INFLUENCE OF THE CONFORMATION OF 2-ACYLPYRROLES ON SPECTROSCOPIC PROPERTIES AND HYDROGEN BOND FORMATION

Załącznik 2B Dr Alina Teresa Dubis University of Białystok Faculty of Biology and Chemisty Institute of Chemistry Department of Natural Product Chemistry Al. J. Piłsudskiego 11/4, 15-443 Białystok, Poland mobile 600-447773, 85-7457800 Documentation for the application for the initiation of the habilitation procedure Scientific achievements presented as a series of nine publications after being granted the PhD degree Scientific achievements as defined in the Act of 1 September 2011 (Journal of Laws No 1165) and of 22 September 2011 (Journal of Laws No 201, Item 1200) are presented under the title: INFLUENCE OF THE CONFORMATION OF 2-ACYLPYRROLES ON SPECTROSCOPIC PROPERTIES AND HYDROGEN BOND FORMATION Białystok 2014

1. SURENAME AND NAME: Alina Teresa Dubis 2. EDUCATION: 1972 1976 I Liceum Ogólnokształcące w Białymstoku 1976 1980 University of Warsaw, Branch in Białystok 3.07.1980 M.SC. in chemistry, teaching specialist, Thesis: "Studia nad wydzielaniem substancji biologicznie czynnych stonki ziemniaczanej (Leptinotarsa decemlineata Say). Identyfikacja frakcji wymrożonych w acetonie Supervisor: dr hab. Janusz Popławski Graduated with Honors 4.03.2004 Ph.D in chemistry, Department of Biology and Chemistry, University of Białystok, Thesis: The Application of IR Spectroscopy, Theoretical Calculation and Bader Theory in Conformational Analysis of -Substituted Pyrroles" Supervisor: dr hab. Sławomir J. Grabowski 1.10.2010-2.07.2011 post graduated study in the field of Management of Research and Development Works completed with very good results; Diploma Thesis: A.T. Dubis, J. Piekutin, Metody badań naukowych z przykładami ich zastosowania chapter in a monograph edited by Professor Bazyli Poskrobko, Experimental Methods, pp.127-145, Wydawnictwo Ekonomia i środowisko, Białystok, 2012. 3. PROFESSIONAL POSITION: 1980-2004 Specialist, Department of Organic Chemistry, Institute of Chemistry, UwB 2004-2007 Research Assistant, Department of Organic Chemistry, Institute of Chemistry, UwB 2007- Adjunct, Department of Chemistry of the Natural Products, Institute of Chemistry, UwB 3

Deputy Director of Institute of Chemistry since 2012 4. INTERNATIONAL FELLOWSHIOS November 1990 June 1991 Rurgers University, Department of Entomology, USA (New Jersey), Prof. Lena B. Brattsten 5. SCIENTIFIC ACHIEVEMENTS Scientific publications:... 39 Before Ph.D:... 17 After Ph.D... 22 Total number of publications from JCR:... 34 Conference messages... 38 Patents... 4 Book chapters:... 2 Review for editors J. Mol. Struct., Vib. Spectroscopy... 10 Total points of MNiSW...935 IF according to the Journal Citation Report (JCR) summarized.... IF =72.43 after PhD...... IF=48.28 average IF per publication... IF= 2.13 Summarized of [H1-H9] directly related to the habilitation thesis... IF=17.64 My Average contribution to the publications [H1-H9] is at the level of... 62,7% 6. CITATION OF THE PUBLICATION DATA FROM WEB OF KNOWLEDGE October,2014: The total number of the publications... 33 Sum of the times cited...376 Sum of times without self-citations...336 Average citation per item... 11.39 H-index... H=12 4

7. INTERNATIONAL COLLABORATION: Prof. dr hab. Sławomir J. Grabowski - Faculty of Chemistry, University of the Basque Country 8. NATIONAL COLLABORATION: Dr Andrzej Łapiński, Institute of Molecular Physics PAS, Poznań Dr hab. Marcin Palusiak, Department of Chemistry, University of Łódź 9. PARTICIPATION IN RESEARCH PROGRAMMES: Research grant: 4T09A 163 22 Project duration : 2002-2004 Project subject: Zastosowanie spektroskopii w podczerwieni, obliczeń ab initio i teorii Badera w badaniach konformacji -podstawionych piroli Head of the project: dr hab. Sławomir J. Grabowski 5

main contractor Research grant: N310 06331/2813 Project duration : 2006-2008 Project subject: Zastosowanie zaawansowanych metod spektralnych w badaniach peraminy jako alternatywnego środka ochrony roślin Head of the project: dr Andrzej Łapiński main contractor grant: NN 20411535 Project duration: 2008-2010 Project subject: Badanie możliwości wykorzystania małych nanocebulek węglowych w bioczujnikach Head of the project: dr Marta Płońska-Brzezińska main contractor grant: OPUS, NCN2011/03/B/ST5/02691 Project duration: 2012-2015 Project subject: Synteza nanocząstek magnetycznych z polimerowymi powłokami chelatującymi Head of the project: dr Agnieszka Wilczewska contractor 10. Received Awards: Rector s Award for Scientific Work, University of Warsaw, Białystok Branch, 1995 Rector s Award of Teaching, University of Białystok 1997 Rector s Award of Organizational Work, University of Białystok, 2001 The Award of the III Multidyscyplinarna Konferencja Nauki o Leku, 2002 for the Best Poster Presentation Hypolipidemic Agents New -Asarone Analogs, Rector s Award for Scientific Work, University of Białystok,2004 Rector s Award of Teaching, University of Białystok, 2005 Rector s Award for Scientific Work, University of Białystok, 2010 Nominated in the pool Lecturer of the year 2010 Kuriera Porannego 6

11. PARTICIPATION IN THE WORK OF THE INTERANATIONAL AND NATIONAL CONFERENCE ORGANIZING COMMITTEES: 55 Zjazd Polskiego Towarzystwa Chemicznego, Białystok, 9-12.IX.1992, patricipation in the Organizing Committee works XXI Międzynarodowa Konferencja Izoprenoidowa, Białowieża, 23-29.IX.2005, patricipation in the Organizing Committee works 55. Zjazd Polskiego Towarzystwa Chemicznego, Białystok, 16-20.IX.2012, the treasurer of the Conference 12. MEMBERSHIP IN SCIENTIFIC ORGANIZATIONS AND SOCIETIES Polish Chemical Society; Physical Organic Chemistry Section 13. CHEMISTRY POPULARIZING ACTIVITIES Chemistry Preparatory Course for high-school students - teaching kinetics and termochemistry (2009-2013) Member of Regional Commmittee of the Polish Chemistry Olympiad PTCh (2001-2010) Co-organizer of Soboty u Chemików (2013-14) meetings with the high-school students Co-organizer of the Podlaski Konkurs Chemiczny for high-school students (2014) 14. ORGANIZATIONAL ACHIEVEMENTS: Participation in work on development of a study programme in Environmental Protection under the TEMPUS project Development and implementation of a new BSc degree in Environmental Protection including chemistry, modern technologies, environmental protection practice and legislation at Universities of Gdańsk and Warsaw Bialystok Branch S_JEP-09615-95 ; 1995-996 Participation in training under the TEMPUS project Development and implementation of a new BSc degree in Environmental Protection including chemistry, modern technologies, environmental protection practice and legislation at Universities of Gdańsk and Warsaw Bialystok Branch S_JEP-09615-95 ; Hertfordshire, 1996 7

The member of the committee for teaching of the Institute of Chemistry Participation in training on the requirements of the National Qualifications Frameworks (2011-12) The member of the Scientific Council of the Institute of Chemistry since 2012 The member of the Faculty Council Committee for the development of the teaching since 2012 Coordinator of the Program Council for teaching since 2012 Preparation of the application to the Competition for funding of basic organizational units of universities in the implementation of systems to improve the quality of education and the National Qualifications Framework KRK2013, MNiSW (spetember 2013) Member of the team preparing the report for the purpose of self-assessment program accreditation of Chemistry (May 2014) 15. TEACHING: I was supervisor of 11 master thesis I was supervisor of 8 bachelor thesis I am supervisor of 1 doctor thesis 16. COURSES FOR STUDENTS OF FACULTY OF BIOLOGY AND CHEMISTRY: Molecular Spectroscopy - lecture 30 h (Chemistry, MSc, 1st year) Molecular Spectroscopy - laboratory (Chemistry, MSc, 1st year) Instrumental Laboratory (Chemistry, MSc, 1st year) Specialization Laboratory I (Chemistry, MSc, 1st year, Environmental Protection, MSc, 1st year) Specialization laboratory II (Chemistry, MSc, 1st year, Environmental Protection, MSc, 2nd year) Specialization Course (Chemistry, Environmental Protection, BSc, 3rd year) The Laboratory of Spectroscopic Methods of Analysis (Environmental Protection, MSc, 1st year) The Laboratory of Chemical Analysis of Environmental Pollution (Environmental Protection, MSc, 1st year) 8

The Laboratory of Organic Chemistry (Chemistry, BSc, 2nd year) The Laboratory of Environmental Monitoring (Environmental Protection, BSc, 3rd t year) The Laboratory of Instrumental Analysis (Environmental Protection, BSc, 3rd year) Specialization Course (Chemistry and Environmental Protection, MSc, 2nd year) Classes were conducted in the dimension of 210-240 hours per year. Evaluation of teaching based on student surveys is contained in the range of 4.5-4.7 on a scale of 5 points. 9

ALL PUBLICATIONS INCLUDED IN THE JOURNAL CITATION REPORTS (JRC) [H1], H[2]...- directly related to the habilitation thesis IF - based on year of publication IF5 - based on 5-year TC - total number of citation PUBLICATIONS BEFORE BEING GRANTED THE PHD DEGREE 1. E. Dubis, E. Maliński, A.T. Dubis, J. Szafranek, J. Nawrot, J. Popławski, J.T. Wróbel Sex-dependent Composition of Cuticular Hydrocarbons of the Colorado Beetle Leptinotarsa decemlineata Say, Comp. Biochem. Physiol., 87A (1987) 839-843. IF5 =2,381 TC = 18 My contribution to [1] is related to collection of the insects and potato leaves, extract preparation, the separation of the hydrocarbon using chromatographic methods, participation in the discussion, preparation of the manuscript. I declare my contribution to be equal to 15 % 2. A.T. Dubis, J.W. Morzycki, J. Popławski The alkali metal reduction of trimethoxybenzenes in hydrocarbon solvents, Journaj f. prakt. Chemie; 333 (1991) 643-650. IF = 0,885 TC = 0 My contribution to [2] is related to formulate a research problem, the synthesis of a sample, interpretation and discussion, preparation of the manuscript I declare my contribution to be equal to 50 % 3. S. J. Grabowski, A.T. Dubis Intramolecular C-H.. O Hydrogen Bonds in the Crystal Structure of Ethyl 3,4,5- Trimethoxybenzoate (ETMB). Polish J. Chem. 69 (1995) 218-222. IF = 0,422 TC =2 10

