Anticancer effect and underlying metabolic remodelling of 4’-methoxy-1-naphthylfenoterol


Danuta Dudzik, Department of Biopharmaceutics and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gdańsk, Poland,
Center for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, San Pablo CEU University, Madrid, Spain
Irving W. Wainer, PAZ Pharmaceuticals, Washington, DC, USA
Wiktoria Struck-Lewicka, Department of Biopharmaceutics and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gdańsk, Poland
Małgorzata Waszczuk-Jankowska, Department of Biopharmaceutics and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gdańsk, Poland
Michał J. Markuszewski, Department of Biopharmaceutics and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gdańsk, Poland
Michel Bernier, National Institute on Aging, National Institutes of Health, Baltimore, Maryland, USA
Coral Barbas, Center for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, San Pablo CEU University, Madrid, Spain
Danuta Siluk, Department of Biopharmaceutics and Pharmacodynamics, Faculty of Pharmacy, Medical University of Gdańsk, Poland

Pancreatic adenocarcinoma (PCA) and colorectal cancer (CRC) are considered two of the most aggressive cancers and are leading causes of cancer-related death worldwide. Despite advances in the field, resistance to chemotherapy results in poor patient survival and remains one of the most significant challenges in long-term management. We studied the novel anticancer effects of 4’-methoxy-1-naphthylfenoterol (MNF), a bi-functional agent that inhibits the pro-oncogenic G protein-coupled receptor GPR55 and activates of the β2-adrenergic receptor in xenograft mouse models of human PANC-1 PCA and CT26 CRC tumors.

Hyphenated mass spectrometry-based untargeted metabolomics approach was applied to study metabolic changes associated with the observed antitumor response of MNF. The target tumors were induced in female BALB/c mice by subcutaneous inoculation with CT26 CRC or PANC-1 PCA tumor cells. The tumor-bearing mice received daily ip injections of either vehicle, (R,R’)-MNF (20 or 30 mg/kg) in CRC or (R,S’)-MNF (40 mg/kg) in PCA models. The treatments significantly reduced tumor growth in each model and prolonged survival.

The changes in plasma and tumor metabolomic profiles associated with the (R,R’)-MNF action in CRC model were evaluated by liquid chromatography mass spectrometry (LC-MS) analysis, whereas changes related to the effect of (R,S’)-MNF in PCA tumors were studied by mass spectrometry liquid chromatography (LC-MS), gas chromatography (GC-MS), and capillary electrophoresis (CE-MS). Raw data acquired was pre-processed, filtered, and corrected for signal drift. A combination of univariate and multivariate statistical analyses was used to evaluate the drug effect on CRC and PCA models. Metabolite annotation was accomplished by searching against several databases including HMDB, KEGG, Metlin, and LipidMaps metabolite databases.

A metabolic-driven elucidation of the anticancer effects of (R,R’)-MNF and (R,S’)-MNF indicate metabolic remodelling related in particular to the class of lipids where glycerolipids and glycerophospholipids were significantly altered in both CRC and PCA models. Prominent changes with +237% increase in ophthalmic acid and +95% increase in its precursor, 2-aminobutyrate, indicated higher oxidative stress under (R,S’)-MNF treatment in PCA model.

The knowledge gained in these studies advanced our understanding of the molecular mechanisms by which MNF inhibits cancer and provide a path towards to optimum use of these agents in CRC, PCA, and other aggressive cancers. The results demonstrate that untargeted metabolomics reveals information locked in metabolites that can lead to scientific breakthroughs and new clinical insights.