001037154 001__ 1037154 001037154 005__ 20250203124522.0 001037154 0247_ $$2doi$$a10.1016/j.ejmech.2024.116606 001037154 0247_ $$2ISSN$$a0009-4374 001037154 0247_ $$2ISSN$$a0223-5234 001037154 0247_ $$2ISSN$$a1768-3254 001037154 0247_ $$2datacite_doi$$a10.34734/FZJ-2025-00499 001037154 0247_ $$2pmid$$a38901105 001037154 0247_ $$2WOS$$aWOS:001347413400001 001037154 037__ $$aFZJ-2025-00499 001037154 082__ $$a610 001037154 1001_ $$0P:(DE-HGF)0$$aBach, Kathrin$$b0 001037154 245__ $$aExtensive targeting of chemical space at the prime side of ketoamide inhibitors of rhomboid proteases by branched substituents empowers their selectivity and potency 001037154 260__ $$aAmsterdam [u.a.]$$bElsevier Science$$c2024 001037154 3367_ $$2DRIVER$$aarticle 001037154 3367_ $$2DataCite$$aOutput Types/Journal article 001037154 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1736844299_548 001037154 3367_ $$2BibTeX$$aARTICLE 001037154 3367_ $$2ORCID$$aJOURNAL_ARTICLE 001037154 3367_ $$00$$2EndNote$$aJournal Article 001037154 520__ $$aRhomboid intramembrane serine proteases have been implicated in several pathologies, and emerge as attractive pharmacological target candidates. The most potent and selective rhomboid inhibitors available to date are peptidyl α-ketoamides, but their selectivity for diverse rhomboid proteases and strategies to modulate it in relevant contexts are poorly understood. This gap, together with the lack of suitable in vitro models, hinders ketoamide development for relevant eukaryotic rhomboid enzymes. Here we explore the structure-activity relationship principles of rhomboid inhibiting ketoamides by medicinal chemistry and enzymatic in vitro and in-cell assays with recombinant rhomboid proteases GlpG, human mitochondrial rhomboid PARL and human RHBDL2. We use X-ray crystallography in lipidic cubic phase to understand the binding mode of one of the best ketoamide inhibitors synthesized here containing a branched terminal substituent bound to GlpG. In addition, to extend the interpretation of the co-crystal structure, we use quantum mechanical calculations and quantify the relative importance of interactions along the inhibitor molecule. These combined experimental analyses implicates that more extensive exploration of chemical space at the prime side is unexpectedly powerful for the selectivity of rhomboid inhibiting ketoamides. Together with variations in the peptide sequence at the non-prime side, or its non-peptidic alternatives, this strategy enables targeted tailoring of potent and selective ketoamides towards diverse rhomboid proteases including disease-relevant ones such as PARL and RHBDL2. 001037154 536__ $$0G:(DE-HGF)POF4-5241$$a5241 - Molecular Information Processing in Cellular Systems (POF4-524)$$cPOF4-524$$fPOF IV$$x0 001037154 588__ $$aDataset connected to CrossRef, Journals: juser.fz-juelich.de 001037154 7001_ $$0P:(DE-HGF)0$$aDohnálek, Jan$$b1 001037154 7001_ $$0P:(DE-HGF)0$$aŠkerlová, Jana$$b2 001037154 7001_ $$0P:(DE-HGF)0$$aKuzmík, Ján$$b3 001037154 7001_ $$0P:(DE-HGF)0$$aPoláchová, Edita$$b4 001037154 7001_ $$0P:(DE-HGF)0$$aStanchev, Stancho$$b5 001037154 7001_ $$0P:(DE-HGF)0$$aMajer, Pavel$$b6 001037154 7001_ $$0P:(DE-HGF)0$$aFanfrlík, Jindřich$$b7 001037154 7001_ $$0P:(DE-HGF)0$$aPecina, Adam$$b8 001037154 7001_ $$0P:(DE-HGF)0$$aŘezáč, Jan$$b9 001037154 7001_ $$0P:(DE-HGF)0$$aLepšík, Martin$$b10 001037154 7001_ $$0P:(DE-Juel1)144613$$aBorshchevskiy, Valentin$$b11 001037154 7001_ $$0P:(DE-HGF)0$$aPolovinkin, Vitaly$$b12 001037154 7001_ $$00000-0003-3677-0907$$aStrisovsky, Kvido$$b13$$eCorresponding author 001037154 773__ $$0PERI:(DE-600)2005170-0$$a10.1016/j.ejmech.2024.116606$$gVol. 275, p. 116606 -$$p116606 -$$tEuropean journal of medicinal chemistry$$v275$$x0009-4374$$y2024 001037154 8564_ $$uhttps://juser.fz-juelich.de/record/1037154/files/1-s2.0-S0223523424004860-main.pdf$$yOpenAccess 001037154 909CO $$ooai:juser.fz-juelich.de:1037154$$pdnbdelivery$$pdriver$$pVDB$$popen_access$$popenaire 001037154 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)144613$$aForschungszentrum Jülich$$b11$$kFZJ 001037154 9131_ $$0G:(DE-HGF)POF4-524$$1G:(DE-HGF)POF4-520$$2G:(DE-HGF)POF4-500$$3G:(DE-HGF)POF4$$4G:(DE-HGF)POF$$9G:(DE-HGF)POF4-5241$$aDE-HGF$$bKey Technologies$$lNatural, Artificial and Cognitive Information Processing$$vMolecular and Cellular Information Processing$$x0 001037154 9141_ $$y2024 001037154 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)1050$$2StatID$$aDBCoverage$$bBIOSIS Previews$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)1190$$2StatID$$aDBCoverage$$bBiological Abstracts$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)0600$$2StatID$$aDBCoverage$$bEbsco Academic Search$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)1200$$2StatID$$aDBCoverage$$bChemical Reactions$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)9905$$2StatID$$aIF >= 5$$bEUR J MED CHEM : 2022$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)1030$$2StatID$$aDBCoverage$$bCurrent Contents - Life Sciences$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)1210$$2StatID$$aDBCoverage$$bIndex Chemicus$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)0113$$2StatID$$aWoS$$bScience Citation Index Expanded$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess 001037154 915__ $$0StatID:(DE-HGF)0030$$2StatID$$aPeer Review$$bASC$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR$$bEUR J MED CHEM : 2022$$d2025-01-07 001037154 915__ $$0StatID:(DE-HGF)0160$$2StatID$$aDBCoverage$$bEssential Science Indicators$$d2025-01-07 001037154 915__ $$0LIC:(DE-HGF)CCBY4$$2HGFVOC$$aCreative Commons Attribution CC BY 4.0 001037154 915__ $$0StatID:(DE-HGF)0420$$2StatID$$aNationallizenz$$d2025-01-07$$wger 001037154 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bClarivate Analytics Master Journal List$$d2025-01-07 001037154 920__ $$lyes 001037154 9201_ $$0I:(DE-Juel1)IBI-7-20200312$$kIBI-7$$lStrukturbiochemie$$x0 001037154 980__ $$ajournal 001037154 980__ $$aVDB 001037154 980__ $$aUNRESTRICTED 001037154 980__ $$aI:(DE-Juel1)IBI-7-20200312 001037154 9801_ $$aFullTexts