Peroxide-Mediated Desulfurization of Phosphorothioate Oligonucleotides and Its Prevention

Desulfurization at the internucleotide phosphorothioate linkage of antisense oligonucleotides (ASOs) in dermatological formulations has been investigated using strong ion exchange chromatography and mass spectroscopy. The formation of phosphate diester linkages appeared to arise from a reaction betw...

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Bibliographic Details
Published in:Journal of pharmaceutical sciences 2005-02, Vol.94 (2), p.341-352
Main Authors: Krotz, Achim H., Mehta, Rahul C., Hardee, Gregory E.
Format: Article
Language:English
Subjects:
antioxidant
Antioxidants - chemistry
antisense oligonucleotide
Biological and medical sciences
Cetomacrogol - chemistry
Chemistry, Pharmaceutical
Chromatography, Ion Exchange
Drug Stability
Excipients - chemistry
formulation
General pharmacology
Mass Spectrometry
Medical sciences
Oligonucleotides - chemistry
Oligonucleotides, Antisense - pharmacology
Peroxides - chemistry
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
polyethylene glycol
Polyethylene Glycols - chemistry
stability
Sulfur - chemistry
Surface-Active Agents - chemistry
Thionucleotides - chemistry
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Summary:Desulfurization at the internucleotide phosphorothioate linkage of antisense oligonucleotides (ASOs) in dermatological formulations has been investigated using strong ion exchange chromatography and mass spectroscopy. The formation of phosphate diester linkages appeared to arise from a reaction between the phosphorothioate oligonucleotide and a potent oxidizing agent. Screening of excipients used in the formulation indicated that the cause of desulfurization was related to the presence of polyethylene glycol-derived nonionic surfactants MYRJ 52 or BRIJ 58. Autoxidation of the polyethylene glycol chain is suggested as the probable origin for the observed incompatibility. The ability of various antioxidants to prevent oxidative degradation of ASO-1 in simple test systems and in oil-in-water emulsions is described. It is found that in test systems both lipophilic and hydrophilic antioxidants are effective. However, in cream formulation (oil-in-water emulsions) of ASO-1 the addition of hydrophilic antioxidants L-cysteine or DL-α-lipoic acid has been shown to be superior in protecting the oligonucleotide from desulfurization upon storage.
ISSN:0022-3549
1520-6017
DOI:10.1002/jps.20235