基因重组人粒细胞集落刺激因子在兔和小鼠中的药物动力学(英文)

目的：研究家兔和小鼠中重组人粒细胞集落刺激因子的药物动力学．方法：用Ｉｏｄｏｇｅｎ法制备１２５ＩｒｈＧＣＳＦ．用分子筛ＨＰＬＣ测定浓度．结果：ｉｖ后浓度时间曲线符合二房室模型．初期和末端Ｔ１／２分别为０２５－０３３ｈ和３２－４６ｈ．ＡＵＣ和剂量接近正比．ｓｃ达峰时间０５９±０２５ｈ，生物利用度为１０．药物主要以原型从尿排泄．给药后泌尿系统放射性最高，胆胃肠道次之，骨髓和脾脏略低或接近血浆．结论：家兔和小鼠ｒｈＧＣＳＦ药物动力学数据为临床试验提供有用参考．     

重组人粒细胞集落刺激因子致急性肾衰竭

1名54岁男性失代偿期肝硬化患者,皮下注射重组人粒细胞集落刺激因子(rhG-CSF)200μg,1次/d。用药后第2天患者出现尿色变深,第4天出现眼睑水肿,肉眼血尿、少尿等症状。BUN由4.8mmol/L升至7.9mmol/L(最高13.9mmol/L),Cr由113μmol/L升至154μmol/L(最高308μmol/L)。停用rhG-CSF,给予还原型谷胱甘肽、硫普罗宁、呋塞米等对症支持治疗。2周后肾功能恢复正常。     

重组人粒细胞集落刺激因子的临床应用

本文回顾近年来相关文献,概述rhG-CSF的临床应用状况.该药可刺激骨髓细胞增加ANC、单核细胞、T淋巴细胞数量,增强ANC的吞噬作用,用于改善肿瘤、白血病患者放化疗后的ANC减少以及因粒细胞缺乏所合并的感染、骨髓移植后的造血恢复、外周血干细胞动员疗效确切.但有些肿瘤细胞能合成G-CSF,G-CSF也能促进某些肿瘤细胞的生长和转移.临床可根据个体情况选择恰当的用药时机、疗程、剂量,并进行药效监控,以达到合理用药.     

人粒细胞集落刺激因子基因重组及在E.coli中的表达和鉴定

将RT－PCR扩增的人粒细胞集落刺激因子（hG－CSF）结构基因与表达载体pJGW1重组，转化含有pGP1－2质粒的E.coliDH5α，进行温度诱导表达。经SDS－PAGE分析，rhG－CSF表达量占菌体总蛋白量的30％以上。将其复性，经CM－52FF离子交换层析纯化，纯化后的rhG－CSF（纯度＞98％）经SDS－PAGE测定分子量约为19kD；经胰酶裂解、反相HPLC分析肽谱具有8条特异肽段；等电聚焦测定PI值约为5．8；免疫印迹实验证实其与标准hG－CSF具有相同的抗原反应特异性；NH2-末端氨基酸分析与文献报道一致，采用小鼠白血病细胞株NFS－60测定活性为I×108IU／mg。     

重组人粒细胞集落刺激因子引起失眠4例

我院自 1998年 1月～ 2 0 0 0年 12月间 ,由血液、内分泌及肿瘤化疗科等 ,应用由日本麒麟啤酒株式会社开发的重组粒细胞集落刺激因子 (商品名惠尔血 ,制造批号 9Z3 B)遇到 4例引起失眠 ,现报导如下 :病例简要1　患者男 ,68岁。三年前无明显诱因出现怕热 ,心慌 ,纳差 ,多汗 ,继双手颤抖 ,体重明显减轻 ,B超提示“结节性甲状腺肿”,诊断为“甲状腺机能亢进症”。予丙基硫氧密啶治疗 ,一年后 ,出现粒细胞进行性下降 ,改服中药效果不理想 ,于2 0 0 0年 8月 5日给碘治疗 ,粒细胞更明显下降 ,最低 15 0 0 /mm3,门诊以“甲亢”及粒细胞减少 ,于 2…     

