![\"\"](\"http:\/\/sites.unica.it\/pe2026\/files\/2025\/09\/cropped-00_Profile_Tondo-Social__Spilla_PE26_1200px.png\")
<\/figure>FPC5 – Giorgio Milli<\/strong><\/p>\n\n\n\n Department of Drug Sciences, University of Pavia, Viale Taramelli 12, Pavia 27100, Italy<\/p>\n\n\n\n giorgio.milli01@universitadipavia.it<\/strong><\/p>\n<\/div><\/div>\n\n\n\n <\/p>\n\n\n\n <\/p>\n","protected":false},"excerpt":{"rendered":" FPC5 – Giorgio Milli Department of Drug Sciences, University of Pavia, Viale Taramelli 12, Pavia 27100, Italy giorgio.milli01@universitadipavia.it Expanding the Chemical Space for LsrK Kinase Modulators: Hit Identification and Biological Validation for AI-2 Quorum-Sensing Quenching and […]<\/p>\n","protected":false},"author":10371,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[],"class_list":["post-1997","post","type-post","status-publish","format-standard","hentry","category-senza-categoria"],"_links":{"self":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts\/1997","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/users\/10371"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/comments?post=1997"}],"version-history":[{"count":6,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts\/1997\/revisions"}],"predecessor-version":[{"id":2146,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/posts\/1997\/revisions\/2146"}],"wp:attachment":[{"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/media?parent=1997"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/categories?post=1997"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sites.unica.it\/pe2026\/wp-json\/wp\/v2\/tags?post=1997"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}Expanding the Chemical Space for LsrK Kinase Modulators: Hit Identification and Biological Validation for AI-2 Quorum-Sensing Quenching and Biofilm Formation Inhibition<\/strong><\/td><\/tr> Milli Giorgio1<\/sup>, Ambrosio Francesca Alessandra2<\/sup>, Bellan Menegussi Elisa3<\/sup>, Pagano Katiuscia4<\/sup>, Ragona Laura4<\/sup>, Costa Giosu\u00e82<\/sup>, Alcaro Stefano2<\/sup>,Pietrocola Giampiero3<\/sup>, Linciano Pasquale1<\/sup>, Collina Simona1<\/sup>.
<\/em>
1<\/sup> <\/em>Department of Drug Sciences, University of Pavia, Viale Taramelli 12, Pavia 27100, Italy
2<\/sup> <\/em>Department of Health Sciences, University \u201cMagna Gr\u00e6cia\u201d of Catanzaro, Viale Europa, 88100 Catanzaro, Italy
3<\/sup> <\/em>Department of Molecular Medicine, Biochemistry Unit, University of Pavia, Viale Taramelli 3\/b, Pavia 27100, Italy
4<\/sup> <\/em>NMR Laboratory, Istituto di Scienze e Tecnologie Chimiche \u201cGiulio Natta\u201d, CNR, via Alfonso Corti, 12, Milano 20133, Italy<\/td><\/tr>Abstract<\/strong>
Antimicrobial resistance (AMR) poses a critical global health crisis by undermining existing treatments and threatening public health. Consequently, identifying novel targets and antimicrobials with innovative mechanisms of action has become essential to combat AMR [1]. In this context, disrupting quorum sensing (QS) and biofilm formation emerges as a promising strategy. LsrK, a kinase that initiates QS and promotes biofilm development through phosphorylation of 4,5-dihydroxy-2,3-pentanedione (DPD) – a ubiquitous signaling molecule in both Gram-positive and Gram-negative bacteria – represents a critical component of QS pathways and its inhibition could be a winning strategy for fighting AMR [2].
To this end, structure-based virtual screening of commercial libraries identified a panel of hit molecules, which were evaluated in biological, following an experimental pathway previously optimized [3], to confirm their ability to inhibit biofilm formation in three bacterial models: Staphylococcus aureus<\/em>, Pseudomonas aeruginosa<\/em>, and Escherichia coli<\/em>. We developed a dedicated synthesis strategy to obtain sufficient quantities of the most promising compounds for comprehensive biological and microbiological evaluations. Their ability to interfere with quorum sensing was assessed in Vibrio harveyi<\/em> reporter assays, while interaction with LsrK was confirmed by biophysical studies, including Saturation Transfer Difference (STD) NMR.
These efforts identified two lead compounds, CzdDPD 8 and CzDPD 10, with a Minimum Biofilm Inhibitory Concentration 50% (MBIC\u2085\u2080) values of 121 and 183 \u03bcg mL\u207b\u00b9 against E. coli<\/em>, and 55 and 166 \u03bcg mL\u207b\u00b9 against P. aeruginosa<\/em>, respectively. Mechanistic studies validated QS disruption and LsrK binding, marking a significant advance against AMR.<\/td><\/tr>References <\/strong>
[1] World Health Organization. Global Antibiotic Resistance Surveillance Report 2025: WHO Global Antimicrobial Resistance and Use Surveillance System (GLASS) \u2013 Summary; World Health Organization, 2025. https:\/\/doi.org\/10.2471\/B09585.
[2] Linciano, P.; Cavalloro, V.; Martino, E.; Kirchmair, J.; Listro, R.; Rossi, D.; Collina, S. Tackling Antimicrobial Resistance with Small Molecules Targeting LsrK: Challenges and Opportunities. J. Med. Chem. 2020<\/strong>, 63 (24), 15243\u201315257. https:\/\/doi.org\/10.1021\/acs.jmedchem.0c01282.
[3] Milli, G.; Pellegrini, A.; Listro, R.; Fasolini, M.; Pagano, K.; Ragona, L.; Pietrocola, G.; Linciano, P.; Collina, S. New LsrK Ligands as AI-2 Quorum Sensing Interfering Compounds against Biofilm Formation. J. Med. Chem. 2024<\/strong>, 67 (20), 18139\u201318156. https:\/\/doi.org\/10.1021\/acs.jmedchem.4c01266.<\/td><\/tr><\/tbody><\/table><\/div><\/figure>\n\n\n\n