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Identification of small molecule inhibitors of the Aurora-A/TPX2 complex
Italia Anna Asteriti1, Frederick Daidone2, Gianni Colotti1
1Institute of Molecular Biology and Pathology, CNR National Research Council, c/o Sapienza University of Rome, 00185, Rome, Italy.
Abstract:
Aurora kinases are a family of cell division regulators that govern the correct assembly of a bipolar mitotic spindle and the fidelity of chromosome segregation. Their overexpression is associated with genomic instability and aneuploidy, and is frequently observed in cancer. Accordingly, competitive inhibitors targeting Aurora kinase activity at the ATP-binding site are being investigated for therapeutic purposes. Despite promising pre-clinical data, these molecules display moderate effects in clinical trials and incomplete selectivity, either against distinct family members, or other kinases. As an alternative approach, protein-protein interaction inhibitors targeting mitotic kinases and their activators can be exploited to achieve increased specificity of action. In this study, a virtual screening of small molecules led to the identification of 25 potential inhibitors of the interaction between Aurora-A and its activator TPX2. In vitro experiments confirmed that 4 hits bind Aurora-A in the low micromolar range and compete for TPX2 binding. Immunofluorescence assays showed that 2 compounds also yield lowered Aurora-A activity and spindle pole defects in cultured osteosarcoma cells. The identified protein-protein interaction inhibitors of the Aurora-A/TPX2 complex might represent lead compounds for further development towards pioneering anti-cancer drugs and provide the proof-of-concept for a new exploitable strategy to target mitotic kinases.
Insights
Researchers identified novel protein-protein interaction inhibitors targeting the Aurora-A/TPX2 complex. These compounds show potential as new anti-cancer drugs by disrupting cell division in cancer cells.
Area of Science:
- Molecular Biology
- Cell Biology
- Oncology
Background:
- Aurora kinases regulate cell division, spindle assembly, and chromosome segregation.
- Overexpression of Aurora kinases is linked to genomic instability, aneuploidy, and cancer.
- Current ATP-binding site inhibitors show limited efficacy and selectivity in clinical trials.
Purpose of the Study:
- To identify novel inhibitors targeting the Aurora-A/TPX2 protein-protein interaction.
- To develop a new therapeutic strategy for cancer by targeting mitotic kinases with increased specificity.
- To provide proof-of-concept for targeting kinase-activator interactions.
Main Methods:
- Virtual screening of small molecules to identify inhibitors of the Aurora-A/TPX2 interaction.
- In vitro binding assays to confirm Aurora-A binding and TPX2 competition.
- Immunofluorescence assays in osteosarcoma cells to assess Aurora-A activity and spindle defects.
Main Results:
- Identified 25 potential inhibitors of the Aurora-A/TPX2 interaction.
- Confirmed 4 compounds bind Aurora-A and compete for TPX2 binding in vitro.
- Demonstrated that 2 compounds reduce Aurora-A activity and cause spindle defects in cancer cells.
Conclusions:
- Protein-protein interaction inhibitors of the Aurora-A/TPX2 complex are viable therapeutic leads.
- This approach offers increased specificity compared to ATP-binding site inhibitors.
- These findings support a novel strategy for developing targeted anti-cancer drugs.