Combinations of Grape Seed and Milk Thistle Extracts Against Lung

葡萄籽原花青素提取物（GSE）和水飞蓟素提取物（MTE）是应用广泛的保健食品 补充剂，以促进心血管（CV）和肝胆健康。GSE和MTE 含有高水平的多酚，结构独特，具有强抗氧化性能， 已显示该药剂对肺癌具有抗癌作用。使用组合的初步数据 的GSE和MTE显示出令人兴奋的，明确的协同抗癌作用，对肺前和癌细胞 体外因此，我们假设，口服白细胞选择性磷脂酶体（LP）， 标准化的GSE和siliphos，一种标准化的MTE，两者都与大豆磷脂复合成磷脂复合物 提高生物利用度，将协同抑制肺癌生长，侵袭，并诱导细胞凋亡， 各种人肺癌异种移植物模型，通过与肺相关的机制的有利调节， 肿瘤发生/促进。为了检验这些假设，我们将确定其药代动力学（PK）和 药效学（PD）。我们还将确定生物标本的实用性，如快照，新鲜冷冻肺 组织匀浆，作为替代模型系统，以监测口服给药的生物利用度和生物活性 将这些药剂施用至靶器官。此外，组合的机械效应 将通过癌症相关、途径特异性基因表达和microRNA进行系统评估 （miRNA）的真实的时间PCR阵列，并与功能意义相关。提出了三个具体目标： 目的1：研究LP与硅磷的最大耐受剂量（MTD）、PK/PD和抗肿瘤作用 在小鼠模型中。将在给予不同剂量组合的裸鼠中进行剂量范围探索研究 以确定MTD。将采集血液和肺样本，以确定GSE、MTE和 代谢物作为GSE和MTE生物利用度的替代标志物。目标1.1。为了确定生物活性， 使用冷冻小鼠肺匀浆与人肺的新型共培养系统的口服LP和硅磷 肿瘤细胞通过共培养对照组的肺组织匀浆，评估肺中的生物活性 （水）与药物处理的具有人肺癌和癌前细胞系的小鼠。不同剂量的影响 组合对共培养细胞中增殖和凋亡的影响将与GSE、MTE和代谢产物相关 程度.目的2：确定LP和硅磷的组合对各种类型的肿瘤的抗癌作用。 人肺癌异种移植小鼠模型。根据MTD结果，将给予不同的剂量组合 通过口服管饲法对携带各种人肺肿瘤异种移植物的小鼠进行长达8周的连续给药， 收集来自每个处理组的血浆、肺组织和肿瘤异种移植物。抗癌效果 将通过肿瘤生长延迟或达到最大肿瘤体积的时间以及增殖（Ki-17）来确定。 67)和凋亡（裂解的半胱天冬酶3）指数，并与各种生物标本中的GSE和MTE水平相关， 以定义与生物活性相关的生理学水平。目标3：查明、描述和联系 GSE联合MTE抗肺癌的分子机制。将评估机械效应 通过比较处理前后的生物活性， 各种样品类型，如通过以下的调节所测量的：1）类花生酸信号传导途径; 2）另外的标志物 炎症和抗肿瘤免疫，白细胞介素（IL）-6，IL-10，IL-12; 3）mir-19 a，-19 b和106 b水平; 4） 癌症相关的、途径特异性基因表达谱; 5）通过miRNA评估的表观遗传谱 表达阵列，和6）常见的癌变生物标志物，如PTEN，IGF 2 R，P53，p27，p21，p16， FHIT和BIRC 5。影响：研究结果将为可行性提供重要见解， GSE和MTE联合治疗肺癌的机制作用，并为临床试验铺平道路 在不久的将来，在推进肺癌的治疗和预防方面具有巨大的潜力。

Grape seed procyanidin extract (GSE), and milk thistle silymarin extract (MTE) are widely used health food supplements to promote cardiovascular (CV) and hepatobiliary health, respectively. Both GSE and MTE contain high levels of polyphenols that are structurally distinct with strong antioxidant properties, and each agent has been shown to exert antineoplastic effects against lung cancer. Preliminary data using combinations of GSE and MTE shows exciting, unequivocal synergistic anticancer effects against lung pre- and cancer cells in vitro. We therefore hypothesize that oral administration of combinations of leucoselect phytosome (LP), a standardized GSE, and siliphos, a standardized MTE, both complexed with soy phospholipids into phytosomes to enhance bioavailability, will synergistically inhibit lung cancer growth, invasion, and induce apoptosis in various human lung cancer xenograft models, via favorable modulations of mechanisms associated with lung tumorigenesis/promotion. To test these hypotheses, we will determine their pharmacokinetics (PK) and pharmacodynamics (PD). We will also determine the utility of biospecimens, such as snap, fresh frozen lung tissue homogenates, as surrogate model systems to monitor the bioavailability and bioactivity of oral administration of these agents to the target organ. Furthermore, the mechanistic effects of the combinations will be assessed systematically with cancer relevant, pathway specific gene expressions and microRNA (miRNA) real time PCR arrays, and correlated to functional significance. Three specific aims are proposed: Aim 1: To determine the maximum tolerated dose (MTD), PK/PD, and anti-cancer effects of LP with siliphos in murine models. A dose range finding study will be conducted in nude mice given varying dose combinations via oral gavage to establish MTD. Blood and lung samples will be obtained to determine PK of GSE, MTE and metabolites as surrogate markers of bioavailability of GSE and MTE. Aim 1.1. To determine the bioactivity of oral LP and siliphos using a novel co-culture system of frozen mouse lung homogenates with human lung neoplastic cells. Bioactivity in the lungs will be assessed by co-culturing lung tissue homogenates from control (water) vs. drug treated mice with human lung cancer and precancerous cell lines. The effects of varying dose combinations on proliferation and apoptosis in co-cultured cells will be correlated to GSE, MTE and metabolites levels. Aim 2: To determine the anti-cancer effects of combinations of LP and siliphos on various types of human lung cancer xenograft mouse models. Based on MTD findings, varying dose combinations will be given via oral gavage to mice bearing a variety of human lung tumor xenografts for up to 8 weeks with serial collections of plasma, lung tissues and tumor xenografts from each treatment group. The anticancer effects will be determined by tumor growth delay or time to reach maximum tumor volume, as well as proliferation (Ki- 67) and apoptotic (cleaved caspase 3) indices, and correlated to GSE & MTE levels in various biospecimens, to define physiologically relevant levels in reference to bioactivity. Aim 3: To identify, characterize and correlate the molecular mechanisms of GSE with MTE against lung cancer. The mechanistic effects will be assessed and correlated systematically and comprehensively, by comparing the bioactivity pre- or post treatment, in various sample types, as measured by modulations of: 1) eicosanoid signaling pathways; 2) additional markers of inflammations and anti-tumor immunity, interleukin (IL)-6, IL-10, IL-12; 3) mir-19a, -19b and 106b levels; 4) cancer relevant, pathway specific gene expression profiles; 5) epigenetic profiles assessed by miRNA expression array, and 6) common biomarkers of cancerization such as PTEN, IGF2R, P53, p27, p21, p16, FHIT, and BIRC5. IMPACT: Findings from the study will provide important insights into the feasibility and mechanistic effects of combinations of GSE and MTE against lung cancer, and pave the way for clinical trials in the near future, with enormous potential in advancing the treatment and prevention of lung cancer.