Li Hong, Black Paul N, Chokshi Aalap, Sandoval-Alvarez Angel, Vatsyayan Ravi, Sealls Whitney, DiRusso Concetta C
Center for Metabolic Disease, Ordway Research Institute, Inc., Albany Medical College, Albany, NY 12208, USA.
J Lipid Res. 2008 Jan;49(1):230-44. doi: 10.1194/jlr.D700015-JLR200. Epub 2007 Oct 10.
Fatty acids are implicated in the development of dyslipidemias, leading to type 2 diabetes and cardiovascular disease. We used a standardized small compound library to screen humanized yeast to identify compounds that inhibit fatty acid transport protein (FATP)-mediated fatty acid uptake into cells. This screening procedure used live yeast cells expressing human FATP2 to identify small compounds that reduced the import of a fluorescent fatty acid analog, 4,4-difluoro-5-methyl-4-bora-3a,4a-diaza-s-indacene-3-dodecanoic acid (C(1)-BODIPY-C(12)). The library used consisted of 2,080 compounds with known biological activities. Of these, approximately 1.8% reduced cell-associated C(1)-BODIPY-C(12) fluorescence and were selected as potential inhibitors of human FATP2-mediated fatty acid uptake. Based on secondary screens, 28 compounds were selected as potential fatty acid uptake inhibitors. Some compounds fell into four groups with similar structural features. The largest group was structurally related to a family of tricyclic, phenothiazine-derived drugs used to treat schizophrenia and related psychiatric disorders, which are also known to cause metabolic side effects, including hypertriglyceridemia. Potential hit compounds were studied for specificity of interaction with human FATP and efficacy in human Caco-2 cells. This study validates this screening system as useful to assess the impact of drugs in preclinical screening for fatty acid uptake.
脂肪酸与血脂异常的发生有关,可导致2型糖尿病和心血管疾病。我们使用标准化的小分子化合物库筛选人源化酵母,以鉴定抑制脂肪酸转运蛋白(FATP)介导的脂肪酸摄取进入细胞的化合物。该筛选程序使用表达人FATP2的活酵母细胞来鉴定能减少荧光脂肪酸类似物4,4-二氟-5-甲基-4-硼-3a,4a-二氮杂-s-茚并[1,2-b]噻吩-3-十二烷酸(C(1)-BODIPY-C(12))摄取的小分子化合物。所使用的化合物库包含2080种具有已知生物活性的化合物。其中,约1.8%的化合物降低了细胞相关的C(1)-BODIPY-C(12)荧光,并被选为人类FATP2介导的脂肪酸摄取的潜在抑制剂。基于二次筛选,28种化合物被选为潜在的脂肪酸摄取抑制剂。一些化合物分为具有相似结构特征的四组。最大的一组在结构上与用于治疗精神分裂症及相关精神障碍的三环吩噻嗪类药物家族有关,这类药物也已知会引起代谢副作用,包括高甘油三酯血症。对潜在的命中化合物进行了与人类FATP相互作用的特异性以及在人Caco-2细胞中的功效研究。这项研究验证了该筛选系统在临床前筛选脂肪酸摄取药物影响方面的实用性。