Harwood H James, Petras Stephen F, Shelly Lorraine D, Zaccaro Lawrence M, Perry David A, Makowski Michael R, Hargrove Diane M, Martin Kelly A, Tracey W Ross, Chapman Justin G, Magee William P, Dalvie Deepak K, Soliman Victor F, Martin William H, Mularski Christian J, Eisenbeis Shane A
Department of Cardiovascular and Metabolic Diseases, Pfizer Global Research and Development, Groton Laboratories, Pfizer Inc., Groton, Connecticut 06340, USA.
J Biol Chem. 2003 Sep 26;278(39):37099-111. doi: 10.1074/jbc.M304481200. Epub 2003 Jul 3.
Inhibition of acetyl-CoA carboxylase (ACC), with its resultant inhibition of fatty acid synthesis and stimulation of fatty acid oxidation, has the potential to favorably affect the multitude of cardiovascular risk factors associated with the metabolic syndrome. To achieve maximal effectiveness, an ACC inhibitor should inhibit both the lipogenic tissue isozyme (ACC1) and the oxidative tissue isozyme (ACC2). Herein, we describe the biochemical and acute physiological properties of CP-610431, an isozyme-nonselective ACC inhibitor identified through high throughput inhibition screening, and CP-640186, an analog with improved metabolic stability. CP-610431 inhibited ACC1 and ACC2 with IC50s of approximately 50 nm. Inhibition was reversible, uncompetitive with respect to ATP, and non-competitive with respect to bicarbonate, acetyl-CoA, and citrate, indicating interaction with the enzymatic carboxyl transfer reaction. CP-610431 also inhibited fatty acid synthesis, triglyceride (TG) synthesis, TG secretion, and apolipoprotein B secretion in HepG2 cells (ACC1) with EC50s of 1.6, 1.8, 3.0, and 5.7 microm, without affecting either cholesterol synthesis or apolipoprotein CIII secretion. CP-640186, also inhibited both isozymes with IC50sof approximately 55 nm but was 2-3 times more potent than CP-610431 in inhibiting HepG2 cell fatty acid and TG synthesis. CP-640186 also stimulated fatty acid oxidation in C2C12 cells (ACC2) and in rat epitrochlearis muscle strips with EC50s of 57 nm and 1.3 microm. In rats, CP-640186 lowered hepatic, soleus muscle, quadriceps muscle, and cardiac muscle malonyl-CoA with ED50s of 55, 6, 15, and 8 mg/kg. Consequently, CP-640186 inhibited fatty acid synthesis in rats, CD1 mice, and ob/ob mice with ED50s of 13, 11, and 4 mg/kg, and stimulated rat whole body fatty acid oxidation with an ED50 of approximately 30 mg/kg. Taken together, These observations indicate that isozyme-nonselective ACC inhibition has the potential to favorably affect risk factors associated with the metabolic syndrome.
抑制乙酰辅酶A羧化酶（ACC），继而抑制脂肪酸合成并刺激脂肪酸氧化，有可能对与代谢综合征相关的多种心血管危险因素产生有利影响。为实现最大疗效，ACC抑制剂应同时抑制生脂组织同工酶（ACC1）和氧化组织同工酶（ACC2）。在此，我们描述了CP - 610431（一种通过高通量抑制筛选鉴定出的非选择性同工酶ACC抑制剂）和CP - 640186（一种代谢稳定性得到改善的类似物）的生化及急性生理特性。CP - 610431抑制ACC1和ACC2的IC50约为50 nM。抑制作用是可逆的，对ATP而言是非竞争性的，对碳酸氢盐、乙酰辅酶A和柠檬酸而言是反竞争性的，表明其与酶促羧基转移反应相互作用。CP - 610431还抑制HepG2细胞（ACC1）中的脂肪酸合成、甘油三酯（TG）合成、TG分泌和载脂蛋白B分泌，EC50分别为1.6、1.8、3. 和5.7 μM，而不影响胆固醇合成或载脂蛋白CIII分泌。CP - 640186也抑制这两种同工酶，IC50约为55 nM，但在抑制HepG2细胞脂肪酸和TG合成方面比CP - 610431强2 - 3倍。CP - 640186还刺激C2C12细胞（ACC2）和大鼠肱三头肌肌条中的脂肪酸氧化，EC50分别为57 nM和1.3 μM。在大鼠中，CP - 640186降低肝脏、比目鱼肌、股四头肌和心肌中的丙二酰辅酶A，ED50分别为55、6、15和8 mg/kg。因此，CP - 640186在大鼠、CD1小鼠和ob/ob小鼠中抑制脂肪酸合成，ED50分别为13、11和4 mg/kg，并以约30 mg/kg的ED50刺激大鼠全身脂肪酸氧化。综上所述，这些观察结果表明非选择性同工酶ACC抑制有可能对与代谢综合征相关的危险因素产生有利影响。
