{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:46:55Z","timestamp":1792784815318,"version":"4.5.0"},"reference-count":47,"publisher":"American Chemical Society (ACS)","issue":"9","license":[{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"Leading Innovative and Entrepreneur Team Introduction Program of Hangzhou","award":["TD2024012"],"award-info":[{"award-number":["TD2024012"]}]},{"name":"New Generation Artificial Intelligence-National Science and Technology Major Project","award":["2026ZD0127803"],"award-info":[{"award-number":["2026ZD0127803"]}]},{"name":"National Natural Science Foundation of China","award":["22678506"],"award-info":[{"award-number":["22678506"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Multicomponent reactions provide powerful modularity for peptide functionalization, but their application to lysine residues remains limited by the low efficiency of direct primary-amine-based activation pathways under peptide-compatible conditions. Here we report a secondary-amine relay strategy for lysine-directed multicomponent functionalization of peptides. In this approach, salicylaldehyde derivatives, arylboronic acids, and a cyclic secondary amine are combined to generate a transient electrophilic intermediate, which is subsequently intercepted by the \u03b5-amine of lysine-containing peptides. By separating electrophile generation from direct lysine activation, this relay protocol improves the efficiency of lysine modification while retaining the modularity of multicomponent chemistry. Mechanistic experiments, isolated intermediate studies, kinetic comparisons, and density functional theory calculations support a pathway involving secondary-amine-mediated intermediate formation followed by lysine capture. The method enables modification of diverse bioactive peptides, incorporation of drug-derived and fluorescent aldehyde or boronic acid components, and lysine\u2013lysine stapling using bifunctional aldehydes. The reaction is further compatible with automated and parallel solid-phase peptide modification workflows, providing rapid access to libraries of lysine-functionalized peptide conjugates. This study provides a practical secondary-amine relay platform for modular peptide functionalization and expands the utility of multicomponent chemistry in automated peptide synthesis.<\/jats:p>","DOI":"10.1021\/jacsau.6c00945","type":"journal-article","created":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T04:02:04Z","timestamp":1788926524000},"page":"5264-5274","source":"Crossref","is-referenced-by-count":0,"title":["Secondary-Amine Relay\nEnables Lysine-Directed Multicomponent\nFunctionalization of Peptides"],"prefix":"10.1021","volume":"6","author":[{"given":"Cheng","family":"Ren","sequence":"first","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]},{"name":"ZJU-Hangzhou Global Scientific and Technological Innovation Center , , ,","place":["Hangzhou, China, 311215"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengyao","family":"Zhang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tiefeng","family":"Jiang","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1150-5114","authenticated-orcid":true,"given":"Feng","family":"Wan","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]},{"name":"ZJU-Hangzhou Global Scientific and Technological Innovation Center , , ,","place":["Hangzhou, China, 311215"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yicheng","family":"Lv","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Yang","sequence":"additional","affiliation":[{"name":"Zhejiang University School of Medicine , , ,","place":["Hangzhou, China, 310058"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3904-0299","authenticated-orcid":true,"given":"Chengxi","family":"Li","sequence":"additional","affiliation":[{"name":"Zhejiang University , , ,","place":["Hangzhou, China, 310027"]},{"name":"ZJU-Hangzhou Global Scientific and Technological Innovation Center , , ,","place":["Hangzhou, China, 311215"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"issue":"3","key":"2026092808095820000_cit1","doi-asserted-by":"publisher","first-page":"1216","DOI":"10.1021\/acs.chemrev.7b00181","article-title":"Lysine acetylation\ngoes global: from epigenetics to metabolism and therapeutics","volume":"118","author":"Ali","year":"2018","journal-title":"Chem. Rev."