IDENTIFICATION OF NEW ACETYL-COA CARBOXYLASE INHIBITORS

描述(申请人提供)：乙酰辅酶A羧基酶(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成为开发治疗肥胖症和2型糖尿病的主要靶点。小鼠的基因敲除研究和使用ACC抑制剂的动物研究已经验证了这种方法。到目前为止，已发现的对真核细胞ACCs最有效的抑制物是天然产物索拉芬。索拉芬已被证明具有与治疗肥胖症和糖尿病的潜力一致的药理特性；然而，由于毒副作用，索拉芬不适合作为治疗药物。此外，由于其复杂的结构，山梨酚不是药物开发的良好起点。我们建议找出新的小分子，这些小分子靶向于山梨醇结合部位，并以高效的方式抑制哺乳动物的ACCs。在第一阶段的研究中，我们克隆并表达了保持高亲和力的人ACC1和ACC2同工酶的生物素羧基酶(BC)结构域，建立了高通量的筛选方法，解决了酵母BC单独的和与山梨酚形成的复合体的X射线晶体结构。在第二阶段研究中，我们将利用这些工具来确定新的抑制剂，并完成下列具体目标：1)将大约2,000,000种商用化合物与沙拉芬结合部位进行分子对接；2)高通量筛选排名最靠前的对接命中和内部小分子文库；3)体外和体内对先导化合物的表征；4)通过迭代合成和测试类似物来优化先导药物；5)确定更多BC结构域的结构和与BC结构域复杂的新先导分子的结构；6)对具有1至3个先导化合物的动物进行疗效研究，以证明其具有治疗肥胖症和2型糖尿病的潜力。该项目将发现抗击肥胖和糖尿病的新药候选药物。这些疾病加在一起，影响了大约35%的美国人，每年造成约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.