Book of Abstracts - Veterinární a farmaceutická univerzita Brno

Transcription

44. KONFERENCE
SYNTÉZA A ANALÝZA LÉČIV 2015
44th CONFERENCE DRUG SYNTHESIS AND ANALYSIS 2015
Organizační výbor:
Organising committee:
doc. PharmDr. Ing. Radka Opatřilová, Ph.D., MBA
prof. RNDr. Jozef Csöllei, CSc.
doc. PharmDr. Josef Jampílek, Ph.D.
PharmDr. Pavlína Marvanová
PharmDr. Tereza Padrtová
Book of Abstracts
2. - 4. ZÁŘÍ 2015
BRNO
2nd – 4th SEPTEMBER 2015
BRNO
1
Dear participants, colleagues, friends
It is pleasing to mention that the agenda of the Conference Synthesis and Analysis
a wide range of very interesting items relating to the properties and consists of the
development of drugs.
Conference Synthesis and Analysis of Drugs is a very important annual gathering
of employees of pharmaceutical universities and research institutions.
This meeting brings together many key partners in the world of drugs.
It is an opportunity to renew contacts and discuss issues of mutual interest.
While the sciences are impressive, more important is the profound impact this
knowledge has for people - the improved quality of life they provide.
We do our best when we go where the science leads us, and base our decisions
on thorough, data-driven analyses. This ensures our work will benefit for health.
Finally, I would like to thank all of you for your support of our mission and our
work. I firmly believe that we can meet the challenges in front of us – especially if we
continue the tradition of science in the service of public health.
I take this opportunity to wish you all the very best for the future.
Assoc. Prof. Ing. PharmDr. Radka Opatřilová, Ph.D., MBA
2
Elektronický sborník 44. Konference Syntéza a analýza léčiv
2. - 4. září 2015
Brno
Vydala Veterinární a Farmaceutická univerzita Brno
ISBN: 978-80-7305-760-2
Abstrakty příspěvků byly redakčně kráceny a formátovány.
Zaslané celé články vyjdou v časopise Česká a Slovenská Farmacie.
Yearbook of 44th Conference Drug Synthesis and Analysis
2th – 4th September 2015
Brno
Published by University of Veterinary and Pharmaceutical sciences Brno
ISBN: 978-80-7305-760-2
Abstracts were shortened and edited by editors.
Articles will be published in Journal Česká a Slovenská Farmacie.
3
PROGRAM OF THE CONFERENCE
Wednesday 2nd September 2015
arrival, accommodation, registration (1 1.00 - 13. 30)
13. 30 - 14. 00
Official opening of the Conference
AULA
University of Veterinary and Pharmaceutical Sciences Brno
Chairmens:
prof. zw. dr hab. inż. Jarosław Polański
doc. PharmDr. Josef Jampílek, Ph.D.
14. 00 - 14. 50
Polanski, J. Chemoinformatics, a tool for drug discovery, an upgrade
for a new century: problems, concepts, definitions
14. 50 - 15. 30
Musiol, R. Close the Gate! Antifungal styrylquinolines as Cdr1
substrates
15. 40 - 16. 00
Stanzel, L. Synthesis and antioxidant activity of phenylcarbamic acid
derivatives acting on cardiovascular system
16. 00 - 16. 30
Coffee break
AULA
4
Chairpersons:
doc. RNDr. Jana Staničová, Ph.D.
prof. Victoriya Georgiyants
16. 30 - 16. 50
Staničová, J. Potential anticancer agent hypericin and its model
compound hypericin: its interaction with DNA
16. 50 - 17. 05
Rišiaňová, L. New derivatives of curcumin with potential anticancer
and antiradical activity
17. 05 - 17. 20
Georgiyants , V. Modern approaches to the directed synthesis of
anticonvulsants
17. 20 - 17. 35
Pietrzyńska, M. Polymer-ceramic monolithic in-needle extraction
device applied for extraction of potential antiresorptive drugs
After the lectures: WELCOME PARTY
http://www.vfu.cz/information-about-university/location-and-university-campus/index.html#changeMapa
5
Thursday 3th September 2015
registration (8. 30 - 11. 00)
Chairmens:
Prof. Dr. Janez Košmrlj
Prof. Dr.Haider Norbert
.
