A human anti-c-Met Fab fragment conjugated with doxorubicin as targeted chemotherapy for hepatocellular carcinoma

• Published in: PloS one Vol. 8; no. 5; p. e63093
• Main Authors: Chen, Ximin, Ding, Guipeng, Gao, Qihe, Sun, Jian, Zhang, Qianqian, Du, Lijian, Qiu, Zhenning, Wang, Changjun, Zheng, Feng, Sun, Bowang, Ni, Jian, Feng, Zhenqing, Zhu, Jin
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 13.05.2013; Public Library of Science (PLoS)
• Subjects: Animal models; Animal tissues; Animals; Anthracyclines; Anticancer properties; Antitumor activity; Binding; Biology; Cancer; Cancer therapies; Carcinoma, Hepatocellular - drug therapy; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - immunology; Carcinoma, Hepatocellular - pathology; Cell Line, Tumor; Cell Survival - drug effects; Chemical synthesis; Chemotherapy; Conjugation; Cytotoxicity; Doxorubicin; Doxorubicin - chemistry; Doxorubicin - pharmacology; Drug delivery systems; Drug Stability; Eye diseases; Gene Expression; Hepatocellular carcinoma; Hepatocytes - drug effects; Hepatocytes - metabolism; Hepatocytes - pathology; High-performance liquid chromatography; Humans; Hydrogen-Ion Concentration; Immunoconjugates - chemistry; Immunoconjugates - genetics; Immunoconjugates - pharmacology; Immunofluorescence; Immunoglobulin Fab Fragments - chemistry; Immunoglobulin Fab Fragments - genetics; Immunoglobulin Fab Fragments - immunology; Kinases; Liquid chromatography; Liver cancer; Liver Neoplasms - drug therapy; Liver Neoplasms - genetics; Liver Neoplasms - immunology; Liver Neoplasms - pathology; Liver Neoplasms, Experimental - drug therapy; Liver Neoplasms, Experimental - genetics; Liver Neoplasms, Experimental - immunology; Liver Neoplasms, Experimental - pathology; Localization; Male; Medical screening; Medicine; Mice; Mice, Nude; Molecular Targeted Therapy; Peptide Library; Phages; Proto-Oncogene Proteins c-met - antagonists & inhibitors; Proto-Oncogene Proteins c-met - genetics; Proto-Oncogene Proteins c-met - immunology; Rodents; Side effects; Toxicity; Tumor cells; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0063093

c-Met is over-expressed in hepatocellular carcinoma(HCC) but is absent or expressed at low levels in normal tissues. Therefore we generated a novel conjugate of a human anti-c-Met Fab fragment (MetFab) with doxorubicin (DOX) and assessed whether it had targeted antitumor activity against HCC and reduced the side-effects of DOX. The MetFab was screened from human phage library, conjugated with DOX via chemical synthesis, and the conjugation MetFab-DOX was confirmed by HPLC. The drug release patterns, the binding efficacy, and cellular distribution of MetFab-DOX were assessed. MetFab-DOX was stable at pH7.2 PBS while release doxorubicin quickly at pH4.0, the binding efficacy of MetFab-DOX was similarly as MetFab, and the cellular distribution of the MetFab-DOX is distinct from free DOX. The cytotoxicity of MetFab-DOX was analyzed by the MTT method and the nude mouse HCC model. The MetFab-DOX demonstrated cytotoxic effects on c-Met expressing-tumor cells, but not on the cells without c-Met expression. MetFab-DOX exerted anti-tumor effect and significantly reduced the side effect of free DOX in mice model. Furthermore, the localization of conjugate was confirmed by immunofluorescence staining of tumor tissue sections and optical tumor imaging, respectively, and the tissue-distribution of drug was compared between free DOX and MetFab-DOX treatment by spectrofluorometer. MetFab-DOX can localize to the tumor tissue, and the concentration of doxorubicin in the tumor was higher after MetFab-DOX administration than after DOX administration. In summary, MetFab-DOX can target c-Met expressing HCC cells effectively and have obvious antitumor activity with decreased side-effects in preclinical models of HCC.