Department of Surgical Oncology, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
Anticancer Agents Med Chem. 2013 May;13(4):546-54. doi: 10.2174/1871520611313040003.
Focal Adhesion Kinase (FAK) is a non-receptor kinase that is overexpressed in many types of tumors and plays a key role in cell adhesion, spreading, motility, proliferation, invasion, angiogenesis, and survival. Recently, FAK has been proposed as a target for cancer therapy, and we performed computer modeling and screening of the National Cancer Institute (NCI) small molecule compounds database to target the ATP-binding site of FAK, K454. More than 140,000 small molecule compounds were docked into the crystal structure of the kinase domain of FAK in 100 different orientations using DOCK5.1 that identified small molecule compounds, targeting the K454 site, called A-compounds. To find the therapeutic efficacy of these compounds, we examined the effect of twenty small molecule compounds on cell viability by MTT assays in different cancer cell lines. One compound, A18 (1,4-bis(diethylamino)-5,8- dihydroxy anthraquinon) was a mitoxantrone derivative and significantly decreased viability in most of the cells comparable to the to the level of FAK kinase inhibitors TAE-226 (Novartis, Inc) and PF-573,228 (Pfizer). The A18 compound specifically blocked autophosphorylation of FAK like TAE-226 and PF-228. ForteBio Octet Binding assay demonstrated that mitoxantrone (1,4-dihydroxy- 5,8-bis[2-(2-hydroxyethylamino) ethylamino] anthracene-9,10-dione directly binds the FAK-kinase domain. In addition, mitoxantrone significantly decreased the viability of breast cancer cells in a dose-dependent manner and inhibited the kinase activity of FAK and Y56/577 FAK phosphorylation at 10-20 μM. Mitoxantrone did not affect phosphorylation of EGFR, but decreased Pyk-2, c-Src, and IGF-1R kinase activities. The data demonstrate that mitoxantrone decreases cancer viability, binds FAK-Kinase domain, inhibits its kinase activity, and also inhibits in vitro kinase activities of Pyk-2 and IGF-1R. Thus, this novel function of the mitoxantrone drug can be critical for future development of anti-cancer agents and FAK-targeted therapy research.
黏着斑激酶（FAK）是一种非受体激酶，在许多类型的肿瘤中过度表达，在细胞黏附、铺展、运动、增殖、侵袭、血管生成和存活中发挥关键作用。最近，FAK 被提议作为癌症治疗的靶点，我们使用 DOCK5.1 对国家癌症研究所（NCI）小分子化合物数据库进行了计算机建模和筛选，以靶向 FAK 的 ATP 结合位点 K454。使用 DOCK5.1 将超过 140000 种小分子化合物以 100 种不同的取向对接入 FAK 激酶结构域的晶体结构中，鉴定出靶向 K454 位点的小分子化合物，称为 A 型化合物。为了找到这些化合物的治疗效果，我们通过 MTT 测定法在不同的癌细胞系中检查了 20 种小分子化合物对细胞活力的影响。一种化合物 A18（1,4-双（二乙基氨基）-5,8-二羟基蒽醌）是米托蒽醌的衍生物，与 FAK 激酶抑制剂 TAE-226（诺华公司）和 PF-573,228（辉瑞公司）相比，它显著降低了大多数细胞的活力。A18 化合物特异性地阻断了 FAK 的自身磷酸化，就像 TAE-226 和 PF-228 一样。ForteBio Octet 结合测定表明，米托蒽醌（1,4-二羟基-5,8-双[2-（2-羟乙基氨基）乙基氨基]蒽-9,10-二酮）直接与 FAK-激酶结构域结合。此外，米托蒽醌以剂量依赖性方式显著降低乳腺癌细胞的活力，并抑制 FAK 的激酶活性和 Y56/577 FAK 磷酸化在 10-20 μM。米托蒽醌不影响 EGFR 的磷酸化，但降低了 Pyk-2、c-Src 和 IGF-1R 激酶的活性。数据表明，米托蒽醌降低了癌症的活力，结合了 FAK 激酶结构域，抑制了其激酶活性，并且还抑制了 Pyk-2 和 IGF-1R 的体外激酶活性。因此，米托蒽醌药物的这种新功能对于未来抗癌药物和 FAK 靶向治疗研究的发展可能至关重要。
