Thioamides Adjacent to the Ionizable Amine Headgroup in Ionizable Lipids Reduce the pKa of Lipid Nanoparticles and Enhance mRNA Transfection Efficiency in Vitro and in Vivo
Yong Chen
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Both authors contributed equally to this work.
Search for more papers by this authorEmily De Lombaerde
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Both authors contributed equally to this work.
Search for more papers by this authorAimée Bugler-Lamb
Laboratory of Myeloid Cell Biology in Tissue Homeostasis and Regeneration, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Laboratory of Myeloid Cell Biology in Tissue Damage and Inflammation, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Biomedical Molecular Biology, Faculty of Science, Ghent University, Belgium
Search for more papers by this authorZifu Zhong
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorMartijn J. Schuijs
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Search for more papers by this authorClaudia M. Brenis Gomez
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Search for more papers by this authorJamie De Baere
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorMark Gontsarik
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorHeleen Lauwers
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorKim Deswarte
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Respiratory Medicine, Ghent University Hospital, Ghent, Belgium
Search for more papers by this authorNiek N. Sanders
Laboratory of Gene Therapy, Ghent University, Ghent, 9820 Belgium
Search for more papers by this authorBart N. Lambrecht
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Respiratory Medicine, Ghent University Hospital, Ghent, Belgium
Search for more papers by this authorMartin Guilliams
Laboratory of Myeloid Cell Biology in Tissue Homeostasis and Regeneration, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Laboratory of Myeloid Cell Biology in Tissue Damage and Inflammation, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Biomedical Molecular Biology, Faculty of Science, Ghent University, Belgium
Search for more papers by this authorCorresponding Author
Bruno G. De Geest
Department of Pharmaceutics, Ghent University, Ghent, Belgium
E-mail: [email protected]
Search for more papers by this authorYong Chen
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Both authors contributed equally to this work.
Search for more papers by this authorEmily De Lombaerde
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Both authors contributed equally to this work.
Search for more papers by this authorAimée Bugler-Lamb
Laboratory of Myeloid Cell Biology in Tissue Homeostasis and Regeneration, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Laboratory of Myeloid Cell Biology in Tissue Damage and Inflammation, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Biomedical Molecular Biology, Faculty of Science, Ghent University, Belgium
Search for more papers by this authorZifu Zhong
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorMartijn J. Schuijs
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Search for more papers by this authorClaudia M. Brenis Gomez
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Search for more papers by this authorJamie De Baere
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorMark Gontsarik
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorHeleen Lauwers
Department of Pharmaceutics, Ghent University, Ghent, Belgium
Search for more papers by this authorKim Deswarte
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Respiratory Medicine, Ghent University Hospital, Ghent, Belgium
Search for more papers by this authorNiek N. Sanders
Laboratory of Gene Therapy, Ghent University, Ghent, 9820 Belgium
Search for more papers by this authorBart N. Lambrecht
Department of Internal Medicine and Pediatrics, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
Laboratory of Mucosal Immunology, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Respiratory Medicine, Ghent University Hospital, Ghent, Belgium
Search for more papers by this authorMartin Guilliams
Laboratory of Myeloid Cell Biology in Tissue Homeostasis and Regeneration, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Laboratory of Myeloid Cell Biology in Tissue Damage and Inflammation, VIB-UGent Center for Inflammation Research, Ghent University, Ghent, Belgium
Department of Biomedical Molecular Biology, Faculty of Science, Ghent University, Belgium
Search for more papers by this authorCorresponding Author
Bruno G. De Geest
Department of Pharmaceutics, Ghent University, Ghent, Belgium
E-mail: [email protected]
Search for more papers by this authorGraphical Abstract
Lipid nanoparticles (LNPs) containing an ionizable lipid with a thioamide adjacent to the tertiary amine have a lower pKa than those containing an analogous lipid with a conventional amide, and exhibit significantly improved mRNA transfection efficiency both in vitro and in vivo.
Abstract
Lipid nanoparticles (LNPs) are currently the most clinically advanced mRNA delivery vectors. However, optimizing LNPs for in vivo applications remains largely empirical. The apparent pKa of LNPs is a predictive factor for in vivo performance, with pKa values between 6 and 7 showing the highest efficacy. Despite this critical role of ionizable lipids in LNPs, the relationship between lipid structure and its influence on LNP pKa remains poorly studied. In this study, we report the design and the synthesis of a novel class of ionizable lipids featuring a thioamide moiety, enabling direct comparison between thioamide-containing (SAM) LNPs and amide-containing (OAM) LNPs. We find that substituting oxygen with sulfur in the amide group significantly decreases the apparent pKa of LNPs, increasing the likelihood of identifying lipids in combinatorial libraries that yield LNPs with a pKa in the desired 6–7 range. The reduction in pKa in LNPs containing SAM lipids, compared with OAM lipids, is attributed to the increased hydrophobicity of the thioamide group. Furthermore, by synthesizing multiple libraries of SAM lipids and varying the ionizable head group, alkyl chains, and linker length, we discovered thioamide lipids with distinct tissue tropism, including lipids that mediate splenic targeting by LNPs.
Conflict of Interests
The authors declare no conflict of interest.
Open Research
Data Availability Statement
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Supporting Information
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Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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