9. 00 - 9. 50
Kosmrlj, J. Chemistry and biological application of selected diazene
derivatives
9. 50 - 10. 30
Haider, N. Structural modifications of the antitumor alkaloid
Luotonin A: cycloaddition reactions as a useful tool
10. 35 - 11. 00
Coffee break
11. 00 - 11. 30
Pazourek, J. HPLC determination of hepatotoxic peptides in water
reservoires with occurence of invasive species of Pectinatella
magnifica
11. 30 - 11. 45
Materiienko , A. Development of spectrophotometric method for
carmoisine determination in quality control of equipment cleaning
12. 00 - 14. 00
Lunch break
6
Doc. Ing. Lucie Cahlíková, Ph.D.
Chairpersons:
Doc. PharmDr. Karel Šmejkal, Ph.D.
14. 00 - 14. 40
Šmejkal, K. Potential therapeutic aplications of prenylated phenols
14. 40 - 15. 15
Cahlíková, L. Alkaloids of family Amaryllidaceae and their
therapeutic potential
15. 20 - 15. 35
Žemlička, M. The possibilities of HPLC with post-column reaction in
the search for plant-derived antioxidants and inhibitors of enzyme
activities
16. 00 - 17. 00
Poster session
SIC (Study and Information Centre), building Nr. 24
18. 30 - 23. 00
GALA EVENING AND AFTER PARTY
SIC, building Nr. 24
Friday 4th September 2015
Free day for trip
7
Lectures
Author
Title of Lecture
L01
CAHLÍKOVÁ, L.
Alkaloids of family Amaryllidaceae and their therapeutic potential
L02
GEORGIYANTS, V.
Modern approaches to the directed synthesis of anticonvulsants
L03
HAIDER, N.
L04
KOSMRLJ, J.
L05
MATERIIENKO, A.
L06
MUSIOL, R.
L07
PAZOUREK, J.
L08
PIETRZYŃSKA, M.
L09
POLANSKI, J.
L10
RIŠIAŇOVÁ, L.
L11
STANIČOVÁ, J.
L12
STANZEL, L.
L13
ŠMEJKAL, K.
L14
ŽEMLIČKA, M.
Structural modifications of the antitumor alkaloid Luotonin A:
cycloaddition reactions as a useful tool
Chemistry and biological application of selected diazene derivatives
Development of spectrophotometric method for carmoisine
determination in quality control of equipment cleaning
Close the Gate! Antifungal styrylquinolines as Cdr1 substrates
HPLC determination of hepatotoxic peptides in water reservoires with
occurence of invasive species of Pectinatella magnifica
Polymer-ceramic monolithic in-needle extraction device applied for
extraction of potential antiresorptive drugs
Chemoinformatics, a tool for drug discovery, an upgrade for a new
century: problems, concepts, definitions
New derivatives of curcumin with potential anticancer and antiradical
activity
Potential anticancer agent hypericin and its model compound hypericin:
its interaction with DNA
Synthesis and antioxidant activity of phenylcarbamic acid derivatives
acting on cardiovascular system
Potential therapeutic applications of prenylated phenols
The possibilities of HPLC with post-column reaction in the search for
plant-derived antioxidants and inhibitors of enzyme activities
8
ALKALOIDS
OF
FAMILY
AMARYLLIDACEAE
AND
THEIR
THERAPEUTIC POTENTIAL
LUCIE CAHLÍKOVÁ
ADINACO Reasearch Group, Department of Pharmaceutical Botany and Ecology, Faculty of Pharmacy, Charles
University, Hradec Králové, Czech Republic; [email protected]
Plants of the family Amaryllidaceae comprise about 85 genera and 1100 species that are distributed
widely in tropical and subtropical regions of the world. Plants of this family have been used for
thousands of years in traditional herbal medicine. The earliest evidence of their therapeutic application
was discovered in the fourth century B.C.E., when Hippocrates of Cos used the oil from daffodil,
Narcissus poeticus L., for the treatment of uterine tumors 1. More than 500 Amaryllidaceae alkaloids
representing 18 skeletal types have been isolated and identified. The representative alkaloids are
lycorine, galanthamine, haemanthamine, pancratistatine, pretazzetine, montanine, narciclassine and
others. Biogenetically, these structures are the result of an intramolecular oxidative coupling of the key
intermediate O-methylnorbelladine, derived from the amino acids L-phenylalanine and L-tyrosine.
