Isolation of the Acetylcholinesterase Inhibitor Ungeremine from Nerine bowdenii by Preparative HPLC Coupled On-Line to a Flow Assay System

In Kyung Rhee; Natalie Appels; Bertil Hofte; Bahadir Karabatak; Cornelis Erkelens; Lucy M. Stark; Lee A. Flippin; Robert Verpoorte

doi:10.1248/bpb.27.1804

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Isolation of the Acetylcholinesterase Inhibitor Ungeremine from Nerine bowdenii by Preparative HPLC Coupled On-Line to a Flow Assay System

In Kyung Rhee, Natalie Appels, Bertil Hofte, Bahadir Karabatak, Cornelis Erkelens, Lucy M. Stark, Lee A. Flippin, Robert Verpoorte

Author information

In Kyung Rhee
Division of Pharmacognosy, Section Metabolomics, Institute of Biology, Leiden University
Natalie Appels
Assay Development Department Screen Tec. BV.
Bertil Hofte
Division of Analytical Chemistry, Leiden/Amsterdam Center for Drug Research, Gorlaeus Laboratories, Leiden University
Bahadir Karabatak
Division of Analytical Chemistry, Leiden/Amsterdam Center for Drug Research, Gorlaeus Laboratories, Leiden University
Cornelis Erkelens
NMR Department, Leiden Institute of Chemistry, Leiden University
Lucy M. Stark
Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute
Lee A. Flippin
Departemtn of Medicinal Chemistry, FibroGen Inc.
Robert Verpoorte
Division of Pharmacognosy, Section Metabolomics, Institute of Biology, Leiden University

Keywords: fluorometric, on-line HPLC, preparative HPLC, acetylcholinesterase inhibitor, ungeremine, Nerine bowdenii

JOURNAL FREE ACCESS

2004 Volume 27 Issue 11 Pages 1804-1809

DOI https://doi.org/10.1248/bpb.27.1804

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Abstract

In an attempt to isolate the active compound while detecting acetylcholinesterase inhibitory activity, we applied a fluorometric flow assay system to an on-line coupled preparative HPLC. The MeOH extract of Nerine bowdenii showed a strong inhibitory peak in the on-line assay, and the active compound was isolated by CPC and HPLC. It was identified as ungeremine by analysis of its ¹H-NMR, 2D-NMR, and NOESY spectra. The assignment of the active N. bowdenii constituent was also confirmed by co-TLC, co-HPLC, and co-¹H-NMR experiments using an authentic sample of synthetic ungeremine. The IC₅₀ value of ungeremine was 0.35 μM, showing stronger activity than galanthamine (2.2 μM).

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