},{"issue":"3","key":"2026092808095820000_cit2","doi-asserted-by":"publisher","first-page":"1480","DOI":"10.1039\/D0CS00556H","article-title":"Peptides as a platform for targeted\ntherapeutics for\ncancer: peptide-drug conjugates (PDCs)","volume":"50","author":"Cooper","year":"2021","journal-title":"Chem.\nSoc. Rev."},{"issue":"9","key":"2026092808095820000_cit3","doi-asserted-by":"publisher","first-page":"3659","DOI":"10.1016\/j.apsb.2023.02.013","article-title":"Research advances in peptide-drug conjugates","volume":"13","author":"Gong","year":"2023","journal-title":"Acta Pharm. Sin. B"},{"issue":"1","key":"2026092808095820000_cit4","doi-asserted-by":"publisher","first-page":"380","DOI":"10.1039\/D3CS00715D","article-title":"Chemical technology\nprinciples for\nselective bioconjugation of proteins and antibodies","volume":"53","author":"Chauhan","year":"2024","journal-title":"Chem. Soc. Rev."},{"key":"2026092808095820000_cit5","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1038\/s41573-024-01080-6","article-title":"Targeting lysine acetylation readers and writers","volume":"24","author":"Zhou","year":"2025","journal-title":"Nat. Rev. Drug Discovery"},{"issue":"28","key":"2026092808095820000_cit6","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2025.218174","article-title":"Peptide-drug\nconjugates in tumor therapy: current advances and future perspectives","volume":"638","author":"Jin","year":"2026","journal-title":"Cancer Lett."},{"issue":"30","key":"2026092808095820000_cit7","doi-asserted-by":"publisher","first-page":"3863","DOI":"10.1021\/acs.biochem.7b00536","article-title":"Residue-specific\npeptide modification:\na chemist\u2019s guide","volume":"56","author":"deGruyter","year":"2017","journal-title":"Biochemistry"},{"issue":"9","key":"2026092808095820000_cit8","doi-asserted-by":"publisher","first-page":"3128","DOI":"10.6023\/cjoc202505023","article-title":"Precise synthesis\nof proteins\/peptides: advances in single-site selective\nchemical modification and automated synthesis technologies","volume":"45","author":"Ren","year":"2025","journal-title":"Chin. J. Org. Chem."},{"issue":"7","key":"2026092808095820000_cit9","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1038\/nchembio.2416","article-title":"Targeting the N terminus for site-selective\nprotein modification","volume":"13","author":"Rosen","year":"2017","journal-title":"Nat. Chem. Biol."},{"issue":"11","key":"2026092808095820000_cit10","doi-asserted-by":"publisher","first-page":"4004","DOI":"10.1021\/jacs.7b12874","article-title":"Chemo-\nand regioselective lysine modification on native proteins","volume":"140","author":"Matos","year":"2018","journal-title":"J. Am. Chem. Soc."},{"issue":"4","key":"2026092808095820000_cit11","doi-asserted-by":"publisher","first-page":"4615","DOI":"10.1021\/jacs.5c20524","article-title":"Near-quantitative\nformation of imines\nin water with allosteric control","volume":"148","author":"Marchetti","year":"2026","journal-title":"J. Am. Chem.\nSoc."},{"issue":"D1","key":"2026092808095820000_cit12","doi-asserted-by":"publisher","first-page":"D451","DOI":"10.1093\/nar\/gkab849","article-title":"CPLM 4.0: an updated\ndatabase with rich annotations for protein lysine modifications","volume":"50","author":"Zhang","year":"2022","journal-title":"Nucleic Acids Res."