud’homme

  2. Optimeos Life Sciences, Princeton, New Jersey, USA

    Robert F. Pagels

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  1. Chester E. Markwalter
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The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.

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Correspondence to Robert K. Prud’homme.

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RKP and RFP have financial interest in Optimeos Life Sciences.

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Markwalter, C.E., Pagels, R.F., Hejazi, A.N. et al. Polymeric Nanocarrier Formulations of Biologics Using Inverse Flash NanoPrecipitation. AAPS J 22, 18 (2020). https://doi.org/10.1208/s12248-019-0405-z

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