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Polysarcosine-Functionalized Lipid Nanoparticles for Therapeutic mRNA Delivery

  • Sara S. Nogueira
    Sara S. Nogueira
    BioNTech RNA Pharmaceuticals GmbH, 55131 Mainz, Germany
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  • , 
  • Anne Schlegel
    Anne Schlegel
    BioNTech RNA Pharmaceuticals GmbH, 55131 Mainz, Germany
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  • , 
  • Konrad Maxeiner
    Konrad Maxeiner
    BioNTech RNA Pharmaceuticals GmbH, 55131 Mainz, Germany
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  • , 
  • Benjamin Weber
    Benjamin Weber
    Institute of Organic Chemistry, Johannes Gutenberg University Mainz, 55128 Mainz, Germany
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  • , 
  • Matthias Barz
    Matthias Barz
    Leiden Academic Center for Drug Research, Leiden University, 2333 CC Leiden, Netherlands
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    Orcidhttp://orcid.org/0000-0002-1749-9034
  • , 
  • Martin A. Schroer
    Martin A. Schroer
    European Molecular Biology Laboratory (EMBL) Hamburg c/o DESY, 22607 Hamburg, Germany
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    Orcidhttp://orcid.org/0000-0002-0747-3965
  • , 
  • Clement E. Blanchet
    Clement E. Blanchet
    European Molecular Biology Laboratory (EMBL) Hamburg c/o DESY, 22607 Hamburg, Germany
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  • Dmitri I. Svergun
    Dmitri I. Svergun
    European Molecular Biology Laboratory (EMBL) Hamburg c/o DESY, 22607 Hamburg, Germany
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  • , 
  • Srinivas Ramishetti
    Srinivas Ramishetti
    Laboratory of Precision NanoMedicine, The Shmunis School of Biomedicine and Cancer Research and Dept. of Materials Sciences and Engineering, Tel Aviv University, Tel Aviv 69978, Israel
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  • , 
  • Dan Peer
    Dan Peer
    Laboratory of Precision NanoMedicine, The Shmunis School of Biomedicine and Cancer Research and Dept. of Materials Sciences and Engineering, Tel Aviv University, Tel Aviv 69978, Israel
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    Orcidhttp://orcid.org/0000-0001-8238-0673
  • , 
  • Peter Langguth
    Peter Langguth
    Department of Pharmaceutics and Biopharmaceutics, Johannes Gutenberg University Mainz, 55099 Mainz, Germany
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  • , 
  • Ugur Sahin
    Ugur Sahin
    BioNTech RNA Pharmaceuticals GmbH, 55131 Mainz, Germany
    TRON - Translational Oncology at the University Medical Center of Johannes Gutenberg University Mainz gGmbH, 55099 Mainz, Germany
    Research Center for Immunotherapy (FZI), University Medical Center at the Johannes Gutenberg University Mainz, 55099 Mainz, Germany
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  • , and 
  • Heinrich Haas*
    Heinrich Haas
    BioNTech RNA Pharmaceuticals GmbH, 55131 Mainz, Germany
    *Email: [email protected]
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    Orcidhttp://orcid.org/0000-0002-5517-5970
Cite this: ACS Appl. Nano Mater. 2020, 3, 11, 10634–10645
Publication Date (Web):September 25, 2020
https://doi.org/10.1021/acsanm.0c01834
Copyright © 2020 American Chemical Society
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    Abstract

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    Polysarcosine (pSar) is a polypeptoid based on the endogenous amino acid sarcosine (N-methylated glycine), which has previously shown potent stealth properties. Here, lipid nanoparticles (LNPs) for therapeutic application of messenger RNA were assembled using pSarcosinylated lipids as a tool for particle engineering. Using pSar lipids with different polymeric chain lengths and molar fractions enabled the control of the physicochemical characteristics of the LNPs, such as particle size, morphology, and internal structure. In combination with a suited ionizable lipid, LNPs were assembled, which displayed high RNA transfection potency with an improved safety profile after intravenous injection. Notably, a higher protein secretion with a reduced immunostimulatory response was observed when compared to systems based on polyethylene glycol (PEG) lipids. pSarcosinylated nanocarriers showed a lower proinflammatory cytokine secretion and reduced complement activation compared to PEGylated LNPs. In summary, the described pSar-based LNPs enable safe and potent delivery of mRNA, thus signifying an excellent basis for the development of PEG-free RNA therapeutics.

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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsanm.0c01834.

    • (Supporting Figure 1) General overview of SAXS data from all systems, (Supporting Figure 2) simulation and experimental data in a linear scale, (Supporting Figure 3) luciferase expression upon IV injection of PEG LNPs, (Supporting Figure 4) effect of pSar chain length on the liver enzyme release profile, (Supporting Figure 5) effect of pSar molar faction on the liver enzyme release profile, (Supporting Figure 6) concentrations of C3a post-incubation of pSar23 and PEG LNPs with human serum, (Supporting Figure 7) luciferase expression of pSar23 and PEG formulations containing DPL14 as the ionizable lipid, (Supporting Table 1) quantitative analysis of the peak as a function of pSar and PEG-DMG molar fraction, (Supporting Table 2) quantitative analysis of the peak as a function of pSar and PEG-DMG molar fraction, (Supporting Table 3) physicochemical characterization of pSar23-LNPs comprising DODMA, Dlin-MC3-DMA, and DPL14 ionizable cationic lipids, and (Supporting Table 4) physicochemical characterization of pSar23 and PEG-DMG LNP containing DPL14 ionizable cationic lipids (PDF)

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