Sonderforschungsbereich 630

    A4 Schirmeister

    Project Part A4

    Proteases as Targets for Drugs against Infectious Diseases

    Prof. Dr. Tanja Schirmeister

    Institute for Pharmacy and Biochemistry, University of Mainz

    The main research topic of TP A4 concerns development of irreversible inhibitors of parasitic, fungal and bacterial cysteine and aspartic proteases. Our primary target enzymes are

    Cysteine proteases: Falcipain 2 and 3 from the malaria pathogen Plasmodium falciparum; rhodesain from Trypanosoma brucei rhodesiense, causing African Trypanosomiasis (sleeping sickness); cruzain from Trypanosoma cruzi, causing Chagas-disease; cathepsins from leishmania.

    Aspartic proteases: SAPs (secreted aspartic proteases) from Candida albicans; plasmepsin II from Plasmodium falciparum. Since playing a major role within the life cycle of the pathogens, these enzymes are promising targets for the development of new anti-infective agents.The classes of inhibitors we are working on include peptidomimetics containing aziridines, epoxides and acceptor-substituted olefins as electrophilic building blocks. Within the project TP A4 new substances are synthesised and tested against the proteases.Cooperating with other SFB projects as well as external partners, in vitro tests with the synthesised substances against the pathogens and human cells are performed (TP of project B, TP Z1).

    Mechanisms of inhibition and possible binding modes, respectively, are elucidated by means of biotinylated inhibitors, QM-, QM/MM-calculations, NMR-spectroscopic experiments and various docking procedures, and, furthermore, by cocrystallisation and structure determination of enzyme-inhibitor-complexes (B- and C-projects).

    With the received information we aim for further optimisation of the inhibitors in respect of activity, selectivity and toxicity. Quantum chemical calculations (cooperation with TP C3, Engels) could already explain the differences in inhibition activity of epoxide and aziridine based cysteine protease inhibitors concerning the dependence on different substituents at the carbon and nitrogen atom of the three-membered ring, respectively, as well as different pH values of the environmental medium. QM/MM-calculations thereby also consider the environment of the protein.In a further project sponge extracts are screened for metabolites with protease inhibiting properties. Goal of these investigations is the identification of new lead structures for protease inhibitors (cooperation with TP A5, Hentschel, A2, Bringmann). Further projects are the development of inhibitors of the InhA of M. tuberculosis (cooperation with TP B7, Kisker, and TP C7, Sotriffer) and of pyrrole-based as well as fumaric acid-based falcipain and rhodesain inhibitors (cooperation with Carsten Schmuck).

    Selected, SFB-relevant Publications

    R. Ettari, L. Tamborini, I. C. Angelo, S. Grasso, T. Schirmeister, L. Lo Presti, C. De Micheli, A. Pinto, P. Conti; Development of rhodesain inhibitors with a 3-bromoisoxazoline warhead. ChemMedChem 2013, 8, 2070-2076. doi:10.1002/cmdc.201300390

    R. Ettari, N. Micale, G. Grazioso, F. Bova, T. Schirmeister, S. Grasso, M. Zappala; Synthesis and molecular modeling studies of derivatives of a highly potent peptidomimetic vinyl ester as falcipain-2 inhibitors. ChemMedChem 2012, 7, 1594-1600. doi:10.1002/cmdc.201200274

    P. Tabares, B. Degel, N. Schaschke, U. Hentschel, T. Schirmeister; Identification of the protease inhibitor miraziridine A in the Red sea sponge Theonella swinhoei. Pharmacognosy Res 2012, 4, 63-66.

    V. Ehmke, J. E. Quinsaat, P. Rivera-Fuentes, C. Heindl, C. Freymond, M. Rottmann, R. Brun, T. Schirmeister, F. Diederich; Tuning and predicting biological affinity: aryl nitriles as cysteine protease inhibitors. Org Biomol Chem 2012.

    P. Tabares, S. M. Pimentel-Elardo, T. Schirmeister, T. Hunig, U. Hentschel; Anti-protease and immunomodulatory activities of bacteria associated with Caribbean sponges. Mar Biotechnol (NY) 2011, 13, 883-892.

