Addressing Undruggable Targets Using Oligonucleotides and Small Molecules (PQ18)

描述(申请人提供)：作为对PQ18的回应，我们建议通过创新地联合使用寡核苷酸和有机小分子来解决通过传统手段无法药物治疗的癌症相关靶点。目前，反义和siRNA寡核苷酸在癌症和其他疾病中的治疗潜力引起了极大的兴趣，多项临床试验正在进行中。然而，无法有效地将这些高极性分子运送到它们在组织内的细胞内作用部位仍然是一个主要障碍。 进步。在这个PQ应用中，我们描述了一种新的寡核苷酸传递的化学生物学策略，这是一种与现在常用的方法完全不同的方法。我们实验室和其他实验室最近的研究表明，寡核苷酸所遵循的细胞摄取和细胞内转运的途径可以对其药理作用产生重大影响。在这里，我们建议使用我们称之为OTECs(‘寡核苷酸转运增强剂化合物’)的小有机分子来操纵寡核苷酸转运途径和随后的药理效应。我们已经利用一组影响早期内体到跨高尔基体运输的小分子，为这一方法建立了原理证明。当使用微摩尔浓度时，这些试剂在细胞培养研究中提供了显着增强的寡核苷酸作用。在PQ18的背景下，我们现在建议：(I)使用OTEC和寡核苷酸来操纵癌症中重要的转录因子的功能；(Ii)测试OTEC化合物是否可以在异种移植瘤中发挥作用；(Iii)使用高通量筛选(HTS)来寻找新的、更有效的OTEC类型的化合物，这些化合物在纳摩尔范围内工作以增强寡核苷酸的作用。这些目标的成功完成将对在癌症治疗中使用寡核苷酸的前景产生变革性的影响，包括解决“不可药物”靶点的能力。 与公共卫生相关：反义、siRNA或剪接开关寡核苷酸可以提供强有力的工具来操纵用传统方法无法下药的治疗靶点。然而，寡核苷酸的药理作用往往是钝化的，因为它们很难进入组织细胞的胞浆和细胞核。在这里，我们追求一种新的方法，通过使用小的有机分子来改变寡核苷酸在细胞内的运输，从而增强它们的药理作用，从而增强寡核苷酸的传递。我成功的这一方法将对寡核苷酸在癌症中的治疗作用产生重大影响。

DESCRIPTION (provided by applicant): In response to PQ18 we propose to address cancer-related targets that are 'undruggable' by conventional means through an innovative conjoint use of oligonucleotides and small organic molecules. There is currently tremendous interest in the therapeutic potential of antisense and siRNA oligonucleotides in cancer and other diseases, with multiple clinical trials underway. However, the inability to effectively deliver these highly polar molecules to their intracellular sites of action within tissues remains a major impediment to progress. In this PQ application we describe a novel chemical biology strategy for oligonucleotide delivery, one that is radically different from the approaches now in common use. Recent studies from our laboratory and others have shown that the pathway of cellular uptake and intracellular trafficking followed by an oligonucleotide can have a major impact on its pharmacological actions. Here we propose to manipulate oligonucleotide trafficking pathways and subsequent pharmacological effects using small organic molecules that we term OTECs ('oligonucleotide trafficking enhancer compounds'). We have already established a proof of principle for this approach using a set of small molecules that affect early endosome to trans-Golgi traffic. These agents provide a marked enhancement of oligonucleotide action in cell culture studies when used at micromolar concentrations. In the context of PQ18 we now propose to: (i) use OTECs and oligonucleotides to manipulate the functions of a transcription factor important in cancer; (ii) test whether OTEC compounds can act in xenograft tumors; (iii) use high throughput screening (HTS) to seek new, more potent OTEC-type compounds that work in the nanomolar range to enhance oligonucleotide action. Successful completion of these aims would have a transformative effect on the prospects for using oligonucleotides in cancer therapy, including the ability to address 'undruggable' targets. PUBLIC HEALTH RELEVANCE: Antisense, siRNA or splice switching oligonucleotides could provide potent tools for manipulating therapeutic targets that are 'undruggable' by conventional means. However, the pharmacological effects of oligonucleotides are often blunted because of their poor delivery into the cytosol and nucleus of tissue cells. Here we pursue a novel approach for enhancing oligonucleotide delivery by using small organic molecules that modify the intracellular trafficking of the oligonucleotides and thus enhance their pharmacological actions. I successful this approach would have a major impact on the therapeutic utility of oligonucleotides in cancer.