},{"issue":"14","key":"2026092808095820000_cit13","doi-asserted-by":"publisher","first-page":"6656","DOI":"10.1021\/acs.chemrev.8b00008","article-title":"Chemical and\nbiochemical perspectives of protein lysine methylation","volume":"118","author":"Luo","year":"2018","journal-title":"Chem. Rev."},{"issue":"6599","key":"2026092808095820000_cit14","doi-asserted-by":"publisher","first-page":"1321","DOI":"10.1126\/science.abn2009","article-title":"A chemoenzymatic\nstrategy for site-selective functionalization of native peptides and\nproteins","volume":"376","author":"Fryszkowska","year":"2022","journal-title":"Science"},{"issue":"5","key":"2026092808095820000_cit15","doi-asserted-by":"publisher","DOI":"10.1016\/j.cclet.2024.110126","article-title":"Recent\nprogress of chemical methods for lysine site-selective modification\nof peptides and proteins","volume":"36","author":"Li","year":"2025","journal-title":"Chin. Chem. Lett."},{"issue":"11","key":"2026092808095820000_cit16","doi-asserted-by":"publisher","first-page":"1008","DOI":"10.1038\/s41557-020-0528-y","article-title":"Lysine\nacylation using conjugating enzymes for site-specific modification\nand ubiquitination of recombinant proteins","volume":"12","author":"Hofmann","year":"2020","journal-title":"Nat. Chem."},{"issue":"9","key":"2026092808095820000_cit17","doi-asserted-by":"publisher","first-page":"1387","DOI":"10.1038\/s41589-025-01869-5","article-title":"Reversible\nhistone\ndeacetylase activity catalyzes lysine acylation","volume":"21","author":"Tsusaka","year":"2025","journal-title":"Nat. Chem. Biol."},{"issue":"11","key":"2026092808095820000_cit18","doi-asserted-by":"publisher","first-page":"1081","DOI":"10.1038\/s41557-021-00765-4","article-title":"A proteome-wide\natlas of lysine-reactive chemistry","volume":"13","author":"Abbasov","year":"2021","journal-title":"Nat. Chem."},{"issue":"5","key":"2026092808095820000_cit19","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202112107","article-title":"Tunable amine-reactive\nelectrophiles for selective profiling of lysine","volume":"61","author":"Tang","year":"2022","journal-title":"Angew. Chem., Int. Ed."},{"issue":"10","key":"2026092808095820000_cit20","doi-asserted-by":"publisher","first-page":"979","DOI":"10.1002\/poc.941","article-title":"Structure-stability correlations\nfor imine formation in aqueous solution","volume":"18","author":"Godoy-Alca\u0301ntar","year":"2005","journal-title":"J.\nPhys. Org. Chem."},{"issue":"6","key":"2026092808095820000_cit21","doi-asserted-by":"publisher","first-page":"2003","DOI":"10.1039\/c2cs15305j","article-title":"Dynamic imine chemistry","volume":"41","author":"Belowich","year":"2012","journal-title":"Chem.\nSoc. Rev."},{"issue":"46","key":"2026092808095820000_cit22","doi-asserted-by":"publisher","first-page":"11108","DOI":"10.1002\/chem.201702088","article-title":"Kinetic selectivity and thermodynamic\nfeatures of competitive imine formation in dynamic covalent chemistry","volume":"23","author":"Kulchat","year":"2017","journal-title":"Chem. - Eur. J."},{"issue":"24","key":"2026092808095820000_cit23","doi-asserted-by":"publisher","first-page":"10299","DOI":"10.1021\/ja303436y","article-title":"Iminoboronates: a new strategy for\nreversible protein modification","volume":"134","author":"Cal","year":"2012","journal-title":"J. Am. Chem.\nSoc."},{"issue":"17","key":"2026092808095820000_cit24","doi-asserted-by":"publisher","first-page":"11840","DOI":"10.1021\/acs.jmedchem.2c00937","article-title":"Chemo- and site-selective lysine\nmodification of peptides and proteins under native conditions using\nthe water-soluble zolinium","volume":"65","author":"Sun","year":"2022","journal-title":"J. Med. Chem."},{"issue":"20","key":"2026092808095820000_cit25","doi-asserted-by":"publisher","first-page":"11245","DOI":"10.1021\/acs.chemrev.9b00214","article-title":"Reactivity and synthetic applications\nof multicomponent Petasis reactions","volume":"119","author":"Wu","year":"2019","journal-title":"Chem. Rev."