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neg. control Cs9-Rhd MM-218Fig. 2. Confocal microscopy of Jurkat cells incubated with Cs9-Rhd or MM-218.in the mixed-lymphocyte assay with human peripheral blood mononuclear cells. Cs9-Rhdand CsA itself were, as expected, immunosuppresive in both assays. Additionally, MM-218has been non-cytotoxic in tested concentrations. The cell-impermeable compound MM-218was also stable to proteolytic degradation as confirmed by an unchanged HPLC-MS profileof this substance during 96 h incubation time at 37°C in the mouse serum. In a chemotacticassay MM-218 completely inhibited activated mouse CD4+ T-cell migration in response toCypA and CypB, but not in response to chemotactic cytokine RANTES. The full advantageof this stable, non-immunosuppresive but potent extracellular Cyp inhibitor was shown inmouse model of asthma, where a drastic reduction of leukocyte numbers only in the lungtissue and airways of asthmatic mice upon intervention with MM-218 was observed [5].Moreover, it was shown that MM-218 intervention significantly reduces hyperreactivity, ina mouse model of acute allergic asthma, as confirmed by measuring the lung airwayresistance and compliance of mice primed with ovalbumin and challenged with increasingdoses of methacholine [5]. The asthmatic mice group treated i.p. with MM-218 showedsimilar airway resistance and lung compliance to those of naïve mice.The important advantage of this novel CsA derivative, MM-218, over CsA itself forthe study of inflammatory diseases is that all effects resulted from inhibiton of differentnuclear transcription factors, calcineurin, intracellular Cyps or other diverse intracellularCsA interaction partners could be completely excluded. The effects observed uponMM-218 intervention in asthmatic mice model could be assigned to inhibition ofextracellular Cyps.In modern drug design, selectivity gains ever-growing scientific attention. As aconsequence of their selectivity, therapeutic agents could act at low concentrations withminimal or no side effects. In particular, selectivity could be achieved even fornon-selective enzyme inhibitors by their targeting to a specific cell population or cellcompartment. In this work, selectivity for extracellular Cyps was achieved by derivatizationof the non-selective CsA molecule with oligo D-Glu peptide. It was clearly shown that thismodification of CsA led to drug improvements in terms of: better solubility, no side effect,lower drug doses, no immunosuppression.It could be concluded that selective targeting of extracellular Cyps is a powerfulapproach to i) study Cyp functions in physiological and pathological processes and ii)develop novel improved anti-inflammatory drugs.AcknowledgmentsSupported by SFB 610 (Deutsche Forschungsgemeinschaft) and AI067254 (National Institutes of Health).References1. Malešević, M., et al. Angew. Chem. Int. Ed. 49, 213-215 (2010).2. Malešević, M., Lücke, C., Jahreis, G. In Peptides 2004, Proceedings of the Third International andTwenty-Eighth European Peptide Symposium, Kenes International, Israel, 2005, p. 391.3. Zhang, Y., et al. J. Biol. Chem. 280, 4842-4850 (2005).4. Fischer, G., et al. Biomed. Biochim. Acta 43, 1101-1111 (1984).5. Balsley, M.A., et al. J. Immunol. Submitted.25