Multicomponent Conjugates of Anticancer Drugs and Monoclonal Antibody with PAMAM Dendrimers to Increase Efficacy of HER-2 Positive Breast Cancer Therapy

Abstract

Purpose Conjugation of nanocarriers with antibodies that bind to specific membrane receptors that are overexpressed in cancer cells enables targeted delivery. In the present study, we developed and synthesised two PAMAM dendrimer-trastuzumab conjugates that carried docetaxel or paclitaxel, specifically targeted to cells which overexpressed HER-2. Methods The 1 H NMR, 13 C NMR, FTIR and RP-HPLC were used to analyse the characteristics of the products and assess their purity. The toxicity of PAMAM-trastuzumab, PAMAM-doc-trastuzumab and PAMAM-ptx-trastuzumab conjugates was determined using MTT assay and compared with free trastuzumab, docetaxel and paclitaxel toward HER-2-positive (SKBR-3) and negative (MCF-7) human breast cancer cell lines. The cellular uptake and internal localisation were studied using flow cytometry and confocal microscopy, respectively. Results The PAMAM-drug-trastuzumab conjugates in particular showed extremely high toxicity toward the HER-2-positive SKBR-3 cells and very low toxicity towards to HER-2-negative MCF-7 cells. As expected, the HER-2-positive SKBR-3 cell line accumulated trastuzumab from both conjugates rapidly; but surprisingly, although a large amount of PAMAM-ptx-trastuzumab conjugate was observed in the HER-2-negative MCF-7 cells. Confocal microscopy confirmed the intracellular localisation of analysed compounds. The key result of fluorescent imaging was the identification of strong selective binding of the PAMAM-doc-trastuzumab conjugate with HER-2-positive SKBR-3 cells only. Conclusions Our results confirm the high selectivity of PAMAM-doc-trastuzumab and PAMAM-ptx-trastuzumab conjugates for HER-2-positive cells, and demonstrate the utility of trastuzumab as a targeting agent. Therefore, the analysed conjugates present an promising approach for the improvement of efficacy of targeted delivery of anticancer drugs such as docetaxel or paclitaxel. Electronic supplementary material The online version of this article (10.1007/s11095-019-2683-7) contains supplementary material, which is available to authorized users.

<p>Trastuzumab Targeted Neratinib Loaded Poly-Amidoamine Dendrimer Nanocapsules for Breast Cancer Therapy</p>

“…This was confirmed by the results of fluorescence intensity quantitative analysis, in which neratinib-loaded-dendrimers-FITC showed statistically significant higher fluorescence intensity compared to that of neratinib-loaded-dendrimers-FITC alone (p ˂ 0.05) ( Table 2). These findings were in agreement with previous studies, 18,21,22 in which cellular uptake and antiproliferative effect of TZ-conjugated dendrimers loaded with docetaxel were significantly higher than for plain dendrimers. 21…”

Targeting Glucose Metabolism to Overcome Resistance to Anticancer Chemotherapy in Breast Cancer

“…More advanced drug designs, such as multicomponent conjugate drugs, demonstrate high efficacy and selectivity in targeting cancer cells. For example, unlike poorly soluble paclitaxel, PAMAM-paclitaxel-trastuzumab is a potent target-based drug, with increased permeability for HER2+ (SKBR-3) BC cells [ 115 ].…”

“…Drugs targeting these enzymes potentially downregulate aerobic glycolysis, lactate production, and drug efflux pumps. PAMAM-PTX is a conjugated drug targeted to mitochondria via GLUT [ 115 ]. ( B ) Targeting PI3K/AKT pathway and G6PDH can reverse cisplatin resistance [ 116 ].…”

A Group of Complexes Based on PAMAM and Quantum Dots Used in Clinical Immunoassays

“…To neutralize the positive charge, we dissolved PAMAM in DMSO and added succinic anhydride (dihydro-2,5-diketotetrahydrofuran) to neutralize the PAMAM amino group. The amino group in PAMAM reacts with succinic anhydride to form amide bonds [6][7][8][9][10]. After the reaction, the product was dialyzed, lyophilized, weighed, and redissolved, the production is nearly neutral and we named the product "N-PAMAM," which is able to conjugate with goat anti-rabbit IgG.…”