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Here, we report the discovery of a stable yet ultrasound-responsive arylurea-dihydrotetrazine (ArU-DHTz) scaffold that undergoes efficient activation under 1 MHz therapeutic ultrasound without requiring exogenous sonosensitizers. Structure\u2013responsiveness studies revealed that activation arises from a unique combination of a monosubstituted arylurea motif and an appropriately tuned DHTz core, while mechanistic investigations identified cavitation-derived hydroxyl radicals as the primary activating species. Ultrasound activation simultaneously triggers cargo release and tetrazine generation, representing, to our knowledge, the first example of ultrasound-triggered tetrazine formation. Leveraging tetrazine as a chemical relay, we developed a \u201cRelease\u2013Relay\u2013Click\u201d strategy that converts a single ultrasound input into dual molecular outputs: therapeutic payload release and downstream click-to-release activation. Representative prodrug D6 showed ultrasound-dependent activation, translating into antitumor efficacy in vitro and in vivo, together with fluorescence self-reporting through tetrazine-mediated bioorthogonal chemistry.<\/jats:p>","DOI":"10.1021\/jacs.6c13439","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T01:54:03Z","timestamp":1788400443000},"page":"39162-39174","source":"Crossref","is-referenced-by-count":0,"title":["Discovery\nof an Ultrasound-Responsive Arylurea-Dihydrotetrazine\nArchitecture for Ultrasound-Programmed Bioorthogonal Chemistry"],"prefix":"10.1021","volume":"148","author":[{"given":"Shuanglong","family":"Chen","sequence":"first","affiliation":[{"name":"China Pharmaceutical University , , , ,","place":["Nanjing, Jiangsu, China, 211198"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaxin","family":"Chang","sequence":"additional","affiliation":[{"name":"China Pharmaceutical University , , , ,","place":["Nanjing, Jiangsu, China, 211198"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingxin","family":"Cheng","sequence":"additional","affiliation":[{"name":"China Pharmaceutical University , , , ,","place":["Nanjing, Jiangsu, China, 211198"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Ban","sequence":"additional","affiliation":[{"name":"China Pharmaceutical University , , , ,","place":["Nanjing, Jiangsu, China, 211198"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0387-7097","authenticated-orcid":true,"given":"Weiwei","family":"Guo","sequence":"additional","affiliation":[{"name":"China Pharmaceutical University , , , ,","place":["Nanjing, Jiangsu, China, 211198"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-8912-6044","authenticated-orcid":true,"given":"Yueqin","family":"Zheng","sequence":"additional","affiliation":[{"name":"China Pharmaceutical University , , , ,","place":["Nanjing, Jiangsu, China, 211198"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"issue":"22","key":"2026091609125868000_cit1","doi-asserted-by":"publisher","first-page":"7737","DOI":"10.1039\/D2CS00889K","article-title":"Bioorthogonal chemistry for prodrug\nactivation in vivo","volume":"52","author":"Fu","year":"2023","journal-title":"Chem. 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