The respiratory influenza viruses A and B remain a significant global health challenge by causing annual seasonal flu, occasional epidemics and pandemics. These threats in combination with the propensity of influenza viruses to develop resistance to commonly used drugs require the continued development of new antiviral agents. This project aims at the identification of novel anti-influenza agents from natural sources targeting two influenza virus proteins, namely the neuraminidase (NA) and the nucleoprotein (NP). These two targets are highly conserved and cover various stages of viral life cycle. Novel agents against NA and NP shall help to overcome resistance and to develop multidrug strategies.

Inhibition of the influenza virus NA impedes the release of newly formed virions from infected cells and causes viral aggregation, thus halting the spread of infection. The high amount of data and information of this established drug target and its ligands will be used as starting point to apply structure-based  computational methods to cope with the flexibility of the NA binding site (Grienke et al. 2010, doi 10.1021/jm901440f), and to scrutinize its ligand-target interactions for the discovery of ligands that overcome the influence of mutational changes conferring drug resistance.

In contrast to NA, the influenza nucleoprotein (NP) only recently emerged as druggable target for potential small molecule therapies. Ligands of this abundantly expressed protein are found to impede influenza A virus replication by triggering the aggregation of NP and inhibition of its nuclear accumulation. Based on previously published compounds interacting on the NP, a ligand-based approach will be the rationale for the target-oriented identification of putative NP-ligands from the rich pool of secondary metabolites.

The selection of natural starting materials for phytochemical investigations will depend on

(i) virtually predicted hits form ligand- and structure-based approaches

(ii) previously determined anti-influenza or anti-viral activities of herbal remedies or constituents thereof (with unknown mechanism of action) and

(iii) empirical knowledge of antiviral herbal remedies form traditional medicine.

Based on these selection criteria, in depth analytical and phytochemical studies will be performed with most promising natural starting materials for each target, primarily from plant origin. Fractions enriched with virtually predicted hits and finally isolated and identified compounds will be provided for assaying on the respective anti-influenza targets and determination of their antiviral efficacy.

We aim to

(i) explain the anti-influenza molecular mechanism of traditionally approved herbal remedies,

(ii) identify naturally derived drug leads for novel NA-inhibitors with lower risk to develop drug resistance, and

(iii) enrich the pool of small molecule ligands for the innovative anti-influenza target NP in an effort to discover agents for a rapidly evolving pathogen.

Virology Group in Jena:

PD Dr. Michaela Schmidtke

Zhongli Xu (PostDoc)

Martina Richter (PhD student)

Elisabeth Walther (PhD student)

Nora Seidel (PhD student)

Anja Hoffmann (PhD student)

Kathrin Bohn (PhD student)

Birgit Jahn (MTA)

Theoretical Chemistry Group in Innsbruck:

Univ. Prof. Dr. Dr. Klaus Liedl

Susanne von Grafenstein (PhD student)

Prof. Dr. Johannes Kirchmair (Professor of Applied Bioinformatics at the Center for Bioinformatics, University of Hamburg)

Medicinal Chemistry Group in Moskow:

Dr. Vadim Makarov

Kick-off Meeting: 'Neuraminidases for Influenza Therapy' and Workshop

in Jena, Germany, on 18./19.07.2012: Program

'Flu Workshop' and Annual Meeting

in Innsbruck, Austria, on 25./26.02.2013: Program

'Flu Workshop' and Annual Meeting

in Innsbruck, Austria, on 30./31.03.2014: Program

'Flu Workshop'

in Jena, Germany, on 16./17.07.2014: Program

'Flu Meeting'

in Vienna, Austria, on 26.05.2015

"Forschungsnewsletter Universität Wien, Oktober 2016"

Die Kraft der Pflanzen

Papers

Grienke, U.; Mair, C.E.; Kirchmair, J.; Schmidtke, M.; Rollinger, J.M. (2018): Discovery of bioactive natural products for the treatment of acute respiratory infections - An integrated approach. Planta Med. Special Issue “Women in Natural Products Science” doi: 10.1055/a-0590-5153