My contribution to [3] is related to the synthesis of a sample, interpretation and discussion, preparation of the manuscript I declare my contribution to be equal to 20 % 4. A.T. Dubis, Z. Łotowski, L. Siergiejczyk, A. Z. Wilczewska, J. W. Morzycki Study of Hydrogen Bonding in Nitro Enamindes. J. Chem. Research (S), (1998) 170-171. IF = 0,522 TC = 3 My contribution to [4] is related to IR spectroscopic investigation, interpretation and discussion, preparation of the manuscript I declare my contribution to be equal to 20 % 5. A.T. Dubis, Z. Łotowski, L. Siergiejczyk, A. Z. Wilczewska, J. W. Morzycki Study of Hydrogen Bonding in Nitro Enamindes. J. Chem. Research (M), (1998) 813-821. IF = 0,522 TC = 3 My contribution to [5] is related to IR spectroscopic investigation, interpretation and discussion, preparation of the manuscript Mój udział procentowy szacuję na 20 %. 6. E.N. Dubis, A.T. Dubis, J. W. Morzycki Comparative Analysis of Plant Cuticular Waxes Using HATR FT-IR Reflection Technique. J. Mol. Struct. 511-512 (1999) 173-179. IF = 0.868 TC = 11 My contribution to [6] is related to formulate a research problem, the choice of research methodology, spectroscopic measurements, interpretation, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 45 % 7. J. Popławski, B. Łozowicka, A.T. Dubis, B. Lachowska, Z. Winiecki, J. Nawrot Feeding-deterrent Activity of -Asarone Isomers Against Some Stored Coleoptera, Pest Manag Sci. 56 (2000) 560-564. IF = 0,642 TC = 12 11

My contribution to [7] is related to formulate a research problem, the choice of research methodology, synthesis of the alpha-asarone derivatives, spectroscopic measurements, interpretation, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 15 % 8. J. Popławski, B. Łozowicka, A.T. Dubis, B. Lachowska, S. Witkowski, J. Cybulski, Z. Chilmonczyk, R. Kaliszan Synthesis and Hypolipidemic and Antiplatelet Activity of Alpha-Asarone Isomers. Journal of Medicinal Chemistry 43 (2000) 3671-3676. IF = 4,139 TC = 29 My contribution to [8] is related to the choice of research methodology, synthesis of the alpha-asarone derivatives, spectroscopic measurements, interpretation, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 15 % 9. L. Siergiejczyk, J. Popławski, A.T. Dubis, B. Lachowska. B. Łozowicka 1 H and 13 C NMR Studies of -Asarone Isomers, Magnetic Resonance in Chemistry, 38 (2000) 1037-1038. IF = 1,006 TC = 11 My contribution to [9] is related to the choice of research methodology, synthesis of the alpha-asarone derivatives, spectroscopic measurements, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 15 % 10. A.T. Dubis, S. J. Grabowski Infrared Spectroscopic and Theoretical Ab Initio Studies on Conformational Isomers of Methyl Pyrrole-2-Carboxylate. J. Mol. Struct. 562 (2001) 107-117. IF = 0,970 TC = 17 My contribution to [10] is related to the choice of research methodology, synthesis of the sample, spectroscopic measurements, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 50 % 12

11. T.M. Krygowski, E. Pindelska, R. Anulewicz-Ostrowska, S.J. Grabowski, A. Dubis Angular group-induced alternation (AGIBA). Part5 Conformational dependence and additivity of the effect: structural studies of 3,5-dimethoxybenzaldehyde and related systems. J. Phys. Org. Chem. 14 (2001) 349-354. IF = 1,330 TC = 8 My contribution to [11] is related to the choice of research methodology, synthesis of the sample, spectroscopic measurements, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 10 % 12. E.N. Dubis, A.T. Dubis, J. Popławski, Determination of the Aromatic Compounds in Plant Cuticular Waxes using FT-IR Spectroscopy. J. Mol. Struct. 596/1-3 (2001) 83-88. IF = 0,970 TC =5 My contribution to [12] is related to the choice of research methodology, synthesis of the aromatic esters, spectroscopic measurements, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 60 % 13. A.T. Dubis, S. J. Grabowski Vibrational Spectrum of Methyl Pyrrole-2-Carboxylate. Spectrochim. Acta A, 58 (2002) 213-215. IF = 1,046 TC =2 My contribution to [13] is related to the choice of research methodology, synthesis of the aromatic esters, spectroscopic measurements, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 60 % 14. A.T. Dubis, S. J. Grabowski Spectroscopic and Theoretical Studies on Monomeric and Dimeric Forms of Methyl Pyrrole-2-Carboxylate. New J. Chem. 26 (2002) 165-169. IF = 2,060 TC =14 13

My contribution to [14] is related to the choice of research methodology, spectroscopic measurements, interpretation, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 50 % 15. M.E. Płońska, A. Dubis, K. Winkler New insights into the electrodeposition and redox properyties of [M(Bipyridyl)3](CLO)4 (M=Co and Fe) films in media of low dielectric constant. J. Electroanal. Chem. 526 (2002) 77-84. IF = 2,027 TC =3 My contribution to [15] is related spectroscopic measurements and discussion of results. I declare my contribution to be equal to 10 % 16. A.T. Dubis, S.J. Grabowski, D. Romanowska, T. Misiaszek, J. Leszczynski, Pyrrole-2-carboxylic acid and its dimers: molecular structure and vibrational spectrum. J. Phys. Chem. A, 106 (2002) 10613-10621. IF = 2,765 TC = 53 My contribution to [16] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 35 % 17. A.T. Dubis, S.J. Grabowski Infrared, Density-Functional Theory, and Atom in Molecules Method Studies on Conformers of Some 2-Substituted 1H-Pyrroles. J. Phys. Chem. A, 107 (2003) 8723-8729. IF = 2,792 TC =14 My contribution to [17] is related to the choice of research methodology, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 65 % 14

PUBLICATIONS AFTER BEING GRANTED THE PHD DEGREE 18. S.J. Grabowski, A. Pfitzner, M. Zabel, A.T. Dubis, M. Palusiak Intramolecular H... H Interactions for the Crystal Structures of [4-((E)-But-1-enyl)-2,6- dimethoxyphenyl]pyridine-3-carboxylate and [4-((E)-Pent-1-enyl)-2,6- dimethoxyphenyl]pyridine-3-carboxylate; DFT Calculations on Modeled Styrene Derivatives. J. Phys. Chem. B. 108 (2004) 1831-1837. IF = 3,834 TC =32 My contribution to [18] is related to the synthesis of the samples, synthesis of the samples, spectroscopic measurements, discussion of results. I declare my contribution to be equal to 20 % 19. [H1] S.J. Grabowski, A.T. Dubis, D. Martynowski, M. Główka, M. Palusiak, J. Leszczynski Crystal and Molecular Structure of Pyrrole-2-carboxylic Acid; -Electron Delocalization of Its Dimers - DFT and MP2 Calculations. J. Phys. Chem. A, 108 (2004) 5815-5822. IF = 2,639 TC =21 My contribution to [19 (H1)] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 35 % 20. [H2] S.J. Grabowski, A.T. Dubis, M. Palusiak, J. Leszczynski Heteronuclear Intermolecular Resonance-Assisted Hydrogen Bonds. The Structure of Pyrrole-2-Carboxamide (PyCa). J. Phys. Chem. B, 110 (2006) 5875-5882. IF = 4,115 TC =17 My contribution to [20 (H2)] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 40 % 15

21. A.J. Rybarczyk-Pirek, A.T. Dubis, S.J. Grabowski, J. Nawrot-Modranka Intramolecular Hydrogen Bond in Crystals of Thiophosphorylbenzopyrane Derivatives X- Ray and FT-IR Studies. Chem. Phys. 320 (2006) 247-258. IF = 1,984 TC =19 My contribution to [21] is related to the IR and NMR spectroscopic measurements,, discussion of results.. I declare my contribution to be equal to 25 % 22. [H3] E. Bilewicz, A.J. Rybarczyk-Pirek, A.T. Dubis, S.J. Grabowski Halogen bonding in crystal structure of 1-methylpyrrol-2-yl trichloromethyl ketone. J. Mol. Struct. 829 (2007) 208-211. IF = 1,486 TC =30 My contribution to [22 (H3)] is related to the choice of research methodology, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 35 % 23. [H4] S.J. Grabowski, M. Palusiak, A.T. Dubis, A. Pfitzner, M. Zabel Inter- and intramolecular hydrogen bonds Structures of 1-methylpyrrole-2-carboxamide and 1-hydroxypyrrole-2-carboxamide. J. Mol. Struct., 844-845 (2007) 173-180. IF =1,486 TC =5 My contribution to [23 (H4)] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 40 % 24. [H5] A.T. Dubis, A. Łapiński Spectroscopic and theoretical study on peramine and some pyrrolopyrazzinone compounds. Vib. Spec. 49 (2009) 265-273. IF = 1,936 TC =2 My contribution to [24 (H5)] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. 16

I declare my contribution to be equal to 90 % 25. [H6] A. Łapiński, A.T. Dubis A DFT/TD-DFT study for the ground and excited states of peramine and some pyrrolopyrazinone compounds. J. Phys. Org. Chem. 22 (2009) 1058-1064. IF = 1,602 TC =1 My contribution to [25 (H6)] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 85 % 26. [H7] A.T. Dubis, M. Domagała, S.J. Grabowski Spectroscopic and Theoretical Studies on Some New pyrrol-2-yl-chloromethyl ketones. New J. Chem. 34 (2010) 556-566. IF = 2,631 TC =3 My contribution to [26 (H7)] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 60 % 27. J. Luszczyn, M.E. Płonska-Brzezinska, A. Palkar, A.T. Dubis, A. Simionescu, D.T. Simionescu, B. Kalska-Szostko, K. Winkler, L. Echegoyen. Small Noncytotoxic Carbon Nano-Onions: First Covalent Functionalization with Biomolecules. Chem. Eur. J. 16 (2010) 4870-4880. IF = 5,476 TC =12 My contribution to [27] is related to the spectroscopic measurements, discussion of results and preparation of the manuscript. I declare my contribution to be equal to 5 % 28. M.E. Plonska-Brzezinska, A.T. Dubis, A. Lapinski, A.Villalta-Cerdas, L. Echegoyen Electrochemical Properties of Oxidized Carbon Nano-Onions: DRIFTS FT-IR and Raman Spectroscopic Analyses. ChemPhysChem, 12 (2011) 2659-2668. IF = 3,412 TC =2 17

