title: Development and Characterization of the Recombinant Human VEGF-EGF Dual-Targeting Fusion Protein as a Drug Delivery System.
abstract: The design, preparation, as well as structural and functional characterizations of the recombinant fusion protein hVEGF-EGF as a dual-functional agent that may target both EGFR (R: receptor) and angiogenesis are reported. hVEGF-EGF was found to bind to EGFR more strongly than did EGF, and to bind to VEGFR similarly to VEGF. Mass spectrometry measurements showed that the sites of DTPA (diethylenetriaminepentaacetic acid) conjugated hVEGF-EGF (for radiolabeling) were the same as those of its parent hEGF and hVEGF proteins. All DTPA-conjugated proteins retained similar binding capacities to their respective receptors as compared to their respective parent proteins. In vitro cell binding studies using BAEC (a bovine aortic endothelial cell) and MDA-MB-231 (a human breast cancer) cells expressing both EGFR and VEGFR confirmed similar results. Treating BAEC cells with hVEGF-EGF induced remarkable phosphorylation of EGFR, VEGFR, and their downstream targets ERK1/2. Nevertheless, the radiolabeled (111)In-DTPA-hVEGF-EGF showed cytotoxicity against MDA-MB-231 cells. Pharmacokinetic studies using (111)In-DTPA-hVEGF-EGF in BALB/c nude mice showed that appreciable tracer activities were accumulated in liver and spleen. In all, this study demonstrated that the fusion protein hVEGF-EGF maintained the biological specificity toward both EGFR and VEGFR and may be a potential candidate as a dual-targeting moiety in developing anticancer drugs.
authors.affiliation: Program in Molecular Medicine, National Yang-Ming University and Academia Sinica , Taipei, Taiwan , 112.
authors.forename: Jia-Je
authors.initials: JJ
authors.lastname: Li
authors.forename: Keng-Li
authors.initials: KL
authors.lastname: Lan
authors.affiliation: Department of Medical Research and Development, Chang Gung Memorial Hospital-Chiayi Branch , Chiayi 613, Taiwan.
authors.forename: Shun-Fu
authors.initials: SF
authors.lastname: Chang
authors.forename: Ya-Fen
authors.initials: YF
authors.lastname: Chen
authors.forename: Wen-Chun
authors.initials: WC
authors.lastname: Tsai
authors.forename: Pei-Hsun
authors.initials: PH
authors.lastname: Chiang
authors.forename: Meng-Han
authors.initials: MH
authors.lastname: Lin
authors.forename: Wolfgang B
authors.initials: WB
authors.lastname: Fischer
authors.forename: Yi-Sheng
authors.initials: YS
authors.lastname: Shih
authors.forename: Sang-Hue
authors.initials: SH
authors.lastname: Yen
authors.forename: Ren-Shyan
authors.initials: RS
authors.lastname: Liu
authors.forename: Yeou-Guang
authors.initials: YG
authors.lastname: Tsay
authors.affiliation: Program in Molecular Medicine, National Yang-Ming University and Academia Sinica , Taipei, Taiwan , 112.
authors.forename: Hsin-Ell
authors.initials: HE
authors.lastname: Wang
authors.forename: Cheng Allen
authors.initials: CA
authors.lastname: Chang
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D002417:Cattle; D002460:Cell Line; D045744:Cell Line, Tumor; D004337:Drug Carriers / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics; D016503:Drug Delivery Systems; D004815:Epidermal Growth Factor / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009369:Neoplasms / Q000188:drug therapy / Q000378:metabolism; D004369:Pentetic Acid / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics
chemical_list: D000970:Antineoplastic Agents; D004337:Drug Carriers; D011993:Recombinant Fusion Proteins; D042461:Vascular Endothelial Growth Factor A; D004815:Epidermal Growth Factor; D004369:Pentetic Acid