Activators of Adipocyte Fatty Acid Oxidation

脂肪细胞脂肪酸氧化激活剂

• 批准号: 6693970
• 负责人: CYDNEY C BROOKS
• 金额: $ 9.99万
• 依托单位: ADIPOGENIX, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6693970
• 关键词: adipocytes; carnitine palmitoyltransferase 1; cell differentiation; clinical research; fatty acids; high throughput technology; human subject; lipid metabolism; mitochondria; oxidation; oxygen consumption; polymerase chain reaction; western blottings

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.

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。现有的药物和饮食治疗肥胖症的效果有限。一种很可能有效治疗肥胖症并具有良好副作用的方法是直接作用于脂肪细胞中脂质代谢的药物。AdipoGenix公司的使命是发现和开发在脂肪细胞水平上起作用的治疗肥胖症、糖尿病和相关代谢紊乱的新疗法。酶肉毒碱棕榈酰转移酶I（CPT I）是细胞中脂肪酸代谢的关键控制点，并提供游离脂肪酸（FFA）氧化为CO2和酯化为甘油三酯（TG）之间的主要开关。氧化和合成之间平衡的轻微变化可能对脂肪储存产生重大影响。丙二酰辅酶A是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作用的化合物可促进CPT I活性，从而促进FFA氧化。我们建议的特点的分化和脂肪库依赖的表达CPT I亚型在人前脂肪细胞，并开发高通量的屏幕，以确定化合物，减轻抑制作用的丙二酰辅酶A对CPT I，从而刺激氧化。将使用相对定量RTPCR和蛋白质印迹法分析分化期间和来自不同解剖学库的人脂肪细胞中CPT I同种型的表达。将开发一种高通量测定法，以测量在存在和不存在抑制剂丙二酰辅酶A的情况下，CPT I在分离的线粒体制剂和透化的人脂肪细胞中的活性。与此同时，将开发一种高通量测定方法，使用基于氧传感微孔板的系统来测量人脂肪细胞中的氧消耗。确定最适合HTS的测定将用于筛选II期的化学文库。这些目标的实现将建立一个可行的HTS测定和二级测定，用于鉴定激活CPT I和增加人脂肪细胞氧化的化合物。此类化合物的开发可导致有效调节脂质代谢和治疗肥胖症的治疗性干预。