Targeted delivery of polyamidoamine-paclitaxel conjugate functionalized with anti-human epidermal growth factor receptor 2 trastuzumab

Abstract

Background Antibody-dendrimer conjugates have the potential to improve the targeting and release of chemotherapeutic drugs at the tumor site while reducing adverse side effects caused by drug accumulation in healthy tissues. In this study, trastuzumab (TMAB), which binds to human epidermal growth factor receptor 2 (HER2), was used as a targeting agent in a TMAB-polyamidoamine (PAMAM) conjugate carrying paclitaxel (PTX) specifically to cells overexpressing HER2. Methods TMAB was covalently linked to a PAMAM dendrimer via bifunctional polyethylene glycol (PEG). PTX was conjugated to PAMAM using succinic anhydride as a cross-linker, yielding TMAB-PEG-PAMAM-PTX. Dynamic light scattering and transmission electron microscopy were used to characterize the conjugates. The cellular uptake and in vivo biodistribution were studied by fluorescence microscopy, flow cytometry, and Carestream In Vivo FX, respectively. Results Nuclear magnetic resonance spectroscopy demonstrated that PEG, PTX, fluorescein isothiocyanate, and cyanine7 were conjugated to PAMAM. Ultraviolet-visible spectroscopy and sodium dodecyl sulfate polyacrylamide gel electrophoresis demonstrated that TMAB was conjugated to PEG-PAMAM. Dynamic light scattering and transmission electron microscopy measurements revealed that the different conjugates ranged in size between 10 and 35 nm and had a spherical shape. In vitro cellular uptake demonstrated that the TMAB-conjugated PAMAM was taken up by HER2-overexpressing BT474 cells more efficiently than MCF-7 cells that expressed lower levels of HER2. Co-localization experiments indicated that TMAB-conjugated PAMAM was located in the cytoplasm. The in vitro cytotoxicity of TMAB-conjugated PAMAM was lower than free PTX due to the slow release of PTX from the conjugate. In vivo targeting further demonstrated that TMAB-conjugated PAMAM accumulated in the BT474 tumor model more efficiently than non-conjugated PAMAM. Conclusion TMAB can serve as an effective targeting agent, and the TMAB-conjugated PAMAM can be exploited as a potential targeted chemotherapeutic drug delivery system for tumors that overexpress HER2.

pH-triggered surface charge-switchable polymer micelles for the co-delivery of paclitaxel/disulfiram and overcoming multidrug resistance in cancer

“…PTX was conjugated to PEG-b-PLL through an SA linker, as reported previously, 28,31 and had some differences. Briefly, PTX (1.0 g, 1.17 mM) was dissolved in 80 mL of dry DMF.…”

Trastuzumab- and Fab′ fragment-modified curcumin PEG-PLGA nanoparticles: preparation and evaluation in vitro and in vivo

“…Currently, nano-drug delivery systems in the field of oncology are attracting significant interest. 1 As a new generation of drug delivery systems, nanoparticles (NPs) can increase the delivery of small molecule anticancer drugs to solid tumours. 2 Drugloaded NPs with a diameter in the range of 200 nm are deposited in solid tumors because Dovepress Dovepress 1832 Duan et al of enhanced permeability and retention, which occurs through passive targeting.…”

“…After thiolation, unreacted Traut's reagent was removed via Sephadex G-25 columns. 1 Compared to TMAB, Fab′ fragments bear sulfhydryl groups that are used directly. 13 To generate Fab′-NPs and TMAB-NPs, Fab′ fragments and thiolated TMAB were mixed with maleimide-functionalized NPs and incubated at room temperature for 16 hours.…”

Octa-arginine modified poly(amidoamine) dendrimers for improved delivery and cytotoxic effect of paclitaxel in cancer

“…The conventional cancer chemotherapy suffers disadvantages such as low solubility, poor permeability, fast clearance, non-specific toxicities, poor biodistribution and emergence of multiple drug resistance due to chronic drug exposure, which leads to meager therapeutic response and patient incompliance [4]. To address the above concerns, nano drug delivery systems have attracted great interest due to the wide range of benefits they offer and significant progress has been achieved in taking these systems into clinic [5]. Various nanosized systems such as liposomes, polymeric micelles, lipid nanoparticles, nano-structured lipid carriers, drug conjugates, dendrimers and nano-suspensions have been proven to efficiently target and deliver the active moieties to cancer either passively or actively [2,3,[6][7][8].…”