Novel Dual Notch/PXR Targeting for Colon Cancer Therapy

用于结肠癌治疗的新型双Notch/PXR靶向

• 批准号: 8627820
• 负责人: Shrikant Anant
• 金额: $ 51.17万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8627820
• 关键词: Adverse drug effect; Affect; Architecture; Binding; Binding Sites; Biological Availability; Biological Markers; Calmodulin; Cancer Etiology; Cell Death; Cell Maintenance; Cells; Cessation of life; Clinical Research; Clinical Trials; Colon Carcinoma; Data; Development; Dose; Drug Industry; Drug Kinetics; Drug Targeting; Drug resistance; Drug toxicity; Endothelial Cells; Epithelial Cells; Fibroblasts; Fluorouracil; Future; Gene Activation; Goals; Growth; HCT116 Cells; In Vitro; Lead; Mediating; Methods; MicroRNAs; Modeling; Morbidity - disease rate; Mus; Neoplasm Metastasis; Notch Signaling Pathway; Oral; Oral Administration; Pathway interactions; Pharmaceutical Preparations; Pharmacodynamics; Phosphorylation; Phosphotransferases; Proteins; Receptor Activation; Receptor Gene; Regimen; Research; Role; Signal Transduction; Stem cells; System; Therapeutic; Therapeutic Agents; Toxic effect; Trans-Activators; Treatment Efficacy; United States; Water; Woman; Work; Xenograft procedure; analog; base; cancer cell; cancer therapy; cell type; chemotherapeutic agent; cis acting element; in vivo; intravenous administration; killings; men; mortality; mouse model; new therapeutic target; notch protein; novel; novel therapeutic intervention; novel therapeutics; pre-clinical; preclinical safety; preclinical study; pregnane X receptor; prevent; promoter; public health relevance; receptor expression; research study; therapeutic target; treatment strategy; tumor; tumor microenvironment; tumor xenograft

SUMMARY Colon cancer remains a leading cause of cancer related morbidity and mortality, both in the US and around the world. Many therapeutic agents and their combinations are being used to inhibit the growth and metastasis of the tumor. However, a couple of significant problems with these strategies are the development of resistance to the drugs and the increased side effects of the drugs. A critical reason for the drug resistance is that directly or indirectly targeted therapeutics through various kinase pathways, they activate the pregnane X receptor (PXR). Hence, novel targeted therapeutics is essential that suppress specific pathways but do not induce PXR. In this regard, we have developed a novel drug MRLTHB and water-soluble analog MRLTHBCD, which inhibits Notch-1 signaling and does not induce PXR. The goal of the current project is to further characterize the drug and generate preclinical data as an oral therapeutic both alone and in combination with 5-fluorouracil (5-FU) for colon cancer. In previous studies, we have determined that downregulating Doublecortin calmodulin-like kinase 1 (DCLK1) suppresses colon cancer xenograft growth, suggesting that targeting DCLK1 would be an efficient strategy for colon cancers. We have now determined that THB and THBCD specifically inhibit DCLK1 kinase activity and but do not affect the kinase activity calmodulin like kinases CAMKII and CAMKIV. In addition, we have determined that the compounds inhibit the growth of colon cancer cells in a novel culture method that we have developed termed "Tumor in a Dish" (TiD) where cancer cells are grown in a three-dimensional culture that includes normal epithelial cells, fibroblasts and endothelial cells. The model creates an in vivo-like tumor microenvironment that provides the necessary cell-cell contact, 3D-architecture, and the influence of different cell types. The observed selective killing of cancer cells in this system suggests that the compounds are highly specific and have good potency. Mechanistically, we have determined that the compounds inhibit the Notch signaling pathway and PXR expression. Based on our preliminary studies, DCLK1 targeting by THB and THBCD resulting in suppression of both Notch signaling and PXR is a valid therapeutic strategy for colon cancers. We aim to continue developing preclinical data in the current application. In aim 1, we propose to determine the role of Notch-1 in PXR expression. In aim 2, we will perform detailed PK/PD studies of the compound and find the optimal dose to perform preclinical studies in xenotransplant and APCmin/+ mouse models. In aim 3, we propose then to continue and determine the effect of the combination of THB and THBCD with 5-fluorouracil to inhibit colon cancer growth. Effect of the compounds on Akt phosphorylation, Notch-1 activation, and DCLK1 and PXR expression in the tumor will be determined. These proposed studies would provide compelling mechanistic evidence for initiating clinical trials for the novel compounds alone and in combination. These studies will also aid in optimizing a targeted chemotherapeutic regimen and identify novel biomarkers for the future clinical studies.

总结 结肠癌仍然是癌症相关发病率和死亡率的主要原因，在美国和世界各地都是如此。 世界许多治疗剂及其组合被用于抑制肿瘤的生长和转移。 肿瘤然而，这些策略的两个重要问题是耐药性的发展 以及药物副作用的增加。耐药性的一个关键原因是， 或通过各种激酶途径的间接靶向治疗剂， （PXR）.因此，新的靶向治疗是必不可少的，抑制特定的途径，但不诱导PXR。 在这方面，我们开发了一种新型药物MRLTHB和水溶性类似物MRLTHBCD，其抑制 Notch-1信号传导，不诱导PXR。当前项目的目标是进一步表征药物 并生成作为单独和与5-氟尿嘧啶（5-FU）组合的口服治疗剂的临床前数据， 结肠癌在以前的研究中，我们已经确定下调双皮质素钙调素样激酶， 1（DCLK 1）抑制结肠癌异种移植物生长，表明靶向DCLK 1将是一种有效的治疗方法。 结肠癌的治疗策略我们现在已经确定THB和THBCD特异性抑制DCLK 1激酶 活性，但不影响钙调蛋白样激酶CAMKII和CAMKIV的激酶活性。另外我们 已经确定，该化合物在一种新的培养方法中抑制结肠癌细胞的生长， 已经开发出一种称为“盘中肿瘤”（TiD）的方法，其中癌细胞在三维培养物中生长 包括正常的上皮细胞、成纤维细胞和内皮细胞。该模型创建了一个体内样肿瘤 微环境提供了必要的细胞-细胞接触，3D结构，以及不同的细胞的影响。 细胞类型。在该系统中观察到的对癌细胞的选择性杀伤表明化合物是高度活性的。 特异性强，效价高。从机理上讲，我们已经确定化合物抑制Notch 信号通路和PXR表达。基于我们的初步研究，THB和 THBCD抑制Notch信号传导和PXR是结肠的有效治疗策略 癌的我们的目标是在当前应用中继续开发临床前数据。在目标1中，我们建议 确定Notch-1在PXR表达中的作用在目标2中，我们将进行详细的PK/PD研究， 化合物，并找到在异种移植和APCmin/+小鼠中进行临床前研究的最佳剂量 模型在目标3中，我们建议继续并确定THB和THBCD组合的效果 用5-氟尿嘧啶来抑制结肠癌的生长。化合物对Akt磷酸化、Notch-1的影响 将确定肿瘤中的DCLK 1和PXR表达。这些拟议的研究将 提供了令人信服的机制证据，用于启动单独的新化合物和 组合.这些研究还将有助于优化靶向化疗方案，并确定新的治疗方案。 生物标志物用于未来的临床研究。