Pashynska V. Mass spectrometric markers and molecular physical mechanisms of action of biologically active substances

Українська версія

Thesis for the degree of Doctor of Science (DSc)

State registration number

0524U000131

Thesis Registration Form

0524U000131.pdf

Applicant for

Vlada Pashynska

Specialization

03.00.02 - Біофізика

Date of defense

08-05-2024

Specialized Academic Board

Д 64.051.13

V.N. Karazin Kharkiv National University

Essay

Pashynska V. A. Mass spectrometric markers and molecular physical mechanisms of action of biologically active substances. – Qualification scientific work for a scientific report. Thesis for the scientific degree Doctor of science in physics and mathematics, specialty 03.00.02 “Biophysics”. – B.I. Verkin Institute for Low Temperature Physics and Engineering of the NAS of Ukraine, V.N. Karazin National University of the Ministry of Education and Science of Ukraine, Kharkiv, 2023 The dissertation is devoted to the actual biophysical problem of disclosure of molecular physical mechanisms of processes which determine the action of biologically active substances on biological systems. A promising methodological approach related to identification of mass spectrometric markers (indicators) of biophysical processes at the molecular level has been developed. Based on the obtained results, the above-mentioned problem of molecular biophysics has been solved in the dissertation for the selected research objects. Namely, the molecular physical mechanisms connected with the biological action of a number of medicines (antimalarial agents of artemisinin type, anti-infective bisquaternary ammonium compounds and rhamnolipids, antituberculosis antibiotic cycloserine, antiviral drug tiloron, cardioprotective agent flokalin) have been proposed. Moreover, the mass spectrometric markers of biologically important molecular physical processes that occur with the participation of biologically active organic components of atmospheric aerosols (in particular, levoglucosan, polyols) have been determined. The molecular physical mechanism, which related to the antimalarial activity of the artemisinin type drugs, has been proposed on the basis of initially determined mass spectrometric markers of formation of stable noncovalent complexes of molecules of these drugs with Fe(III)-heme. In the framework of revealing the molecular physical basis of reported antitumor activity of artemisinin type drugs, noncovalent complexation of artemisinin and dihydroartemisinin with the nucleobases of nucleic acids: adenine, cytosine, and methylthymine, has been experimentally shown for the first time. Due to application of the complex approach, which combines the experimental mass spectrometry method and the DFT calculations, the phenomenon of intermolecular competition between artemisinin agents and molecules of aspirin or vitamin C for noncovalent binding with membrane phospholipids (e.g. dipalmitoylphosphatidylcholine) has been revealed. These competitive processes are proposed for the first time as molecular physical mechanisms of modulating the biological activity of artemisinin type drugs and aspirin or vitamin C when they are used simultaneously. Under study of the molecular mechanisms of modulation of the action of anti-infective bisquaternary ammonium compounds, the formation of stable complexes of dications of decamethoxinum, ethonium or thionium with aspirin has been shown for the first time. At the same time the competition between the medicinal compounds of different groups for binding to dipalmitoylphosphatidylcholine has been revealed. The specific noncovalent complexation of the molecule of antiviral agent tilorone with uridine nucleoside in a polar medium has been revealed for the first time. Such complexation is proposed as a molecular physical mechanism of the antiviral activity of tilorone. Based on the results of the study of intermolecular interactions of recently invented in Ukraine cardioprotective agent flokalin with a number of amino acids, a molecular physical mechanism of the interaction of the drug with cardiac-specific ATP-sensitive membrane channel have been proposed. A complex of mass spectrometric techniques and calculation method AM1 were applied to study bisquaternary ammonium compounds (decamethoxinum, ethonium) in a solvate environment. The obtained results allowed us to determine mass spectrometric markers of structural stability of the agents and to demonstrate that interactions of the bisquaternary dications with the solvate components can seriously effect on the agents stability. A significant result of the dissertation is the development and validation of a technique based on GC/MS method for the determination of biologically active substances (levoglucosan, other monosaccharide anhydrides) in atmospheric aerosols particles. Due to application of the GC/MS technique, the previously undescribed components– polar organic compounds 2-methylthritol and 2-methylerythritol have been identified for the first time in aerosol samples. Key words: biologically active substances, drugs, organic components of atmospheric aerosols, intermolecular interactions, target biomolecules, noncovalent complexes, molecular physical mechanisms of biological action, modulation of drugs activity, solvation, soft ionization mass spectrometry, GC/MS, mass spectrometric markers, quantum mechanical calculations.

