Abstract
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 α-smooth muscle actin (αSMA) 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 αSMA 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.
Footnotes
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This study was supported by SenterNovem Grant TSGE1083 and NWO Science Netherlands Grant R 02-1719, 98-162.
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.106.114496.
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ABBREVIATIONS: HSC, hepatic stellate cell(s); PDGF, platelet-derived growth factor; PDGFR, platelet-derived growth factor receptor; PAP19, phenyl-amino-pyrimidine derivative 19; M6PHSA, mannose 6-phosphate modified human serum albumin; PAP, phenyl-amino-pyrimidine; ULS, universal linkage system; DMF, dimethylformamide; HPLC, high-performance liquid chromatography; PBS, phosphate-buffered saline; BDL, bile duct ligated/ligation; PCR, polymerase chain reaction; αSMA, α-smooth muscle actin; TIMP, tissue inhibitor of metalloproteinase; HSA, human serum albumin, bp, base pairs.
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↵1 Current affiliation: Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, University of Santiago de Compostela, Santiago de Compostela, Spain.
- Received September 22, 2006.
- Accepted March 13, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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