Journal Article

Selective and reversible modification of kinase cysteines with chlorofluoroacetamides

Abe, Yoshito, Okamoto, Kei, Ueda, Tadashi, Tamura, Tomonori, Hamachi, Itaru, Shindo, Naoya, Ono, Mayumi, Tokunaga, Keisuke, Miura, Chizuru, Caaveiro, Jose M. M., Fuchida, Hirokazu, Shibata, Tomohiro, Morimoto, Satoshi, Ojida, Akio, Hatsuyama, Yuji, Sakamoto, Seiichi, Koyanagi, Satoru, Shiroishi, Mitsunori, Ohdo, Shigehiro, Matsunaga, Naoya, Yamaguchi, Yasuchika, Nakao, Takaharu, Kuwata, Keiko, Watari, Kosuke, Sato, Mami

2019

Overview

Irreversible inhibition of disease-associated proteins with small molecules is a powerful approach for achieving increased and sustained pharmacological potency. Here, we introduce α-chlorofluoroacetamide (CFA) as a novel warhead of targeted covalent inhibitor (TCI). Despite weak intrinsic reactivity, CFA-appended quinazoline showed high reactivity toward Cys797 of epidermal growth factor receptor (EGFR). In cells, CFA-quinazoline showed higher target specificity for EGFR than the corresponding Michael acceptors in a wide concentration range (0.1–10 μM). The cysteine adduct of the CFA derivative was susceptible to hydrolysis and reversibly yielded intact thiol but was stable in solvent-sequestered ATP-binding pocket of EGFR. This environment-dependent hydrolysis can potentially reduce off-target protein modification by CFA-based drugs. Oral administration of CFA quinazoline NS-062 significantly suppressed tumor growth in a mouse xenograft model. Further, CFA-appended pyrazolopyrimidine irreversibly inhibited Bruton’s tyrosine kinase with higher target specificity. These results demonstrate the utility of CFA as a new class warheads for TCI. Discovery and exploitation of inherent reaction features of chlorofluoroacetamide (CFA) as a warhead such as low off-target activity and reversible reactivity with cysteine enable specific covalent inhibition of targeted kinases.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 631/154/309 / 631/67 / 631/92/275 / 631/92/613 / Acetamides - chemical synthesis / Acetamides - chemistry / Acetamides - pharmacology / Amino acids / Animals / Antineoplastic Agents / Biochemical Engineering / Biochemistry / Bioorganic Chemistry / Cell Biology / Cell Line / Chemical bonds / Chemistry / Chemistry and Materials Science / Chemistry/Food Science / Cysteine - metabolism / Epidermal growth factor / Epidermal growth factor receptors / ErbB Receptors / Growth factors / Humans / Hydrolysis / Kinases / Lung cancer / Mice / Mice, Nude / Neoplasms / Oral administration / Peptides / Pharmaceutical sciences / Pharmacology / Phosphotransferases - physiology / Protein Kinase Inhibitors - metabolism / Protein Kinase Inhibitors - pharmacology / Protein-tyrosine kinase / Proteins / Pyrimidines - antagonists & inhibitors / Quinazolines - chemical synthesis / Quinazolines - chemistry / Structure-Activity Relationship / Tyrosine / Warheads / Xenograft Model Antitumor Assays / Xenografts / Xenotransplantation