Journal Article

Stereospecific targeting of MTH1 by (S)-crizotinib as an anticancer strategy

Knapp, Stefan, Stukalov, Alexey, Loizou, Joanna I., Pham, Therese, Helleday, Thomas, Superti-Furga, Giulio, Jemth, Ann-Sofie, Sanjiv, Kumar, Colinge, Jacques, Bennett, Keiryn L., Gridling, Manuela, Huber, Kilian V. M., Radic, Branka, Strömberg, Kia, Berglund, Ulrika Warpman, Salah, Eidarus, Elkins, Jonathan M., Göktürk, Camilla

2014

Overview

Activated RAS GTPase signalling is a critical driver of oncogenic transformation and malignant disease. Cellular models of RAS-dependent cancers have been used to identify experimental small molecules, such as SCH51344, but their molecular mechanism of action remains generally unknown. Here, using a chemical proteomic approach, we identify the target of SCH51344 as the human mutT homologue MTH1 (also known as NUDT1), a nucleotide pool sanitizing enzyme. Loss-of-function of MTH1 impaired growth of KRAS tumour cells, whereas MTH1 overexpression mitigated sensitivity towards SCH51344. Searching for more drug-like inhibitors, we identified the kinase inhibitor crizotinib as a nanomolar suppressor of MTH1 activity. Surprisingly, the clinically used ( R )-enantiomer of the drug was inactive, whereas the ( S )-enantiomer selectively inhibited MTH1 catalytic activity. Enzymatic assays, chemical proteomic profiling, kinome-wide activity surveys and MTH1 co-crystal structures of both enantiomers provide a rationale for this remarkable stereospecificity. Disruption of nucleotide pool homeostasis via MTH1 inhibition by ( S )-crizotinib induced an increase in DNA single-strand breaks, activated DNA repair in human colon carcinoma cells, and effectively suppressed tumour growth in animal models. Our results propose ( S )-crizotinib as an attractive chemical entity for further pre-clinical evaluation, and small-molecule inhibitors of MTH1 in general as a promising novel class of anticancer agents. A chemoproteomic screen is used here to identify MTH1 as the target of SCH51344, an experimental RAS-dependent cancer drug; a further search for inhibitors revealed ( S )-crizotinib as a potent MTH1 antagonist, which suppresses tumour growth in animal models of colon cancer, and could be part of a new class of anticancer drugs. MTH1 is Ras-linked target for cancer therapy Mutations in the Ras oncogene are associated with poor prognosis. It was known that overexpression of MTH1, a protein involved in preventing the incorporation of damaged bases into DNA, prevents Ras-induced senescence. In seeking to understand how damaged deoxynucleotides (dNTPs) promote cancer, Thomas Helleday and colleagues found that MTH1 activity is essential for the survival of transformed cells, and isolated two small-molecule MTH1 inhibitors, TH287 and TH588. In the presence of these hydrolase inhibitors, damaged nucleotides are incorporated into DNA only in cancer cells, causing cytotoxicity and eliciting a beneficial response in mouse xenograft cancer models. In a second study, Giulio Superti-Furga and colleagues sought to identify the target of a small molecule, SCH51344, that had been developed for use against Ras -dependent cancers and found that it inactivates MTH1. This allowed them to identify a new potent inhibitor of MTH1 that is enantiomer-selective, ( S )-crizotinib. In the presence of this drug, tumour growth is suppressed in animal models of colon cancer.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 13/1 / 13/106 / 13/44 / 13/51 / 13/89 / 14/63 / 631/154/555 / 631/67/1059/153 / 631/92/613 / 82/58 / Aminoquinolines - pharmacology / Animal growth / Animal models / Animals / Antineoplastic Agents - chemistry / Antineoplastic Agents - pharmacology / Cancer / Colon cancer / Colonic Neoplasms - drug therapy / Colonic Neoplasms - genetics / Colonic Neoplasms - pathology / Crizotinib / Crystallization / Deoxyribonucleic acid / Disease Models, Animal / DNA / DNA Breaks, Single-Stranded - drug effects / DNA Repair / DNA Repair Enzymes - antagonists & inhibitors / DNA Repair Enzymes - biosynthesis / DNA Repair Enzymes - chemistry / DNA Repair Enzymes - metabolism / Enzymes / Experiments / Female / Genetic aspects / Homeostasis / Homeostasis - drug effects / Humanities and Social Sciences / Humans / Inhibitor drugs / Kinases / Life Sciences / Medical prognosis / Mice / Mice, SCID / Models, Molecular / multidisciplinary / Mutation / Nucleotides / Nucleotides - metabolism / Oxidative stress / Phosphoric Monoester Hydrolases - antagonists & inhibitors / Phosphoric Monoester Hydrolases - biosynthesis / Phosphoric Monoester Hydrolases - chemistry / Phosphoric Monoester Hydrolases - metabolism / Physiological aspects / Protein Conformation / Protein Kinase Inhibitors - chemistry / Protein Kinase Inhibitors - pharmacology / Proteomics / Proto-Oncogene Proteins - genetics / Proto-Oncogene Proteins p21(ras) / Pyrazoles - chemistry / Pyrazoles - pharmacology / Pyridines - chemistry / Pyridines - pharmacology / ras Proteins - genetics / Science / Studies / Substrate Specificity / Xenograft Model Antitumor Assays