Hong G, Bazin-Redureau M, Gires P, Scherrmann J M
INSERM U26 Département de Pharmacocinétique, Hôpital Fernad, 75475 Paris, CEDEX 10, France.
Drug Metab Dispos. 1998 Jul;26(7):661-9.
Colchicine-specific goat IgG and Fab fragments were cationized by covalent coupling of hexamethylenediamine. The immunoreactivity of antibodies was not changed following cationization. The interaction of 125I-radiolabeled native (nIgG and nFab) and cationized immunoglobulin G (cIgG) and Fab fragments (cFab) with liver was investigated using isolated perfused rat liver (IPRL) and isolated rat hepatic parenchymal cells (PCs) and nonparenchymal cells (NPCs) in suspension. 125I-cIgG or 125I-cFab were more rapidly cleared from the perfusate than the corresponding native proteins. Both cIgG and cFab declined biexponentially over time in the perfusate. In contrast, the native IgG and Fab decreased monoexponentially. The half-lives of the initial and terminal phases were 5.2 +/- 1.6 min and 355.1 +/- 17.2 min for cIgG and 14.7 +/- 3.4 min and 552.4 +/- 23.7 min for cFab. The terminal half-lives of nIgG (467.4 +/- 11.6 min) and nFab (880.1 +/- 39.6 min) were longer than those of cationized molecules. The biliary protein extraction ratio of cationized IgG and Fab was greater than that of native IgG and Fab: 0.13% (cIgG), 0.02% (nIgG), 0.23% (cFab), and 0.17% (nFab). The uptake of cIgG and cFab by both PCs and NPCs was dose-dependent and was about 6-fold and 8-fold higher than that of their native counterparts, respectively. Throughout the experiment, liver viability was determined, and no toxicity was observed according to physiological analysis (bile flow rate, portal vein pressure, and pH) and biochemical analysis (glucose and hepatic enzymes: alanine transaminase, aspartate transaminase, lactate dehydrogenase) in perfusate.
秋水仙碱特异性山羊IgG和Fab片段通过己二胺的共价偶联进行阳离子化。阳离子化后抗体的免疫反应性未发生改变。使用离体灌注大鼠肝脏（IPRL）以及悬浮状态下的离体大鼠肝实质细胞（PCs）和非实质细胞（NPCs），研究了125I放射性标记的天然（nIgG和nFab）以及阳离子化免疫球蛋白G（cIgG）和Fab片段（cFab）与肝脏的相互作用。125I - cIgG或125I - cFab从灌注液中清除的速度比相应的天然蛋白质更快。cIgG和cFab在灌注液中均随时间呈双指数下降。相比之下，天然IgG和Fab呈单指数下降。cIgG初始和终末阶段的半衰期分别为5.2±1.6分钟和355.1±17.2分钟，cFab分别为14.7±3.4分钟和552.4±23.7分钟。nIgG（467.4±11.6分钟）和nFab（880.1±39.6分钟）的终末半衰期比阳离子化分子的长。阳离子化IgG和Fab的胆汁蛋白提取率高于天然IgG和Fab：0.13%（cIgG），0.02%（nIgG），0.23%（cFab），0.17%（nFab）。PCs和NPCs对cIgG和cFab的摄取均呈剂量依赖性，分别比其天然对应物高约6倍和8倍。在整个实验过程中，测定了肝脏活力，根据灌注液中的生理分析（胆汁流速、门静脉压力和pH值）和生化分析（葡萄糖和肝酶：丙氨酸转氨酶、天冬氨酸转氨酶、乳酸脱氢酶）未观察到毒性。
