Preclinical Development of First-in-Class NDUFS7 Antagonists for the Treatment of Pancreatic Cancer

氧化磷酸化(OXPHOS)在线粒体功能中起重要作用 生物能量学、生物量生产和氧化还原平衡，这些都是肿瘤进展的基本过程， 转移和存活率。因此，靶向OXPHOS对治疗PDAC具有很大的前景。临床用二甲双胍 使用OXPHOS抑制剂，已经证明PDAC患者的存活率增加，这表明OXPHOS 抑制剂可以安全地用于治疗PDAC。关于OXPHOS的基本作用，现在有了更清晰的图景 在PDAC进展中。例如，OXPHOS基因的过度表达与PDAC的预后不良相关 患者对线粒体氧耗的抑制显著延缓了PDAC的进展。药效 在KRAS中消融的耐药PDAC细胞严重依赖OXPHOS，并对OXPHOS敏感 抑制力。胰腺肿瘤启动细胞也表现出对OXPHOS的强烈依赖。重要的是，OXPHOS 抑制剂与吉西他滨具有协同作用，特别是在具有高水平OXPHOS相关基因的细胞中。 遗憾的是，二甲双胍的效价低，选择性差。其他经典的OXPHOS抑制剂包括 鱼藤酮、抗霉素A和寡霉素缺乏选择性。我们最近设计了一系列高效的氧磷酸盐 抑制剂(DX2-201至DX3-236)。我们由ADMET指导的销售线索优化活动产生了200多篇 导致发现具有纳摩尔效力的口服活性OXPHOS抑制剂的类似物，可接受 毒性，合理的器官分布，以推进临床开发。我们描绘了一组105例癌症 细胞系对DX类似物的抗增殖反应，发现我们的化合物深刻地抑制了 7个PDAC细胞系中有5个细胞系增殖。通过产生抗药性克隆，其次是 整个外显子组测序，我们确定NDUFS7为我们的先导化合物DX2-201的6/6的直接靶点 克隆人。我们的综合多组学(rna-seq、bru-seq和基于质谱学的蛋白质组学)研究 发现DX处理后DNA修复基因表达下调。我们的DX化合物也显示出显著的 与标准护理化疗和几种FDA批准的药物的协同作用。我们假设 OXPHOS抑制剂将为PDAC患者提供治疗益处，克服耐药性，并可能 安全有效地与包括免疫检查点抑制剂在内的各种药物联合使用。我们将对此进行测试 通过以下具体目标进行假说：目的1.评估前5种化合物作为单一药物的细胞毒性 与标准化疗以及大型早期临床候选药物相结合 小鼠和人PDAC细胞系的面板。目的2.进行MTD，完整的PK/PD研究，并确定组织 前5种化合物在小鼠体内的分布和积累。目的3.测定TOP5的体内药效 作为单一药物的化合物与来自AIM 1的标准护理药物(S)在原位和 KRAS诱发胰腺癌的GEMM模型。

Oxidative phosphorylation (OXPHOS) plays an important role in mitochondria function by mediating bioenergetics, biomass production, and redox balance, all essential processes for tumor progression, metastasis, and survival. Thus, targeting OXPHOS holds great promise to treat PDAC. Metformin, a clinically used OXPHOS inhibitor, has demonstrated increased survival in PDAC patients, suggesting that OXPHOS inhibitors can safely be used to treat PDAC. A clearer picture is now emerging on the essential role of OXPHOS in PDAC progression. For example, overexpression of OXPHOS genes correlates with poor prognosis in PDAC patients, and suppression of mitochondrial oxygen consumption significantly retards PDAC progression. Drug resistant PDAC cells ablated in KRAS are heavily dependent on OXPHOS and are sensitive to OXPHOS inhibition. Pancreatic-tumor-initiating cells also show strong reliance on OXPHOS. Importantly, OXPHOS inhibitors are synergistic with gemcitabine specifically in cells with high levels of OXPHOS-related genes. Unfortunately, metformin has low potency and poor selectivity. Other classic OXPHOS inhibitors including rotenone, antimycin A, and oligomycin, lack selectivity. We recently designed a series of highly potent OXPHOS inhibitors (DX2-201 to DX3-236). Our ADMET-guided lead optimization campaign generated over 200 novel analogues that led to the discovery of orally active OXPHOS inhibitors with nanomolar potency, acceptable toxicity, and reasonable organ distributions to advance into clinical development. We profiled a set of 105 cancer cell lines for their anti-proliferative response to DX analogues and found that our compounds profoundly inhibit the proliferation of 5 out of 7 PDAC cell lines. Through the generation of drug resistance clones, followed by whole exome sequencing, we identified NDUFS7 as the direct target of our lead compound DX2-201 in 6/6 clones. Our integrated multi-omics (RNA-seq, Bru-seq and mass spectrometry-based proteomics) studies revealed down-regulation of DNA repair genes upon DX treatment. Our DX compounds also show significant synergism with standard-of-care chemotherapy and several FDA-approved drugs. We hypothesize that OXPHOS inhibitors will provide therapeutic benefit to PDAC patients, overcome drug resistance, and could be safely and efficaciously combined with various drugs including immune checkpoint inhibitors. We will test this hypothesis through the following specific aims: Aim 1. To assess cytotoxicity of top 5 compounds as single agent and in combination with standard chemotherapy as well as several early-stage clinical candidates in a large panel of mouse and human PDAC cell lines. Aim 2. To perform MTD, full PK/PD studies, and to determine tissue distribution and accumulation of top 5 compounds in mice. Aim 3. To determine the in vivo efficacy of top 5 compounds as single agents and in combination with standard-of-care drug(s) from Aim 1 in orthotopic and GEMM models of KRAS driven pancreatic cancer.