Activators of Adipocyte Fatty Acid Oxidation

描述（由申请人提供）：肥胖是许多疾病的公认危险因素，包括2型糖尿病和冠心病。现有的治疗肥胖的药物和饮食疗法效果一般。一种很可能对治疗肥胖有效且副作用小的方法是一种直接作用于脂肪细胞脂质代谢的药物。AdipoGenix, Inc.的使命是发现和开发作用于脂肪细胞水平的新疗法，用于治疗肥胖、糖尿病和相关代谢紊乱。肉碱棕榈酰基转移酶I （CPT I）是细胞内脂肪酸代谢的关键控制点，是游离脂肪酸（FFA）氧化成CO2和酯化成甘油三酯（TG）之间的主要开关。氧化和合成之间的平衡稍有变化，就会对脂肪储存产生重大影响。丙二酰辅酶a，一种CPT I的变构抑制剂，调节FFA氧化，从而调节脂肪储存。干扰丙二酰辅酶a对CPT I作用的化合物可以促进CPT I活性，从而促进FFA氧化。我们建议表征人类前脂肪细胞中CPT - 1亚型的分化依赖性和脂肪库依赖性表达，并开发高通量筛选来识别缓解丙二酰辅酶a对CPT - 1的抑制作用从而刺激氧化的化合物。相对定量的RTPCR和Western blotting将用于分析人类脂肪细胞在分化和不同解剖库中CPT I亚型的表达。在存在和不存在抑制剂丙二酰辅酶A的情况下，将开发一种高通量测定方法来测量分离线粒体制剂和渗透性人脂肪细胞中CPT I的活性。同时，将开发一种利用氧传感微板系统测量人体脂肪细胞耗氧量的高通量测定方法。被确定为最适合HTS的试验将用于筛选二期化学文库。实现这些目标将建立一个可行的HTS分析和二级分析，以识别激活CPT I和增加人类脂肪细胞氧化的化合物。这些化合物的开发可能导致有效调节脂质代谢和治疗肥胖的治疗干预。

DESCRIPTION (provided by applicant): Obesity is a well-established risk factor for a number of diseases, including type 2 diabetes and coronary heart disease. Existing drug and dietary treatments for obesity are only modestly effective. An approach that is very likely to be effective in treating obesity and with a good side effect profile is a drug that acts directly on lipid metabolism in the fat cell. The mission of AdipoGenix, Inc. is to discover and develop novel therapeutics acting at the level of the fat cell for treating obesity, diabetes, and related metabolic disorders. The enzyme carnitine palmitoyltransferase I (CPT I) is the critical control point for fatty acid metabolism in the cell and provides the main switch between free fatty acid (FFA) oxidation to CO2 and esterification to triglyceride (TG). A slight change in the balance between oxidation and synthesis could have a major impact on fat stores. Malonyl-CoA, an allosteric inhibitor of CPT I, regulates FFA oxidation and, consequently, fat storage. Compounds that interfere with the effect of malonyl-CoA on CPT I can promote CPT I activity and, thereby, FFA oxidation. We propose to characterize the differentiation- and fat depot-dependent expression of CPT I isoforms in human preadipocytes, and to develop high-throughput screens to identify compounds that relieve the inhibitory effect of malonyl-CoA on CPT I and thereby stimulate oxidation. Relative quantitative RTPCR and Western blotting will be used to analyze expression of CPT I isoforms in human adipocytes during differentiation and from different anatomical depots. A high-throughput assay to measure activity of CPT I in isolated mitochondrial preparations and in permeabilized human adipocytes in the presence and absence of the inhibitor malonyl-CoA will be developed. In parallel, a high-throughput assay to measure oxygen consumption in human adipocytes using an oxygen-sensing microplate-based system will be developed. The assay determined to be most suitable for HTS will be used to screen chemical libraries in Phase II. Achievement of these aims will establish a viable HTS assay and secondary assays for identifying compounds that activate CPT I and increase oxidation in human adipocytes. Development of such compounds may lead to therapeutic interventions that are effective for modulating lipid metabolism and treating obesity.