Characterization of an amino acid based biodegradable surfactant facilitating the incorporation of DNA into lipophilic delivery systems

Julian Dominik Wolf; Markus Kurpiers; Randi Angela Baus; Roman Xaver Götz; Janine Griesser; Barbara Matuszczak; Andreas Bernkop-Schnürch

doi:10.1016/j.jcis.2020.01.088

Characterization of an amino acid based biodegradable surfactant facilitating the incorporation of DNA into lipophilic delivery systems

J Colloid Interface Sci. 2020 Apr 15:566:234-241. doi: 10.1016/j.jcis.2020.01.088. Epub 2020 Jan 24.

Authors

Julian Dominik Wolf¹, Markus Kurpiers¹, Randi Angela Baus², Roman Xaver Götz², Janine Griesser¹, Barbara Matuszczak³, Andreas Bernkop-Schnürch⁴

Affiliations

¹ Thiomatrix Forschungs- und Beratungs GmbH, Research Center Innsbruck, Trientlgasse 65, A-6020 Innsbruck, Austria; Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80-82, A-6020 Innsbruck, Austria.
² Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80-82, A-6020 Innsbruck, Austria.
³ Department of Pharmaceutical Chemistry, Institute of Pharmacy, University of Innsbruck, Innrain 80-82, A-6020 Innsbruck, Austria.
⁴ Thiomatrix Forschungs- und Beratungs GmbH, Research Center Innsbruck, Trientlgasse 65, A-6020 Innsbruck, Austria; Department of Pharmaceutical Technology, Institute of Pharmacy, University of Innsbruck, Innrain 80-82, A-6020 Innsbruck, Austria. Electronic address: andreas.bernkop@uibk.ac.at.

PMID: 32006819
DOI: 10.1016/j.jcis.2020.01.088

Abstract

Hypothesis: Lysine based cationic surfactants are well-tolerated tools for hydrophobic ion pairing (HIP) with DNA and its incorporation into lipophilic delivery systems.

Experiments: Di-Boc-lysine was esterified with 1-hexadecanol and the Boc-residues were cleaved off resulting in hexadecyl lysinate (HL). Subsequently, its Log P_{Octanol/water} and the critical micelle concentration (CMC) were determined. Degradability was evaluated utilizing trypsin and pancreas lipase as well as Caco-2 cells. Afterwards, the viability of Caco-2 cells upon incubation with HL was investigated. Finally, HL was ion-paired with plasmid DNA (pDNA, 6159 bp) and the obtained complex was incorporated into self-emulsifying drug delivery systems (SEDDS) for transfection studies on HEK-293 cells.

Findings: HL was synthesized with a yield of 53% and subsequent characterization revealed a Log P_{Water/Octanol} of 0.05 and a CMC of 2.7 mM. Enzymatic degradation studies showed rapid degradation of HL by isolated enzymes and Caco-2 cells and cell viability experiments revealed no toxic effect of HL even in a concentration of 250 µg·ml^-1 within 24 h. HIP with pDNA was the most efficient in a molar ratio of 6159:1 (HL:pDNA) equalling a charge ratio of 1:1. Formed complexes could be incorporated into SEDDS facilitating successful transfection of HEK-293 cells.

Keywords: Biodegradable; Cationic surfactant; Gene therapy; Hydrophobic ion pairing; SEDDS; Transfection.

MeSH terms

Amino Acids / chemistry
Amino Acids / metabolism*
Caco-2 Cells
DNA / chemistry
DNA / metabolism*
Drug Delivery Systems*
Humans
Hydrophobic and Hydrophilic Interactions
Molecular Structure
Surface-Active Agents / chemistry
Surface-Active Agents / metabolism*

Substances

Amino Acids
Surface-Active Agents
DNA