Novel approaches to plant drug discovery based on high throughput pharmacological screening and genetic manipulation
Novel approaches to plant drug discovery based on high throughput pharmacological screening and genetic manipulation
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DOI：
10.1016/j.lfs.2005.09.013
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发表时间：
2005-12-22
期刊：
影响因子：
6.1
通讯作者：
Falcone, D
中科院分区：
文献类型：
--
作者：
Littleton, J;Rogers, T;Falcone, D
植物是潜在的重要的新的治疗药物的线索，但缓慢的传统方法的研究植物限制了制药行业的热情。为了克服一些缺点，我们已经将高通量药理学筛选（HTPS）应用于粗植物提取物。使用“差异智能筛选”（DSS），在几种密切相关的受体亚型处测量粗提取物中所含的活性谱。然后将该光谱与已知化合物的光谱进行比较。一个独特的光谱表明，提取物值得进一步研究。物种和整个植物的环境库的评估已经证明了这种方法的价值，快速优先级的植物进行调查。此外，植物和植物细胞培养物的基因组和遗传操作可以增加DSS的价值。例如，可以通过使用功能突变的获得来获得植物物种的生物多样性的全基因组潜力，以产生突变克隆培养物的“功能基因组文库”，并通过DSS测试这些培养物的生物活性。可以以这种方式鉴定过量产生不同于野生型植物的活性的克隆。这种“天然产物基因组学”（NPG）策略受到覆盖所有可能的功能获得性突变所需的大量克隆培养物的限制。通过使用表达适当的哺乳动物蛋白的转基因植物可以提高该过程的速度和效率。这些可以被设计成使植物细胞类似于人类细胞的特定形式的毒性。现在，抗性突变克隆的“非自然选择”可用于提供潜在富集有用化合物的亚群。或者，转基因植物细胞可用于“原位筛选”，其中表达与报道构建体连接的哺乳动物受体蛋白，如绿色荧光蛋白。产生这种受体的配体的克隆培养物现在可以在ultra-HTPS中快速目视鉴定。总的来说，我们的目标是使用药理学筛选，功能基因组学方法，使植物药物发现与组合化学竞争。(c)2005年爱思唯尔公司All rights reserved.
Plants are potentially important for novel therapeutic drug leads, but the slowness of conventional methods for investigation of plants limits enthusiasm in the pharmaceutical industry. To overcome some of the drawbacks, we have applied high throughput pharmacological screening (HTPS) to crude plant extracts. Using a "differential smart screen", (DSS) the spectrum of activity contained in a crude extract is measured at several closely related receptor subtypes. This spectrum is then compared to that of known compounds. A unique spectrum suggests that the extract merits further investigation. Evaluation of species and environmental libraries of whole plants has demonstrated the value of this approach for rapid prioritization of plants for investigation. In addition, genomic and genetic manipulation of plants and plant cell cultures can increase the value of DSS. For example, the whole genomic potential of a plant species for biodiversity can be accessed by using gain of function mutations to generate a "functional genomics library" of mutant clonal cultures, and the bioactivity of these cultures tested by DSS. Clones that overproduce activity differing from the wild-type plant can be identified in this way. This "Natural Products Genomics" (NPG) strategy is limited by the massive numbers of clonal cultures that are required to cover all possible gain-of-function mutations. The rapidity and efficiency of this process can be improved by using transgenic plants expressing appropriate mammalian proteins. These may be designed to make the plant cell resemble a human cell for a specific form of toxicity. Now, "unnatural selection" of resistant mutant clones can be used to provide a sub-population potentially enriched in useful compounds. Alternatively, transgenic plant cells can be used for "in situ screening" in which a mammalian receptor protein, linked to a reporter construct, such as green fluorescent protein, is expressed. Clonal cultures that produce ligands for this receptor can now be rapidly identified visually in an ultra-HTPS. Overall, our aim is to use pharmacological screening, together with functional genomic approaches, to make plant drug discovery competitive with combinatorial chemistry. (c) 2005 Elsevier Inc. All rights reserved.