Locked nucleic acid (LNA): Based single-stranded oligonucleotides are not genotoxic
Melanie Guérard1, Zeller Andreas1, Koller Erich1
1Pharmaceutical Sciences, Roche Innovation Center Basel, Pharmaceutical Research and Early Development, F. Hoffmann-La Roche, Ltd, Basel, 4070, Switzerland.
Abstract:
Over the last decade, single stranded oligonucleotides (ON) have gained increased attention as a new drug modality. Because the assessment of genotoxicity risk during early development of pharmaceuticals is essential, we evaluated the potential of locked nucleic acids (LNA)-ONs to induce DNA damage in L5178Y tk+/- cells both with the mouse lymphoma assay (MLA) and the micronucleus test (MNT). Further, the MLA was performed to assess gene and chromosome mutation over 3 and 24h (± metabolic activation). In addition, the MNT was performed to assess, in addition, a potential aneugenic liability. None of the experiments demonstrated a genotoxic effect for the five tested LNA-ONs. We further show data from four proprietary LNA-ONs tested in standard genotoxicity assays in vitro and partially in vivo, which were all negative. In addition, cellular and nuclear uptake of LNA-ONs in L5178Y tk+/- cells was demonstrated. Based on the results presented here as well as in the literature about other representatives of this class, we consider LNA-ONs as generally not DNA reactive and question whether genotoxicity testing of this class of ONs should be required. This is in line with recent recommendation made by the OSWG that extensively assessed the genotoxicity of oligonucleotides. Environ. Mol. Mutagen. 58:112-121, 2017. © 2017 Wiley Periodicals, Inc.
Insights
Locked nucleic acid oligonucleotides (LNA-ONs) show no genotoxic effects in cell-based assays. These findings suggest LNA-ONs are not DNA reactive, potentially reducing the need for extensive genotoxicity testing.
Area of Science:
- Pharmacology and Toxicology
- Molecular Biology
- Genetics
Background:
- Single-stranded oligonucleotides (ONs) are emerging as a significant drug modality.
- Assessing genotoxicity is crucial for early pharmaceutical development.
- Locked nucleic acids (LNA)-ONs represent a novel class of therapeutic oligonucleotides.
Purpose of the Study:
- To evaluate the genotoxic potential of LNA-ONs using established assays.
- To determine if LNA-ONs induce DNA damage, gene mutations, or chromosomal aberrations.
- To assess the necessity of genotoxicity testing for LNA-ONs.
Main Methods:
- Mouse lymphoma assay (MLA) to detect gene and chromosome mutations.
- Micronucleus test (MNT) to assess chromosomal damage and aneugenicity.
- In vitro and in vivo genotoxicity assays for proprietary LNA-ONs.
Main Results:
- No genotoxic effects were observed for five tested LNA-ONs in MLA and MNT.
- Four additional proprietary LNA-ONs also yielded negative results in genotoxicity assays.
- Cellular and nuclear uptake of LNA-ONs in L5178Y tk+/- cells was confirmed.
Conclusions:
- LNA-ONs are considered generally not DNA reactive based on current data.
- The findings support reconsidering mandatory genotoxicity testing for this class of ONs.
- Results align with recent recommendations from the Oligonucleotide Safety Working Group (OSWG).
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