My contribution to [28] is related to the FTIR-DRIFTS spectroscopic measurements, discussion of results. I declare my contribution to be equal to 25 % 29. M.E. Plonska-Brzezinska, A. Lapinski, A.Z. Wilczewska, A.T. Dubis, A.Villalta-Cerdas, K. Winkler, L. Echegoyen The synthesis and characterization of carbon nano-onions produced by solution ozonolysis. Carbon, 49 (2011) 5079-5089. IF = 5,378 TC =6 My contribution to [29] is related to the FTIR-DRIFTS spectroscopic measurements, discussion of results and preparation of the manuscript. I declare my contribution to be equal to 10 % 30. B. Kalska-Szostko, M. Rogowska, A.T. Dubis, K. Szymański Enzymes Immobilization on Fe3O4-goldnanoparticles. Appl. Surf. Sci. 258 (2012) 2783-2787. IF = 2,103 TC =12 My contribution to [30] is related to the FTIR spectroscopic measurements and discussion of results. I declare my contribution to be equal to 10 % 31. M.E. Plonska-Brzezinska, J. Mazurczyk, B. Palys, J. Breczko, A. Lapinski, A.T. Dubis, L. Echegoyen Preparation and Characterization of Composites that contain Small Carbon Nano-Onions and Conducting Polyaniline. Chem. Eur. J. 18 (2012) 2600-2608. IF = 5,47 TC =11 My contribution to [31] is related to the FTIR spectroscopic measurements and discussion of results. I declare my contribution to be equal to 5 % 32. [H8] A.T. Dubis, S. Wojtulewski, K. Filipkowski Spectroscopic and Theoretical Studies on the Aromaticity of Pyrrol-2-yl-carbonyl Conformers. J. Mol. Struct. 1041 (2013) 92-99. 18

IF = 1,404 TC =1 My contribution to [32 (H8)] is related to the choice of research methodology, synthesis of the samples, spectroscopic measurements, theoretical calculations, discussion of results, preparation of the manuscript. I declare my contribution to be equal to 80 % 33. B. Łozowicka, P. Kaczyński, T. Magdziarz, A.T. Dubis Synthesis, antifeedant activity against Coleoptera and 3D QSAR study of alpha-asarone derivatives, SAR QSAR Environ Res. 25(3) (2014) 173-88. IF=1,924 TC=1 My contribution to [33] is related to the synthesis of the samples and spectroscopic measurements. I declare my contribution to be equal to 20 % 34. B. Kalska-Szostko, M. Rogowska, A.T. Dubis, A. Basa Enzyme immobilization on Fe3O4-Silver Nanoparticles. J. Surf. Interfac. Mater. 2 (2014) 69-73. IF = 1,404 TC =0 My contribution to [34] is related to the spectroscopic measurements and discussion of results. I declare my contribution to be equal to 10 % PUBLICATIONS NOT INCLUDED IN THE JCR JOURNAL LIST 35. J. Popławski, S. Lux, S. Witkowski, E.N. Dubis, A.T. Dubis, J.T. Wróbel, J. Dmoch, Preliminary Isolation Study of Colorado Beetle Leptinotarsa Decemlineata Say Body Components and their Influence on Male Behaviour, Bulletin of the Polish Academy of Science 41 (1994) 243-246. My contribution to [35] is related to collection of the insects, extract preparation, the separation of the cuticular waxes using chromatographic methods, participation in the discussion, preparation of the manuscript. I declare my contribution to be equal to 15 % 19

36. A. Łapiński, A.T. Dubis (2008) Zastosowanie zaawansowanych metod spektralnych w badaniach antyfidantu peraminy, jako alternatywnego środka ochrony roślin, Progress in Plant Protection 48(2) (2008) 730-733. My contribution to [36] is related to the FTIR-DRIFTS spectroscopic measurements, discussion of results. I declare my contribution to be equal to 50 % 37. A.T. Dubis, A. Łapiński Własności fizykochemiczne peraminy i jej pochodnych jako alternatywnego środka ochrony roślin. Progress in Plant Protection 48(2) (2008) 715-718. My contribution to [37] is related to the spectroscopic measurements and discussion of results. I declare my contribution to be equal to 70 % 38. M.E. Plonska-Brzezinska, J. Mazurczyk, B. Palys, J. Breczko, A. Lapinski, A.T. Dubis, L. Echegoyen Vibrational spectroscopic study of carbon nano-onions coated with polyaniline. Physica Status Solidi C9, 5 (2012) 1210-1212. TC=1 My contribution to [37] is related to the spectroscopic measurements and discussion of results. I declare my contribution to be equal to 5 % 39. [H9] A.T. Dubis review paper Conformational Preferences of 2-Acylpyrroles in Light of FT-IR and DFT Studies. J. Phys. Chem. Biophys. 4 (2014) 155, doi:10.4172/2161-0398.1000155, open access I declare my contribution to be equal to 100% 20

CHAPTERS IN MONOGRAPHS 1. A.T. Dubis Wyniki analizy metoda FT-IR wybranych fragmentów polepy ze stanowiska 41 w paprotkach Kolonii, gm. Miłki, pow. Gizycko, rozdział w publikacji książkowej Osada z okresu wpływów rzymskich i okresu Wędrówek Ludów w Paprotkach Kolonii Stanowisko 41 w Karinie Wielkich Jezior mazurskich, tom 2 Analizy paleoekologiczne, Podlasko-Mazurska Pracownia Archeologiczna, Białystok 2002, str. 155-157. I declare my contribution to be equal to 100% 2. A.T. Dubis, J. Piekutin Metody eksperymentalne. rozdział w książce Metody badań naukowych z przykładami ich zastosowania pod redakcją Bazylego Poskrobki, Wydawnictwo Ekonomia i środowisko, Białystok, 2012, str.127-145. My contribution is related to the choice of chapter theme, discussion, preparation of the manuscript. I declare my contribution to be equal to 70% INTERNATIONAL AND NATIONAL CONFERENCES BEFORE BEING GRANTED THE PHD DEGREE 1. E. Dubis, E. Maliński, A.T. Dubis, J. Szafranek, J. Nawrot, J. Popławski (1988) Cuticular Hydrocarbons of Larvae of the Colorado Potato Beetle Leptinotarsa decemlineata Say. Proceedings of the International Conference Endocrinological Frontiers in Physiological Insect Ecology 1988, pp. 511-514, Szklarska Poręba, Poland 7-12 IX (poster) 2. E. Dubis., B. Lachowska, A.T. Dubis, J. Popławski, E. Hebanowska, E. Maliński, J. Szafranek, J. Nawrot (1988) A Comparison of the Composition of Surfice Lipids of the Colorado Beetle Leptinotarsa decemlineata Say to that of Potato Leaves Lipid Solanum tuberosum. Proceedings of the Conference, pp. 67-71, Symposium of the Institute of Plant Protection in Poznań, Poland, 10-12 II, (poster) 3. A.T. Dubis, B. Lachowska, E. Dubis, S. Witkowski, J. Popławski (1988) Charakterystyka składu lipidów wewnętrznych chrząszczy stonki ziemniaczanej Leptinotarsa decemlineata (Say). Materiały Zjazdowe, Zeszyt A, 180, Zjazd Naukowy Polskiego Towarzystwa Chemicznego i Stowarzyszenia Inżynierów i Techników Przemysłu Chemicznego, Łódź, 14-17 IX, (poster) 21

4. S. Witkowski, E. Dubis, B. Lachowska, A.T. Dubis, J. Popławski (1988) Wstępna analiza wydzieliny obronnej (reflex bleeding) stonki ziemniaczanej Leptinotarsa decemlineata (Say). Zjazd Naukowy Polskiego Towarzystwa Chemicznego i Stowarzyszenia Inżynierów i Techników Przemysłu Chemicznego, Łódź, 14-17 IX 1988, Materiały Zjazdowe, Zeszyt C, 69, 1988. (poster) 5. A.T. Dubis, B. Lachowska, L. Siergiejczyk, J. Popławski (1988) Synteza biologicznie czynnych izomerów i analogów asaronu 1,2,4-trimetoksy-5- propenylobenzenu. Zjazd Naukowy Polskiego Towarzystwa Chemicznego i Stowarzyszenia Inżynierów i Techników Przemysłu Chemicznego, Łódź, 14-17 IX, (poster) 6. A.T. Dubis, J. Morzycki, J. Popławski (1992) Redukcja trimetoksybenzenów metalami alkalicznymi w rozpuszczalnikach aerotycznych. Zjazd Naukowy Polskiego Towarzystwa Chemicznego i Stowarzyszenia Inżynierów i Techników Przemysłu Chemicznego, Białystok, 9-12 IX, (poster) 7. E.N. Dubis, A.T. Dubis, Z. Winiecki, J. Nawrot, J. Popławski (1993) Właściwości antyfidantne peraminy i jej pochodnych w stosunku do wybranych owadów - szkodników magazynowych. Konferencja Naukowa Jakość Badań w Toksykologii, Supraśl k. Białegostoku, 3-4.VI, (poster) 8. A.T. Dubis, B. Lachowska, B. Łozowicka, L. Siergiejczyk, J. Popławski (1994) Synteza izomerów asaronu, związków obniżających stężenie cholesterolu i triglicerydów we krwi. Jubileuszowe Sympozjum Chemii Organicznej PAN, Warszawa, 17-19.XI, (poster) 9. E. Dubis, A. Dubis, Z. Winiecki, J. Nawrot, J. Popławski (1994) Effects of the Endophyte-associated Alkaloids Peramine and its analogues an Selected Storage Pests. I International Conference on insects Chemical, Physiological an Environmental Aspects, Lądek Zdrój, 26-29.IX, (poster) 10. J. Popławski, B. Łozowcka, A.T. Dubis, B. Lachowska (1995) Synteza izomerów -asaronu o właściwościach hipolipemicznych. II Krakowska Konferencjia Chemii Leków Modelowanie cząsteczkowe w Chemii leków, Kraków, czerwiec, (poster) 11. A.T. Dubis, E.N. Dubis (1998) Comparative analysis of cuticular waxes of potato leaves using FT-IT reflection techniques and HPLC, II International Conference Vibrational Spectroscopy in Material Science, Kraków, Poland, 22-25 X, (poster) 12. E.N. Dubis, A.T. Dubis, J. Popławski (2000) Determination of the Aromatic Compounds in Plant Cuticular Waxes Using FT-IR Spectroscopy, III International Conference Vibrational Spectroscopy in Material Science, Kraków, Poland 23-26 IX, (poster) 22

