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HPLC–UV Monitored Photostability-Test of LE404 and Identification of the Degradation Products via NMR and LC–HRMS

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Abstract

Dibenzoazecines are a new class of potential neuroleptics with a high potential for the treatment of schizophrenia. Initial stress tests indicated that the lead compound, LE404, decomposes when exposed to oxygen and sunlight. In this follow-up study, the influence of oxidative stress and photosensitivity was investigated in accordance with the ICH guidelines. These studies are of great importance for new APIs, as phototoxic and photoallergic reactions pose a significant risk to patients, especially with long-term medication, as it is not possible to completely avoid exposure to sunlight. Furthermore, the identification and prediction of the chemical lability of LE404 can be used to determine suitable storage conditions to prevent compound degradation. The effects of exposure of LE404 to a light source similar in spectrum and intensity to natural sunlight were investigated according to ICH-Q1B. Furthermore, the influence of oxidizing agents was investigated under exclusion of light. Two degradation products were identified. The extent and rate of degradation were continuously monitored using RP-HPLC–UV. Chromatographic separations were performed with a phenomenex™ Gemini 5 µm C18 (250 × 4.60 mm) column and acetonitrile/KH2PO4 buffer (4 mmol L−1, pH 2.5) as mobile phase at 220 nm. The photodegradation product was isolated using semi-preparative RP-HPLC. The oxidation product was obtained by quantitative conversion of LE404 in hydrogen peroxide and subsequent purification by preparative TLC. The structures of both degradation products were elucidated using HR-MS/MS, 1D- and 2D-NMR as well as FT-IR spectroscopy. The characterization of the degradation products serves as the basis for subsequent investigations into their toxicity.

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The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

The authors have no relevant financial or non-financial interests to disclose.

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Authors and Affiliations

  1. Department of Pharmaceutical/Medicinal Chemistry, Institute of Pharmacy, Friedrich-Schiller University Jena, Philosophenweg 14, 07743, Jena, Germany

    Stephanie Zergiebel & Andreas Seeling

  2. Mass Spectrometry Platform, Friedrich Schiller University Jena, Humboldtstraße 8, 07743, Jena, Germany

    Nico Ueberschaar

  3. Department of Functional Interfaces, Leibniz Institute of Photonic Technology (Leibniz-IPHT), Albert-Einstein-Straße 9, 07745, Jena, Germany

    Jonathan Plentz

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  1. Stephanie Zergiebel
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  2. Nico Ueberschaar
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  4. Andreas Seeling
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Contributions

All authors contributed to the study conception and design. All authors read and approved the final manuscript. Conceptualization: S.Z.; Methodology: S.Z.; Formal analysis and investigation: S.Z., N.U.; Visualization: N.U., S.Z.; Writing - original draft preparation: S.Z.; Writing - review and editing: S.Z., A.S., N.U.; Resources: J.P.; Supervision: A.S.

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Correspondence to Andreas Seeling.

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Zergiebel, S., Ueberschaar, N., Plentz, J. et al. HPLC–UV Monitored Photostability-Test of LE404 and Identification of the Degradation Products via NMR and LC–HRMS. Chromatographia 87, 339–349 (2024). https://doi.org/10.1007/s10337-024-04323-1

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  • DOI: https://doi.org/10.1007/s10337-024-04323-1

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