title: Investigation of the transformations of a novel anti-cancer agent combining HPLC, HPLC-MS and direct ESI-HRMS analyses.
abstract: One of the main problems of anti-cancer therapy is an insufficient differentiation between normal and malignant cells by the known anti-proliferant agents. The antibody-directed enzyme prodrug therapy is a promising approach for a selective treatment of cancer, in which a non-toxic prodrug is enzymatically converted into a highly cytotoxic drug at the surface of malignant cells by a targeted antibody-enzyme conjugate. The transformations and the stability of a very promising novel prodrug and its corresponding cytotoxic derivative were now investigated in detail by high-performance liquid chromatography (HPLC)-mass spectrometry (MS). In order to determine the time-dependent DNA alkylation efficiency and the sequence selectivity of the novel compounds, DNA binding studies using direct electrospray-Fourier transform ion cyclotron resonance-MS (ESI-FTICR-MS) have been performed. These measurements were accompanied by HPLC analyses followed by MS of the separated species to confirm the results of the direct ESI-FTICR-MS measurements. The sites of DNA alkylation could be identified unambiguously by the mass spectrometric fragmentation pattern of the alkylated oligodeoxynucleotides as well as by the results of HPLC followed by MS. A combination of all techniques applied led to a better understanding of the mode of action of the new therapeutics and might be used for an estimation of the cytotoxicity of different prodrugs and drugs since the alkylation efficiency correlates with the bioactivity of the compounds in cell culture investigations.
authors.affiliation: Department of Organic and Biomolecular Chemistry, Georg-August-University Göttingen, Tammannstrasse 2, 37077, Göttingen, Germany. ltietze@gwdg.de
authors.forename: Lutz F
authors.initials: LF
authors.lastname: Tietze
authors.forename: Birgit
authors.initials: B
authors.lastname: Krewer
authors.forename: Holm
authors.initials: H
authors.lastname: Frauendorf
mesh_terms: D000478:Alkylation; D018906:Antineoplastic Agents, Alkylating / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004355:Drug Stability; D004926:Escherichia coli / Q000201:enzymology; D005697:Galactosides / Q000032:analysis* / Q000378:metabolism; D008958:Models, Molecular; D011355:Prodrugs / Q000032:analysis* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D018906:Antineoplastic Agents, Alkylating; D005697:Galactosides; D011355:Prodrugs; D004247:DNA; D001616:beta-Galactosidase