13. A.T. Dubis, S.J. Grabowski, D.B. Romanowska, T. Misiaszek, J. Leszczyński (2002) Badania struktury kwasu pirolo-2-karboksylowego przy zastosowaniu spektroskopii w podczerwieni, obliczeń ab initio i teorii Badera, Szkoła Fizykochemii Organicznej "Nowe metody w spektroskopii molekularnej", Karpacz, 10-15.VI, (poster) 14. A.T. Dubis, S.J. Grabowski (2003) Analiza oddziaływań międzycząsteczkowych w dimerach -podstawionych piroli przy zastosowaniu spektroskopii w podczerwieni, obliczeń ab initio i teorii Badera, Szkoła Fizykochemii Organicznej Metody fizykochemiczne badania oddziaływań międzycząsteczkowych w układach biologicznych, Przesieka, 9-14.VI, (poster) 15. A.T. Dubis, S.J. Grabowski (2003) Experimental and theoretical study of cyclic dimers of 2-substituted pyrroles, VII th International Conference on Molecular Spectroscopy, Lądek Zdrój, 11-14.09, (poster) INTERNATIONAL AND NATIONAL CONFERENCES AFTER BEING GRANTED THE PHD DEGREE 16. A.T. Dubis, S.J. Grabowski (2004) Experimental and theoretical study on conformers of some 2-substituted 1Hpyrroles, XXVII European Congress on Molecular Spectroscopy, 5-10 September, Kraków. (oral presentation) 17. M. Palusiak, A.T. Dubis, S.J. Grabowski (2005) Intermolecular resonance Assisted Hydrogen Bonds in Crystals of 1H- and 1-Methylpyrrole-2carboxylic Acid Amide, Konwersatorium Krystalograficzne, Polish Crystallographic Meeting, Wrocław, 30.VI - 1 VII, (poster) 18. A.T. Dubis, S.J. Grabowski (2005) Spectroscopic and Theoretical study on hydrogen Bonded 2-substituted 1H-pyrroles, Structural Organic Chemistry, Central European School on Physical Organic Chemistry, Castle of Czocha, 19-24.06, (poster) 19. M. Palusiak, A.T. Dubis, S.J. Grabowski (2005) Interomolecular resonance assisted hydrogen bonds in crystals of pyrrole-2-carboxylic acid and its derivatives. XVI th International Conference Horizons in Hydrogen Bond research and Graduated School "Hydrogen Bonding and Hydrogen Transfer", Roskilde, Denmark, 29.VIII- 4.IX.2005. (poster) 20. A. Łapiński, A.T. Dubis (2007) FT-IR and Raman spectroscopic study, aided by quantum chemical DFT calculations of the peramine and its derivatives, 4th International Conference on Advanced Vibrational Spectroscopy ICAVS-4, Corfu, Greece, June 10-15, (poster) 23

21. A.T. Dubis, A. Łapiński (2007) Synthesies and spectroscopic study on peramine and its derivatives, 4th International Conference on Advanced Vibrational Spectroscopy ICAVS-4, Corfu, Greece, June 10-15, (poster) 22. A.T. Dubis, A. Łapiński (2008) Spectroscopic studies aided by quantum chemical DFT calculations of the peramine and its derivatives. Central European School on Physical Organic Chemistry, Structure and Properties of Organic Molecules, Karpacz, Poland, 08-12 June, (poster) 23. A. Łapiński, A.T. Dubis (2008) Zastosowanie zaawansowanych metod spektralnych w badaniach antyfidantu - peraminy jako alternatywnego środka ochrony roślin, XLVIII Sesja Naukowa Instytutu Ochrony Roślin, Poznań, 31 styczeń 1 lutego, (poster) 24. A.T. Dubis, A. Łapiński (2008) Własności fizykochemiczne peraminy i jej pochodnych jako alternatywnego środka ochrony roślin. XLVIII Sesja Naukowa Instytutu Ochrony Roślin, Poznań, 31 styczeń 1 lutego 2008, (poster) 25. A. Łapinski, A.T. Dubis, K. Pogorzelec-Glaser (2008) Struktura Krystaliczna i molekularna oraz badaniowa spektralne kryształu molekularnego utworzonego przez 1-(5-chloro-2-oksopentylo)pirolo-2-karboksylan 5-chloro-2-oksopentylu, XVI Ogólnopolska Konferencja Kryształy Molekularne 2008, Poznań-Błażejkowo, 8-12.IX, (komunikat ustny) 26. J. Łuszczyn, M.E. Plonska-Brzezinska, A.T. Dubis, K. Winkler, A. Palkar, A. Simionescu, D.T. Simionescu, L. Echegoyen (2009) Studies of biomolecular interactions in biosensor based on small Carbon Nano- Onions, Central European School on Physical Organic Chemistry - Weak Molecular Interactions, Przesieka, Poland, 2-6 June, (poster) 27. M.E. Płonska-Brzezinska, J. Luszczyn, A.T Dubis, A. Palkar, A. Simionescu, D.T. Simionescu, L. Echegoyen (2009) Studies of biomolecular interactions in biosensor based on small Carbon Nano- Onions, 6 th International Conference on Nanoscience and Nanotechnology, Thessaloniki, Greece, 13-15 July, (poster) 28. M.E. Płonska-Brzezinska, J. Luszczyn, A. Palkar, A.T Dubis, A. Simionescu, D.T. Simionescu, B. Kalska-Szostko, L. Echegoyen (2009) Non-cytotoxic small Carbon Nano-Onions - the first covalent functionalization with biomolecules, European Materials Research Society Meeting, Warsaw, Poland, 14-18 September, (poster) 29. A. Łapiński, M.E. Płońska-Brzezińska, A.T. Dubis, A. Villalta-Cerdas, K. Winkler, A.Z. Wilczewska, L. Echegoyen (2011) Badania Ramana modyfikowanych chemicznie nanocebulek węglowych, IV Poznańskie Seminarium Ramanowskie, Wydział Fizyki UAM, Poznań, 29 kwietnia (oral presenation) 24

30. A.T. Dubis, M.E. Płońska-Brzezińska, A. Łapiński, L. Echegoyen (2011) Zastosowanie spektroskopii oscylacyjnej w badaniach nanocebulek węglowych, XV Ogólnopolskie Sympozjum Zastosowanie Metod Spektroskopowych w badaniu materiałów i związków chemicznych, UAM, Poznań, 25-27 may, (poster) 31. A. Łapiński, M.E. Płońska-Brzezińska, A.T. Dubis, A. Villalta-Cerdas, K. Winkler, A.Z. Wilczewska, L. Echegoyen, Badania Ramana modyfikowanych chemicznie nanocebulek węglowych, XV Ogólnopolskie Sympozjum Zastosowanie Metod Spektroskopowych w badaniu materiałów i związków chemicznych, UAM, Poznań, 25-27 may (oral presentation) 32. A.T. Dubis, L. Siergiejczyk (2011) Badanie aromatyczności alfa-podstawionych piroli przy pomocy spektroskopii NMR, III Spotkanie użytkowników BRUKER, Poznań, 27-28.IX, (poster) 33. A.T. Dubis (2012) Zastosowanie metod spektroskopowych i teoretycznych w badaniu strukturalnych i magnetycznych aspektów aromatyczności 2-podstawionych piroli, 55 Zjazd PTChem i SiTPChem, Bialystok,16-20.IX, (oral presentation) 34. A. Jackowska, A.T. Dubis (2013) Spektroskopowe metody badania fałszerstwa bursztynu. II edycja Ogólnopolskiego Studenckiego Mikrosympozjum Chemików pt. Chemia przyszłość zaczyna się dziś pp.63, Białystok, 17-19 may, (poster) 35. P. Stasiewicz, A.T. Dubis (2013) Badanie międzycząsteczkowych wiązań wodorowych w 2-podstawionych piroli przy zastosowaniu spektroskopii w podczerwieni oraz obliczeń teoretycznych, II edycji Ogólnopolskiego Studenckiego Mikrosympozjum Chemików pt. Chemia przyszłość zaczyna się dziś pp. 92, Białystok, 17-19 may, (poster) 36. E. Jankowska, A.T. Dubis (2013) Zastosowanie obliczeń kwantowo-chemicznych w badaniach wiązań wodorowych, II edycji Ogólnopolskiego Studenckiego Mikrosympozjum Chemików pt. Chemia przyszłość zaczyna się dziś, pp. 64, Białystok, 17-19 may, (poster) 37. A.T. Dubis, P. Stasiewicz, A. Łapiński, K. Pogorzelec-Glaser (2013) Czy w 2-podstawionych pirolach mogą tworzyć się wewnątrzcząsteczkowe wiązania wodorowe? III Konferencja Związki Biologicznie czynne aktywność, struktura, synteza Białystok, 4-6.X, (oral presentation) 17. LECTURES DELIVERED IN BIALYSTOK UNIVERSITY Reflection Techniques in FTIR Spectroscopy, 2006-2008 Molecular Spectroscopy, lecture 30 hours; since 2007 25

18. INVITED LECTURES Analiza konformacyjna alfa-postawionych piroli przy zastosowaniu metod spektroskopowych, Katedra Krystalografii i Krystalochemii Uniwersytetu Łódzkiego, 16 maj 2006 Analiza konformacyjna alfa-postawionych piroli przy zastosowaniu metod spektroskopowych, Pracowni Krystalochemii Wydziału Chemii UW, styczeń 2007 Czy 2-acylopirole mogą tworzyć wewnątrzcząsteczkowe wiązania wodorowe? wykład na zaproszenie Katedry Chemii Teoretycznej i Strukturalnej Uniwersytetu Łódzkiego, 24 październik 2014 19. TRAINING, SCHOOL Participation in the Central European School on Physical Organic Chemistry, Faculty of Chemistry, University of Wrocław, Section of Physical Organic Cehmistry of the Polish Chemical Society Participation in the Raman Seminar on the occasion of the 80-th anniversary of the discovery of the Raman effect. Faculty of Physics, University im. Adama Mickiewicza in Poznan, 18.VI.2008 r. Participation in the Seminar Nowoczesne techniki spektroskopii ramanowskiej: mapowanie i wzmocnienie powierzchniowe (SERS) Faculty of Chemistry, Jagiellonian Univesrity, Kraków, 29.VI.2009 r. Participation in the Spectroscopic Seminar: Zastosowanie metod spektroskopowych w badaniu materiałów i związków chemicznych, Poznań, 2011 20. EXPERTISE (in Polish) Analiza chemiczna polepy w ramach zadania Dolina Węgorapy 2008-2010 (Analiza próbek z Kalu, Wysieczy i Stulichów metodą spektroskopii w podczerwieni FTIR)- ekspertyza wykonana na zlecenie Muzeum Ziemi w Węgorzewie obejmująca analizę spektroskopową IR i Ramana próbek zabytkowych. Badania fizykochemiczne ceramiki zabytkowej z doliny Węgorapy w ramach zadania Dolina Węgorapy 2008-2010. Analiza próbek z Kalu, Wysieczy, Stulichów metoda spektroskopii w podczerwieni FTIR/ATR oraz spektroskopii Ramana, ekspertyza wykonana na zlecenie Muzeum Ziemi w Węgorzewie dotycząca analizy próbek zabytkowej ceramiki ze stanowisk w Kalu, Stulichach i Wysieczy. Analiza zawartości mgieł olejowych na stawiskach pracy metodą spektroskopii w podczerwieni, analizy wykonywane na potrzeby lokalnego otoczenia gospodarczego (ERGONOMIA, ECOCHEM) 2000-2010. 26