重组人粒细胞集落刺激因子致不良反应分析

目的：分析重组人粒细胞集落刺激因子（rhG-CSF）致不良反应的相关情况,为临床药物使用提供参考。方法检索维普、万方、知网等文献库,筛选出有关rhG-CSF报道的文献,归纳分析患者在使用rhG-CSF后出现的不良反应。结果40例ADR患者中,男21例,女19例,性别比例无明显差异。大多数ADR是出现常用剂量的范围内,19例（47.50%）用药后24 h发生ADR,不良反应一般为全身与皮肤的过敏反应。结论rhG-CSF是一种利用重组DNA技术所合成的细胞因子,在临床使用过程中根据患者的情况对药物剂量进行有效控制,提高治疗效果,降低不良反应发生率。     

重组人粒细胞集落刺激因子菌种稳定性研究

菌种的质量及其稳定性在生物工程产品发酵中起着至关重要的作用^[1,2]。本文重点考察了重组人粒细胞集落刺激因子（G－CSF）菌种在LB固体培养基上（4℃条件下）的稳定性，试验表明该菌种可在LB固体培养基上稳定传代三代（60天），从而大大简化了重组人粒细胞集落刺激因子（G－CSF）生产用菌种的制备过程。     

长效化重组人粒细胞集落刺激因子的研究进展

重组人粒细胞集落刺激因子(rhG-CSF)在临床上主要用于辅助治疗癌症患者因放/化疗引起的中性粒细胞减少症,但存在体内稳定性差、半衰期短等问题,需要反复多次给药,容易使患者产生药物耐受性及免疫排斥等不良反应,因此有必要开发长效化rhG-CSF来提高其临床药效。本文综述了近年来利用聚乙二醇(PEG)修饰、融合蛋白以及新剂型等技术开发长效化rhG-CSF的研究进展,展望了其未来发展趋势。     

重组人粒细胞集落刺激因子rhG-CSF稳定性研究

粒细胞集落刺激因子（granulocyte colony stimulating factor G-CFS)主要来源于巨嗜细胞,内皮细胞及纤维母细胞。能高度特异性的刺激中性白细胞系的功能活化因子。为了探索重组人粒细胞集落刺激因子rhG-CSF稳定性,我们在4摄氏度的条件下,不同PH值,不同添加剂的条件下,对其活性的影响。     

重组人粒细胞巨噬细胞集落刺激因子在恒河猴体内的药物动力学

AIM: To examine the pharmacokinetics of i.v. and s.c. recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) in Macaca mulatta. METHODS: Plasma levels of rhGM-CSF were detected with sandwich enzyme-linked immunosorbent assay. RESULTS: Plasma concentration-time curves after i.v. rhGM-CSF in monkeys were best fitted with 3-compartment model. The 1st, 2nd, and 3rd phase T1/2 were 0.05-0.07, 0.14-0.58, and 1.4-4.1 h. Cl and K10 were similar between different doses, respectively. Cmax was 0.93 +/- 0.16 microgram.L-1, Tmax was 2.65 +/- 0.14 h, and elimination T1/2 was 2.5 +/- 0.3 h after s.c. rhGM-CSF. The bioavailability after s.c. rhGM-CSF was 0.61. CONCLUSION: Pharmacokinetics of rhGM-CSF in Macaca mulatta provided a useful index for clinical trial.     