    S. M. Pimentel-Elardo, V. Buback, T. A. Gulder, T. S. Bugni, J. Reppart, G. Bringmann, C. M. Ireland, T. Schirmeister, U. Hentschel; New tetromycin derivatives with anti-trypanosomal and protease inhibitory activities. Mar Drugs 2011, 9, 1682-1697.

    S. Langolf, U. Machon, M. Ehlers, W. Sicking, T. Schirmeister, C. Buchhold, C. Gelhaus, P. J. Rosenthal, C. Schmuck; Development of antitrypanosomal and antiplasmodial nonpeptidic cysteine protease inhibitors based on N-protected-guanidino-furan and -pyrrole building blocks. ChemMedChem 2011, 6, 1581-1586.

    V. Ehmke, C. Heindl, M. Rottmann, C. Freymond, W. B. Schweizer, R. Brun, A. Stich, T. Schirmeister, F. Diederich; Potent and selective inhibition of cysteine proteases from Plasmodium falciparum and Trypanosoma brucei. ChemMedChem 2011, 6, 273-278.

    C. Büchold, Y. Hemberger, C. Heindl, A. Welker, B. Degel, T. Pfeuffer, P. Staib, S. Schneider, P. J. Rosenthal, J. Gut, J. Morschhauser, G. Bringmann, T. Schirmeister; New cis-configured aziridine-2-carboxylates as aspartic acid protease inhibitors. ChemMedChem 2011, 6, 141-152.

    U. Schurigt, C. Schad, C. Glowa, U. Baum, K. Thomale, J. K. Schnitzer, M. Schultheis, N. Schaschke, T. Schirmeister, H. Moll, Aziridine-2,3-dicarboxylate-based cysteine cathepsin inhibitors induce cell death in Leishmania major associated with accumulation of debris in autophagy-related lysosome-like vacuoles. Antimicrob Agents Chemother 2010, 54, 5028-5041

    T. Waag, C. Gelhaus, J. Rath, A. Stich, M. Leippe, T. Schirmeister, Allicin and its derivates are cysteine protease inhibitors with antiparasitic activity. Bioorg Med Chem Lett 2010, 20, 5541-5543

    F. Bova, R. Ettari, N. Micale, C. Carnovale, T. Schirmeister, C. Gelhaus, M. Leippe, S. Grasso, M. Zappala, Constrained peptidomimetics as antiplasmodial falcipain-2 inhibitors. Bioorg Med Chem 2010, 18, 4928-4938

    R. Ettari, M. Zappalà, A. Evers, C, Gelhaus, M. Leippe, T. Schirmeister, N. Micale, S. Grasso, Synthesis of novel peptidomimetics as inhibitors of protozoan cysteine proteases falcipain-2 and rhodesain. Eur J Med Chem 2010, 45, 3228-3233

    A. Paasche, M. Schiller, M. Mladenovic, T. Schirmeister,* B. Engels,* Mechanistic study about the reaction of thiol-containing enzymes with a,b-unsaturated carbonyl substrates by computations and chemoassays. ChemMedChem 2010, 5, 869-880

    S. Ludewig, M. Kossner, M. Schiller, K. Baumann,* T. Schirmeister,* Enzyme kinetics and hit validation in fluorometric protease assays. Curr Top Med Chem 2010, 10, 368-382

    A. Breuning, B. Degel, F. Schulz, C. Büchold, M. Stempka, U. Machon, C. Gelhaus, M. Leippe, M. Leyh, C. Kisker, J. Rath, A. Stich, J. Gut, P. J. Rosenthal, C. Schmuck,* T. Schirmeister,* Michael-acceptor based antiplasmodial and antitrypanosomal cysteine protease inhibitors with unusual peptide sequences. J Med Chem 2010, 53, 1951-1963

    N. Micale, R. Ettari, T. Schirmeister, A. Evers, C. Gelhaus, M. Leippe, M. Zappala, S. Grasso, Novel 2H-isoquinoline-3-ones as antiplasmodial falcipain-2 inhibitors. Bioorg Med Chem 2009, 17, 6505-6511

    U. Machon, C. Büchold, M. Stempka, T. Schirmeister,* C. Gelhaus, M. Leippe, J. Gut, P. J. Rosenthal, C. Kisker, M. Leyh, C. Schmuck*, On-bead screening of a combinatorial fumaric acid derived peptide library yields antiplasmodial cysteine protease inhibitors with unusual peptide sequences. J Med Chem 2009, 52, 5662-5672