Analiza zaniku grup NCO w laminatach foliowych, analizy wykonywane na potrzeby lokalnego otoczenia gospodarczego (PAKPOL, MARPOL ERGONOMIA, EKOCHEM) 2000-2010. 21. PATENTS J. Popławski, A. Dubis, B. Lachowska, B. Łozowicka, J. Cybulski, Z. Chilmończyk, W. Szelejewski, G. Grynkiewicz, 31.03.2005, PL 188701 B1 Sposób otrzymywania /E/-1,2,3-trimetoksy-5-/1 -propenylo/benzenu J. Popławski, A. Dubis, B. Lachowska, B. Łozowicka, J. Cybulski, Z. Chilmończyk, W. Szelejewski, S.Witkowski, 31.03.2005, PL 188702 B1 Sposób otrzymywania /E/-1,2,5-trimetoksy-3-/1 -propenylo/benzenu J. Popławski, A. Dubis, B. Lachowska, B. Łozowicka, J. Cybulski, Z. Chilmończyk, W. Szelejewski, S.Witkowski, 31.03.2005, PL 188703 B1 Sposób otrzymywania /E/-1,2,4-trimetoksy-3-/1 -propenylo/benzenu J. Popławski, A. Dubis, B. Lachowska, B. Łozowicka, K. Kita, S. Kobes, Z. Chilmończyk, J. Cybulski, S. Adamski, 31.10.2006, PL 192464 B1 Nowe pochodne -asaronu i zawierające je środki farmaceutyczne o działaniu hipolipemicznym 27

SELECTED RESEARCH PAPERS FORMING A HABILITATION THESIS [H1]. S.J. Grabowski, A.T. Dubis, D. Martynowski, M. Główka, M. Palusiak, J. Leszczynski Crystal and Molecular Structure of Pyrrole-2-carboxylic Acid; -Electron Delocalization of Its Dimers - DFT and MP2 Calculations. J. Phys. Chem. A, 108 (2004) 5815-5822. IF = 2.639 TC =21 My contribution to [H1] is related to formulate a research problem, the synthesis of a sample, theoretical calculation, interpretation and discussion of results, preparation of the manuscript I declare my contribution to be equal to 35% [H2]. S.J. Grabowski, A.T. Dubis, M. Palusiak, J. Leszczynski Heteronuclear Intermolecular Resonance-Assisted Hydrogen Bonds. The Structure of Pyrrole-2-Carboxamide (PyCa). J. Phys. Chem. B, 110 (2006) 5875-5882. IF = 4.115 TC =17 My contribution to [H2] is related to formulate a research problem, the synthesis of a sample, theoretical calculation, interpretation and discussion of results, preparation of the manuscript I declare my contribution to be equal to 40 % [H3] E. Bilewicz, A.J. Rybarczyk-Pirek, A.T. Dubis, S.J. Grabowski Halogen bonding in crystal structure of 1-methylpyrrol-2-yl trichloromethyl ketone. J. Mol. Struct. 829 (2007) 208-211. IF = 1.486 TC =30 My contribution to [H3] is related to formulate a research problem, the synthesis of a sample, theoretical calculation, interpretation and discussion of results, preparation of the manuscript I declare my contribution to be equal to 35 % [H4] S.J. Grabowski, M. Palusiak, A.T. Dubis, A. Pfitzner, M. Zabel Inter- and intramolecular hydrogen bonds Structures of 1-methylpyrrole-2-carboxamide and 1-hydroxypyrrole-2-carboxamide. J. Mol. Struct., 844-845 (2007) 173-180. IF =1.486 TC =5 28

My contribution to [H4] is related to formulate a research problem, the synthesis of a sample, spectroscopic analysis, theoretical calculation, interpretation and discussion of results, preparation of the manuscript I declare my contribution to be equal to 40 % [H5] A.T. Dubis, A. Łapiński Spectroscopic and theoretical study on peramine and some pyrrolopyrazzinone compounds. Vib. Spec. 49 (2009) 265-273. IF = 1.936 TC =2 My contribution to [H5] is related to formulate a research problem, the synthesis of a sample, theoretical calculation, interpretation and discussion of results, preparation of the manuscript I declare my contribution to be equal to 90 % [H6] A. Łapiński, A.T. Dubis A DFT/TD-DFT study for the ground and excited states of peramine and some pyrrolopyrazinone compounds. J. Phys. Org. Chem. 22 (2009) 1058-1064. IF = 1.602 TC =1 My contribution to [H6] is related to formulate a research problem, the synthesis of a sample, theoretical calculation, interpretation and discussion of results, preparation of the manuscript I declare my contribution to be equal to 85 % [H7] A.T. Dubis, M. Domagała, S.J. Grabowski Spectroscopic and Theoretical Studies on Some New pyrrol-2-yl-chloromethyl ketones. New J. Chem. 34 (2010) 556-566. IF = 2.631 TC =3 My contribution to [H7] is related to formulate a research problem, the synthesis of a sample, theoretical calculation, interpretation and discussion of results, preparation of the manuscript I declare my contribution to be equal to 60 % [H8] A.T. Dubis, S. Wojtulewski, K. Filipkowski Spectroscopic and Theoretical Studies on the Aromaticity of Pyrrol-2-yl-carbonyl Conformers. J. Mol. Struct. 1041 (2013) 92-99. IF = 1.404 TC =1 29

My contribution to [H8] is related to formulate a research problem, theoretical calculation, NICS, HOMA indices calculation, spectroscopic measurements, interpretation and discussion of results, preparation of the manuscript, corresponding author I declare my contribution to be equal to 80 % [H9] A.T. Dubis, Conformational Preferences of 2-Acylpyrroles in Light of FT-IR and DFT Studies. (Review article) J. Phys. Chem. Biophys. 4, (2014) 155, doi:10.4172/2161-0398.1000155, open access I declare my contribution to be equal to 100%. The Aggregate Impact factor for H1-H9 amounts to IF=17.29 30

PRINCIPAL ACHIEVEMENTS IN RESEARCH PAPERS H1-H9 PRESENTED FOR HABILITATION Scientific achievements as defined in the Act of 1 September 2011 (Journal of Laws No 1165) and of 22 September 2011 (Journal of Laws No 201, Item 1200) are presented under the title: INFLUENCE OF THE CONFORMATION OF 2-ACYLPYRROLES ON SPECTROSCOPIC PROPERTIES AND HYDROGEN BOND FORMATION 1. Introduction, aims and objectives The shape of molecules and corresponding charge distribution determine molecular properties and interactions between molecules. They also enable a better understanding of the phenomenon of self-organisation and complementarity of shapes of a molecule and its pharmacological activity. Molecules with the same general formula and the same connectivity differing only in the spatial orientation of their constituent atoms are called stereoisomers. Among them are conformational isomers called conformers which become identical after simple rotation about a single bond. Internal factors, such as the spherical effect of local dipole moments, intramolecular hydrogen bonds, and external factors, such as self-association, association with solvent and lattice structure in the solid phase, determine conformations of the molecule. The influence of 2-substitution on different aspects of the conformational stability of these molecular systems and the usefulness of infrared spectroscopy supported by theoretical calculations in H-bonds and conformational studies were discussed. The purpose of my studies was to investigate the effect of conformation and substituents on the spectroscopic properties of preferred conformers of 2-acylpyrroles (Fig. 1) and hydrogen bonding formation within these systems. The subject of my research are 2-acylpyrroles. This class of compounds is characterised by partial conformational freedom of the carbonyl group. It is noted that the 2- acylpyrrole motif is widespread in nature. 1 31

a) syn b) anti Figure 1. Syn (a) and anti (b) 2-acylpyrroles; R= H, OH, OCH3, NH2, NHCH3, N(CH3)2, CH2Cl, CHCl2, CCl3; R1=H, or CH3. There are many examples of 2-acylpyrrole-based molecules with various biological activities. Representative examples are pyoluteorin and pyrrolomycin D, 2,3 secondary metabolites produced by marine bacteria. There are also certain marine natural products such as nakamuric acid, 4 marinopyrroles 5 or storniamide 6 which show potent activity as inhibitors of multidrug resistance (MDR) 7 (Fig. 2). Among the bioactive compounds of marine origin there are structures based on the 2-acylpyrrole motif such as distamycin and netropsine. 8 Due to promising antibacterial properties, these novel marinopyrroles have recently attracted considerable attention. It is helpful to know the structure of small structural elements which build large molecules of natural origin. 9,10,11,12 32

Figure 2. 2-Acylpyrroles from natural products. The thesis includes H1-H9 scientific papers which present my recent investigations of 2- acylpyrrole conformations, with a particular emphasis on hydrogen bonds being formed within these systems. The influence of 2-substitution on different aspects of stability of these molecular systems and the usefulness of infrared spectroscopy supported by theoretical calculations in H-bonds and conformational studies were presented. [H9] presents an overview of the last decades of investigation into the molecular structure and conformational properties of 2-acylpyrroles. My results are presented on the background of other research. I reviewed relevant literature and included references up to 2014. The central part of the review is discussion of the usefulness of IR spectroscopy as a source of information on molecular spatial structure, and usefulness of theoretical methods to study hydrogen bond interactions. Results based on the structural information collected in the Cambridge Structural Database (CSD) are also presented. 13,14 Structures of 2-acyl compounds were determined by FT-IR, Raman, 1 H, 13 C NMR spectroscopy. Crystal structures were determined by X-ray studies. Theoretical calculations 33

were carried out using Gaussian 15 in the Warsaw Supercomputer Centre (ICM) (G53-7). For the analysed systems the Bader Atoms in Molecules (AIM) theory was applied to find bond and ring critical points and to analyze them in terms of electron densities and their Laplacians. The AIM calculations were carried out using QTAIM. 16 Vibrational spectroscopy is a convenient method to study the conformation of molecules and hydrogen bonds in solution. It can be used in a wide range of concentrations and in solvents of different polarity, which enables observation of conformational changes. The experimental results were correlated with the results obtained using theoretical calculations. Owing to the theoretical calculations of various molecular properties, such as the geometry and spectroscopic properties of individual molecules, the interpretation of the results of the experimental measurements was supported by theory. These studies involved co-operation with Professor Sławomir Grabowski, Faculty of Chemistry, University of the Basque Country, Spain and Prof. J. Leszczynski, Computational Center for Molecular Structure and Interactions, Department of Chemistry, Jackson State University, USA, in the field of theoretical chemistry, and with Dr Marcin Palusiak s group, Department of Crystallography and Crystal Chemistry, University of Łódź, and with Dr Andrzej Łapiński, Institute of Molecular Physics PAS, Poznań. 2. Acknowledgments I wish to express my gratitude to Professor Sławomir Grabowski and Professor Jerzy Leszczynski for showing me the potential of computational methods. I would like to thank Professor Jacek Morzycki for supporting me in the course of my research. The experimental work in the crystallographic studies would not have been possible without the support of crystallographers. I am very grateful to Professor Dr Marek Główka, Dr Dariusz Martynowski, Professor Arno Pfitzner, Dr Manfred Zabel, Dr Marcin Palusiak, Dr Agnieszka Rybarczyk-Pirek and Dr Małgorzata Domagała. Many thanks to Dr Andrzej Łapiński for our fruitful cooperation. 3. Conformation of 2-acylpyrroles Rotation about the C1-C6 bond is possible in 2-acylpyrroles, and there are two stable rotamers of pyrrole-2-yl carbonyl moieties depicted in Figure 1a and 1b. According to the 34