聚乙二醇化重组人粒细胞集落刺激因子的药代动力学与组织分布研究

目的研究聚乙二醇化重组人粒细胞集落刺激因子（PEG-rhG-CSF）在动物体内的药代动力学与组织分布。方法猕猴给予不同剂量（30、100和300μg/kg）的PEG-rhG-CSF,酶联免疫吸附法（ELISA）测定猕猴血浆中PEG-rhG-CSF浓度;[125I]标记示踪法结合分子排阻色潽法观察PEG-rhG-CSF在Wistar大鼠各组织中的分布情况。结果猕猴单次sc PEG-rhG-CSF后血药浓度及系统暴露随给药剂量呈非线性增加。低、中、高剂量组达峰时间（Tmax）为6.67～12.00h,全身清除率（CL）顺序减小,药-时曲线下面积（AUC）与给药剂量不成正比。sc给药的绝对生物利用度为47.5%。与临床使用剂量的rhG-CSF（10μg/kg）相比,猕猴sc中剂量PEG-rhG-CSF后Cmax、AUC分别为（1671±357）μg/L和（45 156±9407（μg.h）/L,均明显高于rhG-CSF组（P〈0.05）;PEG-rhG-CSF组的T1/2β为（13±3）h,较rhG-CSF组的T1/2β（1.52±0.08）h显著增加（P〈0.05）;而CL为（2.3±0.5）ml/（h.kg）,较rhG-CSF组的CL（19.6±2.4）ml/（h.kg）显著降低（P〈0.01）。大鼠sc[125I]PEG-rhG-CSF后血清、肾、肺等组织放射性较高,2 h血清原形药物浓度达峰,肺、肠道、膀胱等组织在给药后分布较慢,8 h放射性达峰值。尿液中主要为小分子[125I]降解代谢产物;未观察到药物与血浆蛋白的结合。结论将rhG-CSF进行PEG化修饰可以显著降低系统清除率,延长体内半衰期,增加药物暴露,可达到长效目的。药物主要分布于血管床,并代谢为小分子从泌尿系统排泄,不易透过血脑屏障。     

Pharmacokinetic comparison of two recombinant human granulocyte colony-stimulating factor after subcutaneous administration in rabbits.

A pharmacokinetic comparison between two formulations (LeukoCIM, CIMAB SA versus Neupogen, Hoffman-La Roche, licensed by Amgen) of recombinant human granulocyte colony-stimulating factor (rhG-CSF) using non-compartmental analysis was performed in male F1 rabbits after a single subcutaneous 11.5 microg/kg dose to help decide whether to conduct further comparability tests. Unlike the absorption phase, a statistical difference was not detected between Neupogen and LeukoCIM for clearance (18.69+/-11.83 versus 28.42+/-12.11 mL/h/kg, P=0.22). In addition, using a multivariate statistical analysis by independent samples test, a significant difference was not found between the two formulations (P=0.88). Finally, the results obtained in this study confirmed the pharmacokinetic comparability between both formulations, supporting the claim for further assessments following the current protocol on biogeneric equivalence.     

Pharmacokinetics of recombinant human granulocyte colony-stimulating factor in the rat. Single and multiple dosing studies.

The pharmacokinetics of recombinant human granulocyte colony-stimulating factor (rhG-CSF) were investigated in male Sprague-Dawley rats at a dose of 5 micrograms/kg. The serum concentrations of rhG-CSF were monitored using a specific sandwich enzyme immunoassay. In single-dose studies, the influence of routes of administration were evaluated. For iv administration, the serum concentration-time data showed the rapid disappearance of rhG-CSF from the systemic blood and a mean residence time (MRT) of 1.341 hr. For sc, im, and ip administration, lower peak serum levels were observed, but after 2 to 3 hr, rhG-CSF levels were higher than those for iv administration. The MRTs after sc, im, and ip injections were 3.918, 2.894, and 3.538 hr, respectively. The serum concentration profiles after extravascular injections showed that an im injection gave slightly faster absorption kinetics of rhG-CSF from the injection site into systemic blood than did sc and ip injections. In multiple-dose studies, rhG-CSF was injected into animals iv and sc at 5 micrograms/kg/day for 7 days. On the day 7 the serum concentration-time profiles after rhG-CSF administration were compared between single and multiple dosing. The AUC after iv multiple dosing decreased by 17.4%, although half-lives and the volume of distribution were not significantly different between single and multiple dosing groups. The AUC after sc multiple dosing decreased by 25.6%; however, the bioavailability and observed maximum serum concentration of rhG-CSF were not significantly different. These results showed that the clearance of rhG-CSF increased after multiple dosing, although the mechanism of increased clearance was not apparent.     

