Oral gene delivery: Strategies to improve stability of pDNA towards intestinal digestion

• Published in: Journal of drug targeting 2006-01, Vol.14 (5), p.311-319
• Main Authors: Loretz, Brigitta, Föger, Florian, Werle, Martin, Bernkop-schnürch, Andreas
• Format: Article
• Language: English
• Subjects: Administration, Oral; Biological and medical sciences; Caco-2 Cells; chitosan; Chitosan - administration & dosage; Chitosan - chemical synthesis; Chitosan - chemistry; Chitosan - pharmacokinetics; Deoxyribonuclease I - antagonists & inhibitors; Deoxyribonuclease I - metabolism; DNA vaccines; DNase inhibitors; Edetic Acid - administration & dosage; Edetic Acid - chemical synthesis; Edetic Acid - chemistry; Edetic Acid - pharmacokinetics; Enzymatic degradation; Gastric Juice - metabolism; gene therapy; General pharmacology; Genetic Therapy - methods; Humans; Intestines - enzymology; Intestines - metabolism; Medical sciences; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Plasmids - administration & dosage; Plasmids - chemistry; Plasmids - pharmacokinetics
• ISSN: 1061-186X; 1029-2330
• DOI: 10.1080/10611860600823766

Purpose: Gastrointestinal (GI) nucleases are responsible for a rapid presystemic degradation of orally administered transgenes. Within the current study, the activity of these degrading enzymes as well as the effect of various nuclease inhibitors on the degradation process were evaluated in order to assess their potential as auxiliary agents in oral gene delivery. Methods: Digestion assays of pDNA with DNaseI and in GI juices were performed in absence and presence of inhibitors. Consequently, a chitosan conjugate with covalently bound ethylendiaminetetraacetic acid disodium salt dihydrat (EDTA) was synthesized and its nuclease inhibitory properties were evaluated. Results: Small intestinal juice was shown to possess a nuclease activity per millilitre corresponding to 0.02 Kunitz units of DNaseI. Inhibition studies revealed that inhibitory activity followed the ranking: EDTA > sodium dodecyl sulfate (SDS) > aurintricarboxylic acid (ATA) > poly (acrylic acid) > cysteine. The chitosan-EDTA conjugate offered good nuclease inhibiting properties. Conclusion: This study determined the nuclease activity of native porcine small intestinal juice as well as enterocytes homogenate. Moreover, several promising strategies to overcome this enzymatic barrier were identified.