Sannikova E P, Bulushova N V, Cheperegin S E, Gubaydullin I I, Chestukhina G G, Ryabichenko V V, Zalunin I A, Kotlova E K, Konstantinova G E, Kubasova T S, Shtil A A, Pokrovsky V S, Yarotsky S V, Efremov B D, Kozlov D G
State Research Institute for Genetics and Selection of Industrial Microorganisms, Moscow, Russia, 117545.
N. N. Blokhin Russian Cancer Research Center, Kashirskoye Shosse 24, Moscow, Russia, 115478.
Mol Biotechnol. 2016 Sep;58(8-9):528-39. doi: 10.1007/s12033-016-9950-1.
The modified asparaginase Was79 was derived from the recombinant wild-type L-asparaginase of Wolinella succinogenes. The Was79 contains the amino acid substitutions V23Q and K24T responsible for the resistance to trypsinolysis and the N-terminal heparin-binding peptide KRKKKGKGLGKKR responsible for the binding to heparin and tumor K562 cells in vitro. When tested on a mouse model of Fischer lymphadenosis L5178Y, therapeutic efficacy of Was79 was significantly higher than that of reference enzymes at all single therapeutic doses used (125-8000 IU/kg). At Was79 single doses of 500-8000 IU/kg, the complete remission rate of 100 % was observed. The Was79 variant can be expressed intracellularly in E. coli as a less immunogenic formyl-methionine-free form at high per cell production levels.
改良型天冬酰胺酶Was79源自琥珀酸沃林氏菌的重组野生型L-天冬酰胺酶。Was79含有负责抗胰蛋白酶水解的氨基酸取代V23Q和K24T，以及负责在体外与肝素和肿瘤K562细胞结合的N端肝素结合肽KRKKKGKGLGKKR。在Fischer淋巴瘤L5178Y小鼠模型上进行测试时，在所有使用的单一治疗剂量（125 - 8000 IU/kg）下，Was79的治疗效果均显著高于参比酶。在Was79单剂量为500 - 8000 IU/kg时，观察到完全缓解率达100%。Was79变体可以在大肠杆菌细胞内以低免疫原性的无甲酰甲硫氨酸形式高水平表达。
