Synthesis and Characterization of PAMAM Dendrimer-Based Multifunctional Nanodevices for Targeting vg src="http://pubs.

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• 作者：Wojciech G. Lesniak ; Muhammed S. T. Kariapper ; Bindu M. Nair ; Wei Tan ; Alan Hutson ; Lajos P. Balogh ; Mohamed K. Khan
• 刊名：Bioconjugate Chemistry
• 出版年：2007
• 出版时间：July 2007
• 年：2007
• 卷：18
• 期：4
• 页码：1148 - 1154
• 全文大小：254K
• 年卷期：v.18,no.4(July 2007)
• ISSN：1520-4812

文摘

We have synthesized a stable and clinically relevant nanodevice (cRGD-BT-ND; ND for short) that exhibitssuperior binding to the biologic target ges/gifchars/alpha.gif" BORDER=0>_vges/gifchars/beta2.gif" BORDER=0 ALIGN="middle">₃ integrins, when either compared to the same free cRGD peptide orto the biotinylated nanodevice without covalently attached peptides (BT-ND). Selective targeting of ges/gifchars/alpha.gif" BORDER=0>_vges/gifchars/beta2.gif" BORDER=0 ALIGN="middle">₃ integrinswas achieved by coupling cyclic cRGD peptides to the nanodevice (ND) surface, while biotin groups (BT) wereused for amplified detection of bound cRGD-BT-ND by anti-biotin antibody or avidin linked to horseradishperoxidase after binding. The synthesis involved the following steps: the amino-terminated ethylenediamine coregeneration 5 poly(amidoamine) (PAMAM_E5.NH₂) dendrimer was first partially acetylated and then biotinylated,and residual primary amine termini were converted to succinamic acid groups (SAH), some of which finallywere conjugated with cRGD peptide residues through the amino group of the lysine side chain. The startingmaterial and all derivatives were extensively characterized by polyacrylamide gel electrophoresis (PAGE), sizeexclusion chromatography (SEC), potentiometric acid-base titration, MALDI-TOF, and NMR. Cytotoxicity ofall dendrimer derivatives was examined in B16F10 melanoma cell cultures using the XTT colorimetric assay forcellular viability. Binding of nanodevices to the biological target was determined using plates coated with humanges/gifchars/alpha.gif" BORDER=0>_vges/gifchars/beta2.gif" BORDER=0 ALIGN="middle">₃ integrin and ges/gifchars/alpha.gif" BORDER=0>_vges/gifchars/beta2.gif" BORDER=0 ALIGN="middle">₃ receptor expressing human dermal microvascular endothelial cells (HDMECs). ThePAMAM_E5.(NHAc)₇₂(NHBT)₈(NHSAH)₃₅(NHSA-cR GD)₄ nanodevice is nontoxic within physiologic concentration ranges and specifically binds to the ges/gifchars/alpha.gif" BORDER=0>_vges/gifchars/beta2.gif" BORDER=0 ALIGN="middle">₃ integrins, apparently much stronger than the cyclic cRGD peptideitself.