These compounds exhibit a diversity of biological activities including antitumor, antiviral, antibacterial,
antifugal, antimalarial, analgesic, acetylcholinesterase (AChE) inhibitory and cytotoxic activities 2. In
terms of bioactivity, the most important alkaloid of the group is galanthamine, a long-acting, selective,
reversible and competitive inhibitor of AChE, which was approved for the clinical management of mild to
moderate Alzheimer´ disease (AD)3. Other important compound of this group lycorine, is one of the
most frequently occurring Amaryllidaceae alkaloids and has been reported as a potent inhibitor of
ascorbic acid synthesis, cell growth and division and organogenesis in higher plants, algae and yeasts.
In addition, lycorine is recognized as a low molecular antiproliferative agent against multidrug resistant
and apoptosis-resistant cancer cells with selective cell type-dependent cytotoxicity in tumor cells by
mitochondrial pathways and inducing apoptosis 2.
1)
Kornienko A, Evidente A. Chem. Rev. 2008, 108, 1982-2014.
2)
Jin Z. Nat. Prod. Rep. 2009; 26, 363-381.
3)
Hampel H, et al. Expert Rev. Neurother. 2015, 15, 83-105.
9
STRUCTURAL MODIFICATIONS OF THE ANTITUMOR ALKALOID
LUOTONIN A: CYCLOADDITION REACTIONS AS A USEFUL TOOL
NORBERT HAIDER
Department of Pharmaceutical Chemistry, Faculty of Life Sciences, University of Vienna, Austria;
[email protected]
Luotonin A is a pentacyclic alkaloid that has been isolated in the late 1990s from the Asian plant,
Peganum nigellastrum Bunge (Zygophyllaceae).1) It has a pyrroloquinazolinoquinoline skeleton and
thus shares some basic structural features with Camptothecin (CPT, cf. Fig. 1), another natural product
which has become well-known because of its pronounced antitumor activity. 2) The latter is based on a
stabilising effect towards the binary complex formed between DNA and the enzyme, topoisomerase I as
an essential step for releasing the supercoiled structure of the nucleic acid, and this stabilisation
effectively prevents DNA replication. While CPT itself has unfavorable pharmacokinetic properties,
some of its derivatives, such as topotecan and irinotecan, have made their way into clinical use as
anticancer agents for the chemotherapy of certain malignancies, 2) although they do not only cause the
typical side effects of most cytotoxic agents, but also suffer from specific adverse effects like bladder
toxicity.3) This problem arises from the labile hydroxylactone structure of CPT-like compounds which
first leads to an undesired early inactivation of the drug by lactone-ring opening at the physiological
blood pH and later (during excretion) to another undesired structural change, namely re-formation of the
lactone in the acidic environment of the urinary bladder. For that reason (together with synthesis-related
aspects), Luotonin A has been receiving increased intrerest as an alternative lead structure for the
development of therapeutically useful topoisomerase I poisons, as it was found to exhibit a similar
biological activity as CPT, albeit with considerably lower potency.4) As shown in Fig. 1, Luotonin A does
not possess the labile lactone structure and, moreover, lacks the chiral center of CPT.
