Oldenlandia diffusa for prostate cancer treatment

描述（由申请方提供）：本探索性提案是对PA-06-400的回应，旨在进行临床前研究，以证明中药白花蛇舌草（OD）治疗前列腺癌的疗效。OD在民间中药中被用作抗癌剂。然而，有有限的临床前研究来研究其功效并鉴定其活性抗癌成分。OD对前列腺癌的影响尚未得到充分研究。我们的初步数据表明，OD抑制前列腺癌细胞的生长，部分通过诱导凋亡，抑制癌细胞的侵袭和血管生成在体外。在这个发展建议中，我们将应用基于细胞功能的体外试验，以确定前列腺癌治疗的OD组分的有效组合。我们假设OD含有生物活性成分，它们以协同或相加的方式相互作用，靶向前列腺癌的生长和转移以及血管生成。具体目标1是鉴定用于抑制前列腺癌细胞的生长和侵袭以及用于抑制体外血管生成的活性OD组分。我们将应用基于细胞功能的分析来鉴定活性成分。正常前列腺上皮细胞将用于评价可能的副作用。当确定副作用较少的活性成分时，将进行系统评价，以制定靶向前列腺癌区室（细胞生长和侵袭）和内皮区室（内皮细胞生长、迁移和管形成）的候选联合方案。具体目的2是验证配制的组合方案对动物模型中前列腺癌治疗的功效。两项动物研究，一项针对雄激素敏感性前列腺肿瘤（LNCaP肿瘤），另一项针对雄激素非依赖性前列腺肿瘤（PC-3或DU 145），将用于确定候选方案的疗效。我们将首先通过使用实验设计来确定有效的联合方案，该实验设计允许通过在癌细胞原位植入后开始治疗方案来确定最大疗效（目的2a）。当针对每种肿瘤表型确定有效的联合方案时，将通过使用“生长延迟”实验设计（目的2b）在其相应的动物模型中证实其疗效。具体目标3是阐明细胞机制，由此组合方案可以以协同方式有效抑制前列腺癌进展。由于联合治疗方案是基于确定的协同/相加细胞功能制定的，因此我们将在动物研究中验证细胞标志物的调节。预计从该探索性方案中获得的结果将成功确定用于前列腺癌治疗的OD组分的有效联合方案，并将提供足够的实验证据支持开发RO 1方案，以研究OD作为前列腺癌治疗的潜在补充方法。拟议项目的目标是确定OD是否含有治疗前列腺癌的活性成分。基于体外细胞功能的生物测定将被应用于鉴定靶向前列腺癌生长和侵袭以及血管生成的活性成分。在临床相关的原位前列腺肿瘤动物模型中，评价靶向前列腺癌和血管生成区室的OD活性组分的配制组合方案对雄激素依赖性和雄激素非依赖性前列腺肿瘤生长的功效。将在体内样品中测定细胞标志物。

DESCRIPTION (provided by applicant): This exploratory proposal, in response to the PA-06-400, is developed to conduct preclinical studies to demonstrate efficacy of the Chinese herb Oldenlandia diffusa (OD) for prostate cancer treatment. OD has been used in folk Chinese medicine as anti-cancer agent. However, there have been limited preclinical studies to investigate its efficacy and to identify its active anti-cancer components. The effect of OD on prostate cancer has not been adequately studied. Our preliminary data showed that OD inhibited the growth of prostate cancer cells in part via induction of apoptosis and inhibited cancer cell invasion and angiogenesis in vitro. In this developmental proposal, we will apply cellular function-based in vitro assays to identify the effective combinations of the OD components for prostate cancer treatment. We hypothesize that OD contains bioactive components that interact, in a synergistic or an additive manner, to target prostate cancer growth and metastasis and angiogenesis. Specific aim 1 is to identify the active OD components for inhibition of growth and invasion of prostate cancer cells and for inhibition of angiogenesis in vitro. We will apply cellular function- based assays to identify the active components. Normal prostatic epithelial cells will be used for evaluation of possible side effect. When the active components with few side effects are identified, a systematic evaluation will be conducted to formulate the candidate combination regimens that target both prostate cancer compartment (cell growth and invasion) and endothelial compartment (endothelial cell growth, migration and tube formation). Specific aim 2 is to verify efficacy of the formulated combination regimens on prostate cancer treatment in animal models. Two animal studies, one for androgen-sensitive prostate tumor (LNCaP tumor) and the other for androgen-independent prostate tumor (PC-3 or DU 145), will be used for determination of efficacy of the candidate regimens. We will first identify the potent combination regimens by using the experimental design that allows determination of maximal efficacy by starting treatment regimens after cancer cells are orthotopically implanted (Aim 2a). When a potent combination regimen is identified for each tumor phenotype, its efficacy will be confirmed in its corresponding animal model by using the "growth delay" experimental design (Aim 2b). Specific aim 3 is to elucidate cellular mechanisms whereby the combination regimens may effectively inhibit prostate cancer progression in a synergistic manner. Because the combination regimens are formulated based on defined synergistic/additive cellular functions, we will verify the modulation of cellular markers in animal studies. It is expected that results derived from this exploratory proposal will successfully identify the potent combination regimens from OD components for prostate cancer treatment, and will provide sufficient experimental evidence to support development of a RO1 proposal to investigate OD as a potential complementary approach for prostate cancer treatment. The goals of the proposed project are to determine if OD contains active components for treatment of prostate cancer. In vitro cellular function-based bioassays will be applied to identify the active components that target prostate cancer growth and invasion and angiogenesis. The formulated combination regimens of OD active components that target both prostate cancer and angiogenesis compartments will be evaluated for their efficacy on the growth of androgen-dependent and androgen-independent prostate tumors in clinically relevant orthotopic prostate tumor animal models. The cellular markers will be determined in the in vivo samples.