Inhibitors of Human 5-Lipoxygenase Potently Interfere With Prostaglandin Transport

Astrid S Kahnt; Carlo Angioni; Tamara Göbel; Bettina Hofmann; Jessica Roos; Svenja D Steinbrink; Florian Rörsch; Dominique Thomas; Gerd Geisslinger; Kai Zacharowski; Sabine Grösch; Dieter Steinhilber; Thorsten J Maier

doi:10.3389/fphar.2021.782584

Inhibitors of Human 5-Lipoxygenase Potently Interfere With Prostaglandin Transport

Front Pharmacol. 2022 Jan 21:12:782584. doi: 10.3389/fphar.2021.782584. eCollection 2021.

Authors

Affiliations

¹ Institute of Pharmaceutical Chemistry, Goethe University, Frankfurt, Germany.
² Pharmazentrum Frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe-University, Frankfurt, Germany.
³ Paul-Ehrlich Institute, Federal Institute for Vaccines and Biomedicines, Langen, Germany.
⁴ Department of Anesthesiology, Intensive Care Medicine and Pain Therapy, University Hospital Frankfurt, Goethe University, Frankfurt, Germany.

Abstract

5-Lipoxygenase (5-LO) is the key enzyme in the formation of pro-inflammatory leukotrienes (LT) which play an important role in a number of inflammatory diseases. Accordingly, 5-LO inhibitors are frequently used to study the role of 5-LO and LT in models of inflammation and cancer. Interestingly, the therapeutic efficacy of these inhibitors is highly variable. Here we show that the frequently used 5-LO inhibitors AA-861, BWA4C, C06, CJ-13,610 and the FDA approved compound zileuton as well as the pan-LO inhibitor nordihydroguaiaretic acid interfere with prostaglandin E₂ (PGE₂) release into the supernatants of cytokine-stimulated (TNFα/IL-1β) HeLa cervix carcinoma, A549 lung cancer as well as HCA-7 colon carcinoma cells with similar potencies compared to their LT inhibitory activities (IC₅₀ values ranging from 0.1-9.1 µM). In addition, AA-861, BWA4C, CJ-13,610 and zileuton concentration-dependently inhibited bacterial lipopolysaccharide triggered prostaglandin (PG) release into human whole blood. Western Blot analysis revealed that inhibition of expression of enzymes involved in PG synthesis was not part of the underlying mechanism. Also, liberation of arachidonic acid which is the substrate for PG synthesis as well as PGH₂ and PGE₂ formation were not impaired by the compounds. However, accumulation of intracellular PGE₂ was found in the inhibitor treated HeLa cells suggesting inhibition of PG export as major mechanism. Further, experiments showed that the PG exporter ATP-binding cassette transporter multidrug resistance protein 4 (MRP-4) is targeted by the inhibitors and may be involved in the 5-LO inhibitor-mediated PGE₂ inhibition. In conclusion, the pharmacological effects of a number of 5-LO inhibitors are compound-specific and involve the potent inhibition of PGE₂ export. Results from experimental models on the role of 5-LO in inflammation and pain using 5-LO inhibitors may be misleading and their use as pharmacological tools in experimental models has to be revisited. In addition, 5-LO inhibitors may serve as new scaffolds for the development of potent prostaglandin export inhibitors.

Keywords: ABC transporter; eicosanoid; inflammation; lipoxygenase inhibitor; multidrug resistance protein 4; prostaglandin.