1
14
A
B
4
N
13
O
2
D
10
N
E
6
7
Luotonin A
N
12 11
3
5
O
C N
N
O
9
8
Camptothecin
OH O
Figure 1. Structures of Luotonin A (with IUPAC numbering scheme) and Camptothecin
Thus, in recent years quite a number of publications have appeared that describe various total
syntheses of Luotonin A as well as some structural modifications of this alkaloid. A comprehensive
2011 review article4) nicely summarises this development. In continuation of previous investigations of
our groups in the field of antitumor-active polycyclic N-heterocycles, we became interested in Luotonin
A as a lead structure for hitherto unexplored derivatives and analogues with potential cytotoxic activity.
10
In particular, variation of ring A of the pentacyclic skeleton in terms of steric and electronic properties
appears highly interesting with regard to an improved activity profile, as suggested by some previous
observations.4)
1)
Ma Z.-Z., Hano Y., Nomura T., Chen Y.-J. Heterocycles 1997; 46, 541–546.
2)
Du W. Tetrahedron 2003; 59, 8649–8687.
3)
Pizzolato J. F., Saltz L. B. Lancet 2003; 361, 2235–2242.
4)
Liang J. L., Cha H. C., Jahng Y. Molecules 2011; 16, 4861–4883.
11
CHEMISTRY AND BIOLOGICAL APPLICATION OF SELECTED
DIAZENE DERIVATIVES
JANEZ KOŠMRLJ
Faculty of Chemistry and Chemical Technology, University of Ljubljana, Slovenia; [email protected]
Cancer is one of the major causes of death in developed countries (1). Despite extensive screening and
investing huge funds for cancer research over the past decade, the decline in cancer mortality rate is
relatively modest. Therefore, to maximize the effectiveness of cancer treatment, new compounds –
potential antitumor drugs – are synthesized and tested.
Cisplatin has been clinically approved to be used to treat solid tumor (1). Although it is one of the most
commonly used and effective agents it causes neuro-, nephro- and ototoxicities as the main side effects
(1). The major obstacle of cisplatin in successful chemotherapy is development of cisplatin-resistance
on tumors, which is the effect of several unrelated mechanisms (1-3). One of those is a result of
cisplatin inactivation by binding to intracellular thiol-containing molecules including glutathione (GSH).
Glutathion-S-transferase (GST) augments the resistance by catalyzing GSH-cisplatin binding4).
Diazenes, compounds with the structure R–N=N–R’, are known for a long time. Some diazenes have
been recognized as biological control agents, the diazene antibiotics (2). These include methyl
phenyldiazenecarboxylate, which inhibited the germination of the fungus Trichoderma viride and
diazenecarboxylic acid bis(N,N-dimethylamide), which halted the growth of Escherichia coli (Figure 1).
The mechanism of action has been studied revealing that these diazenes may oxidize intracellular GSH
into glutathione disulfide (GSSG) 8).
Figure 1. Molecular structures of methyl phenyldiazenecarboxylate (left), diazenecarboxylic acid
bis(N,N-dimethylamide) (middle), and diazenecarboxamides 1 (right).
We have demonstrated that another subclass of diazenes, i.e, diazenecarboxamides 1 (Figure 1) (7),
are cytotoxic for tumor cell lines and their cisplatin (also doxorubicin and vincristine)-resistant sublines
(2-12) The tumor cell lines tested include cervical carcinoma cells (HeLa), glioblastoma cells (A1235),
laryngeal carcinoma cells (HEp2), mammary carcinoma cells (MCF-7), breast adenocarcinoma cells
(SK-BR-3) and others. Diazenecarboxamides were found to be powerful modulators of tumor
intracellular GSH concentration, oxidizing it to GSSG (9, 12, 13). Besides GSH-depleting activity, in
selected examples an activation some alternative cell-death pathways has been observed, indicating
that GSH may not be the only cellular target of diazenecarboxamides (16).
12
In so advanced diseases like cancer a single mono functional targeted drug is unlikely to be superior for
the cure. In this context, combined drugs (so called cocktails) that are capable of affecting several
cellular targets simultaneously have been developed 11). Even further, an alternative approach involves
the use of hybrid molecules. These comprise of two drugs covalently bound into a single entity,
combining pharmacological effects of independently acting drugs . Such hybrid molecules can possess a
synergistic effects that is beyond mere simultaneous administration of two separate agents (12).
