小分子化合物DCZ3301逆转骨髓瘤细胞对硼替佐米耐药的作用及机制研究

Multiple myeloma (MM) is an incurable hematological malignancy. Drug resistance plays a critical role in recurrence and progression of relapsed or refractory MM. Small molecular compound has the advantages in conquering chemoresistance. DCZ3301 is the first aryl guanidine derivative which is similar to sorafenib. The patent application for DCZ3301 has been filed and its patent application number has been authorized by State Intellectual Property Office. In previous study, we found that, as a narrow spectrum compound, DCZ3301 with fewer potential side effects presented significant antitumor activity in bortezomib resistant MM cells, and could synergize with bortezomib. We hypothesized that DCZ3301 could reverse the enhanced chemoresistance of multiple myeloma cells to bortezomib. In the study, molecular biology, cellular biology, proteomics, experimental zoology and other technologies will be applied to explore the effects of DCZ3301 on reversing the enhanced chemoresistance of multiple myeloma cells to bortezomib and its possible mechanism by using bortezomib resistant MM cells as cellular model. It is helpful to provide the theoretical basis and new insights into therapeutic strategies for the treatment of patients with relapsed or refractory multiple myeloma and to gain a targeting small-molecular drug with our own independent intellectual property right. Our investigation achieves well prospects for clinical application.

多发性骨髓瘤（MM）是不可治愈的血液系统恶性肿瘤，细胞耐药在复发、难治MM的复发和进展过程中起关键作用。小分子化合物具有抵抗肿瘤细胞对传统化疗药物耐药的优点。小分子DCZ3301是我们自主合成的首个类似索拉非尼的芳基胍类化合物，已获得国家知识产权局的专利申请号。前期研究结果发现：DCZ3301为窄谱抗癌化合物，潜在的毒副作用小；DCZ3301具有抗耐硼替佐米（BTZ）MM细胞的作用；DCZ3301与BTZ具有协同增效作用。据此提出推论：DCZ3301可逆转MM细胞对BTZ耐药性。我们拟以耐药MM细胞株及原代MM细胞为细胞模型，通过分子生物学、细胞生物学、蛋白质组学和实验动物学等技术，研究DCZ3301逆转MM细胞对BTZ耐药的作用，并阐明其作用的分子机制，将有望获得具有我国自主知识产权的抗MM的靶向小分子药物，为复发、难治MM的治疗提供理论基础和新思路，具有临床应用前景。

DCZ3301是我们自主合成的首个与索拉非尼类似的芳基胍类化合物，与索拉非尼抗癌谱不同，DCZ3301为窄谱抗癌化合物，潜在的毒副作用小。我们发现：1. DCZ3301对多种人类癌细胞系具有很强的细胞毒性，对多发性骨髓瘤（MM）的抗癌活性最强。DCZ3301在MM细胞系及骨髓瘤原代细胞中诱导细胞毒性，而对正常细胞没有细胞毒性作用。DCZ3301可抑制对硼替佐米敏感和耐药的MM细胞增殖，诱导MM细胞发生凋亡，并使细胞周期阻滞于G2/M期。进一步研究其机制，我们发现，DCZ3301能够降低IL-6和VEGF的分泌，抑制ERK1/2、AKT和STAT3信号通路的激活，其中STAT3信号通路的抑制在DCZ3301诱导MM细胞凋亡中起功能性作用。同时体内实验表明，DCZ3301能够减轻对硼替佐米敏感和耐药的MM移植瘤小鼠的肿瘤负荷，且DCZ3301与硼替佐米存在协同作用。DCZ3301可能通过ATM/ATR信号通逆转耐硼替佐米骨髓瘤细胞的对硼替佐米的耐药。2.体外和体内成瘤小鼠模型实验显示DCZ3301可抑制DLBCL细胞系的存活。DCZ3301是通过降低线粒体膜电位诱导细胞阻滞于G2/M期并能诱导细胞凋亡。DCZ3301是通过对AKT、ERK1/2和JAK2/STAT3信号通路的激活剂进行调制发挥了抗肿瘤作用。此外，DCZ3301通过抑制DLBCL中的lck/Yes相关新型蛋白质酪氨酸激酶（lyn）的激活而抑制STAT3磷酸化。在DCZ3301与丙色胺可协调杀伤DLBCL细胞。3.体内异种移植小鼠模型和体外实验显示DCZ3301可抑制T细胞白血病/淋巴瘤细胞的生长。DCZ3301通过下调CDK1、细胞周期蛋白B1和CDC25C的表达而诱导的G2/M细胞周期停滞。DCZ3301也通过降低T细胞白血病/淋巴瘤细胞中的MMP诱导细胞凋亡，而对正常外周血单核细胞（PBMCs）无明显促凋亡作用。此外，DCZ3301诱导的凋亡可能通过半胱天冬酶依赖途径介导。