{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T20:49:23Z","timestamp":1790196563324,"version":"4.1.0"},"reference-count":124,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T00:00:00Z","timestamp":1785715200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["2023.12099.PEX"],"award-info":[{"award-number":["2023.12099.PEX"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["2024.08789.CEECIND"],"award-info":[{"award-number":["2024.08789.CEECIND"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"award":["2023.12099.PEX"],"award-info":[{"award-number":["2023.12099.PEX"]}],"id":[{"id":"https:\/\/ror.org\/00snfqn58","id-type":"ROR","asserted-by":"publisher"}]},{"award":["2024.08789.CEECIND"],"award-info":[{"award-number":["2024.08789.CEECIND"]}],"id":[{"id":"https:\/\/ror.org\/00snfqn58","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000780","name":"EU Horizon Europe research and innovation program, EIC Pathfinder Open network \u201cSMARTdrugs\u201d","doi-asserted-by":"publisher","award":["101129886"],"award-info":[{"award-number":["101129886"]}],"id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Antibody\u2013drug conjugates (ADCs) have emerged as a powerful class of targeted therapeutics in many clinical areas, such as in oncology. Despite their efficacy, the onset of adverse events has been a major drawback in their clinical use. Among other explanations, the clinical performance of the ADCs has been associated with the chemistry of the linker connecting the antibody and payload. Linkers determine plasma stability, intracellular activation, and payload diffusibility, thereby influencing the therapeutic index, off-tumour toxicity, and by-stander activity. Mechanistic insights increasingly show that linker\u2013payload properties govern catabolite permeability and intratumoral distribution, particularly in antigen-heterogeneous settings. Current developments include enzyme-cleavable and tumour-selective linkers, polarity-modulating masking strategies, alternative self-immolative spacers, and dual-trigger systems designed to enhance selectivity and decouple efficacy from toxicity. In parallel, linker behaviour intersects with broader mechanisms of tumour resistance. This review focuses on understanding these processes, which are essential for designing the next generation of linkers capable of improving stability, safety, and long-term therapeutic effectiveness across diverse tumour contexts.<\/jats:p>","DOI":"10.3390\/pharmaceutics18080954","type":"journal-article","created":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T17:26:52Z","timestamp":1785778012000},"page":"954","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Linker Design in Antibody-Drug Conjugates: Balancing Stability and Drug Release"],"prefix":"10.3390","volume":"18","author":[{"given":"Sara N.","family":"Albino","sequence":"first","affiliation":[{"name":"Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028 Lisboa, Portugal"},{"name":"Faculdade de Farm\u00e1cia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-6391-3730","authenticated-orcid":false,"given":"Margarida M.","family":"Domingos","sequence":"additional","affiliation":[{"name":"Funda\u00e7\u00e3o GIMM\u2014Gulbenkian Institute of Molecular Medicine, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teresa R.","family":"Pacheco","sequence":"additional","affiliation":[{"name":"Faculdade de Medicina, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028 Lisboa, Portugal"},{"name":"Funda\u00e7\u00e3o GIMM\u2014Gulbenkian Institute of Molecular Medicine, Universidade de Lisboa, Avenida Professor Egas Moniz, 1649-028 Lisboa, Portugal"},{"name":"Unidade Local de Sa\u00fade Santa Maria, Avenida Professor Egas Moniz, 1649-028 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9967-1821","authenticated-orcid":false,"given":"Ana S.","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Center for Mathematical Studies, Faculdade de Ci\u00eancias, Universidade de Lisboa, C6\u2014Piso 1, Sala 6.1.03, 1749-016 Lisboa, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7891-7562","authenticated-orcid":false,"given":"Miguel A. 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Clin."},{"key":"ref_2","first-page":"7","article-title":"Cancer statistics","volume":"72","author":"Siegel","year":"2022","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1951","DOI":"10.1021\/acs.bioconjchem.3c00374","article-title":"The Evolving Landscape of Antibody-Drug Conjugates: In Depth Analysis of Recent Research Progress","volume":"34","author":"Sasso","year":"2023","journal-title":"Bioconjug. Chem."