Frame Sheelagh, Saladino Chiara, MacKay Craig, Atrash Butrus, Sheldrake Peter, McDonald Edward, Clarke Paul A, Workman Paul, Blake David, Zheleva Daniella
Cyclacel Limited, Dundee, United Kingdom.
Cancer Research UK Cancer Therapeutics Unit, Division of Cancer Therapeutics, The Institute of Cancer Research, London, United Kingdom.
PLoS One. 2020 Jul 9;15(7):e0234103. doi: 10.1371/journal.pone.0234103. eCollection 2020.
Cyclin-dependent kinases (CDKs) contribute to the cancer hallmarks of uncontrolled proliferation and increased survival. As a result, over the last two decades substantial efforts have been directed towards identification and development of pharmaceutical CDK inhibitors. Insights into the biological consequences of CDK inhibition in specific tumor types have led to the successful development of CDK4/6 inhibitors as treatments for certain types of breast cancer. More recently, a new generation of pharmaceutical inhibitors of CDK enzymes that regulate the transcription of key oncogenic and pro-survival proteins, including CDK9, have entered clinical development. Here, we provide the first disclosure of the chemical structure of fadraciclib (CYC065), a CDK inhibitor and clinical candidate designed by further optimization from the aminopurine scaffold of seliciclib. We describe its synthesis and mechanistic characterization. Fadraciclib exhibits improved potency and selectivity for CDK2 and CDK9 compared to seliciclib, and also displays high selectivity across the kinome. We show that the mechanism of action of fadraciclib is consistent with potent inhibition of CDK9-mediated transcription, decreasing levels of RNA polymerase II C-terminal domain serine 2 phosphorylation, the pro-survival protein Myeloid Cell Leukemia 1 (MCL1) and MYC oncoprotein, and inducing rapid apoptosis in cancer cells. This cellular potency and mechanism of action translate to promising anti-cancer activity in human leukemia mouse xenograft models. Studies of leukemia cell line sensitivity identify mixed lineage leukemia (MLL) gene status and the level of B-cell lymphoma 2 (BCL2) family proteins as potential markers for selection of patients with greater sensitivity to fadraciclib. We show that the combination of fadraciclib with BCL2 inhibitors, including venetoclax, is synergistic in leukemic cell models, as predicted from simultaneous inhibition of MCL1 and BCL2 pro-survival pathways. Fadraciclib preclinical pharmacology data support its therapeutic potential in CDK9- or CDK2-dependent cancers and as a rational combination with BCL2 inhibitors in hematological malignancies. Fadraciclib is currently in Phase 1 clinical studies in patients with advanced solid tumors (NCT02552953) and also in combination with venetoclax in patients with relapsed or refractory chronic lymphocytic leukemia (CLL) (NCT03739554) and relapsed refractory acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS) (NCT04017546).
细胞周期蛋白依赖性激酶(CDK)促成了不受控制的增殖和存活率增加等癌症特征。因此,在过去二十年中,人们付出了巨大努力来鉴定和开发CDK药物抑制剂。对特定肿瘤类型中CDK抑制的生物学后果的深入了解,已成功开发出CDK4/6抑制剂用于治疗某些类型的乳腺癌。最近,新一代调节关键致癌和促生存蛋白转录的CDK酶药物抑制剂,包括CDK9抑制剂,已进入临床开发阶段。在此,我们首次披露了法德曲利布(CYC065)的化学结构,它是一种CDK抑制剂和临床候选药物,是在塞利西利布氨基嘌呤骨架的基础上进一步优化设计而成。我们描述了它的合成和作用机制特征。与塞利西利布相比,法德曲利布对CDK2和CDK9表现出更高的效力和选择性,并且在整个激酶组中也显示出高选择性。我们表明,法德曲利布的作用机制与有效抑制CDK9介导的转录一致,降低RNA聚合酶II C末端结构域丝氨酸2磷酸化水平、促生存蛋白髓系细胞白血病1(MCL1)和MYC癌蛋白水平,并诱导癌细胞快速凋亡。这种细胞活性和作用机制在人白血病小鼠异种移植模型中转化为有前景的抗癌活性。白血病细胞系敏感性研究确定混合谱系白血病(MLL)基因状态和B细胞淋巴瘤2(BCL2)家族蛋白水平是选择对法德曲利布更敏感患者的潜在标志物。我们表明,法德曲利布与包括维奈克拉在内的BCL2抑制剂联合使用在白血病细胞模型中具有协同作用,这与同时抑制MCL1和BCL2促生存途径的预测结果一致。法德曲利布的临床前药理学数据支持其在CDK9或CDK2依赖性癌症中的治疗潜力,以及作为血液系统恶性肿瘤中与BCL2抑制剂合理联合使用的潜力。法德曲利布目前正在晚期实体瘤患者中进行1期临床研究(NCT02552953),也正在与维奈克拉联合用于复发或难治性慢性淋巴细胞白血病(CLL)患者(NCT03739554)以及复发难治性急性髓系白血病(AML)或骨髓增生异常综合征(MDS)患者(NCT04017546)。