Department of Chemistry, University of Chicago, Chicago, IL 60637, USA.
Proc Natl Acad Sci U S A. 2010 May 18;107(20):9093-8. doi: 10.1073/pnas.1003721107. Epub 2010 May 3.
Acetyl-CoA carboxylase (ACC) is a key enzyme of fatty acid metabolism with multiple isozymes often expressed in different eukaryotic cellular compartments. ACC-made malonyl-CoA serves as a precursor for fatty acids; it also regulates fatty acid oxidation and feeding behavior in animals. ACC provides an important target for new drugs to treat human diseases. We have developed an inexpensive nonradioactive high-throughput screening system to identify new ACC inhibitors. The screen uses yeast gene-replacement strains depending for growth on cloned human ACC1 and ACC2. In "proof of concept" experiments, growth of such strains was inhibited by compounds known to target human ACCs. The screen is sensitive and robust. Medium-size chemical libraries yielded new specific inhibitors of human ACC2. The target of the best of these inhibitors was confirmed with in vitro enzymatic assays. This compound is a new drug chemotype inhibiting human ACC2 with 2.8 muM IC(50) and having no effect on human ACC1 at 100 muM.
乙酰辅酶 A 羧化酶（ACC）是脂肪酸代谢的关键酶，具有多种同工酶，通常在不同的真核细胞区室中表达。ACC 生成的丙二酰辅酶 A 是脂肪酸的前体; 它还调节动物的脂肪酸氧化和进食行为。ACC 为治疗人类疾病的新药提供了一个重要的靶标。我们开发了一种廉价的非放射性高通量筛选系统，以鉴定新的 ACC 抑制剂。该筛选使用依赖于克隆的人 ACC1 和 ACC2 生长的酵母基因替换菌株。在“概念验证”实验中，已知靶向人 ACC 的化合物抑制了这些菌株的生长。该筛选具有灵敏性和稳健性。中等大小的化学文库产生了新的人类 ACC2 的特异性抑制剂。这些抑制剂中最好的抑制剂的靶标已通过体外酶促测定得到证实。这种化合物是一种新的药物化学类型，以 2.8 μM 的 IC50 抑制人 ACC2，在 100 μM 时对人 ACC1 没有影响。