},{"key":"ref_4","first-page":"124","article-title":"Overview of antibody-drug conjugates nonclinical and clinical toxicities and related contributing factors","volume":"8","author":"Cheng","year":"2025","journal-title":"Antib. Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4677","DOI":"10.1002\/cam4.4052","article-title":"The promising role of antibody drug conjugate in cancer therapy: Combining targeting ability with cytotoxicity effectively","volume":"10","author":"Li","year":"2021","journal-title":"Cancer Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3889","DOI":"10.1016\/j.apsb.2021.03.042","article-title":"Antibody\u2013drug conjugates: Recent advances in linker chemistry","volume":"6","author":"Su","year":"2021","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cavaco, M., Castanho, M.A.R.B., and Neves, V. (2022). The Use of Antibody-Antibiotic Conjugates to Fight Bacterial Infections. Front Microbiol., 13.","DOI":"10.3389\/fmicb.2022.835677"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Pettinato, M.C. (2021). Introduction to Antibody-Drug Conjugates. Antibodies, 10.","DOI":"10.3390\/antib10040042"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Long, R., Zuo, H., Tang, G., Zhang, C., Yue, X., Yang, J., Luo, X., Deng, Y., Qiu, J., and Li, J. (2025). Antibody-drug conjugates in cancer therapy: Applications and future advances. Front. Immunol., 16.","DOI":"10.3389\/fimmu.2025.1516419"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Wei, H., Zhang, Y., Lu, Y., Zou, Y., Zhou, L., Qin, X., and Jiang, Q. (2025). Is ADC a rising star in solid tumor? An umbrella review of systematic reviews and meta-analyses. BMC Cancer, 25.","DOI":"10.1186\/s12885-025-13726-8"},{"key":"ref_11","unstructured":"US Food and Drug Administration (2026, January 13). Available online: https:\/\/www.fda.gov\/."},{"key":"ref_12","unstructured":"(2026, January 13). European Medicines Agency (EMA). Available online: https:\/\/www.ema.europa.eu\/en\/homepage."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1208\/s12248-014-9710-8","article-title":"Antibody Drug Conjugates: Design and Selection of Linker, Payload and Conjugation Chemistry","volume":"17","author":"McCombs","year":"2015","journal-title":"AAPS J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1038\/s41571-023-00850-2","article-title":"Exploring the next generation of antibody\u2013drug conjugates","volume":"21","author":"Tsuchikama","year":"2024","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e202500305","DOI":"10.1002\/cbic.202500305","article-title":"Advanced Antibody\u2013Drug Conjugates Design: Innovation in Linker Chemistry and Site-Specific Conjugation Technologies","volume":"26","author":"Matsuda","year":"2025","journal-title":"ChemBioChem"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Balamkundu, S., and Liu, C.F. (2023). Lysosomal-Cleavable Peptide Linkers in Antibody\u2013Drug Conjugates. Biomedicines, 11.","DOI":"10.20944\/preprints202305.1084.v1"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3526","DOI":"10.1007\/s11095-015-1657-7","article-title":"Current ADC Linker Chemistry","volume":"32","author":"Jain","year":"2015","journal-title":"Pharm. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e202500262","DOI":"10.1002\/cmdc.202500262","article-title":"Linker Design for the Antibody Drug Conjugates: A Comprehensive Review","volume":"20","author":"Lei","year":"2025","journal-title":"ChemMedChem"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e202400109","DOI":"10.1002\/cmdc.202400109","article-title":"The Evolution of Antibody-Drug Conjugates: Toward Accurate DAR and Multi-specificity","volume":"19","author":"Dong","year":"2024","journal-title":"ChemMedChem"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4025","DOI":"10.1016\/j.apsb.2023.06.015","article-title":"Antibody\u2013drug conjugates: Recent advances in payloads","volume":"13","author":"Wang","year":"2023","journal-title":"Acta Pharm. Sin. B."},{"key":"ref_21","first-page":"157","article-title":"A review of conjugation technologies for antibody drug conjugates","volume":"8","author":"Fan","year":"2025","journal-title":"Antib. Ther."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"20525","DOI":"10.1038\/s41598-025-05555-3","article-title":"Linker-GPT: Design of Antibody-drug conjugates linkers with molecular generators and reinforcement learning","volume":"15","author":"Su","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"19871","DOI":"10.1021\/acs.jmedchem.5c01982","article-title":"Recent Advances in Peptide Linkers for Antibody-Drug Conjugates","volume":"68","author":"Li","year":"2025","journal-title":"J. Med. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2220","DOI":"10.1016\/j.apsb.2021.01.017","article-title":"Protease-triggered bioresponsive drug delivery for the targeted theranostics of malignancy","volume":"11","author":"Li","year":"2021","journal-title":"Acta Pharm. Sin. B."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1349","DOI":"10.1111\/cts.12985","article-title":"Estimating peptide half-life in serum from tunable, sequence-related physicochemical properties","volume":"14","author":"Cavaco","year":"2021","journal-title":"Clin. Transl. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1687","DOI":"10.1080\/15384101.2019.1632639","article-title":"\u201cPatchiness\u201d and basic cancer research: Unravelling the proteases","volume":"18","author":"Soond","year":"2019","journal-title":"Cell Cycle"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.cell.2010.03.015","article-title":"Matrix metalloproteinases: Regulators of the tumor microenvironment","volume":"141","author":"Kessenbrock","year":"2010","journal-title":"Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3507","DOI":"10.1039\/D5MD00232J","article-title":"Glycosidase-activated prodrugs of a cytotoxic iron chelator for targeted cancer therapy","volume":"16","author":"Tomar","year":"2025","journal-title":"RSC Med. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"90","DOI":"10.5897\/BMBR11.027","article-title":"Role of proteases in cancer: A review","volume":"7","author":"Rakash","year":"2012","journal-title":"Biotechnol. Mol. Biol. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1126\/science.7233209","article-title":"Lysosomal Cathepsin B: Correlation with Metastatic Potential","volume":"212","author":"Sloane","year":"1981","journal-title":"Science"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1002\/prca.201300105","article-title":"Cathepsin B: Multiple roles in cancer","volume":"8","author":"Aggarwal","year":"2014","journal-title":"Proteom. Clin. Appl."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4361","DOI":"10.1039\/C8CS00676H","article-title":"Cleavable linkers in antibody\u2013drug conjugates","volume":"48","author":"Bargh","year":"2019","journal-title":"Chem. Soc. Rev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"114197","DOI":"10.1016\/j.jconrel.2025.114197","article-title":"Design of cathepsin-sensitive linkers for tumor-selective bioconjugate drug delivery","volume":"387","author":"Giri","year":"2025","journal-title":"J. Control. Release"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1","DOI":"10.2152\/jmi.52.1","article-title":"Involvement of cathepsins in the invasion, metastasis and proliferation of cancer cells","volume":"52","author":"Nomura","year":"2005","journal-title":"J. Med. Investig."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"116329","DOI":"10.1016\/j.ejmech.2024.116329","article-title":"Cathepsin B: The dawn of tumor therapy","volume":"269","author":"Liu","year":"2024","journal-title":"Eur. J. Med. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"575","DOI":"10.3892\/ol.2015.3960","article-title":"Expression profile of cathepsins indicates the potential of cathepsins B and D as prognostic factors in breast cancer patients","volume":"11","author":"Sun","year":"2016","journal-title":"Oncol. Lett."},{"key":"ref_37","first-page":"401","article-title":"Lysosomal Proteases Cathepsins D, B, H, L and Their Inhibitors Stefins A and B in Head and Neck Cancer","volume":"376","author":"Kos","year":"1995","journal-title":"Biol. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1007\/s11307-022-01768-4","article-title":"Cysteine Cathepsins in Breast Cancer: Promising Targets for Fluorescence-Guided Surgery","volume":"25","author":"Linders","year":"2023","journal-title":"Mol. Imaging Biol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Mehrotra, S., Wickremesekera, S.K., Brasch, H.D., Van Schaijik, B., Marsh, R.W., Tan, S.T., and Itinteang, T. (2018). Expression and Localization of Cathepsins B, D and G in Cancer Stem Cells in Liver Metastasis From Colon Adenocarcinoma. Front. Surg., 5.","DOI":"10.3389\/fsurg.2018.00040"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3946320241304911","DOI":"10.1177\/03946320241304911","article-title":"Prognostic impact of matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9) in Egyptian breast cancer patients","volume":"38","author":"Fouad","year":"2024","journal-title":"Int. J. Immunopathol. Pharmacol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.humpath.2007.07.003","article-title":"Expression and prognostic role of MMP2, MMP9, MMP13, and MMP14 matrix metalloproteinases in sinonasal and oral malignant melanomas","volume":"39","author":"Kondratiev","year":"2008","journal-title":"Hum. Pathol."