In
a
combined
(cocktail)
diazenecarboxamide–cisplatin
treatment,
the
GSH-depleting
diazenecarboxamides led to the reversal of the acquired tumor resistances. The synergistic effect with
cisplatin has been demonstrated on different tumor cell lines as well as some cisplatin-resistant
sublines (11,17) Stimulated by this, we prepared and tested platinum complex 2 containing a
diazenecarboxamide ligand that is coordinated to platinum(II) through a pyridine nitrogen atom (Figure
2). The diazenecarboxamide in this complex retained its oxidative properties toward GSH and exhibited
high cytotoxicity against T24 bladder carcinoma cells (12).
Figure 2. Molecular structures of platinum complexes with diazenecarboxamides 2–4.
To increase the efficacy in the preparation of a library of diazenecarboxamide-based ligands, we
developed strategies that are based on copper-catalyzed azide-alkyne cycloaddition protocol, also
known as “Click chemistry (12-18). This allowed an easy access to hybrid molecules that resemble
cisplatin 3 and carboplatin 4 structures (17,26). Unfortunately, the cytotoxic profile of these compounds
against the tumor cell lines was not satisfactory, which was ascribed to their low solubility in aqueous
media. Work is in progress to address this issue.
Ruthenium coordination compounds with redox-active azopyridine ligands have been developed by the
group of Sadler as catalytic anticancer compounds (17). These Ru-azopyridine complexes undergo
activation by reduction inducing redox reactions inside cancer cells. The Ru-azopyridine complexes act
as catalysts for the oxidation of intracellular GSH into GSSG. We surmised that the use of equimolar
amounts of diazenecarboxamides, which are necessary for intracellular GSH oxidation, could be
avoided by the use of ruthenium-diazenecarboxamide complexes. For the latter the oxidant catalytic
anticancer activities could be expected in analogy to the above mentioned Ru-azopyridine complexes.
13
In contrast to the azopyridine ligand, however, diazenecarboxamides possess three different donor
atoms, which makes possible the formation of two different five-membered chelating rings on metal
coordination (N,O- and N,N-) with different properties. Both types of the complexes, N,O- (5), and N,N(6) were easily prepared from N,2-diphenyldiazenecarboxamide and [(Cym)RuCl2]2 as show in Scheme
117).
Scheme 1. Preparation of Ru-diazenecarboxamide complexes 5 and 6.
Indeed, complexes 5 and 6 had completely different properties. Compound 6 exhibit high cytotoxic
activity with IC50 values ranging from 3.4–15.1 M towards the human cervical carcinoma cells (HeLa),
colorectal carcinoma cells (HCT-116), lung carcinoma cells (H460), breast adenocarcinoma cells (MDAMB-231), and laryngeal carcinoma cells (HEp-2) and their carboplatin, cisplatin and curcumin resistant
subline (7T) 34).
Financial support from the Ministry of Education, Science and Sport, Republic of Slovenia, the Slovenian
Research Agency (Grant P1-0230) is acknowledged.
1) Siegel R., Naishadham D., Jemal A. Cancer J. Clin. 2012; 62, 10–29.
2) Boulikas T., Vougiouka M. Oncol. Rep. 2003; 10, 1663–1682.
3) Kelland L. Nature Rev. Cancer 2007; 7, 573–584.
4) Siddik Z. H. Oncogene 2003; 22, 7265–7279.
5) Stewart D. J. Crit. Rev. Oncol./Hematol. 2007; 63, 12–31.
6) Chen H. H. W., Kuo M. T. Metal Based Drugs 2010; 1–6.
7) Košmrlj J., Kočevar M., Polanc S. Synlett 2009; 2217–2235.
8) Kosower E. M., Miyadera T. Glutathione. 6. J. Med. Chem. 1972; 15, 307–312.
9) Osmak M., et al. Neoplasma 1999; 46, 201–206.
10) Osmak M., Bordukalo T., Jernej B., Košmrlj J., Polanc S. Anti-Cancer Drugs 1999; 10, 853–859.
