52nd Conference

Synthesis and Analysis of Drugs 2024

Hradec Králové

BOOK OF ABSTRACTS
Compiled and edited by Andrea Bachtíková and Jan Zitko
Hradec Králové, 2024
Dear colleagues and friends,

The Organizing Committee is pleased to invite you to participate in the 52 nd Conference Synthesis
and Analysis of Drugs (SAL 2024) that will take place in Hradec Králové (Czech Republic) as an onsite
event from September 19th to 20th, 2024. This year’s conference will commemorate the 55th
anniversary of the foundation of the Faculty of Pharmacy in Hradec Králové.

Synthesis and Analysis of Drugs (Syntéza a analýza léčiv) is an annual conference held alternately in
the Czech and the Slovak Republic. It has a long tradition dating back to 1966. The conference covers
all aspects of pharmaceutical chemistry and analysis, including adjoining fields such as biochemistry,
pharmacology, molecular biology, bioorganic and bioinorganic chemistry, and related disciplines.
The conference will be organized under the auspices and on the premises of the Faculty of
Pharmacy, Charles University in Hradec Králové.

I believe the conference will be very pleasant and fruitful. There will be many opportunities to make
new contacts and discuss current challenges in the fields of medicinal chemistry, pharmaceutical
analysis, and other disciplines. Young scientists and PhD students are especially encouraged to
present the results of their work. Dear colleagues, I look forward to meeting you at this traditional
annual conference for experts from the Czech Republic, the Slovak Republic, and many other
countries.

Prof. PharmDr. Martin Doležal, Ph.D.

chairman of the Organising committee
chairman of the Section of Synthetic Drugs, Czech Pharmaceutical Society

1
Members of the Organising committee:
Prof. PharmDr. Martin Doležal, Ph.D.
chairman of the Organising Committee
chairman of the Section of Synthetic Drugs, Czech Pharmaceutical Society

Assoc. Prof. PharmDr. Radim Kučera, Ph.D.,

chairman of the Section of Pharmaceutical Analysis and Bioanalytics, Czech Pharmaceutical Society

Assoc. Prof. PharmDr. Petr Chocholouš, Ph.D.

Assoc. Prof. PharmDr. Martin Krátký, Ph.D.
PharmDr. Marta Kučerová, Ph.D.
Assoc. Prof. PharmDr. Miroslav Miletín, Ph.D.
Assoc. Prof. PharmDr. Hana Sklenářová, Ph.D.
Assoc. Prof. PharmDr. Petra Štěrbová, Ph.D.
Assoc. Prof. PharmDr. Jan Zitko, Ph.D.

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The conference is sponsored by:

3
 POSTERS
BIOORGANIC AND PHARMACEUTICAL CHEMISTRY

4
 P01
SYNTHETIC APPROACH TO UNSYMMETRICAL ABAC-TYPE
PHTHALOCYANINES VIA [3+1] INTERMOLECULAR CYCLIZATION

ABRAHAM, J A., ZIMČÍK, P

Department of Pharmaceutical Chemistry and Pharmaceutical Analysis,, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Phthalocyanines (Pc) are a class of π-conjugated heteroaromatic macrocycles, consisting of four

isoindole rings linked by nitrogen bridges. Upon introducing asymmetry, the molecular structure gets
perturbated electronically allowing a fine-tuning of the physical properties. However, the chemistry
of low-symmetric phthalocyanine has received mediocre attention than that of Pc’s with D4h
symmetry. The mixed condensation reactions using multiple precursors limit the feasibility of the
formation of low-symmetric Pc’s owing to the identical molecular weight and properties of the
product formed.1
Controlled formation of target Pc with a minimal amount of side products could be achieved
by a [3 + 1] approach i.e, a base-promoted condensation of pre-linked trisphthalonitrile (ABA-trimer)
and a free phthalonitrile (C) in the presence of a metal template (M). 2 We have successfully
synthesised the subunit B of ABA-trimer with tert-butylthio linkers at the β-position and propoxy
linkers at the α-position. Correspondingly, the propoxy linkers at the α-position of subunit B are
connected to the α-position of phthalonitrile A, via a sulphur atom constituting the targetted trimer.
Eventually, the ABAC-Pc is obtained via the cyclization reaction of ABA trimer with 4,5-bis(2,6-
bis((1H-imidazol-1-yl)methyl)-4-methylphenoxy)phthalonitrile (C). Similarly, with improved
selectivity and yield, this approach could be pursued for the preparation of low-symmetrical
phthalocyanines having hydrophilic and/or lipophilic moieties attached covalently. Moreover, the
pre-connected ABAtrimer and free phthaonitrile C can be appropriately chosen to generate exotic
phthalocyanines.
The study was supported by ERC-CZ, Project Number: LL2318.

References
1. MACK, J., KOBAYASHI, N.: Chem. Rev., 111, 2011, 281-321.
2. CHAN J, Y, M., NG, D, K, P.: J. Org. Chem., 87, 2022, 7213-7218.

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 P02
IN-CELL ACTIVATABLE PHTHALOCYANINES FOR
PHOTODYNAMIC THERAPY OF CANCER

ALFRED, M.A., ÇAVDARBASHA, N., NOVAKOVA. V.

Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy

in Hradec Králové, Charles University, Czech Republic

e-mail: [email protected]

Conventional chemotherapeutic treatment of Cancer poses a threat to healthy cells, presenting the
need for selective cell death. Although photodynamic therapy1 of cancer is an efficient non-invasive
therapy, an ideal photosensitizer (PS) is still on the lookout. Silicon phthalocyanines are macrocyclic
tetramers with interesting photophysical properties and highly applicative singlet oxygen production
capabilities which can be fine-tuned. Cathepsin B is a lysosomal cysteine protease that plays a pivotal
role in tumour development and its overexpression is associated with tumour environments. Its
dicarboxypeptidase activity makes it cleave a valine-citrulline labile bridge2 at the C-terminus. In this
work we develop an optimized photosensitizer for precision medicine, possessing desired water
solubility, absorption in the biological window of tissues and high singlet oxygen production. It is
modified further with ferrocene to make the PS a PET (photoinduced electron transfer)-controlled
switch for in-cell activation.

Fig 1: Schematic functioning of the antibody-drug conjugate with the cleavable Valine-Citrulline ligand.

The study was supported by ERC-CZ (Project LL2318) and Charles University (SVV 260 666).
References
1. CLAESSENS, C. G., et al.: Chemical Record, 8(2), 2008, 75-97.
2. BARGH, J. D., et al.: Chemical Society Reviews, 48(16), 2019, 4361-4374.

6
 P03
SYNTHESIS, STRUCTURAL CHARACTERIZATION AND
DNA/BSA INTERACTIONS OF NEW PALLADIUM(II) AND PLATINUM(II)
COMPLEXES WITH N-BENZYLPHENOTHIAZINE

ANDREJEVIĆ, T.P.,1 MARKOVIĆ, V.R.,1 AŠANIN, D.P.,2 STEVANOVIĆ, N.LJ.,1 PANTOVIĆ,

B.V.,1 BOGDANOVIĆ, M.G.,3 RODIĆ, M.V.,3 GLIŠIĆ, B.Đ.1

1
Department of Chemistry, Faculty of Science, University of Kragujevac, R. Domanovića 12, 34000 Kragujevac,
Serbia
2
Institute for Information Technologies Kragujevac, University of Kragujevac, Jovana Cvijića bb, 34000 Kragujevac,
Serbia
3
Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovića 3, 21000 Novi Sad, Serbia

e-mail: [email protected]

In this study, we report the synthesis, structural characterization and DNA/BSA interactions of new
palladium(II) and platinum(II) complexes, [PdCl2(N-Bzphtz)2] (1) and [PtCl2(N-Bzphtz)2] (2), where
N-Bzphtz is N-benzylphenothiazine. These complexes were obtained in the reactions of MCl2 (M =
Pd2+ (1) and Pt2+ (2)) with N-Bzphtz ligand in 1 : 2 molar ratio. The reaction was carried out in
acetonitrile at room temperature for the palladium(II) complex, while for the platinum(II) complex,
the reaction mixture was stirred under reflux. The synthesized complexes were characterized by
spectroscopic and electrochemical methods, while their crystal structures were determined by single-
crystal X-ray diffraction analysis. The coordination sphere of the metal ion in these complexes is
occupied by two monodentate N-Bzphtz ligands and two chloride ions. Moreover, the interactions of
the synthesized complexes with calf thymus DNA (ct-DNA) and bovine serum albumin (BSA) were
studied by fluorescence emission spectroscopy to evaluate their binding affinities towards these
biomolecules.

The study was supported by the Science Fund of the Republic of Serbia, Grant No. 7730810, Value-
added biologics through eco-sustainable routes – BioECOLogics. This research has also received
funding from the Ministry of Science, Technological Development and Innovation of the Republic of
Serbia (Agreements No. 451-03-65/2024-03/200122, 451-03-66/2024-03/200122, 451-03-68/2022-
14/200378, 451-03-66/2024-03/200125 and 451-03-65/2024-03/200125).

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 P04
MICELLE FORMATION OF HOMOLOGOUS
3-ALKOXYPHENYLCARBAMOYLOXYETHYL-PYRROLIDINIUM
CHLORIDE IN AQUEOUS SOLUTION

ANDRIAMAINTY F., STOPKOVÁ L., SALANCI E., HRMO E., ČIŽMÁRIK J.

Department of Pharmaceutical chemistry, Faculty of Pharmacy in Bratislava, Comenius University, Slovak republic

e-mail: [email protected]

The investigation of the influence of a homologous series of pyrrolidinoethylesters substituted 3-

alkoxyphenylcarbamic acid1 on the critical micellar concentration (CMC) values in aqueous solution
at laboratory temperature was carried out by measuring the optical density2 at the wavelength 450
nm. The CMC values decrease with increasing the number of carbon atoms nc= 2, 3, 5, 6, 9, and 10
in the hydrophobic chain. The Gibbs free energy, necessary for the transfer of the methylene group
(–CH2) of the 3-alkoxy substituted chain from the aqueous phase to the inner part of the micelle3 was
equal to -0,28 RT.

References
1. ČIŽMÁRIK, J., POLÁŠEK, E., ŠVEC P. et al.: Čes. slov. Farm., 42, 1993, 88‒91.
2. COTTINGHAM, M.G., BAIN, C.D., VAUX, D.J.T.: Lab. Investig., 84, 2004, 523‒529.
3. STOPKOVÁ, L., GALIŠINOVÁ, J., ŠUCHTOVÁ, Z. et al.: Molecules, 23, 2018, 1064.

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 P05
CHROMATOGRAPHIC SEPARATION OF DIBENZOFURAN FROM CLADONIA
STELLARIS AND DIDEPSIDE FROM PSEUDEVERNIA FURFURACEA

BAČKOROVÁ, M.,1 UNGVARSKÁ-MAĽUČKÁ, L.,2 MOLČANOVÁ, L.,3

1
Department of Pharmaceutical Technology, Pharmacognosy and Botany, University of Veterinary Medicine and
Pharmacy in Košice, Slovak Republic
2
Department of Chemistry, Biochemistry and Biophysics, University of Veterinary Medicine and Pharmacy in Košice,
Slovak Republic
3
Department of Natural Drugs, Faculty of Pharmacy in Brno, Masaryk University, Czech Republic

e-mail: [email protected]

Lichens represent symbiotic associations between fungi and algae, or cyanobacteria, and produce
important secondary metabolites. Products of secondary metabolism of lichens include multiple
biological effects, including antibacterial, antiviral, antiproliferative, anti-inflammatory,
antiangiogenic and antioxidant1. The presented study deals with the extraction, isolation and
identification of secondary metabolites present in two lichen species Cladonia stellaris (Opiz) Pouzar
& Vězda (CLA) and Pseudevernia furfuracea (L.) Zopf. (PSE). The methanolic extracts were
evaporated to dryness on a rotary vacuum evaporator. The presence of usnic acid dibenzofuran in the
CLA extract and didepside atranorin in the PSE extract was confirmed using the HPLC method.
Based on the comparison of the chemical shifts of hydrogen and carbon atoms in the 1H and 13C NMR
spectra in the standard and in the CLA extract, we can conclude dibenzofuran usnic acid is present in
the CLA extract. The comparison of the chemical shifts of hydrogen and carbon atoms from the 1D
NMR spectra of the standard shows that didepside atranorin is present in the PSE extract2.

The study was supported by Ministry of Education KEGA, Grant No. 003UVLF-4/2024.

References
1. GOGA, M, et al.: Co-Evolution of Secondary Metabolites, 2020, Springer, 175–209.
2. ELEČKO, J, VILKOVÁ, M., FREŇÁK, R. et al.: Plants, 2022, 11, 1077.

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 P06
PROLYL-tRNA SYNTHETASE INHIBITORS: A VIRTUAL SCREENING AND
RATIONAL DRUG DESIGN APPROACH

BACHTÍKOVÁ, A.,1 JUHÁS, M.,1,2 HOUNGBEDJI, P.,1 ZITKO, J.,1

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Department of Chemistry, Faculty of Science, University of Hradec Králové, Czech Republic

e-mail: [email protected]

The ever-increasing rise in antimicrobial resistance highlights the urgent need to develop
antimicrobials with novel mechanisms of action and calls for innovative research strategies. This
study explores two distinct approaches to identify novel inhibitors of prolyl-tRNA synthetase
(ProRS), a promising antimicrobial target which catalyzes the covalent attachment of proline to its
cognate tRNA, and thus representing an essential step in protein synthesis. The first approach
involves High-Throughput Virtual Screening (HTVS) of a 20-million-compound library from
eMolecules. HTVS exploring several promising hits exhibiting favorable binding interactions, while
some of these compounds demonstrated moderate activity against mycobacterial strains (e.g.,
Mycobacterium tuberculosis H37Ra; MIC = 62.25 μg/mL) and slight activity against several bacterial
strains (e.g., Staphylococcus aureus; MIC = 125 μM). The second approach is based on previously
published 3-aminopyrazine-2-carboxamide derivatives with confirmed activity against
Mycobacterium tuberculosis.2,3 The new derivatives replace the carboxamide linker with 1,2,3-
triazole moiety using click chemistry. So far, 10 derivatives with various substituents on the benzene
ring have been prepared, and their antimicrobial activity is yet to be tested.
The study was supported by Charles University (project GA UK No. 219823), SVV 260 666 and by
“The project National Institute of virology and bacteriology (Programme EXCELES, ID Project No.
LX22NPO5103) - Funded by the European Union - Next Generation EU.“

References
1. BOUZ, G.; ZITKO J.: Bioorg. Chem., 2021, vol. 110, 104806
2. PALLABOTHULA, V.S.K.; KERDA, et al.: Biomolecules, 2022, vol. 12, 1561.
3. PALLABOTHULA, V.S.K.; ABDALRAHMAN, et al: Arch. Pharm., 2024, e2400171

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 P07
BIOSYNTHESIZED SILVER NANOPARTICLES AS ANTIMICROBIALS AND
ANTIOXIDANTS

BEDLOVIČOVÁ, Z.,1 TKÁČIKOVÁ, Ľ.,2 REINEROVÁ, M.,1 ONDUSKOVÁ, N.1

1
Department of Chemistry, Biochemistry and Biophysics, University of Veterinary Medicine and Pharmacy in Košice,
Slovakia
2
Department of Microbiology and Immunology, University of Veterinary Medicine and Pharmacy in Košice, Slovakia

e-mail: [email protected]

Nanoparticles (NPs) represent objects with a size of 1–100 nm. Due to small size and relatively broad
surface, they dispose different properties in comparison to the same material with larger diameters.
Silver nanoparticles (AgNPs) possess broad possibilities of use in various industries from agriculture
to medicine. In the field of medicine, AgNPs are being studied for drug transport, screening of various
diseases, and cancer therapy1. For increasing resistence of pathogens to conventional antibiotics,
AgNPs are also studied as antimicrobial agents2. These facts lead to study of AgNPs as inhibitors of
resistant bacterial strains by multiple mechanisms of action, including oxidative stress, DNA
replication inhibition, or interaction with proteins and enzymes [10, 11].
Our study is focused on the green synthesis of AgNPs using an aqueous medium for extraction. We
also studied the biological activities of prepared AgNPs using a selected set of water extracts of
various plants, namely Stachys recta leaves, Fallopia japonica leaves, and Berberis vulgaris L. fruit.
Namely, antioxidant, and antimicrobial properties were investigated.
The study was supported by project of the Grant Agency of the Ministry of Education, Science,
Research and Sport of the Slovak Republic (2/0112/22).

References
1. JUSTIN PACKIA, J. S, FINUB, J. S., NARAYANAN, A.: Colloids Surfaces B Biointerfaces, 91,
2012, 212–214.
2. SINTUBIN, L., VERSTRAETE, W., BOON, N.: Biotechnolology and Bioengineering, 109, 2012,
2422–2436.
3. TIAN, X., JIANG, X., WELCH, C., et al.: ACS Applied Materials & Interfaces, 10, 2018, 8443–
8450.