},{"key":"ref_42","first-page":"5865","article-title":"The relationship between MMP-2 and MMP-9 expression levels with breast cancer incidence and prognosis","volume":"14","author":"Li","year":"2017","journal-title":"Oncol. Lett."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/s10555-007-9105-8","article-title":"The tumor microenvironment: Regulation by MMP-independent effects of tissue inhibitor of metalloproteinases-2","volume":"27","year":"2008","journal-title":"Cancer Metastasis Rev."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.clbc.2025.12.001","article-title":"ADAM and ADAMTS Proteases in Breast Cancer: Molecular Mechanisms and Therapeutic Implications","volume":"26","author":"Badhai","year":"2026","journal-title":"Clin. Breast Cancer"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1007\/s00262-019-02437-2","article-title":"Proteolytic processing of PD-L1 by ADAM proteases in breast cancer cells","volume":"69","author":"Romero","year":"2020","journal-title":"Cancer Immunol. Immunother."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"111921","DOI":"10.1016\/j.ejmech.2019.111921","article-title":"Therapeutic significance of \u03b2-glucuronidase activity and its inhibitors: A review","volume":"187","author":"Awolade","year":"2020","journal-title":"Eur. J. Med. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1007\/s002100000260","article-title":"Expression and function of \u03b2-glucuronidase in pancreatic cancer: Potential role in drug targeting","volume":"362","author":"Sperker","year":"2000","journal-title":"Naunyn Schmiedebergs Arch. Pharmacol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"9694","DOI":"10.1039\/D2CS00379A","article-title":"Glycosidase activated prodrugs for targeted cancer therapy","volume":"51","author":"Martin","year":"2022","journal-title":"Chem. Soc. Rev."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1104","DOI":"10.1038\/1981104a0","article-title":"Effect of Some Glycosidase Inhibitors on Experimental Tumours in the Mouse","volume":"198","author":"Carr","year":"1963","journal-title":"Nature"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"6421","DOI":"10.1038\/s41388-020-01436-3","article-title":"The cell-surface anchored serine protease TMPRSS13 promotes breast cancer progression and resistance to chemotherapy","volume":"39","author":"Murray","year":"2020","journal-title":"Oncogene"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.bbrc.2014.01.013","article-title":"TMPRSS4 induces cancer cell invasion through pro-uPA processing","volume":"446","author":"Min","year":"2014","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Gu, J., Huang, W., Zhang, J., Wang, X., Tao, T., Yang, L., Zheng, Y., Liu, S., Yang, J., and Zhu, L. (2021). TMPRSS4 Promotes Cell Proliferation and Inhibits Apoptosis in Pancreatic Ductal Adenocarcinoma by Activating ERK1\/2 Signaling Pathway. Front. Oncol., 11.","DOI":"10.3389\/fonc.2021.628353"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"927","DOI":"10.3892\/ijmm.2017.2893","article-title":"Overexpression of TMPRSS4 promotes tumor proliferation and aggressiveness in breast cancer","volume":"39","author":"Li","year":"2017","journal-title":"Int. J. Mol. Med."},{"key":"ref_54","first-page":"59","article-title":"Mechanisms of resistance to antibody-drug conjugates in cancer therapy: Molecular basis and therapeutic strategies","volume":"8","author":"Hao","year":"2025","journal-title":"Cancer Drug Resist."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1186\/s12957-025-03667-8","article-title":"Unravelling the role of ubiquitin-specific proteases in breast carcinoma: Insights into tumour progression and immune microenvironment modulation","volume":"23","author":"Yang","year":"2025","journal-title":"World J. Surg. Oncol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1186\/s13058-014-0461-3","article-title":"Ubiquitin-specific proteases as therapeutic targets for the treatment of breast cancer","volume":"16","author":"Pal","year":"2014","journal-title":"Breast Cancer Res."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"11690","DOI":"10.12998\/wjcc.v10.i32.11690","article-title":"Emerging potential of ubiquitin-specific proteases and ubiquitin-specific proteases inhibitors in breast cancer treatment","volume":"10","author":"Huang","year":"2022","journal-title":"World J. Clin. Cases"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.canlet.2019.02.035","article-title":"Cathepsin B: A sellsword of cancer progression","volume":"449","author":"Panin","year":"2019","journal-title":"Cancer Lett."