title: Development of a protective dermal drug delivery system for therapeutic DNAzymes.
abstract: RNA-cleaving DNAzymes are a potential novel class of nucleic acid-based active pharmaceutical ingredients (API). However, developing an appropriate drug delivery system (DDS) that achieves high bioavailability is challenging. Especially in a dermal application, DNAzymes have to overcome physiological barriers composed of penetration barriers and degrading enzymes. The focus of the present study was the development of a protective and penetration-enhanced dermal DDS that was tailor made for DNAzymes. DNAzyme Dz13 was used as a potential API for topical therapy against actinic keratosis. In the progress of development and selection, different preservatives, submicron emulsions (SMEs) and the physiological pH range were validated with respect to the API's integrity. A physicochemical stable SME of a pharmaceutical grade along with a high API integrity was achieved. Additionally, two developed protective systems, consisting of a liposomal formulation or chitosan-polyplexes, reduced the degradation of Dz13 in vitro. A combination of SME and polyplexes was finally validated at the skin and cellular level by in vitro model systems. Properties of penetration, degradation and distribution were determined. The result was enhanced skin penetration efficiency and increased cellular uptake with a high protective efficiency for DNAzymes due to the developed protective DDS.
authors.affiliation: Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany. Electronic address: Kay.Marquardt@kmub.thm.de.
authors.forename: Kay
authors.initials: K
authors.lastname: Marquardt
authors.affiliation: Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany.
authors.forename: Anna-Carola
authors.initials: AC
authors.lastname: Eicher
authors.affiliation: Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany.
authors.forename: Dorota
authors.initials: D
authors.lastname: Dobler
authors.affiliation: Institute of Medical Physics and Radiation Protection, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany.
authors.forename: Ulf
authors.initials: U
authors.lastname: Mäder
authors.affiliation: Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Schmidts
authors.affiliation: Institute of Laboratory Medicine and Pathobiochemistry, Molecular Diagnostic, Baldinger Strasse, 35043 Marburg, Philipps-University Marburg, Germany.
authors.forename: Harald
authors.initials: H
authors.lastname: Renz
authors.affiliation: Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Wiesenstrasse 14, 35390 Giessen, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Runkel
mesh_terms: D000279:Administration, Cutaneous; D000818:Animals; D002460:Cell Line; D048271:Chitosan / Q000737:chemistry; D003593:Cytoplasm / Q000378:metabolism; D021881:DNA, Catalytic / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D016503:Drug Delivery Systems; D004795:Enzyme Stability; D006801:Humans; D066298:In Vitro Techniques; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism; D055623:Keratosis, Actinic; D008081:Liposomes; D012867:Skin / Q000378:metabolism; D012869:Skin Absorption; D034421:Sus scrofa
chemical_list: D021881:DNA, Catalytic; C485475:Dz13 DNAzyme; D008081:Liposomes; D048271:Chitosan
keywords: Benzoic acid (PubChem CID: 243); Benzyl alcohol (PubChem CID: 244); Chitosan; Chitosan (PubChem CID: 21896651); DNAzyme; DNase; Drug delivery system; Methylparaben (PubChem CID: 7456); Oleyl oleate (PubChem CID: 543844); Phenoxy ethanol (PubChem CID: 31236); Phosphatidylcholin (PubChem CID: 45266626); Polyplex; Propylene glycol (PubChem CID: 1030); Propylparaben (PubChem CID: 7175); Submicron emulsion