Mair, C.E.; Grienke, U.; Wilhelm, A.; Urban, E.; Zehl, M.; Schmidtke, M.; Rollinger, J.M. (2018): Anti-influenza triterpene saponins from the bark of Burkea africana. J. Nat. Prod. 81: 515-523. Special Issue in Honor of Susan Horwitz doi: 10.1021/acs.jnatprod.7b00774

Hoffmann, A.; Richter, M.; von Grafenstein, S.; Walther, E.; Xu, Z.; Schumann, L.; Grienke, U.; Mair, C.E.; Kramer, C.; Rollinger, J.M.; Liedl, K.R.; Schmidtke, M.; Kirchmair, J. (2017): Discovery and characterization of diazenylaryl sulfonic acids as inhibitors of viral and bacterial neuraminidases. Front. Microbiol. 8:205. DOI: 10.3389/fmicb.2017.00205.

Grienke, U.; Richter, M.; Walther, E.; Hoffmann, A.; Kirchmair, J.; Makarov, V.; Nietzsche, S.; Schmidtke, M.; Rollinger, J.M. (2016): Discovery of prenylated flavonoids with dual activity against influenza virus and Streptococcus pneumoniae. Sci. Rep. 6, 27156. DOI: 10.1038/srep27156.

Walther, E.; Xu, Z.; Richter, M.; Kirchmair, J.; Grienke, U.; Rollinger, J.M.; Krumbholz, A.; Saluz, H.P.; Pfister, W.; Sauerbrei, A.; Schmidtke, M. (2016): Dual acting neuraminidase inhibitors open new opportunities to disrupt the lethal synergism between Streptococcus pneumoniae and influenza virus. Front. Microbiol. 7:357. doi: 10.3389/fmicb.2016.00357.

Grienke, U.; Kaserer, T.; Pfluger, F.; Mair, C.E.; Langer, T.; Schuster, D.; Rollinger, J.M. (2015): Accessing biological actions of Ganoderma secondary metabolites by in silico profiling. Phytochemistry 114, 114-124 (Ganoderma special issue). doi: 10.1016/j.phytochem.2014.10.010.

Richter, M.; Schumann, L.; Walther, E.; Hoffmann, A.; Braun, H.; Grienke, U.; Rollinger, J.M.; von Grafenstein, S.; Liedl, K.R.; Kirchmair, J.; Wutzler, P.; Sauerbrei, A.; Schmidtke, M. (2015): Complementary assays helping to overcome challenges for identifying neuraminidase inhibitors. Future Virol. 10, 77-88. DOI: 10.2217/FVL.14.97.

Walther, E.; Richter, M.; Xu, Z.; Kramer, C.; von Grafenstein, S.; Kirchmair, J.; Grienke, U.; Rollinger J.M.; Liedl, K.R.; Slevogt, H.; Sauerbrei, A.; Saluz, H.; Pfister, W.; Schmidtke, M. (2015): Antipneumococcal activity of neuraminidase inhibiting artocarpin. Int. J. Med. Microbiol. 305, 289-297. DOI: 10.1016/j.ijmm.2014.12.004.

von Grafenstein, S.; Wallnoefer, H.G.; Kirchmair, J.; Fuchs, J.E.; Huber, R.G.; Schmidtke, M.; Sauerbrei, A.; Rollinger, J.M.; Liedl, K.R. (2015): Interface dynamics explain assembly dependency of influenza neuraminidase catalytic activity. J. Biomol. Struct. Dyn. 33, 1, 104–120 doi: 10.1080/07391102.2013.855142.

Grienke, U.; Braun, H.; Seidel, N.; Kirchmair, J.; Richter, M.; Krumbholz, A.; von Grafenstein, S.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2014): Computer-guided approach to access the anti-influenza activity of licorice constituents. J. Nat. Prod. (special issue dedicated to Prof. Dr. O. Sticher) 77, 563–570. doi: 10.1021/np400817j.

Grienke, U.; Schmidtke, M.; von Grafenstein, S.; Kirchmair, J.; Liedl, K.R.; Rollinger, J.M. (2012): Influenza neuraminidase: A druggable target for natural compounds. Nat. Prod. Rep. 29, 11-36. doi: 10.1039/C1NP00053E (amongst top ten most accessed articles from the online version of Nat. Prod. Rep. in 2012).