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2344","DOI":"10.7150\/jca.86531","article-title":"The Role of Cathepsin B in Pathophysiologies of Non-tumor and Tumor tissues: A Systematic Review","volume":"14","author":"Wang","year":"2023","journal-title":"J. Cancer"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Niland, S., Riscanevo, A.X., and Eble, J.A. (2021). Matrix Metalloproteinases Shape the Tumor Microenvironment in Cancer Progression. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23010146"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"2722","DOI":"10.1039\/D5SC06664F","article-title":"MMP-cleavable linker platform for tumour-responsive homo- and heterobivalent antibody\u2013drug conjugates","volume":"17","author":"Counsell","year":"2026","journal-title":"Chem. Sci."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1021\/acs.jmedchem.0c01530","article-title":"Peptide-Drug Conjugates with Different Linkers for Cancer Therapy","volume":"64","author":"Alas","year":"2021","journal-title":"J. Med. Chem."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.jconrel.2019.07.004","article-title":"Enzyme responsive drug delivery systems in cancer treatment","volume":"308","author":"Shahriari","year":"2019","journal-title":"J. Control. Release"},{"key":"ref_64","unstructured":"ACROBiosystems (2026, January 16). Proteases for Screening and Validation of ADC Linker. Available online: https:\/\/www.acrobiosystems.com\/insights\/2737."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"25464","DOI":"10.1073\/pnas.2009279117","article-title":"Deep profiling of protease substrate specificity enabled by dual random and scanned human proteome substrate phage libraries","volume":"117","author":"Zhou","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.addr.2015.12.020","article-title":"Proteases in cancer drug delivery","volume":"97","author":"Vandooren","year":"2016","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"479","DOI":"10.2174\/1574892816666210509001139","article-title":"Self-immolative Linkers in Prodrugs and Antibody Drug Conjugates in Cancer Treatment","volume":"16","author":"Edupuganti","year":"2021","journal-title":"Recent Pat. Anticancer Drug Discov."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"6168","DOI":"10.1039\/D4SC01576B","article-title":"A novel bioresponsive self-immolative spacer based on aza-quinone methide reactivity for the controlled release of thiols, phenols, amines, sulfonamides or amides","volume":"15","author":"Ermini","year":"2024","journal-title":"Chem. Sci."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"3435","DOI":"10.1039\/C7OB00121E","article-title":"Syntheses and kinetic studies of cyclisation-based self-immolative spacers","volume":"15","author":"Huvelle","year":"2017","journal-title":"Org. Biomol. Chem."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"e202417651","DOI":"10.1002\/anie.202417651","article-title":"Benzyl Ammonium Carbamates Undergo Two-Step Linker Cleavage and Improve the Properties of Antibody Conjugates","volume":"64","author":"Li","year":"2025","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_71","unstructured":"(2026, January 16). Exploring Self-Immolative Linkers in ADCs: Beyond the Classics\u2014SigutLabs. Available online: https:\/\/sigutlabs.com\/exploring-self-immolative-linkers-in-adcs-beyond-the-classics\/."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"25505","DOI":"10.1021\/acs.jmedchem.5c02681","article-title":"Legumain-Cleavable Top1i Antibody-Drug Conjugates without Self-Immolative Spacers Demonstrate Potent Antitumor Activity","volume":"68","author":"Ojo","year":"2025","journal-title":"J. Med. Chem."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1053\/j.seminhematol.2025.11.004","article-title":"Chemical evolution of antibody\u2013drug conjugates focused on linker design and conjugation strategies in hematology","volume":"62","author":"Park","year":"2025","journal-title":"Semin. Hematol."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"1686","DOI":"10.1002\/anie.202006372","article-title":"The Challenge of Peptide Proteolytic Stability Studies: Scarce Data, Difficult Readability, and the Need for Harmonization","volume":"60","author":"Cavaco","year":"2021","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_75","doi-asserted-by":"crossref","unstructured":"Wells, C.M., Harris, M., Choi, L., Murali, V.P., Guerra, F.D., and Jennings, J.A. (2019). Stimuli-Responsive Drug Release from Smart Polymers. J. Funct. Biomater., 10.","DOI":"10.3390\/jfb10030034"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"16488","DOI":"10.1039\/D3RA00866E","article-title":"Stimuli-responsive nanocarrier delivery systems for Pt-based antitumor complexes: A review","volume":"13","author":"Wang","year":"2023","journal-title":"RSC Adv."