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 P08
SYNTHESIS AND APPLICATION OF NOVEL FLUORESCENT LABELING
TAGS FOR OLIGOSACCHARIDE AND GLYCAN ANALYSIS

BOBÁĽ, P.,1 GREGUŠ, M.,1 PÍŽOVÁ, H.,1 SMOLKOVÁ, D.,2,3 CMELIK, R.,3 LAVICKA, J.,3

1
Department of Chemical Drugs, Faculty of Pharmacy, Masaryk University, Brno, Czech Republic
2
Department of Chemistry, Faculty of Science, Masaryk University, Brno, Czech Republic
3
Department of Fluid Phase Separations, Institute of Analytical Chemistry of the Czech Academy of Sciences, Brno,
Czech Republic

e-mail: [email protected]

Sample preparation is crucial for oligosaccharide and glycan analysis by chromatographic or

electrophoretic methods.[1] Since native oligosaccharides and glycans lack chromophores or
fluorophores, they require molecular modification for optical detection. Adding a charged group also
enhances electrophoretic mobility and ionization efficiency for MS detection.
Standard oligosaccharides and N-linked glycans were derivatized with newly synthesized fluorescent
and MS-active labels. After purification, the labeled compounds were separated by CE or LC and
detected using fluorescence or MS. Novel labels based on 2-phenylpyridine derivatives were created,
allowing tuning of fluorescence and MS properties. Compared to commercially available labels, these
new labels showed higher reaction yields, tunable fluorescence, and better ionization efficiency.
This work showcases the development of novel fluorescent labels with fast reaction kinetics, high
fluorescence yields, and improved MS detectability.

This work supported the study was supported by the Grant Agency of the Czech Republic [22-
00236S].

References
1. SMOLKOVA D., CMELIK R., LAVICKA J., (2023). Trends Anal. Chem., 163, 2023, 117068.

12
 P09
PRENYLATED XANTHONES – IMPORTANT SUBSTANCES OF MACLURA
POMIFERA

DVORSKÁ, M., MACHALOVÁ, I., CHMURA, J., MALANÍK, M., ČULENOVÁ, M.,
ŠVAJDLENKA, E.

Department of Natural Drugs, Faculty of Pharmacy, Masaryk University, Czech Republic

e-mail: [email protected]

Maclura pomifera (Moraceae) is grown in Czech Republic as an ornamental tree. Based on the plant
part, it contains various biologically active substances, especially flavonoids and xanthones,1 often
prenylated. Since prenylation significantly affects the biological activity of the compounds,2 this work
was focused on the isolation and identification of prenylated xanthones, interesting taxonomically as
well as for the reported effects.1
In genus Maclura, they are mainly in the cortex of the root and stem, so alcoholic extracts were
prepared from these plant parts, followed by liquid/liquid extraction and separation of the compounds
by chromatographic methods. Using column chromatography, preparative TLC and HPLC, eight
prenylated xanthones were isolated in quantities sufficient for further study. Identification of the
compounds was carried out using spectroscopic methods. The incorporation of the prenyl substituent
varies from compound to compound, with both a free variant and cyclization to a six or five-
membered ring on the basic skeleton.
Since biological activity varies depending on the structure of the molecule, isolated xanthones will
be subjected to selected biological activity tests. The results will help to assess the relationship
between structure and activity and provide information on the potential therapeutic possibilities of
these interesting compounds.

The study was supported by GAČR project GA23-04655S, Role of prenylation and glycosylation
patterns in anti-inflammatory activity and metabolism of natural phenolic compounds

References
1. DA COSTA, C.T., MARGOLIS, S.A., BENNER JR., B.A., et al.: J. Chromatogr. A, 831, 1999,
167‒178.
2. MUKAI, R.: Biosci. Biotechnol. Biochem., 82, 2018, 207‒215.

13
 P10
MICROWAVE-ASSISTED SYNTHESIS, PURIFICATION AND CYTOTOXIC
ACTIVITY STUDY OF 1,3,5-TRIAZINE DERIVATIVES WITH AMINO ACIDS
ON HT-29 CELL LINE

BODNÁR MIKULOVÁ, M.,1 HANKO, M.,1 HRICOVÍNIOVÁ, J.,2 GREIFOVÁ, G.,2 MIKUŠ,
P.,1

1
Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy, Comenius University
Bratislava, Odbojárov 10, 832 32 Bratislava, Slovakia
2
Department of Cell and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University Bratislava,
Kalinčiakova 8, 832 32 Bratislava, Slovakia

e-mail: [email protected], [email protected]

Over recent years, hybrid molecules incorporating 1,3,5-triazine have shown promising antibacterial,
antiviral, antiphlogistic, anticonvulsive, as well as anticancer activity by inhibiting many enzymatic
pathways.1 From the group of our systematically studied triazine derivatives, 1,3,5-triazines
containing 4-aminobenzenesulfonamide and a pair of identical amino acids with non-polar (Ala, Tyr,
Trp) and polar (Ser, Thr, Asn, Gln) side chains are investigated in the present work. The tested
compounds were synthesized by a newly developed method using microwave irradiation. This
procedure showed significantly shortened reaction time and higher or similar yields and purities of
the products when compared to our conventional reflux conditions2,3. Separation and purification of
the desired derivatives using semi-preparative HPLC methods was performed to obtain the purities
over 98%. Chemical structures and purities were confirmed by NMR, IR spectroscopy and HPLC-
DAD/MS. Then, the cytotoxic activity was screened on human cancer HT-29 cell line using the MTT
assay.

This study was supported by the project VEGA 1/0514/22 and the grant of the Scientific Board of the
Faculty of Pharmacy Comenius University Bratislava No. GVRFaFUK/1/2023.

References
1. MISHRA, C. B., et al.: Med. Res. Rev., 40, 2020, 2485‒2565.
2. MIKUŠ, P., et al.: Bioorg. Chem., 81, 2018, 241‒252.
3. MIKULOVÁ, M. B., et al.: Int. J. Mol. Sci., 22, 2021, 11283.

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 P11
COPPER AND ZINC COMPLEXES OF REDUCED SCHIFF BASES SHOWING
MARKED BIOACTIVITY

HABALA, L.,1 OBOŇOVÁ, B.,1 VALENTOVÁ, J.,1 PAŠKOVÁ, Ľ.,2 HRICOVÍNIOVÁ, J.,2
BILKOVÁ, A.,2 BILKA, F.,2 LITECKÁ, M.3

1
Department of Chemical Theory of Drugs, Faculty of Pharmacy, Comenius University, Bratislava, Slovakia
2
Department of Cell and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University, Slovakia
3
Institute of Inorganic Chemistry of the CAS, Husinec-Řež č.p. 1001, 250 68 Řež, Czech Republic

e-mail: [email protected]

Schiff bases represent a category of compounds with manifold biological effects. The purposeful
modification of their structure by varying the substituents on the amine and/or carbonyl components
allows for directed alteration of their properties. They also act as versatile ligands in metal complexes.
Several reduced Schiff bases containing fluoride substituents and their zinc and copper complexes
1,2
have been prepared and structurally characterized by single-crystal X-ray diffraction.
Benzaldehyde derivatives containing fluorine substituents (as -F or -CF3 groups) were selected as
starting compounds due to the often profound effect of fluorine substituents on biological activity.
The antimicrobial activities of the reduced Schiff bases and their zinc(II) and copper(II) complexes
were evaluated in vitro, with one of the zinc complexes showing excellent antibacterial activity
comparable to the antibiotic ciprofloxacin used as a standard. The cytotoxicity of the products in the
HepG2 cell line was evaluated as well. The copper(II) complexes showed marked cytotoxic activities,
considerably higher than the standard cisplatin. The cytotoxicity depended significantly on the
substitution pattern. Furthermore, the affinity of the complexes towards bovine serum albumin and
calf thymus DNA was established.

The study was supported by the Scientific Grant Agency of Slovak Republic VEGA (Project No.
1/0661/24 and 1/0429/21) and by the Research and Development Agency under the contract No.
APVV-23-0349.
References
1. OBOŇOVÁ, B., HABALA, L., LITECKÁ, M., et al.: Life, 13, 2023, 1516.
2. OBOŇOVÁ, B., VALENTOVÁ, J., LITECKÁ, M., et al.: Int. J. Mol. Sci., in press.

15
 P12
NOVEL SALICYLIC ACID-BASED DERIVATIVES AND TANDEM PEPTIDE
CARRIERS AS TOOLS TO COMBAT INFECTIONS

HORČIC, J.,1 PFLÉGR, V.,1 BŐSZE, S.,2 KRÁTKÝ, M.,1

1
Department of Organic and Bioorganic Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Czech
Republic
2
ELKH Research Group of Peptide Chemistry, Eötvös Loránd University, Budapest, Hungary

e-mail: [email protected]

Infectious diseases are a looming threat to public health worldwide. Tuberculosis, with over
10 million casualties yearly, is one of the biggest. This is further stressed by the growing incidence
of drug resistant strains. Strains resistant to multiple first-line antituberculotics are especially
troubling. Novel antitubercular drugs with unique mechanisms of action are necessary to combat this
epidemic and stop the spread of drug-resistant strains.
Derivates of salicylic acid have exhibited several interesting biological properties including
antimicrobial effects. Based on previously described compounds,1 we prepared two series of
salicylamide derivates containing N-monosubstituted carbamate scaffold and screened them for
antimicrobial activity. One series of compounds showed a broad effect on tested mycobacterial strains
with small variation between individual compounds. Additionally, these compounds showed an
antifungal activity comparable to fluconazole against T. interdigitale. A few of them also exhibited
exceptional activity against G+ bacteria (MIC < 0.1 µmol l-1).
Concurrently we report the synthesis of a novel peptide carriers to further enhance the effectiveness
of small antitubercular molecules. By combining two kinds of cell-penetrating peptides into one
sequence we aim to create a selective and highly efficient intracellular delivery system for new small
molecule drugs.

This work was supported by the project National Institute of Virology and Bacteriology (Programme
EXCELES, ID Project No. LX22NPO5103) – Funded by the European Union – Next Generation EU.
Supported by Ministry of Health of the Czech Republic, grant nr. NW24-05-00539.
References
1. KRÁTKÝ, M., JANĎOUREK, O., BARANYAI, Z., et al.: Eur. J. Med. Chem., 181, 2019, 111578.

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 P13
PYRAZINAMIDE AND 1,2,3-TRIAZOLE: A PROMISING RELATIONSHIP

HOUNGBEDJI, P.1, BOHÁČOVÁ, J.1, TABARESTANI, P.1 ZITKO, J.1,

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Pyrazinamide, a key antituberculotic agent, has been part of the 1st line treatment regimen of
tuberculosis ever since its introduction in 1950.1 Although tuberculosis can be treated, the course is
lengthy (up to 6 months) and often complicated by antibacterial resistance.1 To discover more potent
antituberculotics, this work explores the combination of pyrazinamide with 1,2,3-triazole, a linker
thoroughly reported in literature as part of the structure of many antimycobacterial agents.2 The
backbone of the synthesis of 1,2,3-triazole containing derivatives is so called “click chemistry” – a
reaction combining two molecular building blocks as easily as a key inserted in its lock.3 The specific
click reaction consists of combining starting materials containing terminal alkynes with beforehand
synthetized aromatic azides.3 The reaction is catalysed by Cu(I), obtained by exposing a source of Cu
(II) to a reducing agent, and results in dipolar cyclo addition of azides onto said alkynes.3 This reaction
has high yields, an easy workup, straightforward purification, and obtained compounds present
interesting antimycobacterial activity (M.Tb. H37Ra MIC =15,625 µg.mL-1, M.Tb. H37Rv MIC =
6,25 µg.mL-1, if R= 4-OH and X= 5-Cl).

The study was supported by the project of Specific Academic Research (SVV 260 547) and by the
project National Institute of virology and bacteriology (Programme EXCELES, ID Project No.
LX22NPO5103) - Funded by the European Union - Next Generation EU
References
1. World Health Organization, WHO Consolidated Guidelines on Tuberculosis, Module 5: Management of
Tuberculosis in Children and Adolescents, 2022, Geneva.
2. BOZOROV K, ZHAO J, AISA HA.: 1,2,3-Triazole-containing hybrids as leads in medicinal chemistry: A recent
overview. Bioorg Med Chem. 2019;27(16):3511-3531. doi:10.1016/j.bmc.2019.07.005
3. BARRAL K., MOORHOUSE A.D., MOSES, J.E.: Efficient Conversion of Aromatic Amines into Azides: A One-
Pot Synthesis of Triazole Linkages, Organic Letters 2007 9 (9), 1809-1811 , DOI: 10.1021/ol070527h

17
 P14
SУNTHESIS OF FLUOROQUINOLON HYBRIDS AND THEIR PROSPECTS AS
NEW ANTIMICROBIALS

HRYHORIV, H.1, KOVALENKO, S.2, FILIMONOVA, N.3, PEREKHODA, L.4,

GEORGIYANTS, V.1

1
Department of Pharmaceutical Chemistry, National University of Pharmacy, Kharkiv, Ukraine;
2
Department of Organic Chemistry, V.N. Karazin Kharkiv National University, Kharkiv, Ukraine;
3
Department of Microbiology, Virology and Immunology, National University of Pharmacy, Kharkiv, Ukraine;
4
Department of Medicinal Chemistry, National University of Pharmacy, Kharkiv, Ukraine

e-mail: [email protected]

Overuse and misuse of antibiotics has been leading to the appearance of numerous resistant strains
since the beginning of their utilization in clinical practice. However, it is possible to design new
molecules based on the known ones and the aim of our investigation was to synthesize and study
antibacterial properties of hybridized fluoroquinolones (FQ).
At first, the docking studies were carried out. Their results helped to identify the promising molecules,
mainly among C-7 and C-3 FQ derivatives. Ciprofloxacin and norfloxacin were taken as core
structures. Their C-7 position was modified using 1,2,3-triazole moiety through a developed synthetic
procedure. Then, we studied introduction of the arylsulfonyl moiety into C-3 position with subsequent
hybridization of C-7 and N-1. The ranges of new compounds were obtained with medium yields and
their structures were confirmed using 1H NMR, 13
C NMR, LC/MS spectroscopy and X-Ray
diffraction studies.
FQs hybridized with 1,2,3-triazole moiety revealed moderate antimicrobial and antifungal activities,
and new C-3 substituted arylsulfonyl derivatives showed a bit smaller activity, probably due to their
lower solubility in common solvents. A few hit compounds were identified and selected for further
investigations.

The study was supported by the Ministry of Health of Ukraine from the state budget according to the
topic ‘Molecular design and microbiological screening of innovative derivatives of fluoroquinolone
antibiotics in order to combat resistant strains of microorganisms’ (SRN: 0121U109239).

18
 P15
CHITOSAN-KEFIRAN-BASED FILMS CONTAINING PLANT EXTRACT AS A
POTENTIAL NON-TOXIC AND ANTIBACTERIAL WOUND DRESSING
MATERIALS

CHELMINIAK-DUDKIEWICZ, D.,1 MACHACEK, M.,2 DLUGASZEWSKA, J.,3 MYLKIE, K.,1

SMOLARKIEWICZ-WYCZACHOWSKI, A.,1 ZIEGLER-BOROWSKA, M.,1

1
Department of Biomedical and Polymer Chemistry, Faculty of Chemistry, Nicolaus Copernicus University in Torun,
Poland
2
Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University, Czech Republic
3
Department of Genetics and Pharmaceutical Microbiology, Faculty of Pharmacy, Poznan University of Medical
Sciences, Poland

e-mail: [email protected]

Due to the significant increase in patients struggling with wounds, developing new wound dressings
with good physico-chemical and biological properties is now attracting much more attention1-2. In
this study, we successfully fabricated chitosan and kefiran-based films cross-linked with a new
dialdehyde kefiran obtained by our team. To improve the properties of these materials, we introduced
plant extract into the biopolymer matrix. The test results showed that the obtained films have
antimicrobial, antioxidant, anti-inflammatory, non-hemolysis, and non-toxic material. Moreover,
they can bond with human serum albumin and fibrinogen. Therefore, the presented films have
promising potential for practical wound-dressing applications.

The study was supported by the National Science Centre Poland grant UMO-2022/47/D/NZ7/01821.

References
1. CHELMINIAK-DUDKIEWICZ, D., MACHACEK, M., DLUGASZEWSKA, J., et al.: Int. J.
Biol. Macromol., 253, 2023, 126933.
2. CHELMINIAK-DUDKIEWICZ, D., WUJAK, M., DLUGASZEWSKA, J., et al.: Heliyon, 10,
2024, e35389.

19
 P16
ANTIMICROBIAL DERIVATIVES WITH IMPROVED PHYSICOCHEMICAL
PROPERTIES – A HIT EXPANSION STUDY OF 2-AMINOOXAZOLES

JUHÁS, M., BRÁNYIK, M., LEE, H., ZITKO, J.

Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Antimicrobial drug resistance, combined with the scarcity of new highly active agents, remains a
challenge in the treatment of infectious diseases. Regrettably, little progress has been made over the
years and these issues are expected to persist in future. In our previous study1, we explored a strategy
to address the presence of some PAINS groups, hepatotoxicity or low water solubility common in
drug discovery efforts. Building upon our success, here we present a hit expansion study of the most
promising compounds in the hope of improving their antimicrobial activity while retaining favourable
physico-chemical properties. So far, the synthesis yielded 29 derivatives of general structures as
presented in Figure 1. All derivatives were tested against a panel of microbial species of clinical
importance, including multidrug-resistant clinical isolates. Compounds showed higher activity
against mycobacteria, especially Mycobacterium tuberculosis (best MIC = 0.39 µg/mL). None of the
tested active derivatives were cytotoxic against Hep G2 cell line, thus representing promising
compounds for further development.

Figure 1. Synthetic diagram and general structures of the investigated derivatives

This project was supported by „The project National Institute of virology and bacteriology
(Programme EXCELES, ID Project No. LX22NPO5103) - Funded by the European Union - Next
Generation EU.“
References
1 JUHÁS, M., et al. Pharmaceuticals, 15(5), 2022, 580.