IUPAC terminology rotamers are a set of conformers formed by restricted rotation about a single σ bond. 17 The rotational barrier is the activation energy required to convert one rotamer to another. In the rotation of groups about a bond, the potential energy barrier between two adjacent minima of a molecular entity is a function of the torsion angle. 18,19 If the energy needed to convert one conformer into the other is sufficiently small, free rotation about a single bond may occur and conformers are indistinguishable. At any point in time the majority of particles may be in a more stable conformation and the conformers are in equilibrium. Conformational analysis began to develop since the 1950s when Sir Derek H.R. Barton formed basic principles of conformational analysis, and described the chemical consequences of the spatial structure of molecules. 20,21 These studies led to the explanation of chemical reactions by determining spatial relations between reactive centres and provided the basis for the understanding of reaction mechanisms. Since the adoption of the concept of conformational and rotational isomerism a number of physical methods useful in the study of rotameric equilibria were developed. Among these methods structural methods, spectroscopic methods 22 and theoretical methods can be noted. Because the spatial arrangement of atoms in molecules has a profound effect on the biological function of compounds and the rate of many chemical reactions, 23 conformational studies of molecules are still of interest to researchers. 24,25,26 My studies were related to the analysis of conformation of 2-acylpyrroles (Fig. 1a,b) and analysis of the influence of conformation on spectroscopic properties and the formation of hydrogen bonds within these molecules. The liquid and the solid phase were taken into account in conformational analysis. NMR spectra were recorded in the liquid state only. Using different methods of conformational analysis I took into account the fact that the conformation of molecules in the solid state does not necessarily have to be the same as in the liquid state. The results of spectroscopic analysis were correlated with theoretical calculation results. 15 4. Computational chemistry in conformational analysis Theoretical calculations have developed into an important tool in conformational research and they can greatly support experimental work with quite good accuracy. 27 The use of computational methods in studies of interest to chemists has been made possible by the rapid 35

growth in computing power over the last several decades. Among the most useful parameters are geometrical parameters, relative conformational energies, barriers to internal rotation, 28 vibrational frequencies and chemical shifts for each conformer. Theory shows that the determination of molecular structure involves determination of the geometry of the molecule at the minimum of the potential energy surface. If the barrier is lower than 12-15 kcal/mol, it is accepted that the molecule is not stable at room temperature from the kinetic point of view. Therefore, designation of the geometry of a stable structure requires knowledge of minimum energy values and the rotational barrier for the possible forms. For many molecules the stable form is determined on the basis of existing knowledge. 29,30 The conformation of new structures is determined from energy minima and rotational barriers. In my study of the spectroscopic properties of acylpyrroles a computational method was used. The most stable structure and its vibrational frequencies were predicted. 5. Discussion on the most significant achievements My research focuses on the interdependence between the conformation of 2-acylpyrroles and their spectroscopic properties and hydrogen bond formation. Using theoretical calculations, preferred conformations of 2-acylpyrroles were first found [H1, H2, H4, H7, H8]. I paid particular attention to the energy difference between rotational isomers and the energy barrier separating the syn and anti form [H1, H7, H8, H9]. The next step of the studies was to identify the conformational form of 2-acylpyrroles experimentally. Using vibrational spectroscopy the syn and anti form in the liquid phase was found [H2, H7, H8]. The role of the substituent R group in inter- and intramolecular hydrogen bond formation was analysed [H7, H8, H9]. The stabilisation of the most stable syn form in the liquid state depending on HB formation was also considered. The influence of 2-substitution on different aspects of aromaticity and stability of pyrrol-2-yl carbonyl conformers was also discussed. The vibrational frequency of the pyrrole ring as a measure of p-electron delocalisation over the pyrrole ring was proposed [H8]. The energy of the same rotameric forms of 2-acylpyrroles was calculated [H8, H9]. The calculations show that for all the species studied syn and anti are the most stable forms. The carbonyl group and the N-H or N-CH3 groups are located on the same side of the C2-C6 bond 36

for the syn form. In the anti form the carbonyl group and N-H groups are located on the opposite sides of the C2-C6 bond. In other words, syn and anti rotamers differ in the spatial arrangement of the carbonyl group in respect to the N-H group. Theoretical calculations show that the carbonyl group is in general in the same plane as the pyrrole ring [H8]. For some conformers, such as pyrrole-2-carboxylic acid amide, the carbonyl group is twisted in respect to the pyrrole ring plane. The twist is 1.76 for the syn form and 18.7 for the anti conformer. The anti form of the moiety with N1-CH3 shows the O-C6-C2-N dihedral angle deviating from 0 in a range of 20.6 to 52.3, whereas for syn rotamers (R-NH2, NHCH3 and N,Ndimethylamide) the dihedral angle O-C6-C2-N twist is in a range from 9.2 to 28.5 [H8]. The theoretical calculations (B3LYP/6-311G(d,p)) reveal that the syn forms are more stable than the anti form. The energy difference ΔEsyn/anti=Esyn-Eanti is in a range of 1.06-8.04 kcal/mol (Table 1). For R1=H ΔEsyn/anti is in a range from 1.06 to 5.15; for R1=CH3 ΔEsyn/anti is from 1.93 to 8.08 kcal/mol. For R1=H ΔEsyn/anti is greater than ΔEsyn/anti for R1=CH3 [H8, H9]. The potential energy curve obtained for 2-acylpyrroles was presented in [H9]. The torsional curves computed at B3LYP levels of theory are shown in Figure 3. The angle of rotation is zero for the syn conformation; for the angle of rotation of 180 there is the anti conformer. Fig. 3. Energy profile of pyrrol-2-yl carbaldehyde (2) The most stable syn and anti conformers differ in the spatial arrangement of the carbonyl group about the inverting single bond (C-C). The calculations were performed at the B3LYP/6-311++G(d,p) level of theory. 360 scanning of the dihedral angle N-C-C=O was performed with an increment of 5 [H9]. For all the 2-acylpyrroles studied, the rotational profile obtained from theoretical calculations has a similar shape. The profiles show the existence of two energetically most 37

stable syn and anti conformers corresponding to a value of close to 0 and 180. The highest energies correspond to angles of rotation of 90 and 270 and may be considered transition states for transformation reactions of conformations from the syn to the anti form. The barriers for those reactions vary from 7.03 to 16.21 kcal/mol depending on the R substituent group. Barriers for antisyn reactions vary from 2.07 to 11.94 kcal/mol (Table 1). Barriers for a E syn anti a and E anti syn transformation for pyrrole moieties (R1=CH3) are in a range of 8.98-15.26 kcal/mol and 4.48-11.79 kcal/mol, respectively. These calculations reveal that the syn form is relatively more stable than its anti counterpart. This phenomenon is most likely related to more effective -electron delocalisation in the syn conformation. In the case of 2-acylpyrroles the carbonyl group is directly coupled with the aromatic ring. The rotation of the C=O group has a role in changing the nature of the C2-C6 bond which connects the carbonyl group with the pyrrole ring. The theoretically calculated geometrical parameters of 2-acylpyrroles reveal elongation of the C2-C6 bond for anti compared with syn conformers. These observations suggest relatively larger stability of the syn conformer than its anti counterpart. It can be stated that this phenomenon results from more effective - electron delocalisation. 31 This was very clearly shown by HOMA 32,33,34 and NICS 35 aromaticity indices [H8]. The HOMA index represents aromaticity in relation to the optimal bond length concept. This index is expressed by the following equation: HOMA = 1 α n (R opt R i ) 2 = 1 [α(r opt R ave ) 2 + α n (R ave R i ) 2 ] HOMA = 1 EN GEO where: EN = α(r opt R av ) 2 and GEO = n (R av R i ) 2 for heterocyclic -electron systems the expression for the HOMA term is as follows: 257.7 HOMA1 257.7 1.388 rav rav ri N where N is the number of bonds taken into consideration, rav is averaged bond length n 2 2 R av = 1 R n i=1 i, and Ri is the virtual bond length calculated from the Pauling bond number 33 n i= exp R(1) R(n) c according to the general formula: 38

R n= 1,467 0,1702ln (n) On the basis of these observations, I assumed that the electron donating substituents slightly destabilise the ring system, affecting the aromaticity of the pyrrole ring (Figure 4b). The observed effect may be interpreted as follows: the substitution of the pyrrole ring in position 2 by a group such as -NH2 leads to substantial double bond localisation in the pyrrole ring and in consequence to a decrease in its aromaticity. In the case of chloro derivatives an increase in aromaticity is mainly due to the EN contribution to HOMA. These findings are consistent as the elongation of bond length requires more energy than alternation. 36 a b Figure 4. a) Plot of changes of the HOMA index of syn and anti 2-acylpyrroles in respect to the R group; b) plot of changes of the HOMA index and calculated ring frequencies C=C [H8]. It was suggested based on these observations that the frequency of ring quadrant stretching ring is related with the aromaticity of five-membered rings as measured by HOMA [H8]. For more aromatic moieties lower frequencies ring of ring stretching vibration are observed. There is a correlation with HOMA aromaticity and the frequency of stretching vibration C=C of the pyrrole ring of syn and anti 2-acylpyrrole conformers. This effect is illustrated in Figure 5. 39

Figure 5. Quadrant ring stretching vibration C=C of the pyrrole ring for syn and anti 2-acylpyrroles in relation to the substituent group R [H8]. IR spectroscopy was applied to investigate conformational preferences of 2-acylpyrroles and N-methyl-2-acylpyrroles. It was observed experimentally that the 2-acylpyrroles studied form essentially doublet carbonyl bands. [H7]. The components of the doublet were assigned as being the characteristic absorptions of the syn and anti forms. The spatially crowded pyrrole-2-carboxylic acid amide (R=NHCH3, N(CH3)2) and chloro derivatives of 2-acylpyrroles (R= CCl3) only show a single carbonyl band [H7]. The band asymmetry phenomenon was explained by the formation of the associated forms. The formation of associated forms was the cause of the asymmetry of the low frequency carbonyl band (Fig. 6a). It is noted that 2-substituted pyrroles tend to form dimeric hydrogen bonded species for steric reasons. a b Figure 6. IR spectra of 2-acylpyrroles in diluted cyclohexane solution: a) R1= H; R2= CCl3, CHCl2, CH2Cl; b) R1=CH3; R2= CCl3, CHCl2, CH2Cl [H7]. 40