},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Rahim, M.A., Jan, N., Khan, S., Shah, H., Madni, A., Khan, A., Jabar, A., Khan, S., Elhissi, A., and Hussain, Z. (2021). Recent Advancements in Stimuli Responsive Drug Delivery Platforms for Active and Passive Cancer Targeting. Cancers, 13.","DOI":"10.3390\/cancers13040670"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1038\/s42004-025-01491-5","article-title":"Towards precision medicine using biochemically triggered cleavable conjugation","volume":"8","author":"Parshad","year":"2025","journal-title":"Commun. Chem."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"129901","DOI":"10.1016\/j.ijbiomac.2024.129901","article-title":"Stimuli responsiveness of recent biomacromolecular systems (concept to market): A review","volume":"261","author":"Singh","year":"2024","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1080\/17425247.2021.1828339","article-title":"Multifunctional and stimuli-responsive nanocarriers for targeted therapeutic delivery","volume":"18","author":"Majumder","year":"2021","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1007\/s10462-025-11148-3","article-title":"Bridging machine learning and peptide design for cancer treatment: A comprehensive review","volume":"58","author":"Eslami","year":"2025","journal-title":"Artif. Intell. Rev."},{"key":"ref_82","doi-asserted-by":"crossref","unstructured":"Yan, J., Cai, J., Zhang, B., Wang, Y., Wong, D.F., and Siu, S.W.I. (2022). Recent Progress in the Discovery and Design of Antimicrobial Peptides Using Traditional Machine Learning and Deep Learning. Antibiotics, 11.","DOI":"10.3390\/antibiotics11101451"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1038\/s44222-024-00152-x","article-title":"Machine learning for antimicrobial peptide identification and design","volume":"2","author":"Wan","year":"2024","journal-title":"Nat. Rev. Bioeng."},{"key":"ref_84","doi-asserted-by":"crossref","unstructured":"Mintoo, M., Chakravarty, A., and Tilvawala, R. (2021). N-Terminomics Strategies for Protease Substrates Profiling. Molecules, 26.","DOI":"10.3390\/molecules26154699"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"5264","DOI":"10.1021\/acs.jproteome.1c00289","article-title":"Biochemical Tools for Tracking Proteolysis","volume":"20","author":"Wang","year":"2021","journal-title":"J. Proteome Res."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"4281","DOI":"10.1128\/JVI.02869-12","article-title":"Structural basis of substrate specificity and protease inhibition in Norwalk virus","volume":"87","author":"Muhaxhiri","year":"2013","journal-title":"J. Virol."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"2909","DOI":"10.1111\/itor.13164","article-title":"On the role of metaheuristic optimization in bioinformatics","volume":"30","author":"Calvet","year":"2023","journal-title":"Int. Trans. Oper. Res."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"104896","DOI":"10.1016\/j.compbiomed.2021.104896","article-title":"A novel multi-objective metaheuristic algorithm for protein-peptide docking and benchmarking on the LEADS-PEP dataset","volume":"138","author":"Jafari","year":"2021","journal-title":"Comput. Biol. Med."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"28260","DOI":"10.1038\/s41598-024-70462-y","article-title":"A hybrid metaheuristic algorithm for antimicrobial peptide toxicity prediction","volume":"14","author":"Dao","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_90","doi-asserted-by":"crossref","unstructured":"Boone, K., Wisdom, C., Camarda, K., Spencer, P., and Tamerler, C. (2021). Combining genetic algorithm with machine learning strategies for designing potent antimicrobial peptides. BMC Bioinform., 22.","DOI":"10.1186\/s12859-021-04156-x"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"18124","DOI":"10.1021\/acs.jmedchem.4c01251","article-title":"Exo-Cleavable Linkers: Enhanced Stability and Therapeutic Efficacy in Antibody\u2013Drug Conjugates","volume":"67","author":"Watanabe","year":"2024","journal-title":"J. Med. Chem."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"2471","DOI":"10.1021\/acschembio.9b00608","article-title":"N-Terminomics\/TAILS Profiling of Proteases and Their Substrates in Ulcerative Colitis","volume":"14","author":"Gordon","year":"2019","journal-title":"ACS Chem. Biol."