20
 P17
DESIGN OF NOVEL INHITORS OF PROLYL-TRNA SYNTHETASE WITH
CHALLENGING SYNTHETIC PROCEDURES

KERDA, M.,1 ZITKO, J.,1

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Inhibition of prolyl-tRNA synthetase (PRS) is associated with antitumor activity and suppression of
autoimmune response. One of the unexplored but very promising areas is antimicrobial utilization.
We began with a knowledge of proven inhibitor of human PRS1 and attempted to create scaffold with
higher affinity to the PRS.
This work continues in previous effort to find new inhibitors.2 Our goal is to obtain similar
interactions as ATP, specifically in the ribose binding site. We introduced polar functional group to
our series, which made the synthesis much more challenging. The synthesized compounds will be
tested on inhibition of prolyl-tRNA synthetase to find best achieved pharmacophore. Additionally,
the compounds will be screened on their antimicrobial activity and cytotoxicity.

The study was supported by the Charles University, project GA UK No. 349721, by Ministry of Health
of the Czech Republic, grant nr. NU21-05-00482, by Charles University project SVV 260 666 (Czech
Republic), and by “The project National Institute of Virology and Bacteriology (Programme EXCELES,
ID project no. LX22NPO5103) – Funded by the European Union – Next Generation EU.”

References
1. ADACHI, R., OKADA, K., SKENE, R., ET AL.: Biochem. Biophys. Res. Commun. 488(2), 2017,
393‒399.
2. PALLABOTHULA, V., KERDA, M., JUHÁS, M., et al.: Biomolecules, 12, 2022, 1561.

21
 P18
SELECTIVITY OF N(2)-SUBSTITUTED OXOTRIAZINOINDOLE ALDOSE
REDUCTASE INHIBITORS WITH THE INTERACTION PATTERN IN PRO301-
ARG312 LOOP OF ALDEHYDE REDUCTASE

KOVACIKOVA, L.,1,2 ALMASSY, A.,2 MAJEKOVA, M.,1 ADDOVA, G.,2 BOHAC, A.,2
STEFEK, M.,1

1 Centre of Experimental Medicine SAS, Institute of Exp. Pharmacology and Toxicology, Bratislava, Slovakia
2 Department of Organic Chemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia

e-mail: [email protected]

Inhibition of aldose reductase (ALR2), the first enzyme of the polyol pathway, is a promising
approach in the treatment of diabetic complications. In our previous study, 2-(3-oxotriazinoindol-5-
yl)acetic acid derivatives (OTIs) were identified as ALR2 inhibitors with high efficacy and
selectivity1. Selective inhibition of ALR2 related to ALR1 is crucial due to potential side and
unwanted effects associated with several inhibitors of aldose reductase (ARIs). In this study, a series
of novel N(2)-substituted oxotriazinoindoles was developed with the aim to investigate molecular
interactions within the ALR2 inhibitor binding site. About twice increased inhibition efficacy of the
most efficient derivative 14 (N(2)-CH2CH2COOH) compared to the unsubstituted lead OTI was
obtained, yet at the expense of selectivity relative to anti-target aldehyde reductase (ALR1). To
explain the major drop in selectivity, observed also in other N(2)-substituted derivatives, in silico
molecular modeling approach revealed the role of extra interactions with the residues of Arg309,
Arg312 and Met302 located in the additional C-terminal loop of ALR1 missing in ALR2, which can
prevent or enhance binding in ALR1. These key findings will be used for development of the next
generation of selective OTI inhibitors.

The study was supported by VEGA 2/0008/22, APVV-20-0411, APVV-20-0543, APVV SK-SRB-21-0047 and
APVV SK-FR-22-0017.

1
HLAVAC, M., KOVACIKOVA, L., PRNOVA, S.M. et al.: J. Med. Chem. 63, 2020, 369-81.

22
 P19
A CLICKABLE AZAPHTHALOCYANINE DERIVATIVE FOR
SUPRAMOLECULAR COMPLEXES WITH QUENCHERS

LAPEŠOVÁ, J., ZIMČÍK, P.

Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

For development in the field of photosensitizers, it is desirable to study conventional and

unconventional mechanisms of quenching to design new and improved analyte-activable
fluorescence sensors and “smart” photosensitisers. It was reported, that photoinduced electron
transfer (PET) between a strong electron donor, such as ferrocene, and phthalocyanine acceptor leads
to quenching.1 In our current project, we study the interactions of phthalocyanine close relatives,
azaphthalocyanines, with ferrocene-based quenchers. Azaphthalocyanines contain nitrogen atoms
that increase their electron-deficient character compared to phthalocyanine, what makes them more
susceptible to PET with electron donors. In this project, we test quenching of fluorescence of an
azaphthalocyanine derivative with ferrocene-based quenchers. The structure of both the
azaphthalocyanine, and the quencher is designed to favour their interaction by additional
supramolecular forces (charge-transfer complexes).

1.5×10 6 0.0 mM
Quencher:
Fluorescence [a.u.]

1.0×10 6 40.0 mM

5.0×10 5

0
600 650 700 750 800
Wavelength [nm]

The study was supported by Charles University Grant Agency (reg. No. 170223) and Czech Science
Foundation (23-06177S).
References
1. LAU, J. T. F., LO, P.-C., JIANG, X.-J. et al.: J. Med. Chem. 2014, 57, 4088-4097.

23
 P20
SYNTHESIS AND SOD-MIMETIC ACTIVITIES OF COPPER(II) COMPLEXES
OF SCHIFF BASES DERIVED FROM 4-HYDROXY OR 4-METHOXYSALICYL
ALDEHYDE AND AMINO ACIDS

LINTNEROVÁ, L.,1 VALENTOVÁ, J.1

1
Department of Chemical Theory of Drugs, Faculty of Pharmacy, Comenius University, Slovakia

e-mail: [email protected]

In our study, a wide series of Schiff base ligands derived from 4-hydroxy and 4-methoxysalicyl
aldehyde (2,4-dihydroxybenzaldehyde or 2-hydroxy-4-methoxybenzaldehyde) and amino acids such
as L-serine, L-threonine, β-alanine, γ-aminobutyric and 5-aminovaleric acid was prepared –
altogether 7 ligands. These ligands were used in synthesis of copper(II) complexes using copper(II)
acetate and copper(II) chloride. The SOD mimetic properties of the prepared complexes were
measured by INT (superoxide radical scavenging)1 and ABTS (cation-radical reducing)2 methods.
Complex solutions (1.4 - 2 × 10-4 M in DMSO/water mixtures) were used to measure antiradical
activity calculated as percentage of radical transfer inhibition (INT) up to 57% and as percentage of
radical quenching (ABTS) up to 100%. ABTS method was also used to determine IC50 values of
SOD activity from a series of measurements with decreasing complex concentration. The IC50 were
measured for 12 out of 14 complexes with range 103 – 10 μM with best activities observed for 5-
aminovaleric acid derived ligand complexes.

The study was supported by the grant of Ministry of Education, Research, Development and Youth of
Slovak Republic VEGA 1/0661/24 and Slovak Research and Development Agency APVV-23-0349.

References
1. LINTNEROVÁ, L., VALENTOVÁ, J., HERICH, P., et al.: Monatsh. Chem., 149, 2018, 901–911.
2. ILYASON, I. R., BELOBORODOV, V. L., SELIVANOVA, I. A., et al.: J. Mol. Sci., 21, 2020,
1131.

24
 P21
SYNTHESIS OF DEXRAZOXANE ANALOGUES WITH HYDROXYLATED
LINKER

MACUŠ, M.,1 CEJNAROVÁ E.,1 KARABANOVICH G.,1 ROH J.1

1
Department of Organic and Bioorganic chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Czech
Republic

e-mail: [email protected]

Anthracyclines (ANTs) such as daunorubicin or doxorubicin are very potent anti-cancer

agents widely used in the treatment of various malignancies. However, their clinical usage is limited
by their cardiotoxicity, which can result in severe and sometimes irreversible heart damage that can
lead to heart failure. The mechanism of ANTs cardiotoxicity is primarily based on the interaction and
poisoning of topoisomerase IIβ (TOP2B) in cardiomyocytes.1
The only clinically used drug in the prevention of ANT-induced cardiomyopathy is
dexrazoxane (DEX). Recent data shown that catalytic inhibition of TOP2B is the actual mechanism
of the cardioprotective action of DEX in the prevention of ANTs cardiotoxicity.2
Unfortunately, the solubility of dexrazoxane in water is very low. The aim of this work is to
prepare an analogue of DEX (figure 1) with increased solubility in water by adding a hydroxymethyl
group to the linker. Additionally, the hydroxyl group can be used to prepare prodrugs with enhanced
hydrophilicity. These modifications can improve the solubility and bioavailability of the drug.

Figure 1: Scheme 1: DEX, and methylhydroxy-DEX

The study was supported by GAUK (no. 361422 and 155124)

References
1 Hasinoff, B., B.; Patel, D.; Wu X. Cardiovascular toxicology. 20, 2020, 312-320.
2 Jirkovský E., Jirkovská A., Bavlovič-Piskáčková H. et al. Circulation: Heart Failure 14(11), 2021,
e008209

25
 P22
IN SILICO ASSESSMENT OF SILVER(I) COMPLEX WITH N-
METHYLPHENOTHIAZINE AGAINST SELECTED MICROBIAL STRAINS

MARKOVIĆ, V.R.,1 BRANKOVIĆ, J.,1 PETROVIĆ, V.,1 ANDREJEVIĆ, T.P.,1 STEVANOVIĆ,

N.LJ.,1 PANTOVIĆ, B.V.,1 AŠANIN, D.P.,2 GLIŠIĆ, B.Đ.1

1
Department of Chemistry, Faculty of Science, University of Kragujevac, R. Domanovića 12, 34000 Kragujevac,
Serbia
2
Institute for Information Technologies Kragujevac, University of Kragujevac, Jovana Cvijića bb, 34000 Kragujevac,
Serbia

e-mail: [email protected]

In the present study, the silver(I) complex with N-methylphenothiazine (N-Mephtz),

[Ag(N-Mephtz)4]CF3SO3.1/3H2O, was evaluated in silico against Escherichia coli, Staphylococcus
aureus, Pseudomonas aeruginosa and Candida albicans. For this purpose, prescreening was
performed on thirty relevant protein structures, after which eight of them were highlighted as potential
targets. Based on the obtained binding affinity values, in the case of E. coli, β-ketoacyl-ACP synthase
III (1hnj) and 1,4-dihydroxy-2-naphthoyl-CoA synthase–MenB (3t88) were identified as potential
targets of the [Ag(N-Mephtz)4]CF3SO3∙1/3H2O complex. Among S. aureus protein targets, the most
favorable binding mode of this silver(I) complex was found for tyrosyl-tRNA synthetase (1jij), while
four relevant P. aeruginosa proteins were identified as its potential antimicrobial targets, i.e., (3R)-
hydroxyacyl-ACP dehydratase –FabZ (1u1z), LasR ligand-binding domain (2uv0), 3-
hydroxydecanoyl-Acyl Carrier Protein Dehydratase–FabA (4b0c), and caseinolytic protease ClpP2
(7m1l). Finally, it was shown that the [Ag(N-Mephtz)4]CF3SO3∙1/3H2O complex can interact with
the sterol 14-alpha demethylase (CYP51) with the binding affinity value of -7.2 kcal/mol, which is
slightly higher than that of the antifungal drug fluconazole (-7.0 kcal/mol).

The study was supported by the Science Fund of the Republic of Serbia, Grant No. 7730810, Value-
added biologics through eco-sustainable routes – BioECOLogics. This research has received funding
from the Ministry of Science, Technological Development and Innovation of the Republic of Serbia
(Agreements No. 451-03-65/2024-03/200122, 451-03-66/2024-03/200122 and 451-03-68/2022-
14/200378).

26
 P23
STRUCTURE–ACTIVITY RELATIONSHIP STUDY OF DINITROBENZYL-
1,3,4-OXADIAZOLES, HIGHLY EFFICIENT ANTI-TUBERCULOSIS
COMPOUNDS

MELNIKOVA, I., KARABANOVICH, G., ROH, J.

Department of Organic and Bioorganic Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Czech
Republic

e-mail: [email protected]

Mycobacterium tuberculosis (M.tb.) is a recalcitrant pathogen that is rife around the world, latently
infecting approximately a quarter of the worldwide population. The presence of a nitro group in the
structure of some drugs (delamanid, macozinone) is crucial for their activity against M.tb., despite
the fact that the presence of nitro group can be associated with a toxicity risk, such as hepatotoxicity,
genotoxicity, mutagenicity.
5-Alkyl/aryl-2-(3,5-dinitrobenzylsulfanyl)-1,3,4-oxadiazoles 1 prepared and studied in our group
showed excellent activity against both drug susceptible and drug-resistant M.tb. strains with minimum
inhibitory concentrations as low as 0.03 µM (0.011-0.026 µg/mL) and were highly effective against
non-replicating M.tb. SS18b strain. Lead compounds showed low cytotoxicity against various cell
lines, including isolated human hepatocytes, despite the presence of two nitro groups in the
molecules.1,2
For further development of these potent antimycobacterial agents we focused on replacing one nitro
group of the 3,5-dinitrophenyl moiety with other (electron-withdrawing) substituents:

The study was supported by Program EXCELES, NPO (reg. No. LX22NPO5103).

References
1. KARABANOVICH, G., ZEMANOVÁ, J., SMUTNÝ, T., et al.: J. Med. Chem., 59 (6), 2016,
2362-2380.
2. KARABANOVICH, G., NĚMEČEK, J., VALÁŠKOVÁ, L., et al.: Eur. J. Med. Chem., 126, 2017,
369-383.

27
 P24
ANTIPROLIFERATIVE AND CYTOTOXIC PROPERTIES OF GERANYLATED
FLAVONOIDS FROM PAULOWNIA TOMENTOSA (THUNB.) STEUD. FRUIT IN
HUMAN CANCER CELL LINES

MOLČANOVÁ, L.,1 MELICHAROVÁ, J.,2 KAUEROVÁ, T.,2 KOLLÁR, P.,2 ŠMEJKAL, K.,1

1
Department of Natural Drugs, Faculty of Pharmacy, Masaryk University, Brno, Czech Republic
2
Department of Pharmacology and Toxicology, Faculty of Pharmacy, Masaryk University, Brno, Czech Republic

e-mail: [email protected]

Paulownia tomentosa (Thunb.) Steud. (Paulowniaceae), a traditional Chinese medicinal plant, is a

rich source of secondary metabolites, mainly geranylated flavonoids. These compounds are currently
studied for their promising biological activities, such as antimicrobial, anti-inflammatory,
antioxidant, and cytotoxic.
Our continuous research done on this plant leads to the isolation of a series of geranylated flavonoids
using chromatographic methods. They were obtained from the chloroform portion of the ethanolic
extract of immature fruit of P. tomentosa.1,2
Compounds were tested for potential antiproliferative activity using WST-1 assay in three human
cancer cell lines (DU-145 prostate cancer cell line, THP-1 monocytic leukaemia cell line, and MCF-
7 breast carcinoma cell line). Diplacone, as one of the most potent compounds from that series, was
also shown to induce apoptosis in prostate cancer cell line.

The study was supported by Czech Science Foundation: GA23-04655S (Role of prenylation and
glycosylation patterns in anti-inflammatory activity and metabolism of natural phenolic compounds).

References
1. MOLČANOVÁ, L., KAUEROVÁ, T., DALL'ACQUA, S., et al.: Bioorg. Chem., 111, 2021,
104797.
2. MOLČANOVÁ, L., TREML, J., BREZÁNI, V., et al.: J. Ethnopharmacol., 296, 2022, 115509.

28
 P25
BORONIC ACID FUNCTIONALIZED CHITOSAN COATED-MAGNETIC
NANOPARTICLES FOR GLYCOPROTEIN BINDING

MYLKIE K.,1 SMOLARKIEWICZ-WYCZACHOWSKI A.,1 CHEŁMINIAK-DUDKIEWICZ D.,1

ZIEGLER-BOROWSKA M.,1

1
Department of Biomedical Chemistry and Polymers, Faculty of Chemistry, Nicolaus Copernicus University, Toruń

e-mail: [email protected]

Biomarkers are substances associated with the presence, development, or course of a disease that can
be detected and quantified in the body. Biomarkers include plasma proteins, such as human serum
albumin (HSA) and α-1-acid glycoprotein (AGP).1 Serum HSA and AGP tests are often performed in
diagnostic laboratories to assess patients' health status, diagnose diseases, and monitor the
effectiveness of applied therapies. A significant challenge in working with free, non-immobilized
proteins is the difficulty of separating them from the supernatant. This process typically requires
techniques such as ultrafiltration, ultracentrifugation, and microdialysis. Recently, a substantial
amount of literature has focused on carriers that can be used for the isolation, identification, and
determination of protein concentration in biological samples. One example of such carriers is
magnetic nanoparticles(AGP).2 Many carriers for the immobilization or capture of HSA are well-
known and described in the literature. However, there is a notable lack of research on the binding of
α-1-acid glycoprotein. Therefore, the key goal of my work was to design and synthesize materials
capable of rapid glycoprotein uptake. As a result of the research, magnetic nanoparticles coated with
modified chitosan with free dihydroxyboryl groups were obtained. The obtained material was
characterized (SEM, TEM, ATR-FTIR, DLS) and its ability to bind alpha-1-acid glycoprotein was
tested in an environment of pH 7.4 and 9.0.

References
1. SMITH, SA., WALTERS, N.J., PHARM RES, 36, 2019
2. HOSSEINPOUR, M.N., SALEHZADEH S., RAKHTSHAH J., et. al:Int. J. Biol. Macromol.
, 125, 2018

29
 P26
SYNTHESIS AND PHYSICOCHEMICAL PROPERTIES OF SOME
BENZOFURAN DERIVATIVES

NÁDASKÁ, D., ANDRIAMAINTY, F., ŠANDRIK, R., MALÍK, I.

Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University Bratislava, Slovak Republic

e-mail: [email protected]

Heterocyclic chemistry plays extraordinary roles in the field of current drug design and development.
Compounds containing heterocyclic scaffolds, both of natural and synthetic origin, are (notably)
biologically active and extensively used in clinical practice.1 The benzofuran core might be
considered one of crucial heterocycles, as being incorporated into a structure of many
pharmacologically effective compounds.2 Due to the wide range of beneficial biological activities
connected with this moiety, synthesis, physicochemical and biological parameters determination and
comprehensive structure–activity relationships have deeply attracted the interest of pharmaceutical /
medicinal chemists, culminating in the discovery of several molecules that are effectively used as the
treatment option(s) for many diseases.3 The presented research focused on a multistep synthesis,
spectral characteristics (1H NMR, 13
C NMR, and IR) estimation and several physicochemical
descriptors (solubility in various solvents, melting point values, and thin-layer chromatography
analysis) determination of the compound containing a given privileged scaffold – (variously
substituted)benzofuran-2-yl-(4-phenyl-/4-diphenylmethylpiperazine-1-yl)methanones.

The study was supported by an Excellent Grant of Comenius University Bratislava no.
UK/3033/2024.

References
1. ROTELLA, D. P.: Advances in Heterocyclic Chemistry, 23(1), 2004, 51–58.
2. HIREMATHAD, A., PATIL, M. R., CHETHANA, K. R., et al.: RSC Advances, 5(117), 2015,
96809–96828.
3. CHAND, K, HIREMATHAD, A., SINGH, M., et al.: Pharmacological Reports, 69(2), 2017, 281–
295.

30
 P27
SYNTHESIS AND STABILIZATION OF SILVER NANOPARTICLES BY
ZWITTERIONIC GEMINI SURFACTANTS

OLÁHOVÁ, K.,1 PISÁRČIK, M.,1 ZÁTEKOVÁ, M.,1

1
Department of Chemical Theory of Drugs, Faculty of Pharmacy in Bratislava, Comenius University, Slovak
Republic

e-mail: [email protected]

Dialkylphosphocholines are groups of zwitterionic gemini compounds with a wide spectrum of

biological properties such as cytotoxic, anticandidal and antiamoebal activity. Molecules of
compounds used in our study have two dissymmetric alkyl chains of variable length ranging from 2
to 15 carbon atoms, while the sum of carbon atoms in both chains is being kept constant. These novel
zwitterionics were used for the stabilization of silver nanodispersions. Synthesis and investigations
of physical properties of phosphorus-based surfactants were studied recently1 as well as they were
applied as potent stabilizers of silver nanoparticles (AgNPs).2 In our study, formation and
composition of dialkylphosphocholine-capped AgNPs were analysed by the changes in the UV-VIS
spectra due to the plasmon resonance effect. AgNPs size was determined using dynamic light
scattering method and nanoparticle surface charge was analyzed by zeta potential measurements. As
the results indicate, the ability of dialkylphosphocholine gemini surfactants to efficiently stabilize
AgNPs strongly depends on the molecular structure of the used surfactants, especially on the length
of both alkyl chains in zwitterionic surfactant molecule. The formation of stable zwitterionic
surfactant-capped AgNPs will allow their passage through biological membranes and provides
desired therapeutical effect related effective distribution of silver.
References
1. TIMKO, L., PISÁRČIK, M., MRVA, M., et al.: Synthesis, physicochemical properties and
biological activities of novel alkylphosphocholines with foscarnet moiety. Bioorg. Chem.,
104, 2020, 1‒12.

2. PISÁRČIK, M., LUKÁČ, M., JAMPÍLEK, J., et al.: Silver Nanoparticles Stabilized with
Phosphorus-Containing Heterocyclic Surfactants: Synthesis, Physico-Chemical Properties and
Biological Activity Determination. Nanomaterials, 11, 1883, 2021, 1‒17.

31
 P28
STRUCTURAL CHARACTERIZATION AND CYTOTOXICITY OF SILVER(I)
COMPLEXES WITH N-METHYLPHENOTHIAZINE

PANTOVIĆ, B.V.,1 ANDREJEVIĆ, T.P.,1 STEVANOVIĆ, N.LJ.,1 MARKOVIĆ, V.R.,1 AŠANIN,

D.P.,2 ILIĆ-TOMIĆ, T.,3 NIKODINOVIĆ-RUNIĆ, J.,3 GLIŠIĆ, B.Đ.1

1
Department of Chemistry, Faculty of Science, University of Kragujevac, R. Domanovića 12, 34000 Kragujevac,
Serbia
2
Institute for Information Technologies Kragujevac, University of Kragujevac, Jovana Cvijića bb, 34000 Kragujevac,
Serbia
3
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11042
Belgrade, Serbia

e-mail: [email protected]

In the reaction of equimolar amounts of N-methylphenothiazine (N-Mephtz) and silver(I) salts,

(AgSbF6 for 1 and AgPF6 for 2) carried out in ethanol under reflux for 2 h, two mononuclear silver(I)
complexes were obtained, [Ag(N-Mephtz)4)]SbF6 (1) and [Ag(N-Mephtz)4]PF6 (2). The synthesized
complexes were structurally characterized by spectroscopic (IR, NMR, UV-Vis) methods, while their
crystal structure was determined by single-crystal X-ray diffraction analysis. In these complexes, four
N-Mephtz ligands are monodentately coordinated to the Ag(I) ion through the sulphur atom, forming
a catonic [Ag(N-Mephtz)4]+ species with [SbF6]– and [PF6]– acting as counter anions for 1 and 2,
respectively. The cytotoxicity of the silver(I) complexes was evaluated on human fibroblasts
(MRC5), human colorectal carcinoma (HCT116) and human lung carcinoma (A549) cell lines. The
interactions of complexes 1 and 2 with calf thymus DNA (ct-DNA) and bovine serum albumin (BSA)
were studied to evaluate their binding affinity toward these biomolecules. In this study, we have also
performed fluorescence competition experiments with site markers for BSA to locate the bindig site
of the investigated complexes to this protein.

The study was supported by the Science Fund of the Republic of Serbia, Grant No. 7730810, Value-
added biologics through eco-sustainable routes – BioECOLogics and Ministry of Science,
Technological Development and Innovation of the Republic of Serbia (Agreements No. 451-03-
65/2024-03/200122, 451-03-66/2024-03/200122 and 451-03-68/2022-14/200378, 451-03-66/2024-
03/200042).

32
 P29
LOW-SYMMETRICAL PHTHALOCYANINES
FOR PHOTODYNAMIC THERAPY

RODRIGUES, L. D., NOVAKOVA, V.,

Department of Pharmaceutical Chemistry and Pharmaceutical analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Photodynamic therapy is an effective form of cancer therapy involving a photochemical reaction with
a light activatable molecule – a photosensitizer, light, and molecular oxygen. But optimal
photosensitizer is yet to be developed. Phthalocyanines are aromatic macrocycles having 18 π-
electron and have been shown to be optimal photosensitizers from the spectral and photophysical
point of view1.The macrocylic core can be modified to achieve better targeting, enhanced water
solubility and to have increased singlet oxygen production.
In this project we aim to synthesize low-symmetrical phthalocyanine through the Linstead method
using Mg(BuO)2 as the initiator of the reaction with phthalonitrile bearing alkylsulfanyl or
arylsulfanyl groups of different bulkiness as one of the precursors. Another precursor carries carboxyl
or azide group. Introduction of one such functional group into phthalocyanine can be used for
attaching to a targeting moiety. The alkylsulfanyl groups are known to have better shift in the
absorption spectra and improved singlet oxygen production2.

The study was supported by ERC-CZ Project Number- LL2318

References
1. CLAESSENS, C. G., et al., The Chemical Record, 8(2), 2008, 75-97.
2. NOVAKOVA V., et al., Coordination Chemistry Reviews, 361, 2018, 1-73.

33
 P30
MAGNETIC NANOPARTICLES COATED WITH MODIFIED CHITOSAN AND
HEMOGLOBIN AS POTENTIAL DRUG CARRIERS IN PHOTODYNAMIC
THERAPY (PDT)

SMOLARKIEWICZ-WYCZACHOWSKI, A.,1 KOZERSKA, K.,1 MYLKIE, K.,1 ZIMČÍK,

P.,3 ZIEGLER-BOROWSKA,M.1

1
Department of Biomedical and Polymer Chemistry, Faculty of Chemistry, Nicolaus Copernicus University in Torun,
Republic of Poland
2
Academia Copernicana Inderdysciplinary School. Nicolaus Copernicus University in Torun, Republic of Poland
3
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

The research is focused on finding carriers for photosensitive drugs that can be used in photodynamic
therapy (PDT) that belongs to the established methods of treating cancer1. For this purpose, magnetic
nanoparticles coated with modified chitosan and hemoglobin were designed. The material was
characterized in terms of chemical structure using infrared spectroscopy (ATR-FTIR) and X-Ray
diffraction (XRD). The surface morphology was described based on images from scanning electron
microscopy (SEM) and scanning transmission electron microscopy (STEM). The size of
nanoparticles was determined using dynamic light scattering (DLS). The next step was to attach a
photosensitive drug, chlorin e6, using chemical and physical adsorption. The material was
characterized. The potential photosensitivity of nanoparticles with the drug was tested (based on the
distribution of the singlet oxygen scavenger ADPA), finding that the presence of hemoglobin on the
nanoparticles increases the photosensitive effect. The release profile of chlorin e6 from the material
to which the drug was physically bound was also analyzed. It was noted that the presence of
hemoglobin slowed down the release process of chlorin e6.

References
1. BECHET, D., TRENDS IN BIOTECHNOLOGY, 26, 2008, 612-621.

34
 P31
ANTIMICROBIAL AND ANTITUBERCULAR ACTIVITY OF SILVER(I) AND
GOLD(III) COMPLEXES WITH MICONAZOLE

STEVANOVIĆ, N. LJ.,1 SRIRAM, D.,2 SKARO BOGOJEVIC S.,3 DJURAN, M. I.,4 AND
GLIŠIĆ, B.Đ.1

1
Department of Chemistry, Faculty of Science, University of Kragujevac, R. Domanovića 12, 34000 Kragujevac, Serbia
2
Medicinal Chemistry and Antimycobacterial Research Laboratory, Pharmacy Group, Birla Institute of Technology &
Science-Pilani, Hyderabad, India
3
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, 11042 Belgrade,
Serbia
4
Serbian Academy of Sciences and Arts, Knez Mihailova 35, 11000 Belgrade, Serbia
e-mail: [email protected]

New silver(I) and gold(III) complexes with the antifungal drug miconazole (mcz), [Ag(NO3)(mcz)2]
(1) and [AuCl3(mcz)] (2), were synthesized by the reaction of AgNО3 and K[AuCl4] with an
equimolar amount of mcz in ethanol. The synthesized complexes 1 and 2 were characterized by mass
spectrometry, IR, UV-Vis and 1H NMR spectroscopy. Complex 1 contains two mcz and one
monodentately coordinated nitrate anion. In the case of complex 2, mcz is a monodentate ligand
coordinated to the Au(III) ion via the triazole nitrogen atom, while the remaining coordination sites
of this metal ion are occupied by chloride anions leading to the square-planar arrangement. The
antimicrobial activity of complexes 1 and 2 was investigated against different bacterial and fungal
strains, as well as their cytotoxic activity on the normal human lung fibroblast cell line (MRC-5).
These complexes were assessed for their in vitro antimycobacterial activity against Mycobacterium
tuberculosis (MTB) H37Rv (ATCC 27294) strain. Coordination of mcz to Ag(I) and Au(III) ions led
to enhancement of its activity against Gram-negative Escherichia coli and Pseudomonas aeruginosa
strains, while against the panel of Staphylococcus aureus and Candida species, only complex 2 has
improved activity in respect to mcz. Both complexes 1 and 2 demonstrated good antitubercular
activity, whereby 1 is twice potent than parent mcz drug.

The study was supported by the Ministry of Science, Technological Development and Innovation of
the Republic of Serbia (Agreements No. 451-03-66/2024-03/200042, 451-03-65/2024-03/200122 and
451-03-66/2024-03/200122).

35
 P32
NOVEL ISATINS DERIVATIVES

R. ŠANDRIK 1, D. NÁDASKÁ 1, I. MALÍK 1, F. ANDRIAMAINTY 1

1
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University, Odbojárov 10, 832 32,
Bratislava, Slovak Republic; [email protected]

The research is focused on the synthesis, characterization and study of the biological properties of
isatin derivatives as suitable candidates for new drugs. Isatin and its derivatives belong to the organic
compounds used in chemical practice. They are used primarily in areas such as medicinal chemistry
(e.g. as antibiotics, antimalarials, etc.), in the field of nanotechnology, development of analytical
reagents and dyes, and in the field of synthesis of heterocyclic compounds and stereoselective
procedures. The high application potential of isatin and its derivatives, their occurrence and the
occurrence of their metabolites in plants and in the human body have aroused the great interest of
chemists, doctors and pharmacists in the study of their chemical reactivity. [1] The aim of the work
was the synthesis of new, not yet described in the literature, derivatives of 3-(phenylhydrazono)isatin
with a methylpiperazine substituent in the N-1 position. [2] [3]

The study was supported by APVV-22-0133.

References:
[1] Nath, R.; Pathania, S.; Grover, G.; Akhtar, M. J. J. Mol. Struct. 2020, 1222, 128900.
[2] Pisani, L.; De Palma, A.; Giangregorio, N.; Miniero, D. V.; Pesce, P.; Nicolotti, O.; Campagna,
F.; Altomare, C. D.; Catto, M. Eur. J. Pharm. Sciences 2017, 109, 381-388.
[3] Polychronopoulos, P.; Magiatis, P.; Skaltsounis, A. L.; et al. J. Med. Chem. 2004, 47, 935-946.

36
 P33
SYNTHESIS OF NEW DERIVATIVES WITH CARBAMATE AND
ARYLOXYAMINOPROPANOL PHARMACOPHORE AS CHOLINESTERASE
INHIBITORS WITH β-ANTIADRENERGIC ACTIVITY

UNGVARSKÁ MAĽUČKÁ, L.,1,2 CSÖLLEI, J.2

1
Department of Chemistry, Biochemistry and Biophysics, University of Veterinary Medicine and Pharmacy in Košice,
Slovak Republic
2
Department of Chemical Drugs, Faculty of Pharmacy in Brno, Masaryk University, Czech Republic

e-mail: [email protected]

Today, carbamates make structural and/or functional part of many drugs and prodrugs approved and
marketed for the treatment of various diseases such as cancer, epilepsy, HIV infection, and
Alzheimer’s disease.1,2 β-adrenergic antagonists (β-blockers) with at least one chiral center are an
exceedingly important class of drugs used mostly to treat cardiovascular diseases. At least 70 β-
blockers have been investigated in history. Chemically, these drugs contain in their structure an
aryloxyaminopropanol fragment.3-5 The work deals with the synthesis of new derivatives, which
contain in molecule the above mentioned pharmacophoric groups. It clarifies the structure activity
relationship of selected derivatives which have undergone modification in the aryloxyaminopropanol
chain (replacement Ar-O-CH2(OH)-CH2-NR1R2 for Ar-COO-CH2(OH)-CH2-NR1R2 group). The
basic part of this chain was also chemically modified (-NR1R2); benzylpiperidine was replaced by
various substituted benzylpiperazines. For selected derivatives, anticholinesterase activity were
determined. The effect on β-adrenergic signaling was also tested.

References
1. MATOŠEVIĆ, A., BOSAK, A.: Arh. Hig. Rada Toksikol., 71, 2020, 285‒299.
2. GHOSH, A. K., BRINDISI, M.: J. Med. Chem., 63, 2020, 2751–2788.
3. YANG, Y., WANG, Y., BAO, Z., et al.: Molecules, 17, 2021, 1‒23.
4. OGRODOWCZYK, M., DETTLAFF, K., JELINSKA, A.: Mini Rev. Med. Chem., 16, 2016, 40‒
54.
5. BEKHRADNIA, A. R., EBRAHIMZADEH, M. A.: Med. Chem. Res., 21, 2012, 2571–2578.

37
 P34
STUDY OF PHYSICAL PROPERTIES OF SILVER NANOPARTICLES
STABILIZED BY HEXADECYLPHOSPHOCHOLINE DERIVATIVES

ZÁTEKOVÁ, M.,1 PISÁRČIK, M.,1 OLÁHOVÁ, K.1

1
Department of Chemical Theory of Drugs, Faculty of Pharmacy in Bratislava, Comenius University, Slovak Republic

e-mail: [email protected]

Silver nanoparticles (AgNPs) were prepared by chemical reduction from silver nitrate and stabilized
with a group of zwitterionic surfactant molecules composed of two alkyl chains with unequal variable
length. The hydrophilic part of surfactant molecule contains a positively charged ammonium and a
negatively charged phosphate group. The class of cationic surfactants based on quaternary nitrogen
and phosphorus turned out to be a potent stabiliser of silver nanoparticles.1-3 The presented research
is focused on the utilization of zwitterionic dialkylphosphocholines as stabilizers of AgNPs because
of their smaller toxicity and better biocompatibility. Moreover, zwitterionics are important from the
point of view of biological applications. Besides cytotoxic and antineoplastic activity
alkylphosphocholines also possess antiprotozoal properties. In our study, dialkylphosphocholine-
capped AgNPs are characterized by multiple physical methods such as UV-VIS spectrometry, zeta
potential measurements for surface charge determination, and dynamic light scattering for
hydrodynamic size of AgNPs. Our results indicate that the efficiency of dialkylphosphocholine
gemini surfactants in stabilizing AgNPs strongly depends on surfactant molecular structure,
particularly on the difference in length of both alkyl chains in zwitterionic surfactant molecule.
Moreover, the performed experiments indicate that interchanging the location of both alkyl chains of
unequal length in surfactant molecule dramatically affects their stabilizing efficiency of AgNPs.