Kirchmair, J.; Rollinger, J. M.; Liedl, K. R.; Seidel, N.; Krumbholz, A.; Schmidtke, M. (2011) Identification, biological evaluation and investigations of the binding mode of resistance-braking inhibitors of influenza neuraminidase. Future Med. Chem. 3, 437-450. doi: 10.4155/fmc.10.292

Kirchmair, J.; Distinto, S.; Liedl, K.R.; Markt. P.; Rollinger, J.M., Schuster, D.; Spitzer, G.M.; Wolber, G. (2011) Development of anti-viral agents using molecular modeling and virtual screening techniques. Infect. Disorders-Drug Targets 11, 64-93. doi: 10.2174/187152611794407782

Rollinger, J.M. and Schmidtke, M. (2011) The human rhinovirus - human pathological impact, mechanisms of antirhinoviral agents and strategies for their discovery. Med. Res. Rev. Jan, 31, 42-92. doi: 10.1002/med.20176

Grienke, U.; Schmidtke, M.; Kirchmair, J.; Pfarr, K.; Wutzler, P.; Dürrwald, R.; Wolber, G.; Liedl, K.R.;Stuppner, H.; Rollinger, J.M. (2010): Antiviral potential and molecular insight into neuraminidase inhibiting diarylheptanoids from Alpinia katsumadai. J. Med. Chem. 53, 778-786. doi: 10.1021/jm901440f

Patent

Kirchmair, J.; Schmidtke, M.; Liedl, K.; Rollinger, J.M. (2013): Compounds for the treatment of influenza. Eur. Pat. Appl. EP 2599480 A1 20130605

Talks

Grienke, U.; Richter, M.; Walther, E.; Hoffmann, A.; Kirchmair, J.; Makarov, V.; Nietzsche, S.; Schmidtke, M.; Rollinger, J.M. (2016) Killing two birds with one stone – Prenylated flavonoids disrupt the lethal synergism of influenza A viruses and pneumococci. Oral presentation at the 9th Joint Natural Products Conference in Copenhagen, Denmark. July 25, 2016.

Schmidtke, M. (2015): Inhibitors of influenza virus and pneumococcal neuraminidase from nature. Invited lecture to the Pharma & Food Lecture Series, Faculty of Life Sciences, University of Vienna, Vienna/Austria, May 26, 2015.

Mair, C.E.; Grienke, U.; Draschl, C.; Kramer, C.; Richter, M.; von Grafenstein, S.; Kirchmair, J.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2014): Linking ethnopharmacology with phenotypic and virtual screening for the identification of antiviral agents from natural sources. Oral presentation at the Young Researcher Workshop of the 62nd International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA) in Guimarães, Portugal. August 31, 2014. Abstract: Planta Med. 80, 2014, WS17. DOI: 10.1055/s-0034-1394558.

Rollinger, J.M. (2014): Das Potential von Naturstoffen auf der Suche nach antiviralen Leitstrukturen, invited lecture at the Forum Virologicum, Friedrich Schiller University of Jena/Germany, April 23, 2014.

Grienke, U. (2014): Alternative strategies to access anti-influenza agents from multi-component natural mixtures. Oral presentation at the 'Flu Workshop' in Innsbruck, Austria. March 31, 2014.

Mair, C.E. (2014): Is there a benefit from traditional medicine in a phenotypic antiviral screening? Oral presentation at the 'Flu Workshop' in Innsbruck, Austria. March 31, 2014.

Grienke, U.; Braun, H.; Seidel, N.; Kirchmair, J.; Richter, M.; Krumbholz, A.; von Grafenstein, S.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2014): Shedding light on the anti-influenza effect of Glycyrrhiza glabra roots: Complex multi-component mixtures versus pure constituents. Oral presentation at the 24th Annual Meeting of the Society for Virology in Alpbach/Tyrol, Austria. March 27, 2014.

Walther, E.; Xu, Z.; Grienke, U.; Kirchmair, J.; Rollinger, J.M.; Sauerbrei, A.; Pfister, W.; Schmidtke, M. (2014):Blocking pneumococcus-enhanced influenza virus spread by dual-targeting neuraminidase inhibitors. Oral presentation at the 24th Annual Meeting of the Society for Virology in Alpbach/Tyrol, Austria. March 26, 2014.