},{"key":"ref_93","doi-asserted-by":"crossref","unstructured":"Lualdi, M., Ronci, M., Zilocchi, M., Corno, F., Turilli, E.S., Sponchiado, M., Aceto, A., Alberio, T., and Fasano, M. (2019). Exploring the Mitochondrial Degradome by the TAILS Proteomics Approach in a Cellular Model of Parkinson\u2019s Disease. Front. Aging Neurosci., 11.","DOI":"10.3389\/fnagi.2019.00195"},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"100223","DOI":"10.1016\/j.mcpro.2022.100223","article-title":"TAILS Identifies Candidate Substrates and Biomarkers of ADAMTS7, a Therapeutic Protease Target in Coronary Artery Disease","volume":"21","author":"MacDonald","year":"2022","journal-title":"Mol. Cell Proteom."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1158\/1535-7163.MCT-22-0362","article-title":"An Enzymatically Cleavable Tripeptide Linker for Maximizing the Therapeutic Index of Antibody\u2013Drug Conjugates","volume":"21","author":"Ha","year":"2022","journal-title":"Mol. Cancer Ther."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1038\/s41389-024-00542-1","article-title":"Identification of pancreatic cancer-specific protease substrates for protease-dependent targeted delivery","volume":"13","author":"Slapak","year":"2024","journal-title":"Oncogenesis"},{"key":"ref_97","unstructured":"(2026, January 16). Exo-Linker: Positional Reconfiguration Driving Significant Advances in ADC Stability and Efficacy \u00bb ADC Review. Available online: https:\/\/www.adcreview.com\/exo-linker-positional-reconfiguration-driving-significant-advances-in-adc-stability-and-efficacy\/."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"38696","DOI":"10.1021\/acsomega.5c03755","article-title":"Biological Evaluation of Cleavable Linkers in Exatecan-Based Antibody-Drug Conjugates: A Comparative Study of DXd and Exo-Linker Platforms","volume":"10","author":"Watanabe","year":"2025","journal-title":"ACS Omega"},{"key":"ref_99","unstructured":"(2026, January 16). Exo-Cleavable Linkers: A Promising Architecture for Next-Generation ADCs\u2014SigutLabs. Available online: https:\/\/sigutlabs.com\/exo-cleavable-linkers-next-generation-adcs\/."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"2512","DOI":"10.1038\/s41467-018-04982-3","article-title":"Glutamic acid-valine-citrulline linkers ensure stability and efficacy of antibody-drug conjugates in mice","volume":"9","author":"Anami","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"958","DOI":"10.1158\/1535-7163.MCT-15-1004","article-title":"Molecular Basis of Valine-Citrulline-PABC Linker Instability in Site-Specific ADCs and Its Mitigation by Linker Design","volume":"15","author":"Dorywalska","year":"2016","journal-title":"Mol. Cancer Ther."},{"key":"ref_102","doi-asserted-by":"crossref","unstructured":"Nguyen, T.D., Bordeau, B.M., and Balthasar, J.P. (2023). Mechanisms of ADC Toxicity and Strategies to Increase ADC Tolerability. Cancers, 15.","DOI":"10.3390\/cancers15030713"},{"key":"ref_103","doi-asserted-by":"crossref","unstructured":"Dom\u00ednguez-Oliva, A., Hern\u00e1ndez-\u00c1valos, I., Mart\u00ednez-Burnes, J., Olmos-Hern\u00e1ndez, A., Verduzco-Mendoza, A., and Mota-Rojas, D. (2023). The Importance of Animal Models in Biomedical Research: Current Insights and Applications. Animals, 13.","DOI":"10.3390\/ani13071223"},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"104979","DOI":"10.1016\/j.critrevonc.2025.104979","article-title":"Bystander effect in antibody-drug conjugates: Navigating the fine line in tumor heterogeneity","volume":"216","author":"Wang","year":"2025","journal-title":"Crit. Rev. Oncol. Hematol."},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"4313","DOI":"10.1093\/jxb\/eraf194","article-title":"Four guidelines to identify bona fide protease\u2013substrate pairs","volume":"76","author":"Qi","year":"2025","journal-title":"J. Exp. Bot."},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1016\/j.ccell.2020.03.007","article-title":"Intratumor Heterogeneity: The Rosetta Stone of Therapy Resistance","volume":"37","author":"Marusyk","year":"2020","journal-title":"Cancer Cell"},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/nrclinonc.2017.166","article-title":"Tumour heterogeneity and resistance to cancer therapies","volume":"15","author":"Shaw","year":"2018","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"17","DOI":"10.47248\/chp2502040017","article-title":"Navigating Solid Tumor Heterogeneity: The Promise and Challenges of Antibody-Drug Conjugates","volume":"2","author":"Duhon","year":"2025","journal-title":"Cancer Heterog. Plast."