Anti-Neoplastic Cytotoxicity of Gemcitabine-(C-)-[anti-EGFR] in Dual-combination with Epirubicin-(C-)-[anti-HER2/] against Chemotherapeutic-Resistant Mammary Adenocarcinoma (SKBr-3) and the Complementary Effect of Mebendazole.

AIMS

Delineate the feasibility of simultaneous, dual selective "targeted" chemotherapeutic delivery and determine if this molecular strategy can promote higher levels anti-neoplastic cytotoxicity than if only one covalent immunochemotherapeutic is selectively "targeted" for delivery at a single membrane associated receptor over-expressed by chemotherapeutic-resistant mammary adenocarcinoma.

METHODOLOGY

Gemcitabine and epirubicin were covalently bond to anti-EGFR and anti-HER2/ utilizing a rapid multi-phase synthetic organic chemistry reaction scheme. Determination that 96% or greater gemcitabine or epirubicin content was covalently bond to immunoglobulin fractions following size separation by micro-scale column chromatography was established by methanol precipitation analysis. Residual binding-avidity of gemcitabine-(C-)-[anti-EG-FR] applied in dual-combination with epirubicin-(C-)-[anti-HER2/] was determined by cell-ELIZA utilizing chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) populations. Lack of fragmentation or polymerization was validated by SDS-PAGE/immunodetection/chemiluminescent autoradiography. Anti-neoplastic cytotoxic potency was determined by vitality stain analysis of chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) monolayers known to uniquely over-express EGFR (2 × 10/cell) and HER2/ (1 × 10/cell) receptor complexes. The covalent immunochemotherapeutics gemcitabine-(C-)-[anti-EGFR] and epirubicin-(C-)-[anti-HER2/] were applied simultaneously in dual-combination to determine their capacity to collectively evoke elevated levels of anti-neoplastic cytotoxicity. Lastly, the tubulin/microtubule inhibitor mebendazole evaluated to determine if it's potential to complemented the anti-neoplastic cytotoxic properties of gemcitabine-(C4-)-[anti-EGFR] in dual-combination with epirubicin-(C-)-[].

RESULTS

Dual-combination of gemcitabine-(C-)-[anti-EGFR] with epirubicin-(C-)-[anti-HER2/] produced greater levels of anti-neoplastic cytotoxicity than either of the covalent immunochemotherapeutics alone. The benzimidazole microtubule/tubulin inhibitor, mebendazole complemented the anti-neoplastic cytotoxicity of gemcitabine-(C-)-[anti-EGFR] in dual-combination with epirubicin-(C-)-[].

CONCLUSIONS

The dual-combination of gemcitabine-(C-)-[anti-EGFR] with epirubicin-(C-)-[anti-HER2/] produced higher levels of selectively "targeted" anti-neoplastic cytotoxicity against chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) than either covalent immunochemotherapeutic alone. The benzimidazole tubulin/microtubule inhibitor, mebendazole also possessed anti-neoplastic cytotoxicity against chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3) and complemented the potency and efficacy of gemcitabine-(C-)-[anti-EGFR] in dual-combination with epirubicin-(C-)-[].