Mair, C.E. (2014): Metabolite profiling and anti-influenza screening of micro-fractions from Capsella bursa-pastoris. Oral presentation at the Jena University Hospital Jena/G, Institute for Virology and Antiviral Therapy, Feb. 10, 2014.

Grienke, U.; Richter, M.; Braun, H.; Kirchmair, J.; von Grafenstein, S.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2013): New insights into the anti-influenza activity of licorice constituents. Oral presentation at the Joint Symposium "Trends in natural products research" of the Phytochemical Society of Europe (PSE) and the Austrian Pharmaceutical Society (ÖPhG) at the University Centre Obergurgl/Tyrol, Austria. July 24, 2013.

Mair, C.E.; Grienke, U.; von Grafenstein, S.; Kirchmair, J.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2013): Bioprospecting diverse plant species for antiviral agents against upper respiratory tract infections. Oral presentation at the Joint Symposium "Trends in natural products research" of the Phytochemical Society of Europe (PSE) and the Austrian Pharmaceutical Society (ÖPhG) at the University Centre Obergurgl/Tyrol, Austria. July 24, 2013.

Rollinger, J.M. (2013) Implementation of traditional medicine into structure based approaches for antiviral lead discovery. Invited key lecture at the annual Meeting of the American Society of Pharmacognosy (ASP) in St. Louis, Missouri, July 16, 2013

Grienke, U. (2013) Comparative studies and challenges faced by different flavonoid scaffolds in anti-viral assays. Oral presentation at the 'Flu Workshop' in Innsbruck, Austria. February 26, 2013. 

Grienke, U. (2012) Targeting Inflammation and Influenza: Integration of Computational Methods into Lead Finding From Natural Sources. Award Lecture on the occasion of the Egon Stahl Award in Bronze at the International Congress on Natural Products Research, New York City, July 30, 2012.

Rollinger, J.M. (2012) Integrated data-mining tools for the identification of natural drug leads. Invited guest lecture at the Department of Chemical Engineering of Aristotle University of Thessaloniki, Greece. Thessaloniki, 5^th June 2012

von Grafenstein, S.; Wallnoefer, H.G.; Fuchs, J.E.; Huber, R.G.; Rollinger, J.M.; Kirchmair, J.; Liedl, K.R. (2011) Impact of tetramerization on neuraminidase dynamics and binding site conformations. Oral presentation at the 3rd Life Science Meeting of Innsbruck Universities, Igls, Austria, 23.-24.09.2011

Rollinger, J.M.; Kirchmair, J.; Grienke, U.; Schuster, D.;  Liedl, K. R.; Schmidtke, M. (2011) Common cold and flu: Computational strategies for the identification of antiviral leads from nature. Invited lecture at the 2011 Spring ACS meeting, CINF division "Natural Products/Drug Discovery" in Anaheim, California, 28^th March, 2011.

Rollinger. J.M. (2011) In silico prediction of bioactive phytochemicals: Expectations, realities and examples. Plenary lecture at the 5^th International Conference on Polyphenols and Health, in Sitges/Barcelona, Spain, 19^th October 2011.

Rollinger, J.M. (2011): Combination of ethnopharmacological knowhow with modern in silico tools. Plenary lecture at the 59^th International Congress of the Society for Medicinal Plants and Natural Product Research in Antalya, Turkey, September 5^th, 2011. Abstract: Planta Med. 77, 2011, L2, 1230.

Grienke, U.; Schmidtke, M.; Kirchmair, J.; Stuppner, H.; Liedl, K.R.; Rollinger J.M. (2010) Discovery of novel neuraminidase inhibiting scaffolds from Alpinia katsumadai. Oral presentation at the 58^th International Congress of the Society for Medicinal Plants and Natural Product Research in Berlin, Germany, August 30, 2010. Abstract: Planta Med.76, 2010, SL-8, 1179.

Grienke, U.; Schmidtke, M.; Kirchmair, J.; Liedl, K.R.; Rollinger J.M. (2010) Identification of novel antiinfluenza constituents from Alpinia katsumadai. Oral presentation at the 2nd Life Science Meeting, Igls, Austria, 24.-25.09.2010.

