Local inhibition of liver fibrosis by specific delivery of a platelet-derived growth factor kinase inhibitor to hepatic stellate cells
Local inhibition of liver fibrosis by specific delivery of a platelet-derived growth factor kinase inhibitor to hepatic stellate cells
复制标题
DOI：
10.1124/jpet.106.114496
复制
发表时间：
2007-06-01
影响因子：
3.5
通讯作者：
Kok, Robbert J.
中科院分区：
文献类型：
--
作者：
Gonzalo, Teresa;Beljaars, Leonie;Kok, Robbert J.
肝纤维化的特征在于肝星状细胞（HSC）的过度增殖和活化，血小板衍生生长因子（PDGF）在该过程中起重要作用。因此，通过特异性递送PDGF激酶抑制剂至HSC来抑制肝纤维化可能是一种有吸引力的策略。HSC选择性载体甘露糖-6-磷酸修饰的人血清白蛋白（M6 PHSA）通过基于铂的通用连接系统（ULS）配备酪氨酸激酶抑制剂4-氯-N-[4-甲基-3-（4-吡啶-3-基-嘧啶-2-基氨基）-苯基]-苯甲酰胺（PAP 19）（一种伊马替尼衍生物）。在培养活化的大鼠HSC和来自纤维化大鼠的精确切割的肝切片中评价PAP 19-M6 PHSA的抗纤维化活性。孵育24小时后，通过α-平滑肌肌动蛋白（α SMA）和前胶原1a 1的定量逆转录聚合酶链反应分析，游离抑制剂PAP 19和PAP 19-M6 PHSA均显示出有效活性。接下来，我们检查了胆管结扎（BDL）大鼠中PAP 19-M6 PHSA的器官分布和抗纤维化活性。在BDL后第10天，向雄性Wistar大鼠施用单剂量的PAP 19-M6 PHSA，并在2小时、1天或2天后处死。通过高效液相色谱分析（2小时时注射剂量的30%）定量肝脏中的PAP 19-M6 PHSA的积累，并通过载体染色在肝脏中检测。肝脏药物水平在单次给药后24和48小时持续。此外，与盐水处理的大鼠相比，在这些时间点，PAP 19-M6 PHSA减少了活化HSC的胶原沉积（天狼星红染色）和α SMA染色。因此，我们得出结论，交付的PDGF激酶抑制剂，HSC是一个有前途的技术，以减轻肝纤维化。
Liver fibrosis is characterized by excessive proliferation and activation of hepatic stellate cells (HSC), a process in which platelet-derived growth factor (PDGF) plays an important role. Inhibition of liver fibrosis via specific delivery of a PDGF kinase inhibitor to HSC might therefore be an attractive strategy. The HSC-selective carrier mannose-6-phosphate modified human serum albumin (M6PHSA) was equipped with a tyrosine kinase inhibitor, 4-chloro-N-[4-methyl-3-(4-pyridin-3-yl-pyrimidin-2-ylamino)-phenyl]-benzamide (PAP19) (an imatinib derivative), by means of the platinum-based universal linkage system (ULS). The antifibrotic activity of PAP19-M6PHSA was evaluated in culture-activated rat HSC and precision-cut liver slices from fibrotic rats. After 24-h incubation, both free inhibitor PAP19 and PAP19-M6PHSA showed potent activity, as determined by quantitative reverse transcription-polymerase chain reaction analysis of alpha-smooth muscle actin (alpha SMA) and procollagen 1a1. Next, we examined the organ distribution and antifibrotic activity of PAP19-M6PHSA in bile duct-ligated (BDL) rats. Male Wistar rats at day 10 after BDL were administered a single dose of PAP19-M6PHSA and sacrificed at 2 h, 1 day, or 2 days afterward. The accumulation of PAP19-M6PHSA in the liver was quantified by high-performance liquid chromatography analysis (30% of the injected dose at 2 h) and detected in the liver by staining of the carrier. Liver drug levels were sustained at 24 and 48 h after the single dose. Furthermore, PAP19-M6PHSA reduced collagen deposition (Sirius red staining) and alpha SMA staining of activated HSC at these time points in comparison with saline-treated rats. We therefore conclude that delivery of a PDGF-kinase inhibitor to HSC is a promising technology to attenuate liver fibrogenesis.