Abstract:
Activation of neuropilin-1 (NRP-1) by platelet derived growth factor (PDGF)-C sustains melanoma invasiveness. Therefore, in the search of novel agents capable of reducing melanoma spreading, PDGF-C/NRP-1 interaction was investigated as a potential druggable target. Since the PDGF-C region involved in NRP-1 binding is not yet known, based on the sequence and structural homology between PDGF-C and vascular endothelial growth factor-A (VEGF-A), we hypothesized that the NRP-1 b1 domain region involved in the interaction with VEGF-A might also be required for PDGF-C binding. Hence, this region was selected from the protein crystal structure and used as target in the molecular docking procedure. In the following virtual screening, compounds from a DrugBank database were used as query ligands to identify agents potentially capable of disrupting NRP-1/PDGF-C interaction. Among the top 45 candidates with the highest affinity, five drugs were selected based on the safety profile, lack of hormonal effects, and current availability in the market: the antipsychotic pimozide, antidiabetic gliclazide, antiallergic cromolyn sodium, anticancer tyrosine kinase inhibitor entrectinib, and antihistamine azelastine. Analysis of drug influence on PDGF-C in vitro binding to NRP-1 and PDGF-C induced migration of human melanoma cells expressing NRP-1, indicated gliclazide and entrectinib as the most specific agents that were active at clinically achievable and non-toxic concentrations. Both drugs also reverted PDGF-C ability to stimulate extracellular matrix invasion by melanoma cells resistant to BRAF inhibitors. The inhibitory effect on tumor cell motility involved a decrease of p130Cas phosphorylation, a signal transduction pathway activated by PDGF-C-mediated stimulation of NRP-1.
摘要:
血小板衍生生长因子(PDGF)-C对神经纤毛蛋白-1(NRP-1)的激活维持黑色素瘤的侵袭。因此,寻找能够减少黑素瘤扩散的新型药物,研究了PDGF-C/NRP-1相互作用作为潜在的可成药靶标。由于参与NRP-1结合的PDGF-C区域尚不清楚,基于PDGF-C和血管内皮生长因子-A(VEGF-A)之间的序列和结构同源性,我们假设参与与VEGF-A相互作用的NRP-1b1结构域区也可能是PDGF-C结合所必需的.因此,该区域从蛋白质晶体结构中选择并用作分子对接程序中的靶标。在下面的虚拟筛选中,来自DrugBank数据库的化合物用作查询配体,以鉴定可能破坏NRP-1/PDGF-C相互作用的试剂.在亲和力最高的前45名候选人中,根据安全性选择了五种药物,缺乏荷尔蒙的影响,和目前市场上的可用性:抗精神病药匹莫齐特,抗糖尿病格列齐特,抗过敏色甘酸钠,抗癌酪氨酸激酶抑制剂entrectinib,和抗组胺药物氮卓斯汀.药物对PDGF-C与NRP-1的体外结合和PDGF-C诱导的表达NRP-1的人黑素瘤细胞的迁移的影响的分析表明,格列齐特和恩列替尼是在临床上可达到且无毒的浓度下具有活性的最特异性试剂。两种药物还恢复了PDGF-C刺激对BRAF抑制剂有抗性的黑素瘤细胞侵袭细胞外基质的能力。对肿瘤细胞运动的抑制作用涉及p130Cas磷酸化的减少,PDGF-C介导的NRP-1刺激激活的信号转导途径。