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Updated: Sep 19, 2025

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
Published on: August 17, 2019
Discovery and optimization of oxidative phosphorylation inhibitors from a phenotypic screen
Mahmoud El Shemerly1, Florian Richalet, Dimitri Robay
1Basilea Pharmaceutica International Ltd, Allschwil, Switzerland.
Abstract:
Tumor metabolism and metabolic reprogramming in cancer cells represent a promising area in oncology research, offering new avenues for therapeutic intervention. While the 'Warburg effect' highlights the reliance of many tumors on aerobic glycolysis, emerging evidence indicates that some cancers also depend on mitochondrial oxidative phosphorylation (OXPHOS) for energy production, cancer cell survival, tumor progression, metastasis, and drug resistance. We conducted a high-throughput, differential, phenotypic screening followed by a focused medicinal chemistry campaign, leading to the identification of novel, potent OXPHOS inhibitors. These lead compounds selectively target complex I of the mitochondrial electron transport chain, thereby disrupting ATP production and oxygen consumption in cancer cells. In-vitro studies in breast cancer cell lines, along with published data, suggest that MCT4 expression may serve as a biomarker for drug sensitivity. Notably, low MCT4 expression correlated with higher potency in cell growth assays. The identified compounds exhibited favorable drug-like properties, including good pharmacokinetics and oral bioavailability in mice. Daily oral dosing significantly inhibited tumor growth in two in-vivo breast cancer models with low MCT4 expression levels. This efficacy, however, was accompanied by body weight loss, indicating the need to enhance the therapeutic index through optimization or rational combination therapy strategies. These findings highlight the therapeutic potential of targeting mitochondrial OXPHOS in cancers with defined metabolic dependencies, offering a novel approach for exploiting tumor-specific metabolic vulnerabilities for improved cancer treatment.
Insights
Researchers identified novel inhibitors targeting mitochondrial oxidative phosphorylation (OXPHOS) in cancer cells. These compounds show promise for treating cancers dependent on OXPHOS, offering new therapeutic strategies.
Area of Science:
- Oncology
- Cancer Metabolism
- Mitochondrial Biology
Background:
- Cancer cells exhibit metabolic reprogramming, including aerobic glycolysis (Warburg effect).
- Emerging evidence shows some cancers rely on mitochondrial oxidative phosphorylation (OXPHOS) for survival and progression.
- Targeting OXPHOS presents a novel therapeutic strategy in oncology.
Purpose of the Study:
- To identify and develop novel inhibitors of mitochondrial oxidative phosphorylation (OXPHOS).
- To evaluate the efficacy of OXPHOS inhibitors in preclinical cancer models.
- To explore potential biomarkers for drug sensitivity.
Main Methods:
- High-throughput phenotypic screening and medicinal chemistry campaigns.
- In vitro studies using breast cancer cell lines.
- In vivo efficacy studies in mouse models of breast cancer.
Main Results:
- Novel potent inhibitors targeting Complex I of the mitochondrial electron transport chain were identified.
- Low MCT4 expression correlated with increased drug potency in vitro.
- Oral administration of compounds significantly inhibited tumor growth in vivo in models with low MCT4 expression.
- Compounds demonstrated favorable drug-like properties and oral bioavailability in mice.
Conclusions:
- Targeting mitochondrial OXPHOS is a viable therapeutic strategy for specific cancer types.
- MCT4 expression may serve as a predictive biomarker for OXPHOS inhibitor sensitivity.
- Further optimization is needed to improve the therapeutic index and manage potential side effects.