Schmidtke, M.; Wutzler, P.; Rollinger, J. M. (2009) Target-based discovery of natural inhibitors to combat common cold and flu. Oral presentation at the International Workshop ‘Novel strategies to fight respiratory viral diseases’ in Berlin, Germany, 12-13. Oct. 2009.

Posters

Mair, C.E.; Grienke, U.; Wilhelm, A.; Urban, E.; Zehl, M.; Rollinger, J.M.; Schmidtke, M. (2018): Novel influenza virus inhibitors from the bark of Burkea africana. Poster presentation at the 28th Annual Meeting of the Society for Virology in Würzburg, Germany. March 15, 2018. P200.

Zwirchmayr, J.; Grienke, U.; Richter, M.; Mair, C.E.; Peintner, U.; Schmidtke, M.; Rollinger, J.M. (2018): Unraveling the anti-influenza potential of polypores. Poster presentation at the First Austrian Summit on Natural Products in Seefeld, Austria. January 15, 2018. P9.

Grienke, U.; Kaserer, T.; Pfluger, F.; Mair, C.E.; Schmidtke, M.; Langer, T.; Schuster, D.; Rollinger, J.M. (2015): In silico pharmacological profiling of Ganoderma secondary metabolites to unravel the polypharmacological nature of Reishi. Poster presentation at the 63rd International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA) in Budapest, Hungary. August 24, 2015, PM-124.

Richter, M.; Walther, E.; Bohn, K.; Xu, Z.; Braun, H.; Grienke, U.; Rollinger, J.M.; von Grafenstein, S.; Liedl, K.R.; Kirchmair, J.; Sauerbrei, A.; Schmidtke, M. (2014): Hemagglutination (HA)-based neuraminidase (NA) inhibitory assays to search for novel NA inhibitors (NAI). Poster presentation at the 5th Influenza Conference of the European Scientific Working group on Influenza (ESWI) in Riga, Latvia. September 16, 2014, P101.

Mair, C.E.; Grienke, U.; Draschl, C.; Kramer, C.; Richter, M.; von Grafenstein, S.; Kirchmair, J.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2014): Linking ethnopharmacology with phenotypic and virtual screening for the identification of antiviral agents from natural sources. Poster presentation at the 62nd International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA) in Guimarães, Portugal. September 3, 2014, P2Y12.

Richter, M.; Walther, E.; Bohn, K.; Xu, Z.; Braun, H.; Grienke, U.; Rollinger, J.M.; von Grafenstein, S.; Liedl, K.R.; Kirchmair, J.; Sauerbrei, A.; Schmidtke, M. (2014): Reliability of different assays for identification of novel neuraminidase inhibitors (NAI). Poster presentation at the 24th Annual Meeting of the Society for Virology in Alpbach/Tyrol, Austria. March 26, 2014, P019.

Hoffmann, A.; von Grafenstein, S.; Kirchmair, J.; Liedl, K.R.; Rollinger, J.M.; Schade, D.; Sauerbrei, A.; Schmidtke, M. (2014): Analysis of recombinant influenza virus susceptibility to antiviral agents with respect to inhibitory mechanism studies. Poster presentation at the 24th Annual Meeting of the Society for Virology in Alpbach/Tyrol, Austria. March 26, 2014, P017.

Mair, C.E.; Grienke, U.; von Grafenstein, S.; Kirchmair, J.; Liedl, K.R.; Schmidtke, M.; Rollinger J.M. (2014): Reverse pharmacology: exploiting ethnopharmacological knowledge to identify antiviral agents against ARIs. Poster presentation at the 24th Annual Meeting of the Society for Virology in Alpbach/Tyrol, Austria. March 26, 2014, P008.

Walther, E.; Xu, Z.; Leitzinger, C.; Rollinger, J.M.; Grienke, U.; Kirchmair, J.; Liedl, K.R.; von Grafenstein, S.; Sauerbrei, A.; Pfister, W.; Schmidtke, M. (2013): Characterisation of pneumococcal neuraminidase (NA) and the effect of neuraminidase inhibitors (NAIs) on NA activity, bacterial growth, and biofilm formation. Poster presentation at the Joint Annual Meeting of the German Society for Hygiene and Microbiology (DGHM) and the German Society for Infectious Diseases (DGI) in Rostock, Germany. September 24, 2013, KMP02.

