Division of Hematology, Department of Internal Medicine, Comprehensive Cancer Center, The Ohio State University, Columbus, 455 Wiseman Hall, 400 West 12th Avenue, Columbus, OH, 43210, USA.
Division of Pharmaceutics and Pharmaceutical Chemistry, The Ohio State University, Columbus, OH, USA.
J Hematol Oncol. 2020 Jan 28;13(1):8. doi: 10.1186/s13045-019-0821-7.
Acute myeloid leukemia (AML) is the most common type of adult leukemia. Several studies have demonstrated that oncogenesis in AML is enhanced by kinase signaling pathways such as Src family kinases (SFK) including Src and Lyn, spleen tyrosine kinase (SYK), and bruton's tyrosine kinase (BTK). Recently, the multi-kinase inhibitor ArQule 531 (ARQ 531) has demonstrated potent inhibition of SFK and BTK that translated to improved pre-clinical in vivo activity as compared with the irreversible BTK inhibitor ibrutinib in chronic lymphocytic leukemia (CLL) models. Given the superior activity of ARQ 531 in CLL, and recognition that this molecule has a broad kinase inhibition profile, we pursued its application in pre-clinical models of AML.
The potency of ARQ 531 was examined in vitro using FLT3 wild type and mutated (ITD) AML cell lines and primary samples. The modulation of pro-survival kinases following ARQ 531 treatment was determined using AML cell lines. The effect of SYK expression on ARQ 531 potency was evaluated using a SYK overexpressing cell line (Ba/F3 murine cells) constitutively expressing FLT3-ITD. Finally, the in vivo activity of ARQ 531 was evaluated using MOLM-13 disseminated xenograft model.
Our data demonstrate that ARQ 531 treatment has anti-proliferative activity in vitro and impairs colony formation in AML cell lines and primary AML cells independent of the presence of a FLT3 ITD mutation. We demonstrate decreased phosphorylation of oncogenic kinases targeted by ARQ 531, including SFK (Tyr416), BTK, and fms-related tyrosine kinase 3 (FLT3), ultimately leading to changes in down-stream targets including SYK, STAT5a, and ERK1/2. Based upon in vitro drug synergy data, we examined ARQ 531 in the MOLM-13 AML xenograft model alone and in combination with venetoclax. Despite ARQ 531 having a less favorable pharmacokinetics profile in rodents, we demonstrate modest single agent in vivo activity and synergy with venetoclax.
Our data support consideration of the application of ARQ 531 in combination trials for AML targeting higher drug concentrations in vivo.
急性髓系白血病（AML）是成人中最常见的白血病类型。多项研究表明，癌基因在 AML 中的作用是通过激酶信号通路增强的，如 Src 家族激酶（SFK），包括Src 和 Lyn、脾酪氨酸激酶（SYK）和布鲁顿酪氨酸激酶（BTK）。最近，多激酶抑制剂 ArQule 531（ARQ 531）已被证明能有效抑制 SFK 和 BTK，与慢性淋巴细胞白血病（CLL）模型中的不可逆 BTK 抑制剂 ibrutinib 相比，在体内具有更好的临床前活性。鉴于 ARQ 531 在 CLL 中的优异活性，以及认识到该分子具有广泛的激酶抑制谱，我们在 AML 的临床前模型中探索了其应用。
使用 FLT3 野生型和突变（ITD）AML 细胞系和原代样本在体外检测 ARQ 531 的效力。使用 AML 细胞系测定 ARQ 531 处理后促生存激酶的调节。使用 SYK 过表达细胞系（表达 FLT3-ITD 的 Ba/F3 鼠细胞）评估 SYK 表达对 ARQ 531 效力的影响。最后，使用 MOLM-13 播散性异种移植模型评估 ARQ 531 的体内活性。
我们的数据表明，ARQ 531 治疗在体外具有抗增殖活性，并可损害 AML 细胞系和原代 AML 细胞的集落形成，而与 FLT3 ITD 突变的存在无关。我们证明了 ARQ 531 靶向的致癌激酶磷酸化减少，包括 SFK（Tyr416）、BTK 和 fms 相关酪氨酸激酶 3（FLT3），最终导致下游靶标包括 SYK、STAT5a 和 ERK1/2 的变化。根据体外药物协同数据，我们单独使用 ARQ 531 并与 venetoclax 联合使用在 MOLM-13 AML 异种移植模型中进行了研究。尽管 ARQ 531 在啮齿动物中的药代动力学特征较差，但我们在体内仍显示出适度的单药活性和与 venetoclax 的协同作用。
我们的数据支持考虑将 ARQ 531 应用于 AML 的联合试验，以在体内实现更高的药物浓度。