Official opponents

Galina Dovbeshko
Iryna Doroshenko
Anna Shestopalova

Research papers

Pashynska V. A., Kosevich M. V., Van den Heuvel H., Claeys M. Characterization of noncovalent complexes of antimalarial agents of the artemisinin type and Fe(III)-heme by electrospray ionization mass spectrometry and collisional activation tandem mass spectrometry. J. Am. Soc. Mass Spectrom. 2004. Vol. 15. P. 1181-1190. DOI: https://doi.org/10.1016/j.jasms.2004.04.030.

Pashynska V. A. Mass spectrometric study of intermolecular interactions between the artemisinin-type agents and nucleobases. Біофізичний вісник. 2009. Т. 22(1). С. 20-28.

Pashynska V. A., Kosevich M. V., Van den Heuvel H., Cuyckens F., Claeys M. Study of non-covalent complexes formation between the bisquaternary ammonium antimicrobial agent decamethoxinum and membrane phospholipids by elecrtospray ionization and collision-induced dissociation mass spectrometry. Вісник харківського національного університету ім. Каразіна №637. Біофізичний вісник. 2004. Т. 1-2 (14). С. 123-130.

Pashynska V., Kosevich M., Stepanian S., Adamowicz L. Noncovalent complexes of tetramethylammonium with chlorine anion and 2,5-dihydroxybenzoic acid as models of the interaction of quaternary ammonium biologically active compounds with their molecular targets. A theoretical study. Journal of Molecular Structure: THEOCHEM . 2007. Vol. 815. P.55-62. DOI: https://doi.org/10.1016/j.theochem.2007.03.019.

Pashynska V., Boryak O., Kosevich M., Stepanian S., Adamowicz L. Competition between counterions and active protein sites to bind bisquaternary ammonium groups. A combined mass spectrometry and quantum chemistry model study. Eur. Phys. J. D. 2010. Vol. 58. P.287-296. DOI: https://doi.org/10.1140/epjd/e2010-00125-5.

Pashynska V. A. Mass spectrometric study of rhamnolipid biosurfactants and their interactions with cell membrane phospholipids. Biopоlymers and Cell. 2009. Vol. 25, N 6. P. 504-508. DOI: http://dx.doi.org/10.7124/bc.0007FE.

Pashynska V. A.,. Zholobak N. M, Kosevich M. V., Gomory A., Holubiev P. K., Marynin A. I. Study of intermolecular interactions of antiviral agent tilorone with RNA and nucleosides. Біофізичний вісник. 2018. Т. 39(1). С. 15-26. DOI: https://doi.org/10.26565/2075-3810-2018-39-02.

Pashynska V., Stepanian S., Gomory A., Vekey K., Adamowicz L. New cardioprotective agent flokalin and its supramolecular complexes with target amino acids: An integrated mass-spectrometry and quantum-chemical study. J. Mol. Struc. 2017. Vol. 1146. P. 441-449. DOI: https://doi.org/10.1016/j.molstruc.2017.06.007.

Pashynska V., Stepanian S., Gomory A., Vekey K., Adamowicz L. Competing intermolecular interactions of artemisinin-type agents and aspirin with membrane phospholipids: Combined model mass spectrometry and quantum-chemical study. Chem. Phys. 2015. Vol. 455. P. 81-87. DOI: https://doi.org/10.1016/j.chemphys.2015.04.014.

Pashynska V., Stepanian S., Gömöry Á., Adamowicz L. What are molecular effects of co-administering vitamin C with artemisinin-type antimalarials? A model mass spectrometry and quantum chemical study. J. Mol. Struc. 2021. Vol. 1232. P. 130039. DOI: https://doi.org/10.1016/j.molstruc.2021.130039.

Pashynska V. A., Kosevich M. V., Gomory A., Vekey K. Model mass spectrometric study of competitive interactions of antimicrobial bisquaternary ammonium drugs and aspirin with membrane phospholipids. Biopolymers and Cell. 2013. Vol. 29(2). P. 157-162. DOI: http://dx.doi.org/10.7124/bc.000814.