Grienke, U.; Richter, M.; Braun, H.; Kirchmair, J.; von Grafenstein, S.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2013): New insights into the anti-influenza activity of licorice constituents. Poster presentation at the:

• 61th International Congress of the Society for Medicinal Plant and Natural Product Research in Münster, Germany. September 3, 2013. Abstract: Planta Med. 79, 2013, PG3. DOI: 10.1055/s-0033-1352073
• 5th Life Science Meeting and Annual Meeting of the Austrian Association of Molecular Life Sciences and Biotechnology (ÖGMBT) in Innsbruck, Austria. September 25, 2013, P74.

Mair, C.E.; Grienke, U.; von Grafenstein, S.; Kirchmair, J.; Liedl, K.R.; Schmidtke, M.; Rollinger, J.M. (2013): Bioprospecting diverse plant species for antiviral agents against upper respiratory tract infections. Poster presentation at the:

• 61th International Congress of the Society for Medicinal Plant and Natural Product Research in Münster, Germany. September 4, 2013. Honoured with the Best Poster Award. Abstract: Planta Med. 79, 2013, PN57. DOI: 10.1055/s-0033-1352400
• 5th Life Science Meeting and Annual Meeting of the Austrian Association of Molecular Life Sciences and Biotechnology (ÖGMBT) in Innsbruck, Austria. September 25, 2013, P73.

von Grafenstein, S.; Fuchs, J.E.; Huber, R.G.; Margreiter, M.A.; Mair, C.E.; Grienke, U.; Rollinger, J.M.; Kirchmair, J.; Schmidtke, M.; Liedl, K.R. (2013): Similarity in shape and electrostatics guide virtual screening for novel influenza nucleoprotein ligands. Poster presentation at the:

• Gordon Research Conference – Computer Aided Drug Design in West Dover, VT, United States. July 24&25, 2013.
• 5th Life Science Meeting and Annual Meeting of the Austrian Association of Molecular Life Sciences and Biotechnology (ÖGMBT) in Innsbruck, Austria. September 26, 2013, P174.

Kirchmair, J.; Rollinger, J. M.; Grienke, U.; Seidel, N.; Krumbholz, A.; Schmidtke, M.; Liedl, K. R. (2011) In silico identification of resistance-breaking inhibitors of influenza neuraminidase. Poster presentation at the 9th International Conference on Chemical Structures, Noordwijkerhout, The Netherlands, June 7, 2011.

Schmidtke, M.; Rollinger, J.M.; Grienke, U.; Seidel, N.; Krumbholz, A.; Wutzler, P.; Liedl, K.R.; Kirchmair, J. (2011) Discovery of novel natural neuraminidase inhibitors (NAI) based on in silico screening and antiviral investigations. Poster presentation at the 24^th International Conference on Antiviral Research (ICAR) in Sofia, Bulgaria, May 9, 2011, P78.

von Grafenstein, S.; Wallnöfer, H.G.; Kirchmair, J.; Grienke, U.; Schmidtke, M.; Rollinger, J.M.; Liedl, K.R. (2011): Molecular dynamics simulations of neuraminidase - N1 subtype. Poster presentation at the:

• 25th Molecular Modelling Workshop in Erlangen, Germany. April 4, 2011, P43.
• 6th Meeting of Doctoral students from Innsbruck’s Medicinal University and of the Life Science Disciplines of the Leopold-Franzens-University Innsbruck in Innsbruck, Austria. November 24, 2010, P85.

von Grafenstein, S.; Wallnoefer, H.G.; Fuchs, J.E.; Huber, R.G.; Rollinger, J.M.; Kirchmair, J.; Liedl, K.R. (2011) Impact of tetramerization on neuraminidase dynamics and binding site conformations. Poster presentation at the Joint Meeting of the Austrian and German Pharmaceutical Societies, Innsbruck, 20.09.2011 - 23.09.2011