IDENTIFICATION OF NEW ACETYL-COA CARBOXYLASE INHIBITORS

描述（由申请人提供）：乙酰辅酶A羧化酶（ACC）催化脂肪酸生物合成的第一步，其产物丙二酰辅酶A抑制脂肪酸氧化。这些功能使ACC成为开发治疗肥胖和2型糖尿病的疗法的主要目标。小鼠敲除研究和ACC抑制剂的动物研究已经验证了这种方法。迄今为止鉴定的最有效的真核ACC抑制剂是天然产物soraphen。索拉芬已被证明具有与治疗肥胖和糖尿病的潜力一致的药理学特性;然而，索拉芬由于毒副作用而不适合作为治疗剂。此外，由于其复杂的结构，soraphen不是药物开发的良好起点。我们建议确定新的小分子目标soraphen结合位点，并抑制哺乳动物ACCs的高效力。在第一阶段的研究中，我们克隆和表达的生物素羧化酶（BC）域从人类ACC 1和ACC 2同工酶，保留高亲和力soraphen结合，开发了一种高通量的筛选试验，并解决了X射线晶体结构的酵母BC单独和与soraphen复合。在II期研究中，我们将使用这些工具通过完成以下具体目标来鉴定新的抑制剂：1）将大约2，000，000种市售化合物与索拉芬结合位点进行分子对接; 2）高通量筛选排名靠前的对接命中和内部小分子文库; 3）体外和体内表征先导物; 4）通过类似物的迭代合成和测试进行先导物优化; 5）确定另外的BC结构域和与BC结构域复合的新先导物的结构;和6）在动物中用1至3个先导物进行功效研究，以证明治疗肥胖和2型糖尿病的潜力。该项目将发现用于对抗肥胖和糖尿病的新候选药物。这些疾病影响了大约35%的美国人，每年花费大约2000亿美元。

DESCRIPTION (provided by applicant): Acetyl-CoA carboxylase (ACC) catalyzes the first step in fatty acid biosynthesis and its product, malonyl-CoA, inhibits fatty acid oxidation. These functions make ACC a prime target for the development of therapeutics to treat obesity and type 2 diabetes. Knockout studies in mice, and animal studies with ACC inhibitors, have validated this approach. The most potent inhibitor of eukaryotic ACCs identified to date is the natural product soraphen. Soraphen has been demonstrated to have pharmacological properties consistent with the potential to treat obesity and diabetes; however, soraphen is not suitable as a therapeutic due to toxic side effects. Furthermore, soraphen is not a good starting point for drug development due to its complex structure. We propose to identify novel small molecules that target the soraphen binding site and inhibit mammalian ACCs with high potency. In Phase I research we cloned and expressed biotin carboxylase (BC) domains from the human ACC1 and ACC2 isozymes that retain high affinity soraphen binding, developed a high throughput screening assay, and solved the x-ray crystal structure for yeast BC alone and in complex with soraphen. In Phase II research we will use these tools to identify novel inhibitors by completing the following specific aims: 1) molecular docking of approximately 2,000,000 commercially available compounds to the soraphen binding site; 2) high throughput screening of the top ranked docking hits, and of an in- house small molecule library; 3) in vitro and in vivo characterization of leads; 4) lead optimization through iterative synthesis and testing of analogs; 5) determination of the structures of additional BC domains and of novel leads in complex with the BC domain; and 6) efficacy studies in animals with 1 to 3 leads to demonstrate potential for treating obesity and type 2 diabetes. This project will discover new drug candidates for combating obesity and diabetes. Combined, these diseases affect approximately 35% of Americans and cost approximately $200 billion annually.