Mechanisms of prostate cancer prevention by Korean Angelica

描述（由申请人提供）：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形式，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乙醇提取物的显著前列腺癌生长抑制和化学预防作用，而对宿主小鼠没有任何可观察到的毒性。然而，缺乏活性化学物质和体内分子靶点的机制知识是将预防益处转化为男性的重大障碍。本申请的目的是在两个独立和互补的前列腺癌模型中产生（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提取物发挥体内功效（a）主要通过吡喃香豆素化合物及其代谢产物;（B）通过影响关键细胞过程和分子靶点，这可以通过系统生物学方法有效地分析。我们计划通过追求三个具体目标来测试这些假设：目标1。通过比较纯化的化合物与AGN提取物和吡喃香豆素敲除（KO）提取物在免疫缺陷小鼠的异种移植模型中抑制人前列腺癌生长和转移的功效，确定吡喃香豆素的化学介质作用。目标二。通过与AGN提取物和KO提取物的比较，证实它们在小鼠前列腺转基因腺癌（TRAMP）模型中对原发癌发生的化学预防功效的介导作用。目标3：通过重点分析确定体内疗效的关键细胞指标和分子生物标志物，包括血管生成、凋亡、细胞增殖和侵袭性，并使用前两个目标的合适组织，采用系统生物学方法确定分子靶点。 目标1和2中提出的工作预计将严格评估吡喃香豆素和代谢物作为化学介质对AGN提取物抑制前列腺原发性癌发生和转移的作用，并在小鼠模型中建立其长期安全性特征。这些结果预计将对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点，可以进一步进行机制研究和合理规划男性临床转化。此外，AGN提取物的功效和机制可能可用于预防其他器官部位的癌症。在我们看来，所提出的研究是创新的，因为它代表了对单药/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。采用尖端药物化学方法进行的综合研究（例如，敲除提取物）和系统生物学工具（例如，iTRAQ蛋白质组学）可以帮助将草药疗法转向基于证据的补充和替代医学。

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.