APS:驱虫药帕苯达唑可治疗头颈部鳞状细胞癌

2022-06-24 网络 网络

头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)是起源于口腔、咽部和喉部粘膜上皮的一种上皮性恶性肿瘤,也是头颈部最常见的恶性肿瘤,其五年生存率

头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)是起源于口腔、咽部和喉部粘膜上皮的一种上皮性恶性肿瘤,也是头颈部最常见的恶性肿瘤,其五年生存率低于50%,然而其药物治疗效果非常有限,发现新的有效治疗HNSCC的化合物将有助于改善患者的治疗和预后。

2022年5月,山东大学药学院李敏勇教授团队发表题为“Identification of anthelmintic parbendazole as a therapeutic molecule for HNSCC through connectivity map-based drug repositioning”的文章。该研究利用药物重定位成功将驱虫药帕苯达唑用于头颈部鳞状细胞癌治疗,研究成果发表于Acta Pharmaceutica Sinica B(IF=11.413)。

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研究首先根据GEO、TCGA和GTEx中HNSCC与癌旁组织的基因表达数据,筛选出肿瘤组织与癌旁组织的差异表达基因(134上调,188下调),在cMAP查询得到35个具有治疗潜力的化合物,并依据IC50筛选出最优化合物—帕苯达唑(图1A、B)。

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图1 cMap筛选发现帕苯达唑或可治疗HNSCC

随后,为了确定了帕苯达唑的直接作用靶点,研究人员采用Pull-down+MS的方法,将Biotin标记的帕苯达唑(对照组Biotin)与HN6细胞裂解液孵育后,SDS-PAGE考染后显示的差异条带(55KD和52KD)进行质谱,发现这两个条带与微管蛋白(Tubulin)匹配得分最高(图2A)。随后采用SPR(图2B)、ITC(isothermal titration calorimetry)(图2C)、 TSA(Thermal shift assay)(图2D)对帕苯达唑与Tubulin的互作关系进行了验证。

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图2 帕苯达唑的直接作用靶点的发现与验证

最后,通过细胞实验发现帕苯达唑能够抑制微管蛋白聚合(图3A),诱导细胞凋亡(图3B),阻滞细胞周期并抑制细胞迁移。通过动物实验发现帕苯达唑在斑马鱼和小鼠体内均能抑制肿瘤生长(图3C),毒性低且能与顺铂产生协同抑癌作用。

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图3 帕苯达唑发挥抑制HNSCC的作用

总之,研究根据肿瘤组织与癌旁组织的差异基因结合cMap数据库筛选到帕苯达唑,通过Pull-down+MS发现其直接靶点Tubulin,并采用SPR、ITC、TSA等技术对互作关系进行了验证,最后通过细胞、动物实验验证了帕苯达唑对HNSCC的治疗作用(图4)。

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图4 研究思路模式图

总结与讨论

药物重定位代表了药物研发中的一个相当重要的策略,其在成功率、开发成本和研究时间等方面表现了众多优势,cMAP是其中的一个常见的技术手段。正如本案例的思路,采用cMAP挖掘了已知药物的新用途,即驱虫药帕苯达唑对HNSCC具有积极的治疗作用,为人类癌症提供了一种新的治疗选择。

1) 确定疾病组织/细胞与正常组织/细胞的差异表达基因;

2) 将差异表达基因在cMAP查询筛选出候选化合物(老药)

3) Pull-down/DARTS/CETSA+MS或蛋白芯片等方法筛选药物的直接作用靶点;

4) 细胞+动物实验确定药物改善疾病的效果。

当然,如果能采用临床样本进一步进行验证,或临床验证,其意义与价值更大!

原始出处:

Liang D, Yu C, Ma Z, Yang X, Li Z, Dong X, Qin X, Du L, Li M.Identification of anthelmintic parbendazole as a therapeutic molecule for HNSCC through connectivity map-based drug repositioning.Acta Pharm Sin B . 2022 May;12(5):2429-2442

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    2022-10-28 lixiaol
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    2022-06-26 wshxjq
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