Department of Neurology, New York University School of Medicine, New York, New York, United States of America.
PLoS One. 2011;6(9):e24844. doi: 10.1371/journal.pone.0024844. Epub 2011 Sep 13.
Prion diseases currently have no effective therapy. These illnesses affect both animal and human populations, and are characterized by the conformational change of a normal self protein PrP(C) (C for cellular) to a pathological and infectious conformer, PrP(Sc) (Sc for scrapie). We used a well characterized tissue culture model of prion infection, where mouse neuroblastoma cells (N2a) were infected with 22L PrP(Sc), to screen compounds for anti-prion activity. In a prior study we designed a library of styryl based, potential imaging compounds which were selected for high affinity binding to Alzheimer's disease β-amyloid plaques and good blood-brain barrier permeability. In the current study we screened this library for activity in the N2a/22L tissue culture system. We also tested the anti-prion activity of two clinically used drugs, trimipramine and fluphenazine, in the N2a/22L system. These were selected based on their structural similarity to quinacrine, which was previously reported to have anti-prion activity. All the compounds were also screened for toxicity in tissue culture and their ability to disaggregate amyloid fibrils composed of PrP and β-amyloid synthetic peptides in vitro. Two of the imaging agents, 23I and 59, were found to be both effective at inhibiting prion infection in N2a/22L tissue culture and to be non-toxic. These two compounds, as well as trimipramine and fluphenazine were evaluated in vivo using wild-type CD-1 mice infected peripherally with 139A PrP(Sc). All four agents significantly prolonged the asymptomatic incubation period of prion infection (p<0.0001 log-rank test), as well as significantly reducing the degree of spongiform change, astrocytosis and PrP(Sc) levels in the brains of treated mice. These four compounds can be considered, with further development, as candidates for prion therapy.
朊病毒病目前尚无有效疗法。这些疾病既影响动物也影响人类,其特征是正常的自我蛋白 PrP(C)(C 代表细胞)构象发生变化,转变为病理性和传染性构象,即 PrP(Sc)(Sc 代表瘙痒病)。我们使用了一种经过充分特征描述的朊病毒感染组织培养模型,其中用 22L PrP(Sc)感染鼠神经母细胞瘤细胞(N2a),以筛选具有抗朊病毒活性的化合物。在之前的研究中,我们设计了一个基于芐基的潜在成像化合物库,这些化合物被选为对阿尔茨海默病β-淀粉样斑块具有高亲和力结合和良好的血脑屏障通透性。在当前的研究中,我们在 N2a/22L 组织培养系统中筛选了这个文库的活性。我们还在 N2a/22L 系统中测试了两种临床使用的药物,即三甲丙咪嗪和氟奋乃静的抗朊病毒活性。选择这些药物是基于它们与奎宁的结构相似性,奎宁先前被报道具有抗朊病毒活性。所有化合物还在组织培养中进行了毒性筛选,并在体外测试了它们分解由 PrP 和β-淀粉样肽组成的淀粉样纤维的能力。两种成像剂,23I 和 59,被发现既能有效抑制 N2a/22L 组织培养中的朊病毒感染,又没有毒性。这两种化合物以及三甲丙咪嗪和氟奋乃静在经外周感染 139A PrP(Sc)的野生型 CD-1 小鼠中进行了体内评估。这四种药物都显著延长了朊病毒感染的无症状潜伏期(p<0.0001 log-rank 检验),并显著降低了治疗小鼠大脑中的海绵状变性、星形胶质细胞增生和 PrP(Sc)水平。这四种化合物可以进一步开发,作为朊病毒治疗的候选药物。