Abstract
The clinically approved human epidermal growth factor receptor 2 (HER2)-targeting monoclonal antibodies (mAbs), trastuzumab, and pertuzumab, target domains IV and II, respectively. Trastuzumab is now the standard treatment for HER2-overexpressed breast and gastric cancers, and trastuzumab in combination with pertuzumab showed clinical benefit. However, there still exist patients who do not respond to the therapy. Furthermore, HER2 mutants that cannot be recognized by pertuzumab were found in tumors. Therefore, novel anti-HER2 mAbs and modalities have been desired. In our previous study, we developed a novel anti-HER2 domain I mAb, H(2)Mab-139 (mouse IgG(1), kappa). We herein produced a defucosylated mouse IgG(2a) type of mAb against HER2 (H(2)Mab-139-mG(2a)-f) to enhance antibody-dependent cellular cytotoxicity (ADCC)-mediated antitumor activity. H(2)Mab-139-mG(2a)-f exhibits a high binding affinity in flow cytometry with the dissociation constant (K(D)) determined to be 3.9 × 10(-9) M and 7.7 × 10(-9) M against HER2-overexpressed Chinese hamster ovary (CHO)-K1 (CHO/HER2) and HER2-positive BT-474 cells, respectively. Moreover, we showed that H(2)Mab-139-mG(2a)-f exerted ADCC and complement-dependent cytotoxicity against CHO/HER2 and BT-474 in vitro and exhibited potent antitumor activities in mouse xenograft models. These results indicated that H(2)Mab-139-mG(2a)-f exerts antitumor effects against HER2-positive human breast cancers and is useful as an antibody treatment for HER2-positive human cancers.