Mechanisms of prostate cancer prevention by Korean Angelica

描述(申请人提供)：我们的长期目标是从韩国当归(AGN)中开发无毒、低成本和有效的草药药物，用于预防男性前列腺癌的发生和转移。我们已经在临床前模型中显示了AGN乙醇提取物对宿主小鼠的明显的前列腺癌生长抑制和化学预防作用，而没有任何可观察到的毒性。然而，缺乏对活性化学物质和体内分子靶点的机械知识是将预防益处转化为男性的重大障碍。目前应用的目标是在两个独立和互补的前列腺癌模型中产生关于(1)活性化学物质和(2)细胞和分子靶标的机制知识。我们假设，AGN提取物在体内发挥功效：(A)主要通过吡喃香豆素化合物及其代谢物；(B)通过影响关键的细胞过程和分子靶标，可以通过系统生物学方法有效地描绘出来。我们计划通过追求三个特定的目标来验证这些假说：目的1.通过比较纯化的化合物与AGN提取物和吡喃香豆素基因敲除(KO)提取物在免疫缺陷小鼠异种移植模型中抑制人前列腺癌生长和转移的效果，确定吡喃香豆素的化学中介作用。目的2.通过与AGN提取物和KO提取物的比较，验证它们在转基因小鼠前列腺癌(TRAMP)模型化学预防原发癌变中的中介作用。目的3.通过聚焦分析确定体内效应的关键细胞指标和分子生物标记物，包括血管生成、细胞凋亡、细胞增殖和侵袭性，并利用系统生物学方法确定分子靶点，使用前两个目标的合适组织。AIMS 1和2中提出的工作有望严格评估吡喃香豆素和代谢物作为化学介体对AGN提取物抑制前列腺癌发生和转移的作用，并在小鼠模型中建立它们的长期安全性概况。预计这些结果将对AGN草药产品未来的重点研发工作和质量控制产生积极影响。目的3的结果有望确定候选的体内分子靶点和细胞过程。更好地了解模型系统中的活性化学物质及其分子靶标，可以进一步进行机械性研究，并合理规划男性临床翻译。此外，AGN提取物的功效和机制可能也可用于预防其他器官部位的癌症。在我们看来，这项研究具有创新性，因为它代表了对单剂/单靶点方法的临床前癌症化学预防研究现状的实质性偏离。使用尖端药物化学方法(如基因敲除提取物)和系统生物学工具(如iTRAQ蛋白质组学)进行的综合研究有助于将草药疗法推向循证补充和替代医学。

DESCRIPTION (provided by applicant): Our long-term goal is to develop non-toxic, low-cost and efficacious herbal modalities from Korean Angelica gigas Nakai (AGN) for the prevention of prostate carcinogenesis and metastasis in men. We have shown in preclinical models significant prostate cancer growth-inhibitory and chemopreventive effects of AGN ethanol extracts without any observable toxicity to the host mice. Nevertheless, a lack of mechanistic knowledge of active chemicals and in vivo molecular targets is a significant roadblock to translating the preventive benefits to men. The objective for the current application is to generate mechanistic knowledge of (1) the active chemicals and (2) cellular and molecular targets in two independent and complementary prostate cancer models. We hypothesize that AGN extract exerts in vivo efficacy (a) mainly through pyranocoumarin compounds and their metabolite; (b) by affecting critical cellular processes and molecular targets, which can be effectively profiled through a systems-biology approach. We plan to test these hypotheses by pursuing three specific aims: Aim 1. Establish the chemical mediator role of the pyranocoumarins by comparing the efficacy of purified compounds with AGN extract and pyranocoumarin-knockout (KO) extract to inhibit human prostate cancer growth and metastasis in xenograft models in immunodeficient mice. Aim 2. Validate their mediator role for chemopreventive efficacy against primary carcinogenesis in the transgenic adenocarcinoma of mouse prostate (TRAMP) model by comparing them with AGN extract and KO extract. Aim 3. Determine key cellular indices and molecular biomarkers of in vivo efficacy including angiogenesis, apoptosis, cell proliferation and invasiveness by focused analyses and identify molecular targets with systems-biology approaches, using suitable tissues from the first two aims. The work proposed in Aims 1 and 2 is expected to critically assess the role of pyranocoumarins and metabolites as chemical mediators for the inhibitory efficacy of AGN extract against prostate primary carcinogenesis and metastasis, and establish their long-term safety profiles in mice models. Such results are expected to positively impact future focused R&D efforts and quality control for AGN herbal products. Aim 3 results are expected to identify candidate in vivo molecular targets and cellular processes. Better knowledge of the active chemicals and their molecular targets in model systems enables further mechanistic studies and rational planning for clinical translation in men. Furthermore, the efficacy and mechanisms are likely exportable to the prevention of cancers of other organ sites by AGN extracts. The research proposed is innovative, in our opinion, because it represents a substantial departure from the status quo for preclinical cancer chemoprevention research of single-agent/single-target approach. The comprehensive research with cutting- edge medicinal chemistry methods (e.g., knockout extract) and systems-biology tools (e.g., iTRAQ proteomics) can help moving herbal remedies toward evidence-based complementary and alternative medicine.