References
1. PISÁRČIK, M., LUKÁČ, M., JAMPÍLEK. J. et al.: Nanomaterials, 11, 2021, 1883.
2. PISÁRČIK, M., LUKÁČ, M., JAMPÍLEK. J. et al.: Journal of Molecular Liquids, 314, 2020,
113683.
3. PISÁRČIK, M., JAMPÍLEK. J., LUKÁČ, M. et al.: Molecules, 22, 2017, 1794.

38
 P35
POLYPHARMACOLOGICAL DRUG-LIKE MOLECULES FROM
THE 2,5-DIMETHYL-4-OXO-3,4-DIHYDROTHIENO[2,3-d]PYRIMIDINE-6-
CARBOXAMIDE SERIES

VLASOV, S.V.,1 SEVERINA H. I.,1 BORYSOV, O. V.,2 VLASOVA, O.D.,1 GEORGIYANTS, V.

A.,1 ABU SHARKH, A.I.M.1

1
Department of pharmaceutical chemistry, National University of Pharmacy, Kharkiv, Ukraine
2
Enamine Ltd., Kyiv, Ukraine

e-mail: [email protected]

A considerable number of biologically active molecules are capable of binding to several targets in
the body. Therefore, there is a question modern drug discovery is about modifications of privileged
scaffolds, which are characterized by a wide spectrum of pharmacological activity.
Thieno[2,3-d]pyrimidine heterocyclic system, which is a fragment of the molecules of a large number
of antimicrobial agents, with the potential to interact with bacterial TrmD, is among those scaffolds.
On the other hand, this heterocyclic system is close in structure to purine and may be attractive from
the point of view of interaction with adenosine receptors. In order to broaden the range of potential
biologically active compounds, we conducted the synthesis of 2,5-dimethyl-4-oxo-3,4-
dihydrothieno[2,3-d]pyrimidine-6-carboxamides. The key stage was the preparation of ethyl 2,5-
dimethyl-4-oxo-3,4-dihydrothieno[2,3-d]pyrimidine-6-carboxylate within the one-reactor procedure
using 1,1-dimethoxy-N,N-dimethylmethanamine with futher cyclization of the resulting amidine with
ammonium acetate. Subsequently, the corresponding amides were synthesized from the acid obtained
on the basis of hydrolysis of the ester.
The best parameters of antimicrobial activity were found for unsubstituted benzylamide. At the same
time, the slightly less active at in vitro screening 4-methylbenzylamide showed better placement
parameters in the active site of TrmD isolated from Pseudomonas aeruginosa. It is interesting that
this particular amide also turned out to be the best ligand for the A2A adenosine receptor active site in
comparison to Istradefylline as a reference ligand.
The study was supported by The Ministry of Health Care of Ukraine for the State Budget grant on the
topic “Molecular modeling and synthesis of innovative pyrimidine derivatives as promising agents
for the treatment of neurodegenerative diseases” (State registration number: 0124U002006).

39
PHARMACEUTICAL ANALYSIS AND BIOANALYTICAL CHEMISTRY

40
 P36
DNA/BSA INTERACTIONS OF PALLADIUM(II) COMPLEXES WITH
PHENOTHIAZINE AND N-METHYLPHENOTHIAZINE

AŠANIN, D.P.,1 MARKOVIĆ, V.R.,2 ANDREJEVIĆ, T.P.,2 STEVANOVIĆ, N.LJ.,2 PANTOVIĆ,

B.V.,2 GLIŠIĆ, B.Đ.2

1
Institute for Information Technologies Kragujevac, University of Kragujevac, Jovana Cvijića bb, 34000 Kragujevac,
Serbia
2
Department of Chemistry, Faculty of Science, University of Kragujevac, R. Domanovića 12, 34000 Kragujevac,
Serbia

e-mail: [email protected]

The potential use of palladium-based drugs in the treatment of some types of cancers is well
recognized, whereby the interactions of palladium(II) complexes with biomolecules play an important
role in the mechanism of their antitumor action.1 Considering this, in the present study, we have
investigated the binding affinity of phenothiazine (phtz) and N-methylphenothiazine (N-Mephtz) and
their palladium(II) complexes, trans-[PdCl2(phtz)2] (Pd1) and trans-[PtCl2(N-Mephtz)2] (Pd2) to calf
thymus DNA (ct-DNA) in the presence of an intercalative agent ethidium bromide (EthBr) and minor
groove binder Hoechst 3325 (Hoe) by fluorescence emission spectroscopy. Moreover, fluorescence
competition experiments with site markers, eosin Y, ibuprofen and digitoxin, for bovine serum
albumin (BSA) were performed to determine the binding site of the investigated compounds to this
protein.

The study was supported by the Science Fund of the Republic of Serbia, Grant No. 7730810, Value-
added biologics through eco-sustainable routes – BioECOLogics. This research has received funding
from the Ministry of Science, Technological Development and Innovation of the Republic of Serbia
(Agreements No. 451-03-65/2024-03/200122, 451-03-66/2024-03/200122 and 451-03-68/2022-
14/200378).

References
1. KARAMI, K., MEHVARI, F., RAMEZANZADE, V. et al.: J. Mol. Liq., 362, 2022, 119493.

41
 P37
ANALYSIS OF IMPURITIES AND CONTAMINANTS IN PHARMACEUTICAL
PRODUCTS

BARON, D.,1 PAPOUŠKOVÁ, B.1

1
Department of Analytical Chemistry, Faculty of Science, Palacký University in Olomouc, Czech Republic

e-mail: [email protected]

Identification and accurate determination of naturally occurring impurities and/or contaminants is an

essential part of analyses of pharmaceutical products which has to meet mandatory strict criteria
according to the various regulatory authorities.1
The first part of the presented work focuses on the transfer and further optimization of an HPLC-PDA
method for the determination of impurities in a mucolytic drug. The initial method, obtained from a
commercial laboratory, encountered issues in our laboratory, particularly with insufficient resolution
between the peaks of individual impurities.2 It was discovered that selectivity could be significantly
improved by adding methanol to the mobile phase. The final mobile phase consisted of
methanol/acetonitrile/10 mM ammonium acetate in a 10/30/60 % v/v/v ratio.
The second part of the study was focused on optimizing a chromatographic method for detecting
impurities in pharmaceutical formulations, where the initial HPLC-PDA-MS approach faced
challenges with effective analyte ionization due to an alkaline mobile phase. To resolve this, a UPLC-
PDA-MS/MS method was developed, utilizing an acidic mobile phase that improved ionization in
positive electrospray mode, where separation efficiency and the sensitivity of impurity detection were
significantly enhanced. This approach led to the successful identification of key contaminants,
including 2-hydroxy-methylpropiophenone, which was confirmed through detailed MS analysis and
elemental composition reports. It concluded that the impurities were transferred from printed foil to
the medication during packaging, highlighting the importance of material compatibility in
pharmaceutical production.

References
1. NATH, D., SHARMA, B.: Curr. Pharm. Anal., 15, 2019, 669–680.
2. ICH Q2(R2), 2023. Validation of analytical procedures. International council for harmonisation
of technical requirements for pharmaceuticals for human use, Geneva.

42
 P38
OPTIMIZATION AND VALIDATION OF A RAPID HPLC-ELSD METHOD FOR
CARBOHYDRATES ANALYSIS

CRHA, T., PAZOUREK, J. 1

1
Department of Chemical Drugs, Faculty of Pharmacy Brno, Masaryk University, Czech Republic

e-mail: [email protected]

HPLC in the HILIC (hydrophilic interaction liquid chromatography) mode is often used in pharmacy
to separate and determine polar analytes (in our case, saccharides on a polyol stationary phase).1
However, a conventional UV–detector is unusable due to a lack of chromophores; alternative
detectors, such as a refractometric detector, would require isocratic elution, and therefore, an
evaporative light scattering detector (ELSD) can be used to advantage.2 Nowadays, environmentally
friendly chemistry is quite a topic of discussion. That is why we performed experiments with a shorter
column.3 In this paper, column HALO-Penta HILIC 75x4.6 mm with particle 2.7 µm (AMT) was
used for the optimization of separation mixture of common saccharides present in food and food
beverages (fructose, glucose, saccharose, lactose, maltodextrins). To observe analyte retention,
ribose, and melezitose were added to the saccharide mixture. An HPLC Dionex 3000 (Thermo) with
an ELSD Varian 380-LC (Varian) was used. A multivariable approach in combination with
Artificial Neural Networks (ANNs) was used for optimization. Central composite design with three
parameters: column temperature (10–40°C), mobile phase flow rate (0.2–1 mL.min-1) and gradient
ramp (2-5 %.min-1) was used. Sixteen HPLC methods were created with combinations of the three
inputs. Retention time, peak area, and peak high of ribose, glucose, saccharose, and melezitose were
used as outputs. The best match of predicted outputs with experimentally obtained data was chosen
as optimal chromatographic conditions for our sugar mixture separation. This chromatographic
method was further validated by ICH Q2 (R2) guideline.
The study was supported by an SVV grant (MUNI/A/1452/2023).
References
1. ALPERT, A.: J. Chromatogr. A, 499, 1990, 177–196.
2. CRHA, T., PAZOUREK, J.: Foods, 9, 2020, 1164.
3. LAWLOR, K., CLAUSEN, J., JOHNSTON, A., et al.: J. Chromatogr. A, 1721, 2024, 464803.

43
 P39
OPTIMISATION OF MIDDLE-UP QUANTIFICATION OF INFLIXIMAB IN
PHARMACEUTICAL MATRIX BY CAPILLARY ELECTROPHORESIS-MASS
SPECTROMETRY

HAVLIKOVA, J.,1,2 MARAKOVA, K.,1,2 MIKUS, P.1,2

1
Comenius University Bratislava, Faculty of Pharmacy, Department of Pharmaceutical Analysis and Nuclear Pharmacy,
Bratislava, Slovakia
2
Comenius University Bratislava, Faculty of Pharmacy, Toxicology and Antidoping Centre, Bratislava, Slovakia

e-mail: [email protected]

Infliximab (IFX) is a chimeric mouse-human therapeutic monoclonal antibody (mAb) against tumor
necrosis factor (anti-TNF), commonly used in treatment of inflammatory bowel disease. IFX is,
structurally, a large protein of size approximately 150 kDa consisting of two types of subunits – two
heavy (50 kDa) and two light (25 kDa) chains connected by disulfide bonds. Typically, bottom-up
approach is employed for mAb analysis, where mAbs are enzymatically digested into smaller
peptides and subsequently analysed. This work makes use of an alternative, middle-up (MU)
approach, where the mAb disulfide bonds are reduced by a reducing agent into heavy and light chains.
An MU procedure is used together with capillary electrophoresis-mass spectrometry (CE-MS) for
quantitative analysis of IFX in pharmaceutical matrix. Stock solution of IFX reference standard
(Sigma-Aldrich, US) (1 mg/mL in water) was prepared and IFX disulfide bonds were reduced by
tris(2-carboxyethyl)phosphine (TCEP). The reduced IFX was subsequently analysed by Agilent 7100
CE system coupled to an Agilent 6410 triple quadrupole mass spectrometer (Agilent, Santa Clara,
US). Background electrolyte systems (BGE) containing 0,5 M and 1 M formic acid and acetic acid
in the range of 2 – 4 M were tested together with different separation voltages applied (18 – 25 kV).
Various concentrations of IFX stock solutions (0,01 – 0,5 mg/mL) were analysed to plot a calibration
curve. The obtained results show that the most suitable BGE for analysis of reduced mAb IFX is 1 M
formic acid and the optimal separation voltage to be 25 kV. Furthermore, a calibration curve was
plotted from the acquired data showing that the proposed proof-of-concept quantitative method
requires further optimisation.

The study was supported by VEGA 1/0514/22 and VEGA 1/0483/20.

44
 P40
RAPID DETERMINATION OF NIMESULIDE IN PHARMACEUTICAL
FORMULATIONS BY CAPILLARY ZONE ELECTROPHORESIS (CZE)

HO, C. V.,* PAZOUREK, J.

Department of Chemical Drugs, Faculty of Pharmacy Brno, Masaryk University, Czech Republic

*e-mail: [email protected]

Nimesulide is a relatively COX-2 selective, non-steroidal anti-inflammatory drug (NSAID) with

analgesic and antipyretic properties. Its approved indications are the treatment of acute pain, the
symptomatic treatment of osteoarthritis, and primary dysmenorrhoea in adolescents and adults above
12 years old1. In several countries, nimesulide has been withdrawn from the market due to concerns
about the risk of hepatotoxicity; however, it is still available in many countries, esp. in Europe and
Asia, even as an over-the-counter (OTC) drug.
A rapid method for determining nimesulide by CZE in various dosage forms (powder, tablets, gel)
was developed. Experimental conditions were revised and simplified.2 Under optimized conditions
using sodium salicylate as an internal standard, the run time was less than 5 minutes. Also, the sample
preparation is straightforward: tablets/powder are homogenized in a mortar, suspended in methanol,
and filtrated. In gel samples, the filtration step can be omitted. The method was applied to samples of
three products available in the Czech market: Nimesil (sachets), and Aulin (tablets, gel), including
the content uniformity test (Pharmacopoeia 11.0) of Nimesil sachets.

An example of electropherogram of nimesulide and salicylate under optimized conditions

References
1. KWON, J., KIM, S., YOO, H., LEE, E.: PLOS ONE, 14(1), 2019, e0209264.
2. DOGRUKOL-AK, D., TUNCEL, M., ABOUL-ENEIN, H. Y.: Journal of Separation Science,
24(9), 2001, 743-748.

45
 P41
CHIRAL SEPARATIONS OF NEW POTENTIAL PHARMACOPHORES: NIDO-
[7,8-C2B9H12]- AND [Co(C2B9H11)2]- DERIVATIVES

O. HORÁČEK1, B. GRÜNER2, R. KUČERA1

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Institute of Inorganic Chemistry, Czech Academy of Sciences, Czech Republic

e-mail: [email protected]

Anionic derivatives of nido-[7,8-C2B9H12]- and [Co(C2B9H11)2]- are known for their stability, water
solubility, and low toxicity, finding applications in catalysis, synthetic, and medicinal chemistry.
Despite the vast investigations of these compounds, their chirality is currently underexplored,
hindering the development of new pharmaceuticals. Addressing this knowledge gap is crucial to
understand the possibly different pharmacological activity and toxicity of drugs based on individual
enantiomers of nido-[7,8-C2B9H12]- and [Co(C2B9H11)2]-.
In this presentation, the investigations of the chromatographic behavior of derivatives of nido-[7,8-
C2B9H12]- and [Co(C2B9H11)2]- are discussed using HPLC and SFC using polysaccharide-based
columns and beta-cyclodextrin-based columns. The anionic derivatives of nido-[7,8-C2B9H12]- were
previously unseparated in HPLC due to ionic interactions with the silica-gel surface, which were
mitigated by addition of counterions or chelating agents, achieving chiral separation on the beta-
cyclodextrin-based chiral stationary phase. The zwitterionic derivatives of nido-[7,8-C2B9H12]- were
separated on the same column without the evidence of strong ionic interactions with the surface of
the silica-gel.
After the initial experiments, we tested multiple chiral stationary phases and chromatographic modes
in HPLC and SFC. In general, SFC methods using polysaccharide-based columns are preferred for
zwitterionic and anionic derivatives of [Co(C2B9H11)2]- and zwitterionic nido-[7,8-C2B9H12]-.
However, for anionic derivatives of nido-[7,8-C2B9H12]-, RPLC using a bromated-beta-cyclodextrin-
based chiral stationary phase is the method of choice.
In conclusion, this work lays the foundation for chiral separations of multiple structural types of
derivatives of nido-[7,8-C2B9H12]- and [Co(C2B9H11)2]-, essential for analysis of enantiomeric purity
and development of chiral pharmaceuticals.
This work was supported by the Charles University (SVV 260 666).

46
 P42
RP-HPLC METHOD FOR DETERMINATION OF SALBUTAMOL SULFATE
AFTER APPLICATION FROM A PMDI TYPE OF INHALER INTO THE
PHARMACOPOEIAL GLASS APARATURE

KAPUSTÍKOVÁ, I.,1 PEŤOVSKÁ, D.,1 SÍS, E.,2 BOŠKAJOVÁ, M.,2 PIEŠŤANSKÝ, J., 2,3,4
GALBA, J.,1,5 MOLITORISOVÁ, M.2

1
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University in Bratislava, Slovakia
2
Department of Galenic Pharmacy, Faculty of Pharmacy, Comenius University in Bratislava, Slovakia
3
Toxicologic and Antidoping Centre, Faculty of Pharmacy, Comenius University in Bratislava, Slovakia
4
Institute of Neuroimmunology, Slovak Academy of Sciences, Slovakia
5
Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Slovakia

e-mail: [email protected]

This work presents the study of the release of antiasthmatic salbutamol sulfate from the pMDI type
of inhaler into the pharmacopoeial glass apparatus, which represents a model of human respiratory
tract, while the physiological inhalation is imitated by a vacuum pump. The aim of the experiment
was to determine the amount of the drug trapped in the inhaler and three compartments of the glass
apparatus after application of one dose. 25 applications were analyzed, while a trend was observed
when the least amount of salbutamol was captured in the inhaler (2.9 %), then in the upper separation
chamber (10.9 %), the throat (38.6 %), and the largest portion of drug dose was captured in the lower
separation chamber (47.6 %), which represents lower respiratory tract and lungs, and thus the main
site of drug’s therapeutic effect.
Quantitative analysis was carried out using Dionex UltiMate 3000 System. A Symmetry® C18
column (4.6 × 250 mm, 5 μm) and a mobile phase consisting of methanol and aqueous solution of
phosphoric acid (7.3 mmol L−1) in a volume ratio of 70:30 were used. The flow rate of the mobile
phase was 1 mL min−1, the column temperature was maintained at 40 °C and detection was performed
at a wavelength of 229 nm. As part of the method validation, linearity, detection limit, quantification
limit, intraday and interday precision, accuracy, specificity, selectivity and robustness were evaluated.

