高比活人睫状神经营养因子突变体基因的构建及其在巴斯德毕赤酵母中的表达  

Construction of superagonist mutein of human CNTF and its expression inPichia pastoris

作　　者：赵洪亮[1] 薛冲[1] 熊向华[1] 张伟[1] 刘志敏[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《微生物学报》2005年第3期392-396,共5页Acta Microbiologica Sinica

基　　金：国家"8 63计划"( 2 0 0 2AA2Z3 45B)~~

摘　　要：人睫状神经营养因子(hCNTF)及其突变体有望成为治疗肥胖症的新型药物。为了减少hCNTF的副反应,提高其疗效,在hCNTF四重突变体AX15 (R13K)的基础上引入S16 5D Q16 6H突变,构建了高比活的DH_AX15 (R13K)突变体。体外和体内实验表明DH_AX15 (R13K)的活性约是AX15 (R13K)的5倍。同时体内实验还发现DH_AX15(R13K)的作用比AX15 (R13K)更为持久。这种更为持久的作用可能是由于活性提高而非半衰期延长引起的。高比活的hCNTF突变体一方面可以在保证疗效的前提下减少蛋白用量,减少副反应;另一方面可以在不增加副反应的前提下增加最大耐受剂量,提高疗效。Human ciliary neurotrophic factor (hCNTF) and its derivatives are promising therapeutics for obesity associated with diabetes. To reduce its side effects and increase its efficacy, superagonist mutein of human CNTF was constructed by the introduction of S165D/Q166H mutation into AX15(R13K), which is a mutein of naturally occurring hCNTF, with improved biological activity, stability, solubility and KEX2 resistance.In vitro TF_1 cell survival assay andin vivo antiobesity tests showed DH_AX(R13K) was about 5 fold more potent than AX15(R13K). It was further demonstrated that the antiobesity effect of DH_AX15(R13K) was more durable than that of AX15(R13K). The more durable effects of DH_AX15(R13K) is ascribed to its higher specific activity, but not to its prolonged half-life. The superagonist mutein of human CNTF would have an improved side effect profile and thus have superior therapeutic potential.

关 键 词：睫状神经营养因子 高比活突变体 巴斯德毕赤酵母 

分 类 号：Q78[生物学—分子生物学]