设计低毒性的CRM1抑制剂

Recent research shows that toxicity of potent broad-spectrum anti-cancer CRM1 (nuclear export factor Chromosomal Region Maintenance 1) inhibitors might be related to its covalent binding property. With the knowledge of several CRM1 structures, I plan to design and optimize the first non-covalent CRM1 inhibitor, a 10 amino acids peptide. With two to three rounds of protein crystallization, structure determination, mutation design and binding affinity measurement, peptide’s potency will be accessed by cancer cells, and its toxicity by immortalized cells. Potent and low toxicity candidate will injected to leukemia mouse model to further check its potency and toxicity in vivo. The newly designed peptide will be the first non-covalent CRM1 inhibitor ever known, playing important role in studying influence of covalent binding on toxicity. This study will also provide theoretical foundation for future design of non-covalent, less toxic small molecule anti-cancer drugs and provide a starting template for small molecule design. I have published relevant preliminary research has been published on PNAS (2013), Blood (2012) and Leukemia (2013) as first author or co-first author.

近几年的研究显示，具有强效广谱抗肿瘤活性的CRM1（出核因子Chromosomal Region Maintenance 1）抑制剂的毒性很可能与其共价结合的方式有关。申请人将根据已知的的结构知识，优化设计全新的非共价结合的CRM1抑制剂，即一种约10个氨基酸的多肽。在两到三轮的蛋白质结晶，结构解析，突变设计和亲和力测定试验后，通过细胞实验评估其抗癌效果及毒性。特效低毒的多肽将被注入白血病小鼠体内做进一步测试。全新设计的非共价结合的CRM1抑制剂，将被用来评估共价结合对毒性的影响，也为将来设计出非共价结合的，更低毒的小分子抗肿瘤药物提供理论基础和模型起点。相关的基础研究，申请者已经以第一作者或者共同第一作者发表在PNAS（2013），Blood（2012）和Leukemia（2013）等期刊上。

核运输蛋白CRM1控制着很多肿瘤发生发展的通路，是个少有的肿瘤特异性靶点，特别是对晚期的耐药肿瘤。我们通过3轮设计和优化现存的一个多肽，综合运用结构生物学，细胞生物学，生物物理和生物化学等多种手段，成功得到了皮摩尔级别的非共价CRM1抑制剂。优化后的多肽在细胞内与CRM1共定位增强，能更有力地抑制出核和杀死肿瘤细胞。受限于多肽较差的穿膜能力，目前我们正在动物上采用基因疗法测试多肽的抗肿瘤活性以及毒副作用。本项目论证了非共价CRM1抑制剂的可行性，为进一步设计优化小分子抑制剂提供了理论基础和结构依据。多肽抑制剂或者今后研发的小分子抑制剂将可能成为一个广谱抗晚期肿瘤的临床一线药物，具有潜在巨大的社会和经济效益。