Bioreductive prodrugs for the targeting of epigenetic enzymes

Abstract: Epigenetic proteins play a crucial role in the regulation of gene expression and thereby influence the phenotype of the cell. The lysine-specific histone demethylase (LSD1) and histone deacetylases (HDACs) are enzymes that can remove methyl and acetyl groups from the ε-amino group of lysine residues in histones, respectively. The deregulation of these enzymes is correlated with several human malignancies. Even though potent and selective inhibitors for LSD1 and HDACs have been developed for the treatment of cancer, clinical efficacy is impaired by the observed side effects. In this project, prodrugs for a site-specific drug release were developed that are activated by the NADH-dependent bacterial nitroreductase NfsB (NTR) and subsequently release the LSD1 or HDAC inhibitor. The NTR, which is introduced into the cancer cell by virus-mediated transfection, reduces nitroaryl systems to the corresponding hydroxylamine, thereby triggering the disassembly of the prodrug that results in the release of the inhibitor.

The aim of the project was the generation of prodrugs of the inhibitors that are selectively reduced by the NTR and do not display a biological effect themselves. Therefore, diverse nitroaryl systems were linked to specific inhibitors, including primary and secondary amines, as well as hydroxamic acids. For the measurement of the activation by the NTR and the subsequent release of inhibitor, different assay systems were developed. This includes the kinetic measurement of NADH consumption by the NTR and the quantification of released inhibitor by derivatisation with a fluorophore and following HPLC analysis. Also, the influence of the reactive side product on the inhibitor release was determined. Several prodrugs were identified that are efficiently activated by the NTR to release the inhibitor, which consequently inhibits the target enzyme LSD1 or HDAC. Cellular experiments confirmed the specific inhibition by the prodrugs in transfected cells, demonstrating their suitability for further approaches to selectively target cell-types, organs or tissues. Furthermore, the developed negative controls were used to distinguish the effects of the released inhibitor from the effects of the side product.

In addition, luminescent probes were developed that can be used for the detection of cellular NTR activity. The cellular assay also enables the testing of different methods to target the NTR gene towards the cell. Together with the prodrugs for LSD1 and HDAC inhibitors, a prodrug-system for the targeted inhibition of epigenetic enzymes was realized and can now be applied in different targeting strategies

Angaben zum Objekt

• Standort: Deutsche Nationalbibliothek Frankfurt am Main
• Umfang: Online-Ressource
• Sprache: Englisch
• Anmerkungen: Universität Freiburg, Dissertation, 2020

Klassifikation und Themen

• Schlagwort: Prodrugs ; Enzymes ; Pro-Pharmakon ; Pharmazeutische Chemie ; Epigenetik ; Synthese ; Enzym

Beteiligte, Orts- und Zeitangaben

• Ereignis: Veröffentlichung
• (wo): Freiburg
• (wer): Universität
• (wann): 2020
• Urheber: Herrlinger, Eva-Maria
• Beteiligte Personen und Organisationen: Jung, Manfred ; Albert-Ludwigs-Universität Freiburg. Fakultät für Chemie und Pharmazie

Weitere Informationen

• DOI: 10.6094/UNIFR/167042
• URN: urn:nbn:de:bsz:25-freidok-1670426
• Rechteinformation: Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Letzte Aktualisierung: 25.03.2025, 13:48 MEZ