This work was supported by the Slovak Research and Development Agency under the Contract no.
APVV-23-0508.

47
 P43
DETERMINATION OF RIFAMPICIN AND ITS METABOLITES IN CELL
MEDIUM

LOCHMAN, L.,1 SMUTNY, T.,2 PAVEK, P.,2 KUCERA, R.,1

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University, Czech
Republic

e-mail: [email protected]

Rifampicin (RIF) is a macrocyclic antibiotic used in the treatment of tuberculosis. In addition, RIF is
known to induce cytochrome P450 enzymes, especially the 3A4 isoform. Due to this property, RIF
is used as a model compound in pharmacological studies. RIF is primarily metabolized into 25-
desacetyl RIF through enzymatic deacetylation and into RIF quinone via nonenzymatic
autooxidation.1 Other metabolites or degradation products, such as 3-formyl RIF and RIF N-oxide,
have also been reported.2 Several HPLC methods for detecting RIF and RIF quinone in human plasma
or urine have been published3. Still, there remains a need for a straightforward LC-UV method for
the determination of RIF and its metabolites.
In our study, we developed an LC-UV method for the detection of RIF and its metabolites. The
separation was performed on an ACE Excel3 C18-AR column (3 µm, 100 × 3 mm). The mobile phase
consisted of ammonium acetate (30 mM, pH 6.30) (A) and methanol (B). The gradient was as follows:
0-9 min (65 → 90% B), 9-11 min (90% B), 11-11.1 min (90 → 65% B), and 11.1-15 min (65% B).
This chromatographic method was effectively used for the quantification of RIF and its metabolites
in a pharmacological study examining the potential of RIF and its derivatives to activate the pregnane
X receptor. Additionally, we will discuss the stability of RIF and RIF quinone in cell medium, both
in the presence and absence of cells.
The study was supported by SVV project (260 666, Czech Republic).

References
1. SHI, F., LI, H., PAN, L., et al.: J. Pharm. Biomed. Anal., 132, 2017, 17–23.
2. SUTRADHAR, I., ZAMAN, M. H.: J. Pharm. Biomed. Anal., 197, 2021, 1‒4.
3. MISHRA, P., PAWAR, R. P., BOSE, D., et al.: Bioanalysis, 11, 2019, 713‒726.

48
 P44
BRIDGING MICROSAMPLING WITH MICROEXTRACTION FOR
DOXORUBICIN AND DOXORUBICINOL DETERMINATION

REGULI, A.,1 BAVLOVIČ PISKÁČKOVÁ, H1., LENČOVÁ, O.,2

KOLLÁROVÁ-BRÁZDOVÁ, P.,2 ŠTĚRBA, M.,2 ŠTĚRBOVÁ-KOVAŘÍKOVÁ, P.,1

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University, Czech Republic

e-mail: [email protected]

Volumetric Absorptive Microsampling (VAMS) facilitates repeated blood collection from a single
animal, minimizing overall animal use in preclinical studies. VAMS devices stand out by relying less
on hematocrit, elevating precision, and making sample collection more straightforward than
conventional dried blood spots. [1] Electromembrane extraction (EME) is a relatively new extraction
technique based on hollow fiber microextraction. The application of direct current voltage in EME
accelerates analyte isolation, leading to increased recoveries within a shorter time, and it also
facilitates the use of a 96-well format. [2]
This study aimed to establish a methodology for assay of doxorubicin (DOX) and its metabolite,
doxorubicinol (DOXol), from blood absorbed onto VAMS tips using EME followed by UHPLC-
MS/MS analysis. The primary focus was on optimizing sample preparation. Various EME parameters
(supported liquid membranes, voltage, time, donor and acceptor solutions) were systematically
explored to attain optimal recovery. After optimization, the method was fully validated. Additionally,
protein precipitation, as a conventional extraction method, was developed and compared to EME.
The method’s applicability was confirmed by analyzing VAMS samples taken after DOX
administration in pharmacokinetic studies involving mice and rabbits.

The study was supported by Charles University (GAUK 232223 and SVV 260666).

References
1. PROTTI, M., MANDRIOLI, R., MERCOLINI, L.: Anal. Chim. Acta, 1046, 2019, 32-47.
2. HUANG, C., CHEN, Z., GJELSTAD, A., et al.: Trends Analyt. Chem., 95, 2017, 47-56.

49
 P45
EXTRACTS FROM THE MEDICINAL MUSHROOM CORDYCEPS SINENSIS
AND THEIR ABILITY TO REDUCE SILVER IONS

UHRINOVÁ, A., UNGVARSKÁ MAĽUČKÁ, L.

Department of Chemistry, Biochemistry and Biophysics, University of Veterinary Medicine and Pharmacy in Košice,
Slovak Republic

e-mail: [email protected]

An entomopathogenic fungus, Cordyceps sp. has been known to have numerous pharmacological and
therapeutic implications, especially, in terms of human health making it a suitable candidate for
ethno-pharmacological use.1 An ever growing list of symptoms remedied using Cordyceps sinensis
include respiratory, renal, liver, nervous system, cardiovascular diseases, cancerous tumors,
decreased libido and even stress, fatigue and aging.2 Silver nanoparticles have attracted vast research
interest because of their potential functions in many fields such as biomedical, environmental,
electronic, catalysis, and antimicrobial areas. Silver nanoparticles prepared from Cordyceps sinesis
extracts exhibit outstanding antibacterial activity, which is needed for many medical and biological
applications.3
This work deals with the preparation of silver nanoparticles using extracts prepared from the fungus
Cordyceps sinensis. The extracts were prepared from samples of the fungus that were cultured on two
different substrates; pea and maize sown. Maceration and ultrasonication were used in the preparation
of the extracts. The formation of silver nanoparticles was monitored spectrophotometrically in the
region of 400-500 nm using 10.0 mmol L-1 AgNO3 solution and elevated temperature over a period
of 1 hour. As shown from the measurements, for all extracts, there was a reduction of silver ions by
the polysaccharides present in the Cordyceps sinensis extracts, which was reflected by a significant
increase in absorbance in the measured area. As revealed from the measurements, the change of
substrate during the preparation of the extracts is an important factor affecting the formation of silver
nanoparticles.
References
1. TULI, H. S., SANDHU, S. S., SHARMA, A. K.: Biotech., 4, 2014, 1‒12.
2. XIN CHEN, P., WANG, S., NIE, S. et al.: J. Funct. Foods, 5, 2013, 550‒569.
3. YANG, X., WU, J. Y.: Materials, 15, 2022, 5620.

50
PLENARY LECTURES

51
 PL-1
THE DESIGN OF PDT CHROMOPHORES

RÜDIGER FAUST

University of Kassel, Institute for Chemistry and CINSaT – Center for Interdisciplinary Nanostructure Science and
Technology, Heinrich-Plett-Str. 40, 34132 Kassel, Germany, [email protected]

Photodynamic tumour therapy (PDT) is a minimally invasive treatment for various forms of cancer
that utilizes the ability of some chromophores to produce cytotoxic singlet oxygen 1O2 when
stimulated by light of a suitable wavelength. While PDT has certain advantages over conventional
therapies (e.g. it is a local therapy with limited dark toxicity) it also has limits, inter alia with respect
to the oxygen environment: Many tumors tend to develop in hypoxic regions, which significantly
reduces the efficiency of photosensitization. In addition, PDT itself further increases the hypoxic state
of the tissue.
We are currently exploring concepts for a delayed for singlet oxygen provision in the design of
molecular catch-and-release systems. At the core of these concepts lies the covalent attachment of
pyridone units on photosensitizing chromophores such as subphthalocyanines or the so-called
BOIMPY chromophores shown in the Figure below. The photosensitization process generates a
pyridone endoperoxide from which singlet oxygen can be released by a thermal process. presentation
will describe our synthetic entry into this field and will highlight recent progress of this ongoing
research.

3O
2

PYR EPO

Δ
PS

52
 PL-2
AUTOMATION OF MATRIX PRECIPITATION FOR FOOD ANALYSIS

HORSTKOTTE, B.1, FIKAROVÁ, K.1, YILIDIRIM, S.2, NOVÁKOVÁ, L.1,

PILAŘOVÁ V1., SKLENAŘOVÁ, H.1

1
Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Karadeniz Technical University, Faculty of Pharmacy, Department of Analytical Chemistry, Farabi Street, 61080,
Trabzon, Turkiye

e-mail: [email protected]

Matrix precipitation is a frequent preparative step used before the analysis food and biological
samples by liquid chromatography. Inefficient removal of particles or precipitable matrix components
(salts, proteins, fibers, …) can cause column blockage or inhibit solid phase extractions. The typically
manual protocols differ mostly by the inducing agent, e.g., chaotropic organics, metal salts, or aprotic
solvents. This is followed by sample filtration or centrifugation to separate the formed precipitate.
Herein, we show centrifugation-less deproteination based on homogeneous liquid-liquid
microextraction, automated by the Lab-In-Syringe technique [1,2].
The developed methodology was applied to milk and serum using selected pharmaceuticals as model
analytes [3,4] and coupled online to HPLC and LC-MS/MS, respectively with quantitative analyte
recovery, high operational reproducibility, and operation in parallel to the running chromatographic
separation. System design, operation principle, method optimization, technical challenges, and
application are shown together with an outlook on current research.

The study was supported by the project EFSA-CDN co-funded by ERDF

(No. CZ.02.1.01/0.0/0.0/16_019/0000841).

References
1. HORSTKOTTE, B., SOLICH, P.: Molecules, 25, 2020, 1612.
2. HORSTKOTTE, B., SUÁREZ, R., SOLICH, P., et al.: Anal. Chim. Acta, 788, 2013, 52‒60.
3. FIKAROVA, K., MACHIÁN D, YILDIRIM, S., et al.: Anal. Chim. Acta, 1233, 2022, 340507.
4. YILDIRIM, S., FIKAROVÁ, K., PILAŘOVÁ, V., et al.: Anal. Chim. Acta, 1251, 2023,

53
 PL-3
DEVEOPMENT OF THE PECAM1 COVALENT BLOCKER AS A NOVEL
THERAPIUTIC APPROACH FOR ANTI-AUTOIMMUNE DISORDERS
TREATMENT

GETTER, T.,1 MARGALIT, R.,2 KAHREMANY-HOFFMAN, S.,1 LEVY-NISSIM, L.,1 BLUM, E.,1
KHAZANOV, N.,1 LAHAV, R.,2 ZILBER, S.,3 SENDEROWITZ, H.,1 BRADFIELD, P.,4 IMHOF, B.
A.,4,5 ALPERT, G.,2 GRUZMAN A.1

1
Department of Chemistry, Faculty of Exact Sciences, Bar-Ilan University, Ramat-Gan, Israel
2
AltA-ZuZ LTD, Ness-Tziona, Israel
3
Department of Pathology, Shaarey-Zedek Hospital, Jerusalem, Israel
4
“MesenFlow LTD”, Geneva, Switzerland
+ 5
Department of Pathology
2 and Immunology, Geneva University, Switzerland.

e-mail: [email protected]

Leukocyte transendothelial migration from the blood to the tissues is one

+
of the most important steps in launching an inflammatory immune
response (its cellular component). However, misregulation of this
process can lead to devastating autoimmune diseases. We designed a
3
molecule (GT-73) with an ability of covalent binding to active form of
PECAM1 (CD31) (Platelet and Endothelial Cell Adhesion Molecule 1).
GT-73 PECAM1 is one of the key players in the regulation of the leukocyte
penetration througth the endothelial barrier. We showed that in vitro GT-73 completely blocked
H2SO4, reflex 5 h. b) K2CO3, KI, CH3CN, 70O C 5 h. c) EtOH/H2O 1:1,
human monocyte, T-lymphocytes, B-lymphocytes and neutrophils transendothelial migration,
barbituric acid, reflex overnight.
without any toxic effects on immune or endothelial cells (IC50=2.4 µM). In vivo, GT-73 exhibited
significant therapeutic effects in inflammatory bowel disease (IBD)/Crohn’s disease, multiple
sclerosis, fatty liver disease, acute respiratory distress syndrome, lupus and rheumatoid arthritis
mouse models. GT-73 was active in pharmacological relevant doses (10-30 mg/kg) and in several
routes of administration and almoust complitlyb elimnated the symptoms of thouse disorders in mice.
The compound was not active in atopic dermatitis and diabetis type one mouse models. A detailed
acute and chronic toxicity profile, including GLP toxicity of GT-73 did not reveal any toxic effects1-
3
.
54
The study was supported by “AltA-ZuZ LTD”, Israel; “MesenFlow LTD”, Switzerland; and
“Silverskate Bio LTD“, Israel/Japan.

References
1. GETTER, T., MARGALIT, R., KAHREMANY, S., et al.: Novel inhibitors of leukocyte
transendothelial migration. Bioorg Chem., 92, 2019, 103250.
2. BLUM, E., MARGALIT, R., LEVY-NISSIM, L., et al.: A potent leukocyte transmigration
blocker: GT-73 showed a protective effect against LPS-induced ARDS in mice. Molecules.
Special Issue: “Chemical Biology” 26, 2021, 4583.
3. GRUZMAN A, GETTER T, IMHOF B, et al.: Novel barbituric acid-based leucocyte
transmigration inhibitors as drug candidates for treating inflammatory diseases, autoimmune
diseases and cancer. Patent WO2019043706A1. The patent was licensing to the company:
“Silverskate Bio LTD”.

55
 PL-4
GENE THERAPY FOR CYSTIC FIBROSIS

MALÍK, I., NÁDASKÁ, D., ŠANDRIK, R., ANDRIAMAINTY, F.

Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University Bratislava, Slovak Republic

e-mail: [email protected]

Recent advancements in the field of genetic engineering with the development of highly efficient
biomedical technology tools and strategies notably renewed the enthusiasm for gene therapy of
multiple severe (difficult-to-treat) diseases / disorders or rare diseases including the ones
characterized by an alteration at the gene level.1 Cystic fibrosis (CF) is a life-shortening inherited
autosomal recessive disease caused by various mutations in the cystic fibrosis transmembrane
regulator (CFTR) gene encoding a cystic fibrosis transmembrane conductance regulator (CFTR)
protein.2 Versatile therapeutic interventions for CF have been proposed, developed or approved for
clinical use generally focused on i) non-gene therapy strategies aiming small molecule drugs (CFTR
modulators) and their suitable combinations which improve trafficking and processing of the mutated
CFTR protein, and ii) gene therapy strategies including the use of antisense oligonucleotides, RNA
interference technology for regulating CFTR gene expression (modulation of gene expression
approaches by interaction with cellular mRNA), CRISPR-based gene editing strategies, base editors
as well as prime editors (genome editing technologies).3–5 Current lecture briefly considered several
aspects of both non-gene and gene therapeutical tools with special attention paid to the latter ones –
their enormous potential in addressing the genetic root of the disease, promises, challenges, pittfalls
and future directions.
References
1. SAYED, N., ALLAWADHI, P., KHURANA, A., et al.: Life Sci., 294, 2022, art. no. 120375, 1–
21.
2. CUTTING, G.: Nat. Rev. Genet. 16, 2015, 45–56.
3. BERGERON, CH., CANTIN, A. M.: Int. J. Mol. Sci., 22, 2021, art. no. 6193, 1–21.
4. LEE, J.-A., CHO, A., HUANG, E. N., et al.: J. Transl. Med., 19, 2021, art. no. 452, 1–15.
5. WANG, G.: Cells, 12, 2023, art. no. 1555, 1–15.
This research was supported by the Project APVV-22-0133.

56
SHORT COMMUNICATIONS

57
 SC-1
J-DIMERS OF PHTHALOCYANINES

ZIMČÍK, P., BEDNÁRIK, Š., DEMUTH, J., NOVÁKOVÁ, V.

Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Phthalocyanines (Pcs) are macrocyclic dyes that tend to form aggregates based on the π-π stacking of
the planar core. Such aggregates are called H-aggregates and typically are characterized by blue-
shifted Q-band and non-fluorescent nature. On the other hand, several papers have reported formation
of slipped J-dimers with retained fluorescence and red-shifted absorption Q-band, however, the
examples are extremely rare.1-3 Our work has been focused on various aza-analogues of Pc (AzaPcs)
that formed the slipped J-dimers on the basis of coordination of peripheral substitutes (various
dialkylamino substituents or heteroaryls) with central cation (Zn(II) or Mg(II)). Unequivocal
dependence between dimerization constant and bulkiness of peripheral binding sites was observed.
The J-dimers can be disassembled to monomers with changes to shape and intensity of fluorescence
and also in production of singlet oxygen using various external stimuli – increased temperature,
presence of external coordinating ligand (see figure below) or protonization of the binding site by
acids.

