Small Molecule Inhibitors of the Inflammatory Cytokine Oncostatin M

项目摘要/摘要 该项目将合成和研究小分子抑制物(SMI)，这些小分子抑制物结合并抑制细胞外信号转导。 炎性细胞因子，抑癌素M(OSM)，作为FDA批准的药物长期计划的一部分 用于预防乳腺癌转移和其他OSM相关疾病的早期保护。2021年，超过 预计美国将新增28.1万例浸润性乳腺癌病例。五年 被诊断为局部疾病的患者存活率为99%，而远处疾病患者的存活率降至27% 转移，强调了对预防早期转移的治疗的需要，当 干预可能是最有益的。与这一提议相关的是，OSM信号促进了许多转移- 相关事件，包括上皮细胞向间充质细胞的转变和肿瘤细胞的分离、迁移和 入侵。重要的是，OSM信号与乳腺癌患者预后不良有关。 乳腺癌患者肿瘤组织OSM水平高与生存率显著下降相关 与低OSM水平的人相比。还测定了两组患者的OSM血清浓度 非转移性和转移性乳腺癌患者的血清水平显著高于健康人 个人(分别是3.8倍和4.9倍)。对于这一提议，初步实验使用了高 通过计算筛选出与OSM结合的约165万种化合物。这些化合物带有 对体外抑制OSM信号的最佳预测结合特性进行了测试。确定了两个SMI 作为这项提议的先导化合物。指导拟议实验的中心假设是 基于铅SMI的结构优化，通过重要结合相互作用的知识促进， 将导致对OSM信号的增强抑制。为了检验这一假设，我们提出了两个具体目标： 1)设计并合成了基于SMI-10和SMI-2的OSM小分子抑制剂库。 26个支架和2)确定抑制细胞内OSM信号并显示最小细胞数的最佳主要SMI 毒性。在目标1中，将求解SMI/OSM络合物的核磁共振溶液结构，然后与 开发结构-活动模型的计算建模，该模型将用于设计和合成新的 SMIS。在目标2中，基于高结合亲和力、高体外抑制OSM介导的SMI将被优先考虑 在几个乳腺癌细胞系中的信号通路，以及低细胞毒性。该项目的最终目标是 鉴定与OSM具有纳米分子结合亲和力并提高抑制活性的新化合物。建议数 研究将有助于满足开发转移性乳腺癌新疗法的迫切需要--通常 被描述为“不治之症”。因为OSM与其他癌症和炎症相关 疾病(如脓毒症、类风湿性关节炎和囊性纤维化)，这项研究为新的类别奠定了基础 抗炎药。

PROJECT SUMMARY/ABSTRACT This project will synthesize and study small molecules inhibitors (SMIs) that bind to and inhibit signaling of the inflammatory cytokine, oncostatin M (OSM) as part of a long-term program to generate an FDA-approved drug for early protection against breast tumor metastasis and other OSM-associated diseases. In 2021, more than 281,000 new cases of invasive breast cancer are expected to be diagnosed in the United States. The five-year survival rate is 99% for patients diagnosed with localized disease but drops to 27% for those with distant metastases, underscoring a need for therapeutics that protect against the early stages of metastasis, when intervention could be most beneficial. Relevant to this proposal, OSM signaling promotes many metastatic- related events, including an epithelial to mesenchymal transition and tumor cell detachment, migration, and invasion. Importantly, OSM signaling is associated with a poor prognosis for breast cancer patients. Breast cancer patients with high levels of tumor tissue OSM correlated with a significant decrease in survival compared to those with low OSM levels. It was also determined that serum concentrations of OSM from both non-metastatic and metastatic breast cancer patients was significantly higher than levels in serum from healthy individuals (3.8-fold and 4.9-fold, respectively). For this proposal, preliminary experiments used a high throughput computational screen of ~1.65 million compounds for binding to OSM. Those compounds with the best predicted binding properties were tested for in vitro inhibition of OSM signaling. Two SMIs were identified as lead compounds for this proposal. The central hypothesis guiding the proposed experiments is that structural optimization based on lead SMIs, facilitated by knowledge of important binding interactions, will result in enhanced inhibition of OSM signaling. To test the hypothesis, two specific aims are proposed: 1) Design and synthesize a focused library of small molecule inhibitors of OSM based on the SMI-10 and SMI- 26 scaffolds and 2) Identify optimal lead SMIs that suppress OSM signaling in cells and display minimal cellular toxicity. In Aim 1, the NMR solution structure of an SMI/OSM complex will be solved and then combined with computational modeling to develop a structure-activity model that will be used to design and synthesize new SMIs. In Aim 2, SMIs will be prioritized based on high binding affinity, high in vitro inhibition of OSM-mediated signaling pathways in several breast cancer cell lines, and low cellular toxicity. The end goal of the project is to identify new compounds with nanomolar binding affinity to OSM and improved inhibitory activity. The proposed research will assist in fulfilling the critical need to develop novel treatments for metastatic breast cancer—often described as an “incurable” disease. Since OSM has been linked to other cancers and inflammatory-related diseases (e.g., sepsis, rheumatoid arthritis, and cystic fibrosis), this research lays the groundwork for a new class of anti-inflammatory drugs.