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To overcome bacterium\u2010induced chemotherapeutic drug resistance and potentiate antitumor immunity, herein a novel molecule Biotin\u2010Lys(SA\u2010Cip\u2010OH)\u2010Lys(SA\u2010CPT)\u2010Phe\u2010Phe\u2010Nap (<jats:bold>Biotin\u2010Cip\u2010CPT\u2010Nap<\/jats:bold>) is rationally designed containing four functional motifs (i.e., a biotin motif for targeting, Phe\u2010Phe(\u2010Nap) motif for self\u2010assembly, ciprofloxacin derivative (Cip\u2010OH) motif for antibacterial effect, and camptothecin (CPT) motif for chemotherapy). Using the designed molecule, a novel strategy of intracellular enzymatic nanofiber formation and synergistic antibacterium\u2010enhanced chemotherapy and immunotherapy is achieved. Under endocytosis mediated by highly expressed biotin receptor in colorectal cancer cell membrane and the catalysis of highly expressed carboxylesterase in the cytoplasm, this novel molecule can be transformed into <jats:bold>Biotin\u2010Nap<\/jats:bold>, which self\u2010assembled into nanofibers. Meanwhile, antibiotic Cip\u2010OH and chemotherapeutic drug CPT are released, overcoming bacterium\u2010induced drug resistance and enhancing the therapeutic efficacy of immunotherapy towards colorectal cancer. This work offers a feasible strategy for the design of novel multifunctional prodrugs to improve the efficiency of colorectal cancer treatment.<\/jats:p>","DOI":"10.1002\/adma.202312153","type":"journal-article","created":{"date-parts":[[2024,3,6]],"date-time":"2024-03-06T01:54:45Z","timestamp":1709690085000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Three\u2010in\u2010One Peptide Prodrug with Targeting, Assembly and Release Properties for Overcoming Bacterium\u2010Induced Drug Resistance and Potentiating Anti\u2010Cancer Immune Response"],"prefix":"10.1002","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-3767-237X","authenticated-orcid":false,"given":"Ge","family":"Gao","sequence":"first","affiliation":[{"name":"Cancer Institute Xuzhou Medical University  209 Tongshan Road Xuzhou Jiangsu 221004 China"},{"name":"Center of Clinical Oncology The Affiliated Hospital of Xuzhou Medical University 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