The work was supported by Czech Science Foundation, grant Nr. 23-06177S
References
1. KAMEYAMA, K., MORISUE, A., SATAKE, A., et al..: Angew. Chem. Int. Ed., 44, 2005, 4763-
4766.
2. GARCÍA-IGLESIAS, M., PEUNTINGER, K., KAHNT, J., et al..: J. Am. Chem. Soc., 135, 2013,
19311-19318.
3. DEMUTH, J., MILETIN, M., KUCERA, R., et al.: Org. Chem. Front., 7, 2020, 445-456.
 SC-2
ASSEMBLY OF PHTHALOCYANINE DERIVATES INTO J-DIMERS

BEDNARIK, S. 1, DEMUTH, J. 1, NOVAKOVA, V. 1, ZIMCIK, P. 1,

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Phthalocyanines (Pcs) are synthetic macrocyclic dyes formed by four isoindoline units connected by
azomethine bridges structurally close to porphyrins. Due to their extended 18 π-conjugated system,
they show unique photophysical properties, and they have been largely investigated in various fields,
such as fluorescence sensors or photosensitizers in photodynamic therapy (PDT).1 However, these
properties are related mostly to the monomeric form of the Pcs only. Aggregation of the Pcs is usually
an unfavorable phenomenon. The planar Pc core tends to aggregate due to π-π stacking interactions.
The most common H-type aggregates (Figure 1c) align molecules into a sandwich-like arrangement,
resulting in increased absorption at blue-shifted wavelengths and strongly decreased fluorescence
emission. However, J-aggregates (Figure 1d) give rise to red-shifted absorption bands and retain
fluorescent properties.2 In this work, we synthesized unsymmetrical Pc derivatives containing one
ligand (coordinating moiety, e.g., pyridyl) that formed slipped J-dimers upon coordination to the
central cation of the second Pc molecule in non-coordinating solvents.
0.3

0. 26 M
Absorbance

0.2 Pyrazine

0.1 0.0 M
Pyrazine

0.0
400 500 600 700 800
Wavelength, nm

The study was supported by Charles University (SVV 260 666).

References
1. NOVAKOVA, V., DONZELLO PIA, M., ERCOLANI, C., et al.: Coord. Chem. Rev., 361, 2018,
1-73.
2. NOVAKOVA, V., HLADIK, P., FILANDROVA, T., et al.: Phys. Chem. Chem. Phys., 16, 2014,
5440-5446.
 SC-3
DERIVATIVES OF 3-AMINOPYRAZINE-2-CARBOXAMIDE AS INHIBITORS
OF PROLYL-tRNA SYNTHETASE

ZITKO, J. 1, PALLABOTHULA, V. S. K.1

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic

e-mail: [email protected]

Prolyl-tRNA synthetase (ProRS), a member of the aminoacyl-tRNA synthetases (aaRSs) family, is

essential for the growth and survival of all cells. The aaRSs are part of both eukaryotes and
prokaryotes, but their significant evolutionary divergence offers the possibility to develop selective
inhibitors. 3-Aminopyrazine-2-carboxamide derivative (1) is a reported ATP-competitive inhibitor of
human ProRS (hsProRS).1 A nitrogen isostere 2 (NCP26) has a strong potential for the treatment of
multiple myeloma2, but also as an antimalarial agent.3 In our laboratory, we prepared several series
of structurally related derivatives (3) and based on the type of the linker and substituent R obtained
compounds with affinity to hsProRS,4 or compounds with antimycobacterial activity (with prpbable
binding to mycobacterial ProRS).5,6 The structure-activity relationships will be discussed.

This research was supported by the Ministry of Health of the Czech Republic, grant nr. NU21-05-
00482.
References
1. ADACHI, R.; OKADA, K.; SKENE, R. et al.: Biochem. Biophys. Res. Commun., 488,
2017, 393–399.
2. KURATA, K.; JAMES-BOTT, A., et al. Blood Cancer Journal, 13. 2023, 12.
3. TYE, M. A.; PAYNE, N. C. et al. Nat Commun, 13, 2022, 4976.
4. PANG, L.; WEEKS, S.D.; JUHÁS, M. et al. Int. J. Mol. Sci., 22, 2021, 7793.
5. PALLABOTHULA, V. S. K.; KERDA, M.; JUHÁS, M. et al. Biomolecules, 12, 2022,
1561.
6. PALLABOTHULA, V. S. K.; ABDALRAHMAN, N. T.; MORI, M. et al. Arch. Pharm.,
357, 2024, e2400171.
 SC-4
AUTOMATION OF LONG-TERM PROCESSES

SKLENÁŘOVÁ, H.1

1
Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Czech Republic

e-mail: [email protected]

Advantages of universal manipulation of sample and reagents were applied in the past for the
liberation of drugs using Franz diffusion unit(s) in a sequential injection (SIA) system. In this talk,
an overview on recent development in this field will be given with respect to two areas where
automated monitoring can significantly deepen knowledge based on the real-time information.
Kinetic profiles of active substances trapped in nanofiber materials made of different polymers and
produced by means of different technologies were examined.1
Similar flow-programming setup based on the SIA concept was proposed for on-line monitoring of
cell membrane permeation.2 The permeation kinetics of the fluorescent marker rhodamine 123 was
studied in a fully automated mode by sampling the apical and basolateral compartments of a 3D
printed permeation module. The effect of the P-glycoprotein transporter inhibitor on the efficiency of
a marker transport across a cell monolayer was investigated in detail. Finally, a 3D printed module
was used for the automation of toxicity assays based on the release of Metridia luciferase upon
interaction with toxic substances and thus changes in cell membrane permeability were studied.3
The study was supported by the project New Technologies for Translational Research in
Pharmaceutical Sciences (NETPHARM, project ID CZ.02.01.01/00/22_008/0004607), co-funded by
the European Union.

References
1. HÁKOVÁ, M., MODEBELU, U., ERBEN, J., MATYSOVÁ, L., ŠATÍNSKÝ, D., ŠVEC, F.,
SKLENÁŘOVÁ, H.: Talanta, 269, 2024, 125415.
2. SKLENÁŘOVÁ, H., ROSECKÁ, M., HORSTKOTTE, B., PÁVEK, P., MIRÓ, M., SOLICH. P.:
Anal. Chim. Acta, 1153, 2021, 338296.
3. AGUINAGA MARTÍNEZ, M.V., JOZIČOVÁ, N., DUŠEK, J., HORSTKOTTE, B., PÁVEK, P.,
MIRÓ, M., SKLENÁŘOVÁ: Talanta, 245, 2022, 123465.
 SC-5
ANALYSIS OF SELECTED DRUGS IN WHOLE BLOOD COLLECTED WITH A
VOLUMETRIC ABSORPTIVE MICROSAMPLING DEVICE

ŠTĚRBOVÁ-KOVAŘÍKOVÁ, P.1, ADAM REGULI, A.1, DUDÁŠOVÁ, P.1, BAVLOVIČ

PISKÁČKOVÁ, H.1, LENČOVÁ,O.2, KOLLÁROVÁ-BRÁZDOVÁ, P.2, ŠTĚRBA, M. 2

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University, Czech Republic

e-mail: [email protected]

Preclinical research plays a crucial role in developing new drugs. However, ethical concerns about
using animals in research have led to the 3Rs (replacement, reduction, and refinement) principles.
Microsampling aligns with the 3Rs by reducing the number of animals needed for a study via
collecting low blood volume from a single animal multiple times, which is particularly beneficial for
small rodents. It also provides the advantage of gentle sample collection and improved data quality.
Volumetric absorptive microsampling (VAMS) is a reliable method for collecting precise whole
blood volume and minimizes the impact of hematocrit on the assay. Analytes are commonly extracted
from VAMS tips by direct desorption, but microextractions have a high potential for this purpose
and offer an ethical and environmentally friendly sample cleanup solution. This presentation focuses
on developing various sample extraction protocols to isolate selected drugs from whole blood
microsamples collected with VAMS devices (10 µl, Mitra®, Neoteryx) for preclinical studies. After
isolation, the extracts are analyzed using UHPLC-MS/MS. Doxorubicin, a clinically used anticancer
drug, and its metabolites doxorubicinol, which are commonly used in preclinical cancer/cardio-
oncology studies, were isolated from the VAMS tips using electromembrane extraction in a 96-well
plate format. The cardioprotective agents, dexrazoxane, and its more potent analog ICRF-193 were
extracted from whole blood microsamples using ultrasonic-assisted desorption to organic solvent.
The reliability of the developed procedures was demonstrated through the analysis of real samples
from PK studies with athymic nude mice.
The study was supported by Czech Science Foundation, (No. 23-06558S) and /NETPHARM
(ID CZ.02.01.01/00/22_008/0004607), co-funded by the European Union.
 SC-6
HPLC AND SFC AS USEFUL TOOLS FOR CHIRAL SEPARATION OF
CARBORANES - PROMISING COMPOUNDS IN DRUG DEVELOPMENT

KUČERA, R.,1 HORÁČEK, O.,1 GRÜNER, B.,2

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Institute of Inorganic Chemistry, Academy of Sciences of the Czech Republic, Řež, Czech Republic

e-mail: [email protected]

Boron cluster compounds represent a distinctive category of highly stable abiotic molecules. In
medicinal chemistry, these clusters are being investigated as innovative isosteres of phenyl rings in
medicinal chemistry.1 Unfortunately, the chirality of boron clusters has been largely overlooked for
the past sixty years. Our research group has recently developed efficient methods for enantiomeric
purity evaluation of these compounds. We conducted a thorough investigation into the primary factors
hindering the successful chiral separations of anionic carboranes in HPLC. Our findings indicate that
cyclodextrin-based stationary phases are effective in separating enantiomers of anionic carboranes.
Notably, we have achieved the enantioseparation of 7,8-dicarba-nido-undecaborate(1-) ions in HPLC
for the first time.2 Our research also demonstrates that polysaccharide-based chiral selectors can
facilitate the chiral separation of anions and zwitterions under reversed-phase conditions. Moreover,
SFC has emerged as a valuable technique for rapid chiral baseline separations, capable of completing
the process within 10 minutes. This method substantially outperforms HPLC in terms of the number
of separated chiral carboranes, resolution, and analysis time. 3,4
In summary, our findings provide essential insights into the development of chiral methods for
anionic and zwitterionic carboranes, with significant implications for drug development.
Furthermore, these results have broader applications in any field where the chirality of these
intriguing molecules is a concern.
References
1. Lesnikowski, Z. J.: Expert. Opin. Drug Discov. 1, 2016, 569-578.
2. Horáček, O., Papajová-Janetková, M., Grüner, B., et al.: Talanta, 222, 2021, 121652.
3. Horáček, O., Marvalová, J., Štilcová, K. et al.: J. Chromatogr. A., 1672, 2022, 463051.
4. Horáček, O., Nováková, L., Tüzün E., et al.: Anal. Chem., 94, 2022, 17551-17558
 SC-7
DEXRAZOXANE - NEW INSIGHT INTO THE MECHANISM OF ACTION OF
THIS CLINICALLY USED DRUG

KARABANOVICH, G.,1 ŠTĚRBOVÁ-KOVAŘÍKOVÁ, P.,1 ŠIMŮNEK, T.,1 ŠTĚRBA, M.,2

ROH, J.1
1
Faculty of Pharmacy in Hradec Králové, Charles University, Akademika Heyrovského 1203, 500 03 Hradec Králové,
Czech Republic
2
Faculty of Medicine in Hradec Králové, Charles University, Šimkova 870/13, 500 03 Hradec Králové, Czech Republic

e-mail: [email protected]

Anthracyclines (ANTs) like doxorubicin (DOX), daunorubicin (DAU) and epirubicin rank among the
most effective anticancer drugs. However, their clinical use is hampered by cardiotoxicity.
Particularly feared are chronic forms, which are largely irreversible and manifest themselves as a
congestive heart failure and dilated cardiomyopathy. Moreover, mechanisms of ANT cardiotoxicity
are not fully elucidated.
Dexrazoxane (DEX) is the only cardioprotective drug approved for clinical use against ANT
cardiotoxicity. Its cardioprotective effects have been well established in experimental settings, but,
surprisingly, its mechanism of action is still not fully understood. Moreover, the absence of any other
cardioprotective agent active against chronic ANT cardiotoxicity in vivo complicates the elucidation
of DEX mechanism of action and thus blocks the rational development of new potent cardioprotective
drug. Traditionally, DEX has been considered as a pro-drug of metal chelating agent ADR-925 which
should prevent ANT-induced oxidative stress and myocardial damage by iron chelation with
subsequent disruption of ROS formation. But DEX is also catalytic inhibitor of topoisomerase II,
which is also a target of ANTs action.
The goal of our research project was to elucidate the mechanism of action of DEX, to investigate its
structure-activity relationships and to develop another effective cardioprotective agent against ANT-
induced cardiotoxicity based on the data obtained.

This work was supported by the NTER-EXCELLENCE project (reg. no. LUAUS24335) and by the
project OncoPharm (CZ.02.01.01/00/23_021/0008442).
 SC-8
DEVELOPMENT OF COVALENT MODULATORS OF MACROMOLECULES:
NOVEL ALKYLATING AGENTS AGAINST GLIOBLASTOMA

ROTHSTEIN A., TRIFONOV L., VISKIND O., AFRI M., KORSHIN E. E., AND GRUZMAN A.

1
Department of Chemistry, Faculty of Exact Sciences in Ramat Gan, Bar Ilan University, Israel

e-mail: [email protected]

Glioblastomas represent 15% of brain tumors; it is the most common and aggressive cancer beginning
within the brain. There is no cure for GBM. Untreated gliobastoma patients have a median survival
of 3 months. Currently the only FDA approved drug for use against GBM is Temozolomide (TMZ)
which has been found to increase life span by a few months when paired with tumor resection and
radiotherapy. TMZ is only useful in high doses, and GBM often becomes resistant to the drug over
time.
Michael acceptors are potential DNA alkylators that have been used for cancer chemotherapy. Based
on the structure of TMZ we designed and synthesized a set of novel, structurally simple, tetrazole-
containing cyclic and acyclic Michael acceptors as potential drug candidates using a combination of
novel and classical synthetic methods. Some of the prepared tetrazole-containing Michael acceptors
exhibited significant anticancer activity in vitro exceeding TMZ potency by a factor of 10 with
considerably lower toxicity toward normal cells. These compounds can be used as candidates for
further development of anti- glioblastoma drugs. SAR (structure activity relationship) studies were
conducted for optimization of activity and specificity. However, the biological molecular mechanism
of action is still unclear and extensive biological studies are required.

This study was supported by Bar-Ilan University President PhD fellowship and by Adama Ltd,
through the Adama Scholarship.
 SC-9
REPLACEMENT OF NITRO FUNCTION BY FREE BORONIC ACID IN NON-
STEROIDAL ANTI-ANDROGENS

ŠLECHTA P.,1 VITÁK, R.,2, KOUCKÁ, K.,1 BERKOVÁ, M.,1 ŽĎÁROVÁ, D.,1 PETRÍKOVÁ,
A.,1 KUNEŠ, J.,4 KUBÍČEK, V.,1 DOLEŽAL, M.,1 KUČERA, R.,2 KUČEROVÁ-
CHLUPÁČOVÁ, M.1

1
Department of Pharmaceutical Chemistry and Pharmaceutical Analysis, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic
2
Department of Pharmacology and Toxicology, Faculty of Medicine in Pilsen, Charles University, Czech Republic
3
Department of Biophysics and Physical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University,
Czech Republic
4
Department of Organic and Bioorganic Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Czech
Republic

e-mail: [email protected]

With the aim to find new compounds for androgen deprivation treatment to combat prostate cancer,
a series of new potential flutamide-like antiandrogens was designed and synthesized. In this work we
present the possibility of replacement the characteristic feature of nitro group by a boronic acid
functionality, in addition, the compounds were variated in the acyl part, in the linker as well as in the
substitution of the benzene ring in the general scaffold 4-nitro-3-trifluoromethylanilide of non-
steroidal androgens. Compound 4-(5-chloropyrazine-2-carboxamido)-2-fluorophenylboronic acid
exhibited higher activity against LAPC-4 than the standards flutamide and bicalutamide and even
lower toxicity against non-cancerous cell line HK-2. The initial in-silico study did not support the
covalent bonding to androgen receptor, which was confirmed in an NMR binding experiment.
The structure-activity relationships obtained in this study might show directions for further research
in the field of non-steroidal antiandrogens.

