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

 描述(申请人提供)：炎症性疾病州是导致死亡和残疾的主要原因，也是美国医疗保健的主要成本。由于非类固醇抗炎药的不良反应，死亡和住院的情况普遍存在。TAI-LCx(姜黄抗炎亲脂成分X)是一种从姜黄(Curuma Longa L.)中未知的药理活性成分中发现的新型抗炎药物，已显示出作为一种更安全、更具成本效益的治疗方法的高潜力。TAI-LCx不同于姜黄素和姜黄素类化合物，后者已被大量研究和很好地表征。通过对多种炎症介质的微妙作用，同时避免COX途径，TAI-LCx有望减少临床并发症，并降低现有治疗方法的惊人经济成本。拟议研究的假设是，TAI-LCx中的一个成分(或一小群小分子)具有特定的、有效的和临床相关的抗炎(AI)活性。因此，拟议研究的目的是检验可以确定改进和优化的TAI-LCx提取方法的假设，纯化活性成分(S)并确定其分子式(从而能够在第二阶段完成结构测定)，并进一步表征 TAI的作用，以确定姜黄的其他成分是否可能与TAI-LCx协同参与产生迄今检测到的AI特性。拟议项目的具体目标如下。目的1：优化超临界流体萃取TAI-LC_x的工艺条件，并对其有效成分(S)进行表征。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，是一项潜在的专利创新。对超临界流体萃取方法进行优化，然后对TAI-LCx中的活性成分进行纯化，将有助于放大和生产50克TAI-LCx。超临界萃取法将比最初(在原始专利中)用于产生活性TAI-LCx的溶剂型萃取法更具体、更高效、更环保、更具成本效益。目的#2：测定泰来香的化学成分，鉴定有效成分(S)。进一步的药物开发将需要详细的成分信息。成功的超临界萃取物和含有TAI-LCx活性的后续纯化样品将通过高分辨率质谱仪进行分析，以确定萃取物中所有组分的分子式和活性成分(假定)的结构一致性(S)。目的#3：探讨TAI-LCx对COX-1、NFkB、AP-1、IL-2、IL-6等其他抗炎途径的影响。这将展示治疗意义和应用，并提供与FDA批准的现有疗法的比较，以及提供产品差异化，并有可能增加有关炎症途径的知识。

 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.