},{"key":"ref_109","doi-asserted-by":"crossref","unstructured":"Honda, T., and Zhu, G.D. (2026). From Unmet Medical Need to Drug Candidate: A Translational Therapeutic Development Roadmap Illustrated by Dual-Payload Antibody\u2013Drug Conjugates. Biomolecules, 16.","DOI":"10.3390\/biom16071052"},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"3528","DOI":"10.1038\/s41467-021-23793-7","article-title":"Antibody-drug conjugates with dual payloads for combating breast tumor heterogeneity and drug resistance","volume":"12","author":"Yamazaki","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1038\/s41557-024-01507-y","article-title":"Homogeneous multi-payload antibody-drug conjugates","volume":"16","author":"Journeaux","year":"2024","journal-title":"Nat. Chem."},{"key":"ref_112","doi-asserted-by":"crossref","first-page":"2498162","DOI":"10.1080\/19420862.2025.2498162","article-title":"Homogeneous antibody-drug conjugates with dual payloads: Potential, methods and considerations","volume":"17","author":"Wen","year":"2025","journal-title":"MAbs"},{"key":"ref_113","unstructured":"(2026, January 16). CMC Regulatory Considerations for ADCs\u2014BOC Sciences. Available online: https:\/\/www.bocsci.com\/blog\/cmc-regulatory-considerations-for-adcs\/."},{"key":"ref_114","unstructured":"(2026, January 16). Navigating CMC Complexities in ADC Development. Available online: https:\/\/premier-research.com\/perspectives\/the-process-is-the-product-navigating-cmc-complexities-in-adc-development\/."},{"key":"ref_115","unstructured":"BioAgilytix (2026, January 16). Bioanalytical Strategies for CMC Characterization of Antibody-Drug Conjugates. Available online: https:\/\/www.bioagilytix.com\/resources\/videos\/bioanalytical-strategies-for-cmc-characterization-of-antibody-drug-conjugates\/."},{"key":"ref_116","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1208\/s12249-026-03457-9","article-title":"Antibody-Drug Conjugates Drug Product Formulation and Process Development, Scalability and Stability Considerations","volume":"27","author":"Sorret","year":"2026","journal-title":"AAPS PharmSciTech"},{"key":"ref_117","doi-asserted-by":"crossref","first-page":"197","DOI":"10.12793\/tcp.2025.33.e21","article-title":"A review of clinical pharmacology considerations in antibody-drug conjugates approved by the US Food and Drug Administration between 2000 and 2025","volume":"33","author":"Koo","year":"2025","journal-title":"Transl. Clin. Pharmacol."},{"key":"ref_118","unstructured":"FDA (2026, July 28). Clinical Pharmacology Considerations for Antibody-Drug Conjugates Guidance for Industry, Available online: https:\/\/www.fda.gov\/regulatory-information\/search-fda-guidance-documents\/clinical-pharmacology-considerations-antibody-drug-conjugates-guidance-industry."},{"key":"ref_119","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1186\/s12943-025-02489-2","article-title":"Antibody-drug conjugates in cancer therapy: Current landscape, challenges, and future directions","volume":"24","author":"Chen","year":"2025","journal-title":"Mol. Cancer"},{"key":"ref_120","doi-asserted-by":"crossref","first-page":"104865","DOI":"10.1016\/j.critrevonc.2025.104865","article-title":"Safety assessment of antibody-drug conjugates: Nonclinical toxicology, non-active ingredients, and environmental risk considerations","volume":"215","author":"Liu","year":"2025","journal-title":"Crit. Rev. Oncol. Hematol."},{"key":"ref_121","doi-asserted-by":"crossref","first-page":"e172156","DOI":"10.1172\/JCI172156","article-title":"Antibody-drug conjugates in breast cancer: Overcoming resistance and boosting immune response","volume":"133","author":"Chang","year":"2023","journal-title":"J. Clin. Investig."},{"key":"ref_122","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1016\/j.apsb.2024.12.036","article-title":"Resistance to antibody\u2013drug conjugates: A review","volume":"15","author":"Li","year":"2025","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_123","doi-asserted-by":"crossref","unstructured":"Sheyi, R., de la Torre, B.G., and Albericio, F. (2022). Linkers: An Assurance for Controlled Delivery of Antibody-Drug Conjugate. Pharmaceutics, 14.","DOI":"10.3390\/pharmaceutics14020396"},{"key":"ref_124","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1186\/s12951-025-03277-2","article-title":"Current progress and remaining challenges of peptide\u2013drug conjugates (PDCs): Next generation of antibody-drug conjugates (ADCs)?","volume":"23","author":"Wang","year":"2025","journal-title":"J. Nanobiotechnol."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/8\/954\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T17:27:36Z","timestamp":1785778056000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/8\/954"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,3]]},"references-count":124,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["pharmaceutics18080954"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics18080954","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,3]]}}}