},{"issue":"1","key":"2026092808095820000_cit26","doi-asserted-by":"publisher","DOI":"10.1007\/s41061-025-00491-2","article-title":"The Petasis reaction: applications and organic synthesis\u2500A\ncomprehensive review","volume":"383","author":"Pandit","year":"2025","journal-title":"Top. Curr. Chem."},{"issue":"34","key":"2026092808095820000_cit27","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202307782","article-title":"Tryptophan in multicomponent\nPetasis reactions for\npeptide stapling and late-stage functionalization","volume":"62","author":"Krajcovicova","year":"2023","journal-title":"Angew. Chem., Int. Ed."},{"issue":"16","key":"2026092808095820000_cit28","doi-asserted-by":"publisher","DOI":"10.1002\/cmdc.202400148","article-title":"Ideas behind\nthe tryptophan-mediated Petasis reaction (TMPR) concept for peptide\nstapling","volume":"19","author":"Krajcovicova","year":"2024","journal-title":"ChemMedChem"},{"issue":"71","key":"2026092808095820000_cit29","doi-asserted-by":"publisher","first-page":"10567","DOI":"10.1039\/C9CC05934B","article-title":"Conformationally\nstable peptide macrocycles assembled using the Petasis Borono-Mannich\nreaction","volume":"55","author":"Yamaguchi","year":"2019","journal-title":"Chem. Commun."},{"issue":"1","key":"2026092808095820000_cit30","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1039\/C9SC04697F","article-title":"Secondary amine selective\nPetasis (SASP) bioconjugation","volume":"11","author":"Sim","year":"2020","journal-title":"Chem. Sci."},{"issue":"10","key":"2026092808095820000_cit31","doi-asserted-by":"publisher","first-page":"3679","DOI":"10.1021\/jo062586z","article-title":"Nucleophilicities\nof primary and\nsecondary amines in water","volume":"72","author":"Brotzel","year":"2007","journal-title":"J. Org. Chem."},{"issue":"36","key":"2026092808095820000_cit32","doi-asserted-by":"publisher","first-page":"6379","DOI":"10.1002\/ejoc.200900925","article-title":"Nucleophilic\nreactivities of primary and secondary amines in acetonitrile","volume":"2009","author":"Kanzian","year":"2009","journal-title":"Eur. J. Org. Chem."},{"issue":"9","key":"2026092808095820000_cit33","doi-asserted-by":"publisher","first-page":"2700","DOI":"10.1002\/anie.201812620","article-title":"Introducing the\nPetasis reaction for late-stage multicomponent diversification, labeling,\nand stapling of peptides","volume":"58","author":"Ricardo","year":"2019","journal-title":"Angew. Chem., Int.\nEd."},{"issue":"1","key":"2026092808095820000_cit34","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1002\/cjoc.201090033","article-title":"Theoretical study on the mechanism of the Petasis-type Boronic Mannich\nreaction of organoboronic acids, amines, and \u03b1-hydroxy aldehydes","volume":"28","author":"Tao","year":"2010","journal-title":"Chin. J. Chem."},{"issue":"93","key":"2026092808095820000_cit35","doi-asserted-by":"publisher","first-page":"76337","DOI":"10.1039\/C5RA16678K","article-title":"Insights\non the Petasis Borono-Mannich multicomponent\nreaction mechanism","volume":"5","author":"Souza","year":"2015","journal-title":"RSC Adv."},{"issue":"25","key":"2026092808095820000_cit36","doi-asserted-by":"publisher","first-page":"4063","DOI":"10.1021\/ol006710r","article-title":"2H-chromenes from salicylaldehydes\nby a catalytic Petasis reaction","volume":"2","author":"Wang","year":"2000","journal-title":"Org. Lett."},{"issue":"12","key":"2026092808095820000_cit37","doi-asserted-by":"publisher","first-page":"1859","DOI":"10.1002\/ejoc.200900056","article-title":"Water: a suitable\nmedium for the Petasis Borono-Mannich reaction","volume":"2009","author":"Candeias","year":"2009","journal-title":"Eur. J. Org. Chem."},{"issue":"1","key":"2026092808095820000_cit38","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1021\/jo0006627","article-title":"Alkylation of amino\nacids and glutathione in water by o-quinone methide.\nReactivity and selectivity","volume":"66","author":"Modica","year":"2001","journal-title":"J. Org. Chem."