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成为治疗肥胖症和2型糖尿病的主要目标。小鼠的基因敲除研究和ACC抑制剂的动物研究证实了这种方法。迄今为止鉴定出的最有效的真核ACCs抑制剂是天然产物sorphen。Soraphen已被证明具有与治疗肥胖和糖尿病的潜力一致的药理特性；然而，由于其毒副作用，sorphenen不适合作为治疗药物。此外，索拉芬由于其复杂的结构而不是药物开发的良好起点。我们建议鉴定新的靶向soraphen结合位点的小分子，并有效抑制哺乳动物acc。在I期研究中，我们从人类ACC1和ACC2同工酶中克隆并表达了保留高亲和力sorphenen结合的生物素羧化酶（BC）结构域，开发了高通量筛选试验，并解决了酵母BC单独和与sorphenen复合的x射线晶体结构。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识别新的抑制剂：1)将大约200万种市售化合物与soraphen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，以及内部小分子文库；3)引线的体外和体内表征；4)通过迭代合成和测试类似物进行先导优化；5)确定附加BC结构域和与BC结构域配合的新型引线的结构；6)在动物身上进行的1到3个导联的疗效研究证明了治疗肥胖症和2型糖尿病的潜力。这个项目将发现对抗肥胖和糖尿病的新候选药物。加起来，这些疾病影响了大约35%的美国人，每年造成大约2000亿美元的损失。

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.