Transforming growth factor (TGF)-β triggers the epithelial-to-mesenchymal transition (EMT) of cancer cells via well-orchestrated crosstalk between Smad and non-Smad signaling pathways, including Wnt/β-catenin. Since EMT-induced motility and invasion play a critical role in cancer metastasis, EMT-related molecules are emerging as novel targets of anti-cancer therapies. Traf2- and Nck-interacting kinase (TNIK) has recently been considered as a first-in-class anti-cancer target molecule to regulate Wnt signaling pathway, but pharmacologic inhibition of its EMT activity has not yet been studied. Here, using 5-(4-methylbenzamido)-2-(phenylamino)thiazole-4-carboxamide (KY-05009) with TNIK-inhibitory activity, its efficacy to inhibit EMT in cancer cells was validated. The molecular docking/binding study revealed the binding of KY-05009 in the hinge region of TNIK, and the inhibitory activity of KY-05009 against TNIK was confirmed by an ATP competition assay (Ki, 100 nM). In A549 cells, KY-05009 significantly and strongly inhibited the TGF-β-activated EMT through the attenuation of Smad and non-Smad signaling pathways, including the Wnt, NF-κB, FAK-Src-paxillin-related focal adhesion, and MAP kinases (ERK and JNK) signaling pathways. Continuing efforts to identify and validate potential therapeutic targets associated with EMT, such as TNIK, provide new and improved therapies for treating and/or preventing EMT-based disorders, such as cancer metastasis and fibrosis.

中文翻译：

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一种新型氨基噻唑 KY-05009 具有抑制 Traf2 和 Nck 相互作用激酶 (TNIK) 的潜力，可减弱人肺腺癌 A549 细胞中 TGF-β1 介导的上皮间质转化。

转化生长因子 (TGF)-β 通过 Smad 和非 Smad 信号通路（包括 Wnt/β-catenin）之间精心安排的串扰触发癌细胞的上皮间质转化 (EMT)。由于 EMT 诱导的运动和侵袭在癌症转移中起关键作用，因此 EMT 相关分子正在成为抗癌治疗的新靶点。Traf2 和 Nck 相互作用激酶（TNIK）最近被认为是调节 Wnt 信号通路的一流抗癌靶分子，但尚未研究其 EMT 活性的药理学抑制。在这里，使用具有 TNIK 抑制活性的 5-(4-methylbenzamido)-2-(phenylamino)thiazole-4-carboxamide (KY-05009)，验证了其抑制癌细胞 EMT 的功效。分子对接/结合研究揭示了 KY-05009 在 TNIK 铰链区的结合，并且 KY-05009 对 TNIK 的抑制活性通过 ATP 竞争测定 (Ki, 100 nM) 得到证实。在 A549 细胞中，KY-05009 通过减弱 Smad 和非 Smad 信号通路，包括 Wnt、NF-κB、FAK-Src-桩蛋白相关的粘着斑和 MAP，显着且强烈地抑制 TGF-β 激活的 EMT激酶（ERK 和 JNK）信号通路。继续努力识别和验证与 EMT 相关的潜在治疗靶点，例如 TNIK，为治疗和/或预防基于 EMT 的疾病（例如癌症转移和纤维化）提供新的和改进的疗法。KY-05009 通过减弱 Smad 和非 Smad 信号通路，包括 Wnt、NF-κB、FAK-Src-桩蛋白相关粘着斑和 MAP 激酶（ERK 和JNK) 信号通路。继续努力识别和验证与 EMT 相关的潜在治疗靶点，例如 TNIK，为治疗和/或预防基于 EMT 的疾病（例如癌症转移和纤维化）提供新的和改进的疗法。KY-05009 通过减弱 Smad 和非 Smad 信号通路，包括 Wnt、NF-κB、FAK-Src-桩蛋白相关粘着斑和 MAP 激酶（ERK 和JNK) 信号通路。继续努力识别和验证与 EMT 相关的潜在治疗靶点，例如 TNIK，为治疗和/或预防基于 EMT 的疾病（例如癌症转移和纤维化）提供新的和改进的疗法。