},{"issue":"23","key":"2026092808095820000_cit39","doi-asserted-by":"publisher","first-page":"1940","DOI":"10.1080\/00397911.2016.1239740","article-title":"C-C bond cleavage:\nMetal-free-catalyzed reaction of Betti bases with various heterocycles\nunder microwave irradiation","volume":"46","author":"Deb","year":"2016","journal-title":"Syn. Commun."},{"issue":"27","key":"2026092808095820000_cit40","doi-asserted-by":"publisher","first-page":"5367","DOI":"10.1016\/S0040-4020(02)00496-9","article-title":"o-Quinone methides:\nintermediates underdeveloped and underutilized in organic synthesis","volume":"58","author":"Van De Water","year":"2002","journal-title":"Tetrahedron"},{"issue":"99","key":"2026092808095820000_cit41","doi-asserted-by":"publisher","first-page":"55924","DOI":"10.1039\/C4RA11444B","article-title":"ortho-Quinone methide (o-QM): a highly reactive, ephemeral\nand versatile intermediate in organic synthesis","volume":"4","author":"Singh","year":"2014","journal-title":"RSC Adv."},{"issue":"21","key":"2026092808095820000_cit42","doi-asserted-by":"publisher","first-page":"10145","DOI":"10.1021\/acs.joc.6b01367","article-title":"Emerging roles of in situ generated quinone methides\nin metal-free catalysis","volume":"81","author":"Jaworski","year":"2016","journal-title":"J. Org. Chem."},{"issue":"28","key":"2026092808095820000_cit43","doi-asserted-by":"publisher","first-page":"30476","DOI":"10.1021\/jacs.6c09909","article-title":"A modular PMDA linker enables lysine-selective\ncyclization of unprotected peptides and automated macrocycle assembly","volume":"148","author":"Wu","year":"2026","journal-title":"J. Am. Chem. Soc."},{"issue":"27","key":"2026092808095820000_cit44","doi-asserted-by":"publisher","DOI":"10.1002\/anie.7454058","article-title":"SPECTRAL: an intelligent and ultra-sensitive\nphotonic hydrogel platform\nfor biomarker-based cancer prediction","volume":"65","author":"Qin","year":"2026","journal-title":"Angew.\nChem., Int. Ed."},{"issue":"5","key":"2026092808095820000_cit45","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1038\/s41557-021-00662-w","article-title":"Automated\nsynthesis of prexasertib and derivatives enabled by continuous-flow\nsolid-phase synthesis","volume":"13","author":"Liu","year":"2021","journal-title":"Nat. Chem."},{"issue":"1","key":"2026092808095820000_cit46","doi-asserted-by":"publisher","first-page":"986","DOI":"10.1021\/jacs.5c16902","article-title":"Automated rapid synthesis of high-purity\nhead-to-tail cyclic peptides via a diaminonicotinic acid scaffold","volume":"148","author":"Wan","year":"2026","journal-title":"J. Am. Chem. Soc."},{"key":"2026092808095820000_cit47","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-026-69441-w","article-title":"CycloPepper:\na machine learning platform for predicting cyclization outcomes and\noptimizing synthesis of therapeutic cyclopeptides","volume":"17","author":"Pan","year":"2026","journal-title":"Nat. Commun."}],"container-title":["JACS Au"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/jacsau.6c00945","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jaaucr\/article-pdf\/6\/9\/5264\/68454122\/jacsau.6c00945.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jaaucr\/article-pdf\/6\/9\/5264\/68454122\/jacsau.6c00945.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T12:31:42Z","timestamp":1790598702000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jaaucr\/article\/6\/9\/5264\/5426692\/Secondary-Amine-Relay-Enables-Lysine-Directed"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,8]]},"references-count":47,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9,8]]},"published-print":{"date-parts":[[2026,9,28]]}},"URL":"https:\/\/doi.org\/10.1021\/jacsau.6c00945","relation":{},"ISSN":["2691-3704"],"issn-type":[{"value":"2691-3704","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,8]]}}}