The study was supported by Charles University project SVV 260 666 (Czech Republic).
“COOPERATIO” program, research area Pharmaceutical Sciences.
References
1. SAMPSON, N., NEUWIRT, H., PUHR, M., et all.: Endocrine-Related Cancer, 20 (2), 2013, R49-
R64.
 SC-10
DESIGNING AND OPTIMIZING NRF2 ACTIVATORS FOR ENHANCED
DERMO-PROTECTION IN OXIDATIVE STRESSRELATED PATHOLOGIES

NISSIM, L.,1 EZGI SARAÇ, B.,2 KAHREAMANY, C.,1 COHEN, D.,3 KARAASLAN, C.,2
GRUZMAN, A.,1

1
Department of Chemistry, Faculty of Exact Science in Ramat Gan, Bar Ilan University, Israel
2
Department of Microbiology, Faculty of Life Science in Ankara, Hacettepe University, Turkey
3
Skin Research Institue, The Dead-Sea & Arava Science Center in Massada, Ben Gurion University, Israel

e-mail: [email protected]

Nrf2 (nuclear factor erythroid 2-related factor 2) is a crucial transcription factor regulating the cellular
defense response under oxidative stress. Anti-oxidative protection ability of Nrf2 in human
pathological conditions, such as Alzheimer’s disease, Parkinson’s disease, psoriasis, acute dermatitis,
multiple sclerosis, macular degeneration etc. has been widely investigated. Among all human tissues,
skin is extremely sensitive to oxidative stress. The high sensitivity to oxidative stress is related to
exposure to environmental harmful factors such as ultraviolet light (UV) and the subsequent
production of reactive oxygen species (ROS). Taking into consideration that oxidative stress is a key
event in the pathogenesis of skin damage and inflammation, this pathological condition may therefore
be a suitable target for therapeutic intervention. The activity of Nrf2 is tightly regulated by a negative
regulator termed Keap1 (Kelch ECH-associating protein 1). The Keap1-Nrf2- ARE complex is
responsible for cytoprotective effects against endogenous and exogenous stress caused by ROS. Nrf2
binds to ARE (antioxidant response element) in the regulatory regions of target genes, and Keap1, a
repressor protein, binds to Nrf2 and promotes its degradation by the proteasome. To enhance Nrf2
activity, disruption of the Keap1−Nrf2 interactions at the protein−protein interface is necessary.
Thus, we hypothesized that compounds that interact with Keap1 at its putative binding site with Nrf2
have the potential to disrupt Nrf2 binding. In this way, free Nrf2 might transmigrate to nuclei and
lead to an activation of the transcription of anti-oxidative stress response genes and the expression of
the corresponding proteins that will course neuro- and dermo-protective effect.
We developed two Nrf2 activators that showed very significant antioxidant in vitro and ex vivo effects
in neurons, keratinocytes, and human skin samples. Although both compounds have good efficacy
and potency in the pharmacological concentrations, they contain a metabolically unstable imine bond
as well as potentially unstable phenol groups. To increase their metabolic stability, we designed and
synthesized a series of their derivatives with increased metabolic stability containing amide bond
instead of the imine one, as well as acetyl protected hydroxyl groups instead of the phenols. This
increased metabolic stability and lipophilicity thereby improved membrane and skin penetration
while retaining an ideal balance between the polar and unipolar groups thereby resulting in
structurally and functionally optimized compounds which exhibited significant protective effects.
 SC-11
NOVEL HETEROCYCLIC COMPOUNDS AS ANTIMICROBIAL AGENTS

BRULÍKOVÁ, L.

Department of Organic Chemistry, Faculty of Science, Palacký University, Czech Republic

e-mail: [email protected]

Heterocyclic compounds represent the most diverse scaffolds with unique structural properties that
offer various pharmacological activities. Current advances in synthetic methodologies enable the
effective synthesis of highly functionalized heterocycles to fine-tune their chemical and biological
properties. During the last few years, we have been involved in synthesizing compounds with
potential antituberculosis effects.1-4 We have developed several promising compounds against
Mycobacterium tuberculosis inhibiting the mycobacterial ATP synthase1,2 or mycobacterial virulence
factor Zmp1.3,4 During the development of these compounds, we also came across interesting
heterocyclic compounds exhibiting nanomolar activities against the parasite Trypanosoma brucei.
We believe our results might open up new avenues for designing selective antimicrobial agents
addressing the urgent need for novel selective and effective antibiotics in the face of antibiotic
resistance.

The study was supported by grants JG_2019_002 and IGA PrF 2024 028 from Palacký University in
Olomouc, FWO grant no. G066619N, project ELIXIR CZ no. LM2022123 from the Ministry of
Education, Youth and Sports of the Czech Republic.

References
1. CHASÁK, J., VAN MOLL, L., MATHEEUSSEN, A. et al.: ACS Omega, 9, 2024, 34808–34828.
2. CHASÁK, J., OORTS, L., ŠLACHTOVÁ, V. et al.: Bioorg. Med. Chem., 95, 2023, 117504.
3. DAK, M.; ŠLACHTOVÁ, V.; ŠEBELA, M. et al.: Eur. J. Med. Chem., 244, 2022, 114831.
4. ŠLACHTOVÁ, V.; ŠEBELA, M.; TORFS, E. et al.: Eur. J. Med. Chem., 185, 2020, 111812.
 SC-12
DEVELOPMENT OF DINITROPHENYL-CONTAINING ANTITUBERCULOSIS
AGENTS

KARABANOVICH, G.,1 STOLAŘÍKOVÁ, J.,2 MIKUŠOVÁ, K.,3 PÁVEK, P.,4 ROH, J.1
1
Department of Organic and Bioorganic Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, Czech
Republic
2
Department of Bacteriology and Mycology, Regional Institute of Public Health, Ostrava, Czech Republic
3
Department of Biochemistry, Faculty of Natural Science, Comenius University in Bratislava, Slovakia
4
Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University, Czech
Republic

e-mail: [email protected]

Tuberculosis (TB) remains one of the deadliest diseases in the world, and the situation has not
changed significantly in recent years. There were 1.45 million deaths and 10 million new TB cases
in 2018, and 1.3 million deaths and 10.6 million new TB cases in 2022.1 However, the growing
concern is related to the increasing number of drug-resistant TB cases, with MDR-TB rising from
187,000 in 2018 to 410,000 in 2022. Therefore, the demand for new drugs with a novel mechanism
of action remains high.
Recently, our group discovered 3,5-dinitrobenzylsulfanyl-1,3,4-oxadiazoles, which showed high
activities against both drug-susceptible and drug-resistant Mycobacterium tuberculosis strains in both
replicating and non-replicating states, had low toxicity against proliferating cell lines and isolated
human hepatocytes, and didn’t inhibit the growth of bacteria and fungi.2 Intensive study of the
structure-activity relationships has shown that the position of both nitro groups on the benzyl moiety
as well as the position of the dinitro-phenyl fragment in the molecular scaffold can have a crucial
influence on the activity of the compounds as well as on their mechanism of action.3

The study was supported by Czech Health Research Council (project NU21-05-00446)
References
1. https://www.who.int/teams/global-tuberculosis-programme/tb-reports .
2. KARABANOVICH, G., ZEMANOVÁ, J., SMUTNÝ, T., et al.: J. Med. Chem., 59, 2016, 2362-
2380.
3. KARABANOVICH, G., FABIÁNOVÁ, V., VOCAT, A., et al.: J. Med. Chem., 67, 2024, 81-109.
 SC-13
SYNTHESIS AND BIOLOGICAL ACTIVITY OF STYRYLQUINOLINE
DERIVATIVES

CIEŚLIK, W.,1 SZCZEPANIAK, J.,2 KRASOWSKA, A.,2 JAMPILEK, J.,1,3 MUSIOŁ, R.,1

1
Institute of Chemistry, University of Silesia in Katowice, 75 Pułku Piechoty 1a, 41-500 Chorzów, Poland
2
Department of Biotransformation, Faculty of Biotechnology, University of Wrocław, Joliot-Curie 14a, 50-383
Wrocław, Poland
3
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University, Odbojarov
10, 83232 Bratislava, Slovakia

e-mail: [email protected]

Advances in medical sciences have significantly contributed to the progress in the treatment of many
diseases; however, there remains an urgent need to explore new therapeutic approaches. We are
observing an alarming increase in bacterial and fungal infections. Therefore, the search for new drugs
remains one of the primary challenges in contemporary science.
In our research group, we have been focusing for years on the discovery of new, active derivatives
that exhibit strong antifungal and antibacterial properties, along with high selectivity towards normal
cells. These efforts have led to the development of a series of styrylquinoline derivatives. We
conducted in vitro screening of synthesized compounds against various fungal and bacterial strains.1
Subsequently, the antifungal activity of selected compounds was further evaluated by determining
the MIC values for a wild-type Candida albicans strain and cdrΔ single and double mutants. It was
found that cells with CDR1 deletion exhibited a significant increase in sensitivity to styrylquinolines,
suggesting that these compounds are substrates of Cdr1p. This indicates a potential mechanism of
action as inhibitors of this transporter. Additionally, synergistic effects with fluconazole were
investigated.2,3

References
1. CIEŚLIK, W., MUSIOŁ, R., NYCZ, J., et al.: Bioorg. Med. Chem., 20, 2012, 6960–6968.
2. SZCZEPANIAK, J., CIEŚLIK, W., ROMANOWICZ, A., et al.: Int. J. Antimicrob. Ag., 50, 2017,
171‒176.
3. CIEŚLIK, W., SZCZEPANIAK, J., KRASOWSKA, A., et al.: Molecules, 25, 2020, 1‒12.
 SC-14
NOVEL INHIBITORS OF HUMAN CARBONIC ANHYDRASE

HAVRANKOVA, E.,1 ŠELIGOVA, K.,1 RAVASZOVA, K.,2 HOFROVA, A.,2

1
Department of Chemical Drugs, Faculty of Pharmacy, Masaryk University Brno, Czech Republic
2
Department of Chemistry, Faculty of Science, Masaryk University Brno, Czech Republic

e-mail: [email protected]

Carbonic anhydrases (CA, EC 4.2.1.1) are metalloenzymes catalyzing the reversible hydration of
CO2, thereby affecting the pH and related physiological processes in various organisms. In human
bodies, 15 different isoforms of CAs can be found, including two tumor-associated: hCA IX and hCA
XII (human CA). These two isoforms are responsible for up-regulated metabolism, growth, and
survival of cancer cells. They are involved in spreading metastasis, and correlated with resistance
towards chemotherapy or radiotherapy.1 Given the above, it is clear that CA inhibitors are very
promising compounds in cancer treatment. However, for now, their crucial limitation is their
selectivity towards specific isoenzymes.

A new series of inhibitors of tumor-associated CA was designed and synthesized, specifically 1,3,5-
triazines containing aminobenzenesulfonamide, piperazine, aniline, and other structural moieties.
New compounds' inhibitory activity and selectivity were determined against hCA IX, hCA XII, and
hCA II.

The study was supported by grant number MUNI/G/1002/2021.

References
1. PASTOREKOVA, S., GILLIES, R.J.: Cancer Metastasis Rev., 38, 2019, 65-67.
 SC-15
VITAMIN D SUPPLEMENTATION WITH THE INFLUENCE OF ARTI (CASE
REPORT)

DĚDEČKOVÁ, E.1, ŠIMÁNEK, V.2, KUČERA ,R. 1, 2, KRÁLOVÁ, M. 1, TOPOLČAN, O. 2

1
Department of Pharmacology and Toxicology, Faculty of Medicine in Pilsen, Charles University, Czech Republic
2
Department of Immunochemistry, University Hospital and Faculty of Medicine in Pilsen, Charles University, Czech
Republic

e-mail: [email protected]

Low serum 25-hydroxyvitamin D concentrations are associated with a higher susceptibility to acute
respiratory tract infections (ARTIs). The aim of this case study is to present the association between
vitamin D levels, supplementation and the incidence of ARTIs.
A 23-year-old female patient with vitamin D deficiency successfully increased her vitamin D level
from 45.60 nmol/l to 85.91 nmol/l (reference range 75-200 nmol/l) by supplementation. However,
surprisingly, in the following period, instead of the expected level of 120 nmol/l, a decrease in vitamin
D level to 70.04 nmol/l was observed, although the patient continued taking supplementation. Further
investigations revealed that the patient had been suffering from common symptoms of acute
respiratory tract infection during the time of supplementation. Determination of total vitamin D levels
was performed with the chemiluminescence kit ACCESS 25(OH) Vitamin D on a Unicel® DxI 800
instrument.
This case study demonstrates the complex relationship between vitamin D levels, proper
supplementation and ARTI. The observed decline in vitamin D levels during supplementation and
ongoing acute respiratory tract infection suggests that respiratory infections may significantly affect
vitamin D metabolism.

The study was supported by the Ministry of Health, Czech Republic conceptual development of
research organization (Faculty Hospital in Pilsen-FNPl, 00669806), BBMRI-CZ: Biobank net-work
– a versatile platform for the research of the etiopathogenesis of diseases
Z.02.1.01/0.0/0.0/16_013/000167 and LM2015089, and by the “Cooperatio” Program, research
area Pharmaceutical Sciences.
 SC-16
INHIBITORY EFFECTS OF SOME CHALCONES ON ACHE ENZYME

D. ÖZMEN ÖZGÜN1, R. SAĞLAMTAŞ2, S. YAKUT3, A.S. AYDIN4, H.İ. GÜL4

1
Ağrı İbrahim Çeçen University, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Ağrı, Türkiye
2
Ağrı İbrahim Çeçen Unıversity, Vocational School of Health Services, Central Research and Application Laboratory,
Ağrı, Türkiye
3
1Ağrı İbrahim Çeçen Unıversity, Faculty of Pharmacy, Graduate Student, Ağrı, Türkiye
4
Ataturk Unıversity, Faculty of Pharmacy, Department of Pharmaceutical Chemistry, Erzurum, Türkiye

Natural compounds are essential in the discovery of new therapeutic agents. Chalcone derivatives are
one of the most studied flavonoid-derived structures.1 Under acidic or basic conditions, chalcone (1,3-
diaryl-2-propenone) derivatives can be classically synthesized by Claisen Schmidt condensation.2-4
Chalcone and its analogs have therapeutic importance, such as anticancer, anti-inflammatory,
antioxidant, antimicrobial, antituberculosis, antimalarial, and antiallergic properties. Reports indicate
that it possesses numerous essential bioactivities.1 Researchers have also investigated some
antipsychotic phenothiazines as potential cancer therapeutics. For instance, studies have found
promising activity of chlorpromazine against endometrial, lung, colorectal, and breast cancer.5 In
addition to neuroleptic effects, phenothiazine derivatives have pharmacological effects such as
antiemetic, antihistamine, analgesic, and anthelmintic.6 Additionally, it has essential bioactivities
such as anticancer,7 antibacterial,8 cholinesterase inhibitor,9 antimalarial,10 antituberculosis,11 and
anti-HIV.12 Importantly, AChE is a promising tumor suppressor.13 Given this information, we aimed
to synthesize five chalcones bearing phenothiazine rings, elucidate their chemical structures, and
investigate their inhibitory activities on the AChE enzyme.

This research project was supported by TUBITAK-2209-A (1919B012214350). We thank TUBITAK

for their support in this study.

References

[1] Mahapatra, DK, Bharti, SK, Asati, V. 2015. Anti-cancer chalcones: Structural and molecular
target perspectives. Eur J Med Chem 98: 69-114.
[2] Bisi, A, Meli, M, Gobbi, S, Rampa, A, Tolomeo, M, Dusonchet, L. 2008. Multidrug resistance
reverting activity and antitumor profile of new phenothiazine derivatives. Bioorg Med Chem 16:
6474-6482.
[3] Dimmock, JR, Das, U, Gul, HI, Kawase, M, Sakagami, H, Barath, Z, Ocsovsky, I, Molnar, J.
2005. 3-Arylidene-1-(4-nitrophenylmethylene)-3,4-dihydro-1H-naphthalen-2-ones and related
compounds displaying selective toxicity and reversal of multidrug resistance in neoplastic cells.
Bioorg Med Chem Lett 15: 1633-1636.
[4] Dimmock, JR, Kandepu, NM, Hetherington, M, Quail, JW, Pugazhenthi, U, Sudom, AM,
Chamankhah, M, Rose, P, Pass, E, Allen, TM, Halleran, S, Szydlowski, J, Mutus, B, Tannous, M,
Manavathu, EK, Myers, TG, De Clercq, E, Balzarini, J. 1998. Cytotoxic activities of Mannich bases
of chalcones and related compounds. J Med Chem 41: 1014-1026.
[5] Sarhan, M. O., Fayadh, N. A., Elhaggar, R., & Zaghary, W. (2023). Antitumor Potential and
Possible Targets of Phenothiazine Derivatives: A Recent Review. Egyptian Journal of Chemistry,
66(13), 1001-1014.
[6] Pluta, K, Morak-Mlodawska, B, Jelen, M. 2011. Recent progress in biological activities of
synthesized phenothiazines. Eur J Med Chem 46: 3179-3189.
[7] Molnar, A, Amaral, L, Molnar, J. 2003. Antiplasmid effect of promethazine in mixed bacterial
cultures. Int J Antimicrob Agents 22: 217-222.
[8] Amaral, L, Kristiansen, JE. 2001. Phenothiazines: potential management of Creutzfeldt-Jacob
disease and its variants. Int J Antimicrob Agents 18: 411-417.
[9] Darvesh, S, McDonald, RS, Darvesh, KV, Mataija, D, Conrad, S, Gomez, G, Walsh, R, Martin,
E. 2007. Selective reversible inhibition of human butyrylcholinesterase by aryl amide derivatives of
phenothiazine. Bioorg Med Chem 15: 6367-6378.
[10] Dominguez, JN, Lopez, S, Charris, J, Iarruso, L, Lobo, G, Semenov, A, Olson, JE, Rosenthal,
PJ. 1997. Synthesis and antimalarial effects of phenothiazine inhibitors of a Plasmodium falciparum
cysteine protease. J Med Chem 40: 2726-2732.
[11] Bate, AB, Kalin, JH, Fooksman, EM, Amorose, EL, Price, CM, Williams, HM, Rodig, MJ,
Mitchell, MO, Cho, SH, Wang, Y, Franzblau, SG. 2007. Synthesis and antitubercular activity of
quaternized promazine and promethazine derivatives. Bioorg Med Chem Lett 17: 1346-1348.
[12] Viveiros, M, Martins, M, Couto, I, Kristiansen, JE, Molnar, J, Amaral, L. 2005. The in vitro
activity of phenothiazines against Mycobacterium avium: potential of thioridazine for therapy of the
co-infected AIDS patient. In Vivo 19: 733-736.
[13] Xi, H. J., Wu, R. P., Liu, J. J., Zhang, L. J., & Li, Z. S. (2015). Role of acetylcholinesterase in
lung cancer. Thoracic cancer, 6(4), 390-398.
 SC-17
PHENOTHIAZINE BASED D-A SYSTEMS: FROM STRUCTURAL AND
PHOTOPHYSICAL TO BIOLOGICAL PROPERTIES

SŁODEK, A.

University of Silesia in Katowice

Abstract not available at the time of publication.

THANK YOU AND SEE YOU NEXT YEAR!