    S. Donat, K. Streker, T. Schirmeister, S. Rakette, T. Stehle, M. Liebeke, M. Lalk, K. Ohlsen, Transcriptome and functional analysis of the eukaryotic-type ser/thr kinase PknB in Staphylococcus aureus. J Bacteriol 2009, 191, 4056-4069

    R. Ettari, N. Micale, T. Schirmeister, C. Gelhaus, M. Leippe, E. Nizi, M. E. Di Francesco, S. Grasso, M. Zappalà, Novel peptidomimetics containing a vinyl ester moiety as highly potent and selective falcipain-2 inhibitors. J Med Chem 2009, 52, 2157-2160

    V. Buback, M. Mladenovic, B. Engels, T. Schirmeister, Rational design of improved aziridine-based inhibitors of cysteine proteases. J Phys Chem B 2009, 113, 5282-5289

    M.-A. Dude, U. Kaeppler, M. Herb, M. Schiller, F. Schulz, B. Vedder, S. Heppner, G. Pradel, J. Gut, P. J. Rosenthal, T. Schirmeister, M. Leippe, C. Gelhaus, Synthesis and evaluation of non-peptidic cysteine protease inhibitors of P. falciparum derived from etacrynic acid. Molecules 2009, 14, 19-35

    M. Mladenovic, R. F. Fink, W. Thiel, T. Schirmeister, B. Engels, Atomistic insights into the inhibition of cysteine proteases: First QM/MM calculations clarifying the stereoselectivity of epoxide-based inhibitors. J Phys Chem B 2008, 112, 11798-11808

    M. Mladenovic, R. F. Fink, W. Thiel, T. Schirmeister, B. Engels, On the origin of the stabilization of the zwitterionic resting state of cysteine proteases and its activation upon substrate binding: a theoretical study. J Am Chem Soc 2008, 130, 8696-8705

    R. Ettari, E. Nizi, M. E. Di Francesco, N. Micale, S. Grasso, M. Zappala, R. Vicik, T. Schirmeister, Nonpeptidic vinyl- and allyl phosphonates as falcipain-2 active site-directed Michael acceptors. ChemMedChem 2008, 3, 1030-1033

    M. Mladenovic, K. Junold, R. F. Fink, W. Thiel, T. Schirmeister, B. Engels, Atomistic insights into the inhibition of cysteine proteases: First QM/MM calculations clarifying the regiospecificity and the inhibition potency of epoxide- and aziridine-based inhibitors. J Phys Chem B 2008, 112, 5458-5469

    I. Rupp, R. Bosse, T. Schirmeister, G. Pradel, Effect of protease inhibitors on exflagellation in Plasmodium falciparum. Mol Biochem Parasitol 2008, 158, 208-212

    R. Ettari, E. Nizi, M. E. Di Francesco, M.-A. Dude, G. Pradel, R. Vicik, T. Schirmeister, N. Micale, S. Grasso, M. Zappala, Development of peptidomimetics with a vinyl sulfone warhead as irreversible falcipain-2 inhibitors J Med Chem 2008, 51, 988-996

    B. Degel, P. Staib, S. Rohrer, J. Scheiber, E. Martina, C. Büchold, K. Baumann, J. Morschhäuser, T. Schirmeister, Cis-configured aziridines are new pseudo-irreversible dual-mode inhibitors of Candida albicans secreted aspartic protease 2. ChemMedChem 2008, 3, 302-315

    P. Staib, J. Blaß-Warmuth, U. Lermann, B. Degel, R. Würzner, M. Monod, T. Schirmeister, J. Morschhäuser, Tetracycline-inducible expression of individual Secreted Aspartic Proteases in Candida albicans allows isoenzyme-specific inhibitor screening. Antimicrob Agents Chemother 2008, 52, 146-156

    F. Schulz, C. Gelhaus, B. Degel, R. Vicik, S. Heppner, A. Breuning, M. Leippe, J. Gut, P. J. Rosenthal, T. Schirmeister, Screening of protease inhibitors as antiplasmodial agents. Part I: Aziridines and epoxides. ChemMedChem 2007, 2, 1214-1224