以微管蛋白和PARP-1为双靶点的新型抗肿瘤化合物的设计、合成及生物活性评价

Tubulin inhibitors (such as paclitaxel) are widely used in clinical treatment for a variety of malignant tumors, but are easy to produce the drug resistance. Multiple studies have demonstrated that overexpression of poly(ADP-ribose) polymerase 1 (PARP-1) is associated closely with cancer drug resistance to tubulin inhibitors. Accordingly, we get the idea that dual inhibitors of tubulin and PARP-1 possibly have better antitumor activity and is less prone to drug resistance (at present there are no such reports). We have discovered a molecule (D1) derived from quinazolinone scaffold by a computer model, which effectively inhibits tubulin and PARP-1. The bioassay of compound D1 certified that it has showed strong antitumor activity against human lung squamous carcinoma, human malignant melanoma, and human non-small cell lung cancer, especially for the drug-resistant cancer cell line. Based on our previous work, we decide to further optimize the lead compound to increase its activity and selectivity, using computer-aided design methods such as active site analysis, drug-likeness prediction and virtual screening. We will also undertake further study on qualitative and quantitative structure-activity relationship, in vitro and in vivo antitumor activity and mechanism. We hope to lay a solid foundation for the development of dual new anticancer drugs.

微管蛋白抑制剂（如紫杉醇）广泛用于临床多种恶性肿瘤的治疗，但易产生耐药性。多项研究表明多聚ADP核糖聚合酶-1（PARP-1）的过量表达与微管蛋白抑制剂的肿瘤耐药性密切相关。基于此，我们提出科学设想：微管蛋白和PARP-1双靶点抑制剂可能具有更强的抗肿瘤活性且不易产生耐药（目前国际上尚未有该类抑制剂相关报道）。我们前期采用计算机叠合方法发现了具有双重抑制活性的喹唑啉酮类骨架分子D1；活性测试证实，该化合物对肺鳞癌、黑色素瘤和非小细胞肺癌均显示了良好的抗肿瘤活性，尤其对耐药肿瘤细胞系仍然显示了较好的活性。本项目在前期研究基础上，采用基于受体的活性位点分析、类药性预测、双靶点虚拟筛选等计算机辅助设计方法，进一步优化苗头化合物结构，以提高微管蛋白和PARP-1的选择性和抗肿瘤活性，并进行深入的定性、定量构效关系研究，体内、外抗肿瘤活性及其作用机制研究，为获得双靶向的新型抗肿瘤药物奠定坚实基础。

微管蛋白和多聚ADP核糖聚合酶-1（PARP-1）通常在多种肿瘤中高表达。因此，以微管蛋白和PARP-1为靶点的双靶点抑制剂有望成为治疗肿瘤的理想药物。然而，这种以微管蛋白和PARP-1为靶点的抑制剂尚未见报道。本项目我们基于微管蛋白和PARP-1的结晶结构生成了的药效团模型，对SPECS和Vitas-M数据库进行了潜在双靶向抑制剂的虚拟筛选，并最终获得了10个化合物用于生物活性评价。在这些化合物中，TP-3不仅对微管蛋白和PARP-1显示了有效地抑制活性，而且体外对多种肿瘤细胞（MDA-MB-231、HepG2和MDA-MB-231耐药株）显示了较好的抗肿瘤活性。在机制研究中，TP-3可以有效地干扰MDA-MB-231细胞微管蛋白聚合和微管组装。它还通过引起MDA-MB-231细胞的G2/M期阻滞。此外，TP-3剂量依赖性地诱导MDA-MB-231细胞凋亡。在HUVEC细胞中，TP-3破坏了毛细血管样管的形成，表明其作为血管破坏剂的潜力。重要的是，TP-3在乳腺癌异种移植小鼠模型中显著抑制肿瘤生长且无明显毒性。这项研究表明TP-3可能作为一种潜在的化疗药物用于治疗乳腺癌。