My attempts focused on the investigation of hydrogen bond formation using theoretical calculations. The experimental C=O frequencies of syn, anti and dimeric forms were compared with the theoretically predicted carbonyl frequencies. Having considered these calculations, it was observed that anti and syn conformers are present in diluted solution whereas the dimeric form occurs in concentrated solution [H1, H7]. Figure 7. 2-acylpyrrole cyclic dimers; R1= H; R2= OH, CH2Cl, CHCl2, CCl3 [H7]. The formation of associated forms gives rise to an additional C=O and N-H band (Figure 8a, b). a B Figure 8. IR spectra of 2-acylpyrrole in cyclohexane: a) R1= H; R2= CCl3, b) R1=H; R2= CCl3, CHCl2, CH2Cl [H7]. These bands are slightly red-shifted compared with the C=O(C=O) and N-H (N-H) band of the free syn or anti form. This phenomenon is a result of weakening of the dimer C=O and N-H bond in comparison with the monomer. Theoretical calculations also predict that the C=O band of the anti form lies at a higher frequency than the C=O band of the syn form (Table 1). 41

Table 1. Experimental and calculated (B3LYP/6-311++(d,p)) IR stretching frequencies of C=O, C=C and N-H groups for syn and anti pyrrol-2-yl carbonyl conformers (1-38). E is the difference between the energies of the syn and anti form. HOMA 49 is the aromaticity index. syn anti Eksperymentalne i obliczone częstości IR [cm -1 ] Eksp. Obl.* Eksp. Obl. Eksp. Obl. N-H N-H C=O C=O C=C C=C E syn/anti [kcal/mol] Bariera rotacyjna E syn anti [kcal/mol] Bariera rotacyjna E anti syn [kcal/mol] Indeks HOMA [a.u.] 1 pirol 3485 3600 1532 b 1540 0.854 2 R1=H, R=H 3460 3565 1659 d 1685 1552 b 1553 0.923 3.84 16.21 10.38 3 R1=H, R=H 3576 1712 1554 0.911 4 R1=H, R=OH 3465 3570 1671 d 1723 1555 c 1556 11.3 0.921 1.11 11.94 5 R1=H, R=OH 3587 1754 1561 (13.03) 0.916 3465 b 6 R1=H, R=OCH3 3571 1719 d 1554 1705 b 3480 b 1557 c 1555 1.06 12.35 0.917 11.36 (1.10) (12.43) 7 R1=H, R=OCH3 3589 1701 d 1736 1559 0.913 1644 c 8 R1=H, R=NH2 3415 3559 1603 c 1683 1552 c 1550 0.917 3336 c 4.91 10.17 6.60 9 R1=H, R=NH2 3582 1710 1560 0.882 10 R1=H, R=NHCH3 3571 1665 1558 0.914 6.02 8.75 3.33 11 R1=H, R=NHCH3 3581 1690 1548 0.875 1548 12 R1=H, R=N(CH3)2 3552 1612 1623 a 3450 1547 b 1544 0.911 4.95 7.03 2.07 13 R1=H, R2=N(CH3)2 3587 1697 1560 0.886 14 R1=H, R=CH2Cl 3562 1662 d 1546 1695 a 3458 d 1545 c 1546 0.923 5.15( 3.1) 13.9 10.8 15 R1=H, R=CH2Cl 3594 1683 d 1714 1552 0.899 16 R1=H, R=CHCl2 3561 1658 d 1543 1692 a 3457 d 1542 c 1546 0.927 4.19 (2.4) 13.89 11.45 17 R1=H, R=CHCl2 3562 1683 d 1710 1548 0.921 1538 a 18 R1=H, R=CCl3 3563 1670 b 1687 1539 b 1542 0.926 3456 d 1538 c 2.55 12.85 10,33 (2.51) 1679 d 19 R1=H, R=CCl3 3583 1681 b 1708 1545 0.922 20 metylopirol 1517 0.865 21 R1=CH3, R=H 1689 1528 b 1531 4.52 15.31 0.914 1670 11.79 22 R1= CH3, R=H 1694 1533 (3.47) (15.26) 0.906 1530 d 23 R1= CH3, R2=OH 1670 1724 1536 b 1531 0.913 1.93 11.19 9.25 24 R1= CH3, R=OH 1734 1536 0.904 25 R1= CH3, R=OCH3 1706 1532 b 1531 0.910 1711 1.93 10.37 8.44 26 R1= CH3, R=OCH3 1717 1537 0.902 27 R1= CH3, R=NH2 1683 1529 c 1528 0.908 1639 c 5.54 9.67 7.21 28 R1= CH3, R=NH2 1695 1537 0.882 29 R1= CH3, R=NHCH3 1667 1537 0.908 4.32 8.98 4.65 30 R1= CH3, R=NHCH3 1677 1542 0.877 1537 31 R1= CH3, R=N(CH3)2 1635 a 1630 d 1536 b 1529 0.902 3.85 13.51 32 R1= CH3, R=N(CH3)2 1660 1542 0.870 33 R1= CH3, R=CH2Cl 1662 d 1527 a 1694 1526 1522 c 6.61 0.909 12.74 6.67 (5.68) 34 R1= CH3, R=CH2Cl 1683 d 1693 1527 0.886 35 R1= CH3, R=CHCl2 1658 d 1526 a 1692 1524 c 1526 6.64 0.905 13.5 6.59 (6.97) 36 R1= CH3, R=CHCl2 1683 d 1690 1527 0.889 1524 37 R1= CH3, R=CCl3 1690 a 1522 c 1526 8.08 0.899 11.62 4.48 (8.04) 38 R1= CH3, R=CCl3 1681 d 1692 1522 0.878 42

FSD-IR 37,38 was used to extract individual components from the overlapping band of N-H and C=O groups. In such a way separation of the overlapping bands was achieved. FSD-IR was applied for separating anti, syn and dimeric overlapping carbonyl bands [H7]. Figure 9. IR spectra of 2-acylpyrroles in cyclohexane: a) R1= H; R2= CCl3, b) R1=H; R2= CCl3, CHCl2, CH2Cl [H7]. It was found that conformational equilibrium depends on solvent polarity. 39 When two conformers have very different dipole moments, spectra in solvents with different polarities reveal variable intensities of some bands. It is noted that a method is available to discern the anti and the syn rotamer. 40 The greater stability of the syn form over the anti rotamer for 2- acylpyrroles in a low polarity solvent is due to its lower dipole moment. The moments for the pyrrole ring and the oxo group are mutually opposing which leads to much lower dipole moments of the syn forms [H2]. Infrared spectra of N-methyl 2-acylchloropyrroles were investigated in acetonitrile and nonpolar solution to distinguish the anti and the syn rotamers [H7]. In such a way the AsynC=O/AantiC=O ratio of integrated intensities of the carbonyl bands was determined. The ratio is lower in acetonitrile solution compared to the value in less polar cyclohexane solution. It means that the population of the more polar anti form increases with higher solvent polarity (Figure 9) [H7]. Further confirmation of dimer formation was provided by comparing experimental and theoretically calculated spectra of monomers (the syn and anti form) and the dimeric form [H2]. 43

Conformational studies of 2-acylpyrroles using NMR spectroscopy provided qualitative or quantitative data. On the basis of the chemical shift values the conformers were identified. The 1 H and 13 C NMR spectra of pyrrole-2-carboxamide enable observation of syn and anti conformers in DMSO solution. (Figure 10). The population of the molecular forms was calculated from the integral intensity of the carbon signals in the 13 C NMR spectra. The syn/anti ratio calculated from 13 C NMR on the basis of 1H decoupling without NOE 41 is 3:1 [H2]. Figure 10. 13 C NMR spectra of pyrrole-2-carboxylic acid amide in DMSO solution [H2]. It was calculated from the integral intensity of the carbon signals in the 13 C NMR spectra. The syn/anti ratio calculated from 13 C NMR on the basis of 1H decoupling without NOE is 3:1 [H2]. The chemical shift is a useful tool for measuring ring currents and studying aromaticity of the systems under investigation. For the s-cis form of 2-acylpyrroles, a correlation between the 1 H chemical shift of the H5 pyrrole proton (H5) and the GEO part of the HOMA index was found (Figure 11). The π-electron delocalisation of the homogenous set of 2- acylpyrrole conformers was evaluated on the basis of 5-H chemical shifts [H8]. 44

Figure 11. Plot of HOMA vs. d5-h chemical shift for the syn conformers of 2-substituted pyrroles. 2-acylpyrroles were studied by experimental FT-IR as well as by theoretical methods. The theoretical calculations of vibrational frequencies performed at different levels of approximation showed that centrosymmetric dimers (Fig. 8) formed with two equivalent N H O bonds between the N H pyrrole ring and the C=O carboxyl group [H7, H9]. The strength of the hydrogen bond within cyclic dimers was determined by theoretical calculations (Table 2). Binding energies for the analysed complexes were computed as a difference between the total energy of the complex and the energies of the isolated monomers and corrected for the basis set superposition error (BSSE) via the standard counterpoise method (counterpoise correction) of Boys and Bernardi. 42 45

Table 2. Binding energies, C=O modes and topological parameters of HB interactions for the 2-acylpyrrole dimers, B3LYP/6-311++G(d,p) and B3LYP/6-311+G* (in parentheses). R=H R=OH R=CH3 R=CH2Cl R=CHCl2 R=CCl3 C=O [cm -1 ] (mon-dim) 17 21 34 28 25 EHB [kcal/mol] -6.9-5.9 (-5.7) -6.23-6.45-6.63-5.26 BCP [au] 0.030 0.0263 0.0283 0.0278 0.0282 0.0261 RCP [au] 0.0029 0.0030 0.0036 0.0036 0.0036 H8 H9 H7 H7 H7 H7 R=NH2 R1=H R=NH2 R1=CH3 R=NHCH3 R=OH syn anti syn anti syn anti syn anti C=O (mon-dim) 10 25 7.5 15.6 (53) (60) EHB [kcal/mol] -6.78-6.96-6.01-6.57-6.98-7.61-8.0 (-7.7) -8.5 (-8.3) BCP [au] 0.0303 0.0298 0.0314 0.0483 0.0498 0.0081 0.0081 RCP [au] 0.0048 0.0049 0.0058 (0.008) (0.008) H2 H2 H9 H9 H9 H9 H1 H1 The crystal structure of pyrrole-2-carboxylic acid determined by single-crystal X-ray diffraction proved that both C=O H-O and N-H O=C syn dimers exist in the solid state. Pyrrole- 2-carboxylic acid has a proton donating and proton accepting COOH carboxylic group as well as an N H proton donating bond. All of them are involved in hydrogen bond interactions in the crystal structure of the acid where centrosymmetric dimers are formed with two equivalent O H... O bonds between the carboxylic groups and N H... O bonds between the N H 46

pyrrole ring and the C=O carbonyl group. Hence, according to Etter grafs designations, the following H-bond motifs were detected in the crystal structure of the acid: R2 2 (8) and R2 2 (10) (Figure 13). The energy of the H-bonds within the dimers (C=O H-N) has a medium value (Table 2): 5.9 kcal/mol. The H O contact length is 1.880 Å, and the N-H O angle is 163.7. According to theoretical calculations dimers linked through two C=O H-O bonds are more stable than those formed through N-H O=C bonds. Additionally, the O-H O bond within the dimer formed by the syn conformers is stronger than the O-H O bond within the anti dimer. EHB is 8.0 kcal/mol. The H O contact length is 1.661 Å and the O-H O angle is 179.7 [H1, H9]. B3LYP/6-311++G** calculations for centrosymmetric dimers of both N-methylpyrrole-2-carboxamide conformers connected through equivalent N H... O hydrogen bonds were carried out. The calculated binding energy of the N-H O intermolecular HB is 11.49 and 12.67 kcal/mol for the syn and anti dimer, respectively [H4]. Other commonly accepted criteria for the description of hydrogen bonds were introduced by Koch and Popelier. 43 They proposed that hydrogen bonds can be characterised based on various descriptors originating from Bader s quantum theory of Atoms In Molecules (AIM). 16 There are eight effects for charge density being the necessary criteria for the existence of inter- or intramolecular hydrogen bonds. Among them there are the bond critical point (BCP) for H Y (where Y is a proton acceptor centre within the X-H Y hydrogen bond) and the ring critical points (RCP) existing due to the formation of an intramolecular hydrogen bond or due to the formation of a dimer (Figure 12b). 47