Kasian N. A., Pashynska V. A., Vashchenko O. V., Krasnikova A. O., Gomory A., Kosevich M. V., Lisetski L. N. Probing of the combined effect of bisquaternary ammonium antimicrobial agents and acetylsalicylic acid on model phospholipid membranes: differential scanning calorimetry and mass spectrometry studies. Molecular BioSystems. 2014. Vol.10. P. 3155-3162. DOI: 10.1039/c4mb00420e.

Pashynska V. A., Kosevich M. V., Gomory A. Mass spectrometry study of noncovalent complexes formation of antibiotic cycloserine wіth N-acetyl-D-glucosamine and ascorbic acid. Біофізичний вісник. 2020. Vol. 43. P. 103-110. DOI: https://doi.org/10.26565/2075-3810-2020-43-11.

Pashynska V. A., Kosevich M. V., Van den Heuvel H., Claeys M. The effect of cone voltage on electrospray mass spectra of the bisquaternary ammonium salt decamethoxinum. Rapid Commun. Mass Spectrom. 2006. Vol. 20(5). P. 755-763.

Пашинская В. А., Косевич М. В., Степаньян С. Г. Квантовомеханическое исследование структуры гидратированного бисчетвертичного аммониевого соединения декаметоксина. Вісн. Харк. Ун-ту N 49. Біофізичний вісник. 2000. Т. 2(7). С. 29-34.

Pashynska V. A., Kosevich M. V., Gomory A., Vekey K., Claeys M., Chagovets V. V., Pokrovskiy V. A. Variable Electrospray Ionization and Matrix-Assisted Laser Desorption/Ionization Mass Spectra of the Bisquaternery Ammonium Salt Ethonium. Mass Spectrometry & Purification Techniques. 2015. Vol. 1:103. P. 1-9. DOI: 10.4172/2469-9861.1000103.

Kosevich M. V., Boryak O. A., Chagovets V. V., Pashynska V. A., Orlov V. V., Stepanian S. G., Shelkovsky V. S. “Wet chemistry” and crystallochemistry reasons for acidic matrix suppression by quaternary ammonium salts under matrix-assisted laser desorption/ionization conditions. Rapid Commun. Mass Spectrom. 2007. Vol. 21(11). P. 1813-1819. DOI: https://doi.org/10.1002/rcm.3020.

Vashchenko O. V., Pashynska V. A., Kosevich M. V., Panikarskaya V. D., Lisetski L. N. Modulation of bisquaternary ammonium agents affect on model biomembranes by complex formation with an organic anion. Biopolymers and Cell. 2010. Vol. 26, N 6. P. 472-477. DOI: http://dx.doi.org/10.7124/bc.000176.

Pashynska V., Vermeylen R., Vas G., Maenhaut W., Claeys M. Development of a gas chromatography/ion trap mass spectrometry method for determination of levoglucosan and saccharidic compounds in atmospheric aerosols. Application to urban aerosols. J. Mass Spectrom. 2002. Vol. 37. P.1249-1257. DOI: https://doi.org/10.1002/jms.391.

Claeys M., Graham B., Vas G., Wang W., Vermeylen R., Pashynska V., Cafmeyer J., Guyon P., Andreae M., Artaxo P., Maenhaut W. Formation of secondary organic aerosols through photooxidation of isoprene. Science. 2004. Vol. 303. P. 1173-1176. DOI: 10.1126/science.1092805.

Decesari S., Fuzzi S., Facchini M.C., Mircea M., Emblico L., Cavalli F., Maenhaut W., Chi X., Schkolnik G., Falkovich A., Rudich Y., Claeys M., Pashynska V., Vas G., Kourtchev I., Vermeylen R., Hoffer A., Andreae M.O., Tagliavini E., Moretti F., Artaxo P. Characterization of the organic composition of aerosols from Rondônia, Brazil, during the LBA-SMOCC 2002 experiment and its representation through model compounds. Atmos. Сhem. Phys. 2006. Vol. 6. P. 375-402. DOI: https://doi.org/10.5194/acp-6-375-2006.

Claeys M., Kourtchev I., Pashynska V., Vas G., Vermeylen R., Wang W., Cafmeyer J., Chi X., Artaxo P., Andreao M.O., Maenhaut W. Polar organic marker compounds in atmospheric aerosols during the LBA-SMOCC 2002 biomass burning experiment in Rondônia, Brasil: sources and source processes, time series, diel variations and size distributions. Atmos. Chem.Phys. 2010. Vol.10. P. 9319-9331. DOI: https://doi.org/10.5194/acp-10-9319-2010.

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