Extraction and Characterization of a Safer, Cost-effective Anti-inflammatory

 描述（由申请人提供）：炎症性疾病状态是美国死亡和残疾的主要原因，也是医疗保健的主要成本。由于NSAID的不良反应，死亡和住院的情况很普遍。TAI-LCx（姜黄抗炎-亲脂组分X）是在姜黄（Curcuma longa L.）的以前未被认识的药理活性成分中发现的新型抗炎剂，并且已经显示出作为更安全、更成本有效的治疗的高潜力。TAI-LCx与姜黄素和姜黄素类化合物不同，后者已被大量研究和充分表征。TAI-LCx对多种炎症介质有微妙的作用，同时避免了考克斯通路，有望减少临床并发症，降低现有治疗的惊人经济成本。拟议研究的假设是，TAI-LCx中的一种成分（或一小组小分子）具有特异性，强效和临床相关的抗炎（AI）活性。因此，所提出的研究的目的是检验以下假设，即可以鉴定用于提取TAI-LCx的改进和优化的方法，以将活性成分纯化至均匀并确定其分子式（从而使得能够在第II阶段努力完成结构测定），并进一步表征 TAI-LCx的AI作用，以确定姜黄的其他成分是否可能与TAI-LCx协同参与产生迄今为止检测到的AI特性。拟议项目的具体目标如下。目的#1：通过超临界流体萃取（SFE）优化TAI-LCx萃取并表征活性成分。这将增加 样品纯度和商业可扩展性，并导致更好地表征活性成分，是一个潜在的专利创新。优化SFE方法，然后纯化TAI-LCx中的活性成分，将能够扩大规模并生产>50 g TAI-LCx。SFE方法将比最初（在原始专利中）用于产生活性TAI-LCx的基于溶剂的提取方法更具体，更有效，更环保，更具成本效益。目的#2：测定TAI-LCx的组分并鉴别活性化合物。进一步的药物开发将需要详细的成分信息。将通过高分辨率质谱法分析来自成功SFE提取和后续纯化的含有TAI-LCx活性的样品，以确定提取物中所有组分的分子式和活性成分的（推定）结构特性。目的#3：确定TAI-LCx对其他抗炎途径如考克斯-1、NF κ B、AP-1和白细胞介素（IL-2、IL-6）的作用。这将证明其治疗意义和应用，并提供与现有FDA批准的疗法的比较，并提供产品差异化，并可能增加有关炎症途径的知识体系。

 DESCRIPTION (provided by applicant): Inflammatory disease states are leading causes of death and disability and a major cost to healthcare in the United States. There is an epidemic of death and hospitalizations due to adverse effects from NSAIDs. TAI- LCx (Turmeric Anti-Inflammatory-Lipophilic Component X) is a novel anti-inflammatory discovered in previously unrecognized, pharmacologically-active constituents of turmeric (Curcuma longa L.), and has demonstrated high potential as a safer, more cost-effective treatment. TAI-LCx is distinct from curcumin and curcuminoids that have been heavily researched and well characterized. With a nuanced action on multiple inflammatory mediators while avoiding the COX pathway, TAI-LCx is expected to reduce clinical complications and reduce the staggering economic costs of existing treatments. The hypothesis of the proposed research is that one constituent (or a small group of small molecules) within TAI-LCx possesses specific, potent and clinically relevant anti-inflammatory (AI) activity. The purpose of the proposed research, therefore, is to test the hypothesis that improved and optimized methods for extraction of TAI-LCx can be identified, to purify to homogeneity the active constituent(s) and determine its molecular formula (thereby enabling in phase II efforts to complete structural determination), and to further characterize the AI action of TAI-LCx, to determine whether additional constituents of turmeric might be synergistically involved with TAI-LCx in producing the AI properties detected to date. The Specific Aims of the proposed project are as follows. Aim # 1: Optimize TAI-LCx extraction via supercritical fluid extraction (SFE) and characterize the active constituent(s). This will increase sample purity and commercial scalability, and lead to better characterization of the active principle, and is potentially a patentable innovation. Optimization of the SFE method followed by purification of the active constituents in TAI-LCx will enable scale-up and production of >50 g of TAI-LCx. The SFE method will be more specific, more efficient, more environmentally friendly, and more cost-effective than the solvent-based extraction method used originally (in the original patent) to generate active TAI-LCx. Aim # 2: Determine the constituents of TAI-LCx and identify the active compound(s). Further drug development will require detailed composition information. Samples from successful SFE extractions and subsequent purification containing TAI-LCx activity will be analyzed by high resolution mass spectrometry to determine the molecular formula of all components within the extracts and the (putative) structural identity of the active principle(s). Aim # 3: Determine the effect of TAI-LCx on other anti-inflammatory pathways, such as COX-1, NFkB, AP-1, and interleukins (IL-2, IL-6). This will demonstrate therapeutic significance and application, and provide a comparison with existing FDA approved therapies, as well as provide product differentiation and potentially add to the body of knowledge concerning inflammatory pathways.