[Construction of superagonist mutein of human CNTF and its expression in Pichia pastoris].
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 and in 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.
Zhao
Hong-liang
HL
Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China. hlzhao@vip.sina.com
Xue
Chong
C
Xiong
Xiang-hua
XH
Zhang
Wei
W
Liu
Zhi-min
ZM
D000818:Animals; D019440:Anti-Obesity Agents / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D020934:Ciliary Neurotrophic Factor / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D006801:Humans; D051379:Mice; D008815:Mice, Inbred Strains; D009154:Mutation; D010843:Pichia / Q000235:genetics* / Q000378:metabolism
D019440:Anti-Obesity Agents; D020934:Ciliary Neurotrophic Factor