Target Identification of Kinase Inhibitor Alisertib (MLN8237) by Using DNA‐Programmed Affinity Labeling

Accurate identification of the molecular targets of bioactive small molecules is a highly important yet challenging task in biomedical research. Previously, a method named DPAL (DNA‐programmed affinity labeling) for labeling and identifying the cellular targets of small molecules and nucleic acids was developed. Herein, DPAL is applied for the target identification of Alisertib (MLN8237), which is a highly specific aurora kinase A (AKA) inhibitor and a drug candidate being tested in clinical trials for cancer treatment. Apart from the well‐established target of AKA, several potential new targets of MLN8237 were identified. Among them, p38 mitogen‐activated protein kinase (p38) and laminin receptor (LAMR) were validated to be implicated in the anticancer activities of MLN8237. Interestingly, these new targets were not identified with non‐DNA‐based affinity probes. This work may facilitate an understanding of the molecular basis of the efficacy and side effects of MLN8237 as a clinical drug candidate. On the other hand, this work has also demonstrated that the method of DPAL could be a useful tool for target identification of bioactive small molecules. Lighting‐up targets: New molecular targets have been identified for Alisertib (MLN8237), which is an aurora kinase A (AKA) inhibitor and an anticancer drug candidate, through the application of DPAL (DNA‐programmed affinity labeling; see figure). The DPAL method could be a useful tool for target identification of bioactive small molecules.