In vitro and in vivo hematopoietic effect of mutant human granulocyte colony-stimulating factor.
In vitro and in vivo hematopoietic effect of mutant human granulocyte colony-stimulating factor.
复制标题
突变型人粒细胞集落刺激因子的体外和体内造血作用。
DOI：
10.1182/blood.v75.9.1788.1788
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发表时间：
1990
期刊：
影响因子：
20.3
通讯作者：
T. Oka
中科院分区：
文献类型：
--
作者：
M. Okabe;M. Asano;T. Kuga;Y. Komatsu;M. Yamasaki;Y. Yokoo;S. Itoh;M. Morimoto;T. Oka
用各种基因突变技术，合成了约100种人重组粒细胞集落刺激因子（rhG-CSF）的衍生物，筛选出N端5个氨基酸被替换的KW-2228，并与rhG-CSF进行了生物学和理化性质的比较。KW-2228在小鼠和/或人骨髓祖细胞中的比活性通过软琼脂中的集落形成单位测定和液体培养中的细胞增殖测定显示出比完整的rhG-CSF高2至4倍。KW-2228通过单次静脉注射给药于正常C3 H/He小鼠时，显示出增加外周中性粒细胞计数的效力。观察到总白细胞计数和中性粒细胞增加，在低剂量（0.5 - 1.0 μ g/小鼠）下8 - 12小时达到峰值水平，在高剂量（5 - 10 μ g/小鼠）下维持最高水平24 - 30小时。KW-2228的粒细胞生成作用通过单疗程（每日一次，持续10天）或多疗程（每日两次注射，持续5天，然后在一个周期中停止9天，总共3个周期）的几个剂量来检查。KW-2228的活性高于rhG-CSF，尤其是在多次疗程的次优剂量下。此外，发现KW-2228在物理化学和生物学上比完整的rhG-CSF更稳定，特别是在56 ℃的热条件下和在37 ℃的人血浆中，表明具有蛋白酶抗性。药代动力学研究表明，正常小鼠静脉注射KW-2228蛋白后，测定其生物活性的血浆浓度在60分钟内保持高于完整rhG-CSF的水平。这些结果表明，KW-2228可能显示出上级完整的rhG-CSF的体内造血作用，这是由于其对祖细胞的高比活性，以及由于其在宿主动物中改善的物理化学、生物学和药代动力学稳定性。
About 100 derivatives of human recombinant granulocyte colony-stimulating factor (rhG-CSF) were created by various gene-mutagenic techniques, and KW-2228, in which amino acids were replaced at five positions of N-terminal region of intact rhG-CSF, was picked up and evaluated for its biologic and physicochemical properties in comparison with intact rhG-CSF. KW-2228 showed two to four times higher specific activity than that of intact rhG-CSF in mouse and/or human bone marrow progenitor cells by colony-forming unit assay in soft agar, and by cell-proliferation assay in liquid culture. KW-2228 showed a potency to increase peripheral neutrophil counts when it was administered to normal C3H/He mice by single intravenous injection. Increase of total leukocyte count and neutrophils was observed, with peak level at 8 to 12 hours at low doses (0.5 to 1.0 micrograms/mouse), and the highest level was maintained for 24 to 30 hours at high doses (5 to 10 micrograms/mouse). The granulopoietic effect of KW-2228 was examined by several doses of single course (once daily for 10 days) or multiple courses (twice daily injection for 5 days followed by cessation for 9 days on one cycle, 3 cycles in total) of treatment. KW-2228 showed higher activity than that of rhG-CSF, especially at sub-optimal doses of multiple courses of treatment. Furthermore, KW-2228 was found to be more stable physicochemically and biologically than intact rhG-CSF, especially under thermal conditions at 56 degrees C and in the human plasma at 37 degrees C, suggesting a protease resistancy. Pharmacokinetic study showed that plasma concentration of KW-2228 assayed for its bioactivity maintained a higher level than that of intact rhG-CSF for 60 minutes after intravenous injection of this protein to normal mice. Those results suggest that KW-2228 might show a superior in vivo hematopoietic effect to intact rhG-CSF due to its high specific activity to progenitor cells, and also due to its improved physicochemical, biologic, and pharmacokinetic stability in host animals.
影响因子：
56.9
作者：
SOUZA, LM;BOONE, TC;WELTE, K
通讯作者：
WELTE, K