Wang K K, Nath R, Posner A, Raser K J, Buroker-Kilgore M, Hajimohammadreza I, Probert A W, Marcoux F W, Ye Q, Takano E, Hatanaka M, Maki M, Caner H, Collins J L, Fergus A, Lee K S, Lunney E A, Hays S J, Yuen P
Department of Neuroscience Therapeutics, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, MI 48105, USA.
Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6687-92. doi: 10.1073/pnas.93.13.6687.
Overactivation of calcium-activated neutral protease (calpain) has been implicated in the pathophysiology of several degenerative conditions, including stroke, myocardial ischemia, neuromuscular degeneration, and cataract formation. Alpha-mercaptoacrylate derivatives (exemplified by PD150606), with potent and selective inhibitory actions against calpain, have been identified. PD150606 exhibits the following characteristics: (i) Ki values for mu- and m-calpains of 0.21 microM and 0.37 microM, respectively, (ii) high specificity for calpains relative to other proteases, (iii) uncompetitive inhibition with respect to substrate, and (iv) it does not shield calpain against inactivation by the active-site inhibitor trans-(epoxysuccinyl)-L-leucyl-amido-3-methylbutane, suggesting a nonactive site action for PD150606. The recombinant calcium-binding domain from each of the large or small subunits of mu-calpain was found to interact with PD150606. In low micromolar range, PD15O6O6 inhibited calpain activity in two intact cell systems. The neuroprotective effects of this class of compound were also demonstrated by the ability of PD150606 to attenuate hypoxic/hypoglycemic injury to cerebrocortical neurons in culture and excitotoxic injury to Purkinje cells in cerebellar slices.
钙激活中性蛋白酶(钙蛋白酶)的过度激活与多种退行性疾病的病理生理过程有关,包括中风、心肌缺血、神经肌肉变性和白内障形成。已鉴定出α-巯基丙烯酸酯衍生物(以PD150606为例),其对钙蛋白酶具有强效和选择性抑制作用。PD150606具有以下特性:(i)对μ-钙蛋白酶和m-钙蛋白酶的Ki值分别为0.21微摩尔和0.37微摩尔,(ii)相对于其他蛋白酶对钙蛋白酶具有高特异性,(iii)对底物的非竞争性抑制,以及(iv)它不会保护钙蛋白酶免受活性位点抑制剂反式-(环氧琥珀酰基)-L-亮氨酰-氨基-3-甲基丁烷的失活作用,这表明PD150606具有非活性位点作用。发现来自μ-钙蛋白酶大亚基或小亚基的重组钙结合结构域与PD150606相互作用。在低微摩尔范围内,PD15O6O6在两个完整细胞系统中抑制钙蛋白酶活性。PD150606减轻培养的大脑皮质神经元的缺氧/低血糖损伤以及小脑切片中浦肯野细胞的兴奋毒性损伤的能力也证明了这类化合物的神经保护作用。