Design, synthesis and biological evaluation of novel indolin-2-one derivatives as potent second-generation TRKs inhibitors

Tropomyosin receptor kinases (TRKs) are effective targets for anti-cancer drug discovery. The first-generation type I TRKs inhibitors, larotrectinib and entrectinib, exhibit durable disease control in the clinic. The emergence of acquired resistance mediated by secondary mutations in the TRKs domain significantly reduces the therapeutic efficacy of these two drugs, indicating an unmet clinical need. In this study, we designed a potent and orally bioavailable Trk Inhibitor, compound 24b, using a molecular hybridization strategy. Compound 24b exhibited significant inhibitory potency against multiple TRK mutants in both biochemical and cellular assays. Furthermore, compound 24b induced Apoptosis of Ba/F3-TRKA^G595R and Ba/F3-TRKA^G667C cells in a dose-dependent manner. Additionally, compound 24b exhibited moderate kinase selectivity. In vitro stability revealed that compound 24b showed excellent plasma stability (t_1/2 > 289.1 min) and moderate liver microsomal stability (t_1/2 = 44.3 min). Pharmacokinetic studies have revealed that compound 24b is an orally bioavailable Trk Inhibitor with a good oral bioavailability of 116.07%. These results indicate that compound 24b be used as a lead molecule for further modifications to overcome drug-resistant mutants of TRK.

Anticancer; Drug resistance; NTRK gene Fusion; TRK inhibitors.