A Novel GTP-Binding Inhibitor, FX2149, Attenuates LRRK2 Toxicity in Parkinson's Disease Models
A Novel GTP-Binding Inhibitor, FX2149, Attenuates LRRK2 Toxicity in Parkinson's Disease Models
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DOI：
10.1371/journal.pone.0122461
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发表时间：
2015-03-27
期刊：
影响因子：
3.7
通讯作者：
Smith, Wanli W.
中科院分区：
文献类型：
--
作者：
Li, Tianxia;He, Xinhua;Smith, Wanli W.
富含亮氨酸的重复蛋白-2(LRRK2)是一种同时具有GTP结合和激酶活性的细胞质蛋白，已成为治疗帕金森病(PD)的极具前景的药物靶点。LRRK2中与PD相关的大多数突变使其GTP结合和蛋白激酶活性失调，这可能是导致神经变性的原因之一。虽然大多数已知的LRRK2抑制剂都是针对激酶结构域开发的，但我们最近发现了第一个LRRK2 GTP结合抑制剂68，它不仅在体外高效地抑制LRRK2 GTP结合和激酶活性，而且还能减少神经退行性变。然而，68的体内效应很低，因为它的大脑渗透有限。为了解决这个问题，我们在这里报道了一种新的68类似物FX2149的设计和合成，旨在提高体内疗效。体外药理实验表明，FX2149在10 nM的浓度下对LRRK2 GTP结合活性的抑制程度接近90%。此外，FX2149在50-200 nM的浓度下对突变型LRRK2诱导的SH-SY5Y细胞的神经变性具有保护作用。重要的是，FX2149为10 mg/kg(i.p.)通过小鼠脑内LRRK2 GTP结合和磷酸化实验，显示出与68(20 mg/kg)相当的脑抑制作用。此外，FX2149 10 mg/kg(i.p.)减轻脂多糖(LPS)诱导的小鼠神经炎症模型中小胶质细胞的激活和LRRK2上调，与20 mg/kg的68相当。我们的结果突出了一种新的GTP结合抑制剂，它具有更好的脑疗效，为进一步了解PD的发病机制和治疗研究提供了新的先导化合物。
Leucine-rich repeat kinase-2 (LRRK2), a cytoplasmic protein containing both GTP binding and kinase activities, has emerged as a highly promising drug target for Parkinson's disease (PD). The majority of PD-linked mutations in LRRK2 dysregulate its GTP binding and kinase activities, which may contribute to neurodegeneration. While most known LRRK2 inhibitors are developed to target the kinase domain, we have recently identified the first LRRK2 GTP binding inhibitor, 68, which not only inhibits LRRK2 GTP binding and kinase activities with high potency in vitro, but also reduces neurodegeneration. However, the in vivo effects of 68 are low due to its limited brain penetration. To address this problem, we reported herein the design and synthesis of a novel analog of 68, FX2149, aimed at increasing the in vivo efficacy. Pharmacological characterization of FX2149 exhibited inhibition of LRRK2 GTP binding activity by similar to 90% at a concentration of 10 nM using in vitro assays. Furthermore, FX2149 protected against mutant LRRK2-induced neurodegeneration in SH-SY5Y cells at 50-200 nM concentrations. Importantly, FX2149 at 10 mg/kg (i.p.) showed significant brain inhibition efficacy equivalent to that of 68 at 20 mg/kg (i.p.), determined by mouse brain LRRK2 GTP binding and phosphorylation assays. Furthermore, FX2149 at 10 mg/kg (i.p.) attenuated lipopolysaccharide (LPS)-induced microglia activation and LRRK2 upregulation in a mouse neuroinflammation model comparable to 68 at 20 mg/kg (i.p.). Our results highlight a novel GTP binding inhibitor with better brain efficacy, which represents a new lead compound for further understanding PD pathogenesis and therapeutic studies.