Pharmacokinetics of glycosylated recombinant human granulocyte colony-stimulating factor (lenograstim) in healthy male volunteers

AIMS: The aim of this open, randomised, crossover, parallel-group study was to compare the pharmacokinetics and neutrophil responses of lenograstim when administered subcutaneously (s.c.) and intravenously (i.v.). METHODS: A total of 27 healthy male volunteers was recruited. Lenograstim doses (0.5, 2, 5, or 10 microg kg(-1)) were administered s.c. or i.v. once-daily for 5 days, and then, after a 10-day washout period, vice versa for a further 5 days. Lenograstim concentrations and absolute neutrophil counts (ANCs) were measured predosing and postdosing on days 1 and 5. RESULTS: Maximum serum concentrations of lenograstim were higher following i.v. dosing (mean 5.2-185.5 vs 0.7-30.0 ng ml(-1) after s.c. dosing on day 1) and attained sooner (median 0.5-0.8 vs 4.7-8.7 h on day 1). However, apparent elimination half-lives of lenograstim were longer following s.c. dosing (mean 2.3-3.3 vs 0.8-1.2 h after i.v. dosing on days 1 and 5). ANCs increased in a dose-dependent manner with both routes of lenograstim, but more prolonged rises and higher ANC peaks were attained following s.c. doses. ANCs peaked on day 6 following 5 microg kg(-1) s.c. doses (mean peak=26.3x10(9) cells l(-1)), but on day 2 after 5 microg kg(-1) i.v. doses (mean peak = 12.4 x 10(9) cells l(-1)). Irrespective of route, the most common adverse events were headaches and back/spine pain; at doses of up to 5 microg kg(-1) these were mild and generally well tolerated. CONCLUSIONS: While supporting the use of both s.c. and i.v. administered lenograstim to treat neutropenia, these results demonstrate that neutrophil responses are more sustained and prolonged with the s.c. route.     

Differences in bone responses to recombinant human granulocyte colony-stimulating factor between mice and rats

The authors previously demonstrated that high doses of recombinant human granulocyte colony-stimulating factor (rhG-CSF) induce bone changes characterized by accelerated osteoclastic bone resorption and osteogenesis due to intramembranous ossification in rats. As a basis for future analysis of the mechanisms of rhG-CSF-induced bone changes, the present study was undertaken to determine whether the bone changes observed in rats are also induced in mice. The experiment was conducted under the conditions that clearly induce bone changes in rats. Namely, 4- or 6-week-old mice received a subcutaneous injection of rhG-CSF (1,000 or 5,000 microg/kg) for 14 or 28 days, and then the femur and tibia were evaluated histopathologically. A marked increase in peripheral blood leukocyte counts, prominent splenomegaly, and a marked increase in the number of granulopoietic cells in the bone marrow, all of which are related to the major pharmacological activity of G-CSF, was observed in the rhG-CSF-treated mice. The histopathological changes observed in the rat bone, such as accelerated osteoclastic bone resorption and osteogenesis due to intramembranous ossification, were not detected in mice. These results suggest that there is a species-related difference between mice and rats in the response of the bone to rhG-CSF treatment, and that the reactivity in mice is lower than in rats.     

Pharmacokinetics and pharmacological effect of recombinant human granulocyte colony-stimulating factor conjugated to poly(styrene-co-maleic acid) in rats.

A new derivative of recombinant human granulocyte colony-stimulating factor (rhG-CSF) has been synthesized by conjugating rhG-CSF to poly(styrene-co-maleic acid) (poly(styrene-co-maleic acid)-rhG-CSF) to try to avoid glomerular filtration and thus potentiate the neutrophil-proliferating activity of rhG-CSF. Poly(styrene-co-maleic acid)-rhG-CSF was highly bound to bovine serum albumin (BSA) and the molecular weight of the poly(styrene-co-maleic acid)-rhG-CSF-BSA complex was estimated to be about 90000 by gel filtration. Intravenous administration of poly(styrene-co-maleic acid)-rhG-CSF to normal rats resulted in a dose-dependent increase in neutrophil count. The neutrophil-proliferating activity of poly(styrene-co-maleic acid)-rhG-CSF was about 10 times greater than that of rhG-CSF. After intravenous injection at a dose of 5 microg protein kg(-1) the total clearance of rhG-CSF fell from 71.0 to 32.1 mLh(-1) kg(-1) following poly(styrene-co-maleic acid) modification. An isolated perfusion study in rat kidney showed that the filtered fraction of rhG-CSF was reduced by conjugation with poly(styrene-co-maleic acid). These results suggest that poly(styrene-co-maleic acid)-conjugation can potentiate the neutrophil-proliferating activity of rhG-CSF by reducing, at least in part, its renal clearance.     

Effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on allogeneic bone marrow transplantation in mice.

The effects of recombinant human granulocyte colony stimulating factor (rhG-CSF) on the survival rate and hemopoiesis after allogeneic bone marrow transplantation (BMT) was examined. Specific pathogen-free (SPF) C3H mice, kept in a flexible isolator for germ-free animals, received 10Gy of total body irradiation (TBI), and an allogeneic BMT 24 hours later. The 14 day survival rate was 33%. When the mice were received rhG-CSF (30 micrograms/kg/day subcutaneously) daily for 2 weeks following BMT, the 14 day survival rate rose to 79%. The effect of rhG-CSF treatment on hemopoiesis in BMT mice resulted in higher neutrophil counts in the peripheral blood by day 14 post-BMT. Administration of rhG-CSF after allogeneic BMT benefited survival rate and neutrophil recovery without any apparent complications.     

Pharmacokinetics of recombinant human basic fibroblast growth factor in rabbits and mice serum and rabbits aqueous humor

AIM: To study the pharmacokinetics of recombinant human basic fibroblast growth factor (rhbFGF) in rabbits and mice after iv and postocular administration, and the changes of rhbFGF in rabbits aqueous humor after postocular administration. METHODS: After iv or postocular administration three doses of rhbFGF in rabbits and mice, rhbFGF concentration in serum and rabbit aqueous humor was determined by enzyme-linked immunosorbent assay. RESULTS: Serum concentration-time data of rabbits after iv administration of rhbFGF 1, 2, and 4 μg/kg were fitted to bi-exponential equations with half-lives of 0.9, 0.9, and 0.6 min for T1/2α and 7, 8, and 4.7 min for T1/2β. Plasma concentration-time data of mice after iv administration of rhbFGF 2.5, 5 and 10 μg/kg were fitted to bi exponential equations with half-lives of 0.4, 0.6, and 0.9 min for T1/2α　and 6, 5, and 7 min for T1/2β.The AUCs were linearly correlated to doses in both cases (rrabbit=0.997, rmouse=0.999). The serum concentrations of rhbFGF were very l     

The influence of recombinant human granulocyte colony-stimulating factor on granulopoiesis in mice recovering from cyclophosphamide treatment

Recombinant human granulocyte colony-stimulating factor (rhG-CSF) was evaluated for its effects on granulopoiesis recovery in mice pretreated with cyclophosphamide. The cytotoxic agent was administered on days 0 and 7. rhG-CSF therapy (injected from days 8 to 28) accelerated the recovery and maintained an increased level of peripheral blood neutrophils. Marrow granuloid precursors depression, due the second dose of cyclophosphamide, was prevented by rhG-CSF, but this hemopoietic growth factor was unable to increase these cells in a similar way to that observed in normal mice. This suggests a decreased granuloid marrow proliferation capacity in cyclophosphamide treated mice. In contrast, in the spleen, rhG-CSF highly increased granuloid precursors. However, the contribution of spleen to granuloid recovery was scarce. Mice receiving rhG-CSF after cyclophosphamide demonstrated a biphasic recovery pattern in bone marrow GM-CFU population. A first rebound of GM-CFUs was followed by a nadir and then a second recovery where GM-CFU progenitor cells were significantly increased was observed. On the other hand, rhG-CSF therapy highly increased the spleen GM-CFU numbers at days 24 to 28. Although rhG-CSF increased splenic granulopoiesis, this result shows that the bulk of granulopoiesis recovery due to rhG-CSF therapy in cyclophosphamide pretreated mice occurred in the marrow.