Sajad Yaghoubi1, Tohid Gharibi2,3, Mohammad Hossein Karimi4, Muhammad Sadeqi Nezhad5,6, Alexander Seifalian7, Reza Tavakkol8, Nader Bagheri9, Asiyeh Dezhkam10, Meghdad Abdollahpour-Alitappeh11. 1. Department of Clinical Microbiology, Iranshahr University of Medical Sciences, Iranshahr, Iran. 2. Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. 3. Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran. 4. Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. 5. Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Gorgan, Iran. 6. Stem Cells and Regenerative Medicine Center of Excellence, Tehran University of Medical Sciences, Tehran, Iran. 7. Nanotechnology and Regenerative Medicine Commercialization Centre (Ltd), The London BioScience Innovation Centre, London, UK. 8. Department of Nursing, School of Nursing, Larestan University of Medical Sciences, Larestan, Iran. 9. Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran. 10. Department of Midwifery, School of Nursing and Midwifery, Iranshahr University of Medical Sciences, Iranshahr, Iran. 11. Cellular and Molecular Biology Research Center, Larestan University of Medical Sciences, Larestan, Iran. Abdollahpour1983@yahoo.com.
Abstract
BACKGROUND: Trastuzumab, a humanized monoclonal antibody targeting Human Epidermal growth factor Receptor 2 (HER2), is a therapeutic option used for the treatment of patients with HER2-overexpressing breast cancers. The primary purpose of the present study was to establish a trastuzumab-based antibody drug conjugate (ADC) to enhance the biopharmaceutical profile of trastuzumab. METHODS: In this study, trastuzumab was linked to the microtubule-disrupting agent monomethyl auristatin E (MMAE) through a peptide linker. Following conjugation, MMAE-trastuzumab ADCs were characterized using SDS-PAGE, UV/VIS, and cell-based ELISA. The inhibitory effects of the ADCs were measured on MDA-MB-453 (HER2-positive cells) and HEK-293 (HER2-negative cells) using in vitro cell cytotoxicity and colony formation assays. RESULTS: Our findings showed that approximately 3.4 MMAE payloads were conjugated to trastuzumab. MMAE-trastuzumab ADCs produced six bands, including H2L2, H2L, HL, H2, H, and L in non-reducing SDS-PAGE. The conjugates exhibited the same binding ability to MDA-MB-453 as unconjugated trastuzumab. The MTT assay showed a significant improvement in the trastuzumab activity following MMAE conjugation, representing a higher antitumor activity as compared with unconjugated trastuzumab. Furthermore, ADCs were capable of potentially inhibiting colony formation in HER2-positive cells, as compared with trastuzumab. CONCLUSION: MMAE-trastuzumab ADCs represent a promising therapeutic strategy to treat HER2-positive breast cancer.
BACKGROUND:Trastuzumab, a humanized monoclonal antibody targeting Human Epidermal growth factor Receptor 2 (HER2), is a therapeutic option used for the treatment of patients with HER2-overexpressing breast cancers. The primary purpose of the present study was to establish a trastuzumab-based antibody drug conjugate (ADC) to enhance the biopharmaceutical profile of trastuzumab. METHODS: In this study, trastuzumab was linked to the microtubule-disrupting agent monomethyl auristatin E (MMAE) through a peptide linker. Following conjugation, MMAE-trastuzumab ADCs were characterized using SDS-PAGE, UV/VIS, and cell-based ELISA. The inhibitory effects of the ADCs were measured on MDA-MB-453 (HER2-positive cells) and HEK-293 (HER2-negative cells) using in vitro cellcytotoxicity and colony formation assays. RESULTS: Our findings showed that approximately 3.4 MMAE payloads were conjugated to trastuzumab. MMAE-trastuzumab ADCs produced six bands, including H2L2, H2L, HL, H2, H, and L in non-reducing SDS-PAGE. The conjugates exhibited the same binding ability to MDA-MB-453 as unconjugated trastuzumab. The MTT assay showed a significant improvement in the trastuzumab activity following MMAE conjugation, representing a higher antitumor activity as compared with unconjugated trastuzumab. Furthermore, ADCs were capable of potentially inhibiting colony formation in HER2-positive cells, as compared with trastuzumab. CONCLUSION:MMAE-trastuzumab ADCs represent a promising therapeutic strategy to treat HER2-positive breast cancer.
Entities:
Keywords:
Antibody drug conjugate (ADC); Breast cancer; Human epidermal growth factor receptor 2 (HER2); MMAE-trastuzumab; Targeted therapy
Authors: Rashmi K Murthy; Sherene Loi; Alicia Okines; Elisavet Paplomata; Erika Hamilton; Sara A Hurvitz; Nancy U Lin; Virginia Borges; Vandana Abramson; Carey Anders; Philippe L Bedard; Mafalda Oliveira; Erik Jakobsen; Thomas Bachelot; Shlomit S Shachar; Volkmar Müller; Sofia Braga; Francois P Duhoux; Richard Greil; David Cameron; Lisa A Carey; Giuseppe Curigliano; Karen Gelmon; Gabriel Hortobagyi; Ian Krop; Sibylle Loibl; Mark Pegram; Dennis Slamon; M Corinna Palanca-Wessels; Luke Walker; Wentao Feng; Eric P Winer Journal: N Engl J Med Date: 2019-12-11 Impact factor: 91.245
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