PO.CL07.04 · 临床研究
AOH1996及下一代caPCNA抑制剂与靶向及化疗药物联合的抗肿瘤协同作用
Antitumor synergy of AOH1996 and next-generation caPCNA Inhibitors in combination with targeted and chemotherapeutic agents
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：AOH1996是一种小分子，选择性靶向增殖细胞核抗原的癌症相关异构体（caPCNA），通过抑制转录-复制冲突的有效解决来破坏DNA复制和修复，并促进DNA链断裂。基于其有前景的临床前疗效，我们也在开发和测试具有改善效力和成药性的下一代caPCNA抑制剂。我们已开发出两种新型抑制剂，相对于母体化合物展现出高达10倍的细胞效力，以及一种与母体化合物效力相似但溶解度高2.4倍的抑制剂。鉴于PCNA在癌细胞赖以生存的诸多过程中发挥多样化作用，例如协调DNA合成、损伤耐受、细胞周期控制、染色质组装和应激反应信号传导，难以预测哪些其他抗癌药物会与AOH1996及下一代抑制剂有效联合。为此，我们用AOH1996及三种下一代抑制剂筛选了一个包含179种抗癌药物的组合，以鉴定可能表现出协同作用的潜在组合。
方法：使用AOH1996及下一代抑制剂与179种不同抗癌药物的药物组合进行了系统性联合药物筛选。协同作用评估：我们使用人神经母细胞瘤细胞系SK-N-AS进行了初始筛选。我们采用Chou-Talalay法进行药物联合分析来评估组合的协同作用。对于有利的药物组合，我们测试了额外的细胞系，包括所鉴定药物在治疗方案中会被临床使用的癌症细胞类型。机制研究：我们通过流式细胞术分析细胞周期效应和凋亡。我们还通过免疫印迹和免疫荧光测量gammaH2AX、切割的PARP和磷酸化Chk1，以评估DNA损伤和检查点激活。结果：AOH1996及其下一代类似物均与某些DNA损伤和修复抑制药物（如顺铂）表现出协同作用。酪氨酸激酶抑制剂（TKIs）尤其是EGFR TKI与AOH1996及下一代抑制剂协同良好。线粒体凋亡通路抑制剂（如venetoclax）也表现出协同作用。
结论：AOH1996及其下一代类似物与靶向治疗和DNA修复抑制剂表现出强效的协同活性，支持PCNA抑制作为一种增强治疗疗效和克服耐药的有前景策略。这些发现为AOH1996联合方案的临床开发以及具有卓越效力和耐受性的caPCNA靶向类似物的持续优化提供了临床前依据。
查看英文原文 English abstract
Background: AOH1996 is a small molecule that selectively targets the cancer-associated isoform of proliferating cell nuclear antigen (caPCNA), disrupting DNA replication and repair, and promoting DNA strand breaks through inhibition of effective resolution of transcription-replication conflicts. Building on its promising preclinical efficacy, we are also developing and testing next-generation caPCNA inhibitors with improved potency and druglikeness. We have developed two new inhibitors that exhibit up to 10-fold higher cellular potency relative to the parent compound, and an inhibitor with similar potency as the parent compound but 2.4-fold more soluble. Given PCNA's diverse role in processes that cancer cells depend upon for survival, e.g., coordinating DNA synthesis, damage tolerance, cell cycle control, chromatin assembly, and stress response signaling, it is hard to predict what other anticancer drugs would combine productively with AOH1996 and next-generation inhibitors. To that end, we screened a panel of 179 anticancer drugs with AOH1996 and three next-generation inhibitors to identify potential combinations that exhibit synergy.
Methods: A systematic combination drug screen was performed using AOH1996 and next-generation inhibitors and a drug panel of 179 different anticancer drugs. Synergy evaluation: We performed initial screens using the human neuroblastoma cell line SK-N-AS. We assessed the synergy of the combinations using the Chou-Talalay method for drug combination analysis. For favorable drug combinations, we tested additional cell lines including cancer cell types where the drug identified would be used clinically in a treatment regimen. Mechanistic studies: We analyzed cell cycle effects and apoptosis by flow cytometry. We also assessed DNA damage and checkpoint activation by using immunoblotting and immunofluorescence to measure gammaH2AX, cleaved PARP, and phospho-Chk1. Results: Both AOH1996 and its next-generation analogs demonstrated synergy with some DNA-damaging and repair inhibiting agents such as cisplatin. Tyrosine kinase inhibitors (TKIs) in particular EGFR TKIs synergized well with AOH1996 and next-generation inhibitors. Inhibitors of the mitochondrial apoptosis pathway (e.g., venetoclax) also synergized as well.
Conclusions: AOH1996 and its next-generation analogs exhibit potent synergistic activity with targeted therapies and DNA repair inhibitors, supporting PCNA inhibition as a promising strategy to enhance therapeutic efficacy and overcome resistance. These findings provide a preclinical rationale for clinical development of AOH1996 combination regimens, as well as continued optimization of caPCNA-targeting analogs with superior potency and tolerability.
利益披露 Disclosure
R. G. Lingeman, None.