Figure 12. a) Syn and anti conformers of 2-acylpyrrole; b) conformation of different types of 2-acylpyrrole dimers, where R = OH, NH2, NHCH3. The dimer represents combination of a 2-acylpyrrole pair found in many aggregates presented in Table 2; small circles correspond to ring critical points (RCP). The value of electron density at the BCP (H.. Y) should be within a range of 0.002 0.040 a.u. The corresponding Laplacian of electron density at the BCPs ( 2 BCP) should be within a range of 0.024 0.139 a.u. Additionally there are energetic descriptors of BCPs such as electron energy density at the BCP (HC) and its components, potential electron energy density (VC) and kinetic electron energy density (GC). According to Rozas et al. 44 weak H-bonds are characterised by 2 BCP > 0 and HBCP > 0. The studies of the nature of the HB using QTAIM theory revealed that acid dimers are more stable than cyclic dimers formed by N-H... O=C bonds [H1]. The dimers formed by N-H O=C amide bonds are more stable than cyclic dimers formed by N(1)-H O=C bonds [H2, H9]. Because the 2-acylpyrroles prefer formation of cyclic dimers in concentrated solutions and in the solid state, I analysed the N-H.. O=C hydrogen bond [H1, H2, H4] weak interaction of a C- H... O type and the halogen bond [H3]. On this basis it was found that the N-H O=C interaction plays a key role in the stabilisation of 2-acylpyrrole conformation. The X-ray studies were very valuable in conformational analysis and hydrogen bond detection. Pyrrole-2-carboxylic acid has a proton donating and proton accepting COOH carboxylic group as well as an N H proton donating bond. All of them are involved in hydrogen bond interactions in the crystal structure of the acid where centrosymmetric dimers are formed with two equivalent O H... O bonds between the carboxylic groups and N H... O bonds between the N H pyrrole ring and the C=O carbonyl group. Hence, according to Etter grafs 48

designations 45, the following H-bond motifs were detected in the crystal structure of the acid: R2 2 (8) and R2 2 (10) (Fig. 13). According to this designation the (C=O H-O-C=O H-O) motif is formed by hydrogen bonds with a centre of inversion. Such patterns R2 2 (8) exist in the crystal structure with two O-H O bonds between the oxygen atoms and the O-H donors of the carboxylic groups as well as between the oxygen atoms of the carboxylic groups and the C-H donors C-H O and O-H N. 46 These structural motifs fit well to the structural models predicted by modelling of the acid dimer structure using DFT results (B3LYP/6-311+G(d) level of theory) as well as based on infrared and Raman spectroscopic studies. For the acid dimer connected by two C=O O-H hydrogen bonds intermolecular resonance-assisted hydrogen bonds (RAHBs) 47 were detected. The real molecular structure of the pyrrole-2-carboxylic acid dimer may be considered a mixture of tautomeric forms. Their effective mixing was supported by the process of -electron delocalisation [H1]. Syn conformers of pyrrole-2-carboxylic acid create a network of cyclic dimers formed through C=O H-O interactions, connected by an additional network of N-H O=C hydrogen bonds with slightly lower energy (Fig. 13). Figure 13. H-bond motifs detected in the crystal structure of the pyrrole-2-carboxylic acid. Dotted lines show hydrogen bonds [H1]. The analysis of the crystal structure of certain 2-acylpyrroles leads to a conclusion that the energetically more stable syn form is present in the solid state. For crystal structures of pyrrole-2-carbamide there is only the syn conformer in crystals with bifurcated N-H O hydrogen bonds. [H2] 49

Figure 14. Arrangement of molecules in the crystal of pyrrole-2-carbamide; H-bonds and HB motifs are shown: R2 2 (8), R2 2 (10) and R4 2 (8) [H2] In the crystal structure of pyrrole-2-carbamide there are heteronuclear hydrogen bonds. 48 There are three H-bond motifs corresponding to three interactions of the O atom: R2 2 (8), R2 2 (10) and R4 2 (8). The N-H O hydrogen bonds as a result of interactions through the amide groups are the strongest in amide crystals since N O distances are the shortest. DFT calculations and X-ray analyses indicate that N-H O hydrogen bonds are resonance-assisted. [H2]. Figure 15. Network of intermolecular contacts in the crystal structure of N-methylpyrrole-2-carboxamide. N-H O interactions are shown with dotted lines [H4]. 50

The crystal structure of N-methylpyrrole-2-carboxamide (Fig. 15) also shows the existence of syn conformation. Figure 14 shows the arrangement of molecules in crystals [H4]. There are R2 2 (8) H-bonded motifs with two N H O hydrogen bonds equivalent owing to symmetry constraints. The 2-acylpyrroles for which two conformers exist in solution as a rule crystallise exclusively in the most stable syn conformation. In the crystals without strong proton donors unconventional hydrogen bonding CH... O 49, 50 plays an important role. An example of this interaction is illustrated in [H1] and [H3]. The existence of C5-H5 O2 interactions in the crystal structure of the acid was also found [Fig. 16b]. The H O distance is 1.94 Å, lower than van der Waals radii, 51 and the C-H O angle is 138 [H1]. For N-methylpyrrole-2-carboxamide there is also an extra intramolecular C H O interaction (Figure 14). There is no classic proton donating group, and the C-H group is a proton donor [H4]. The existence of the C Cl... O halogen bond 52,53 was found in the crystal structure of 1-methylpyrrol-2-yl trichloromethyl ketone [H3]. The halogen atom connected with the carbon atom (C Cl) acts as a Lewis acid. The O-atom of the carbonyl group acts as an acceptor centre. Cl O distances are 3.047 Å. This is slightly less than the corresponding sum of van der Waals radii of 3.27 Å, according to Bondi. 51 This means that the halogen bond and the hydrogen bond affect the arrangement of the molecules in the crystal and influence their conformation. a b Figure 16. a) Network of intermolecular contacts in the crystal structure of 1-methylpyrrol-2-yl trichloromethyl ketone [H3]. Arrangement of molecules in the crystal of pyrrole-2-carboxylic acid; C-H O contacts are shown with dotted lines [H1]. 51

The analysis of the crystal structure of some 2-acylpyrroles leads to the conclusion that the energetically more stable syn form is present in the solid state. The most stable form is the syn conformer since it enables the formation of a hydrogen bond between the pyrrole N- H and C=O groups. For all of the above crystal structures of 2-acypyrrole derivatives a crucial role of carbonyl group hydrogen bonds influencing the arrangement of molecules in crystals is seen [H1, H3]. The analysis of crystallographic data of 2-acylpyrroles indicates that the hydrogen bonds have a locking effect on conformation about the C-C bonds. Cambridge Structural Database (CSD) searches on 2-acylpyrroles reveal that there are also some other instances of 2-acylpyrroles which form dimers in the solid phase by N-H O=C interactions. Among the 2-acylpyrrole dimers the mean d(n)h...o distance is 2.018 Å and differs generally by 0.05-0.1 Å. The mean dn...o distance is 2.875 Å and differs generally by 0.1 Å while the mean dc=o bond length is 1.22 Å. It is noted that the longest C=O bond was observed for compounds in which R is NH2, OH, N(CH3)2 [H9]. The shortest dc=o was observed for electron-accepting substituents such as R = OCH3, OC2H5 [H8]. The results of the analysis of the conformation of the 2-acylpyrroles are extremely valuable in the discussion of the conformation of peramine and its derivatives, which was the subject of research under the State Committee for Scientific Research grant. Peramine 54 belongs to a group of secondary metabolites produced by perennial ryegrass fungal endophytes 55 (Figure 17). N O NCH 3 NH HBr N NH 2 H Peramine fungal endophytes Figure 17. Peramine - bioprotective secondary metabolites produced by fungal endophytes Peramine and its derivatives are antifeedant agents which protect plants against pests by discouraging feeding.56 Because of the great potential importance of peramine as a modern 52

and environmentally friendly plant protection agent, conformational analysis with spectroscopic properties of these pyrrolopyrazine alkaloids was performed [H5, H6].57,58 On the basis of the stretching C=C and C=O modes and 1 H NMR spectra of peramine derivatives it has been found that pyrrolopyrazinone rings are conjugated [H5]. Furthermore, I found that the oscillation modes of carbonyl and C=C groups are interrelated thus giving rise to C=O and C=C absorption bands, a product of both vibrations (Fig. 18a). On the basis of theoretical calculations the most stable rotamers were identified. Theoretically predicted IR and Raman as well as electronic spectra for these selected rotamers were in excellent agreement with experimental data. Using VEDA 4 (Vibrational Energy Distribution Analysis), 59 analysis of the predicted spectra with potential energy distribution of normal modes was performed. 60 a) b) Figure 18. a) Experimental and calculated IR spectra of 3-(3-chloropropyl)-2-methylpyrrole[1,2-a]pyrazin-1- (2H)-one. Atomic displacements of C=O and C=C in vibrational modes: out-of-phase and in phase ; b) layered structure of 3-(3-chloropropyl)-2-methylpyrrole[1,2-a]pyrazin-1-(2H)-one viewed along the a axis. This method was found to be useful in the analysis of vibrational properties of peramine and its derivatives [H5, H6]. On the basis of both FT-IR and 1H NMR spectroscopy it is found that pyrrolopyrazinone rings are conjugated. X-ray analysis reveals two rings arranged in one plane and a side arm oriented in the ring plane (Fig. 18b). Dihedral angle N1 C6 C9 C10 between 53