11) Osmak M., Bordukalo T., Ambriović Ristov A., Jernej B., Košmrlj J., Polanc S. Neoplasma 2000; 47, 390–395.
12) Moskatelo D., Benjak A., Laketa V., Polanc S., Košmrlj J., Osmak M. Chemother. 2002; 48, 36–41.
13) Moskatelo D., Polanc S., Košmrlj J., Vuković L., Osmak M. Pharmacol. Toxicol. 2002; 91, 258–263.
14) Čimbora T., Bombek S., Polanc S., Osmak M. Toxicol. in Vitro 2003; 17, 159–164.
15) Čimbora-Zovko T., Bombek S., Košmrlj J., Kovačić L., Polanc S., Katalinić A., Osmak M. Drug Devel. Res.
2004; 61, 95–100.
16) Jakopec S., Dubravčić K., Polanc S., Košmrlj J., Osmak M. Toxicol. in Vitro 2006; 20, 217–226.
14
17) Jakopec S., Dubravčić K., Brozović A., Polanc S., Osmak M. Cell Biol. Toxicol. 2006; 22, 61–71.
18) Martin-Kleiner I., et al. Toxicol. in Vitro 2007; 21, 1453–1459.
19) Vajs J., et al. Acta Chim. Slov. 2013; 60, 842–852.
20) Gediya L. K., Njar V. C. O Expert Opin. Drug Discov. 2009; 4, 1099–1111.
21) Meunier B. Acc. Chem. Res. 2008; 41, 69–77.
22) Monti E., et al. J. Med. Chem. 2005; 48, 857–866.
23) Müller-Schiffmann A., et al. Angew. Chem. Int. Ed. 2010; 49, 8743–8746.
24) Grabner S., Košmrlj J., Bukovec N., Čemažar, M. J. Inorg. Biochem. 2003; 95, 105–112.
25) Urankar D., Košmrlj J. J. Comb. Chem. 2008; 10, 981–985.
26) Urankar D., Pevec A., Košmrlj J Eur. J. Inorg. Chem. 2011; 1921–1929.
27) Urankar D., Košmrlj J. Inorg. Chim. Acta 2010; 363, 3817–3822.
28) Urankar D., Steinbücher M., Kosjek J., Košmrlj J. Tetrahedron 2010; 66, 2602–2613.
29) Urankar D., et al. Inorg. Chem. 2010; 49, 4820–4829.
30) Urankar D., Pevec A., Turel I., Košmrlj J. Cryst. Growth Des. 2010; 10, 4920–4927.
31) Bratsos I., et al. Dalton Trans. 2011; 40, 5188–5199.
32) Stojanović N., et al. Acta Chim. Slov. 2013; 60, 368–374.
33) Dougan S. J., et al. PNAS 2008; 105, 11628–11633.
34) Sommer M. G., et al. Chem. Eur. J. 2014; 20, 17296–17299.
15
DEVELOPMENT OF SPECTROPHOTOMETRIC METHOD FOR
CARMOISINE
DETERMINATION
IN
QUALITY
CONTROL
OF
EQUIPMENT CLEANING
ANNA MATERIIENKO, VOLODYMYR GRUDKO, VICTORIYA GEORGIYANTS
Department
of
Pharmaceutical
Chemistry,
The
National
University
of
Pharmacy,
Kharkiv,
Ukraine;
[email protected]
Pharmaceutical product can be contaminated by other active pharmaceutical ingredients, by cleaning
agents, or by other materials. The cleaning processes in the pharmaceutical industry have been
regulated by the implementation of the Good Manufacturing Practice rules. Cleaning procedure
includes sampling and testing for allowable trace amounts of active pharmaceutical ingredients or
excipients that were a part of the previous preparation 1, 2).
There are two methods of sampling that are considered to be acceptable: direct surface sampling
(swab method) and indirect sampling (use of the rinse solutions). The acceptance limits of
contaminants and residues are determined in the International Conference on Harmonisation (ICH)
guidelines on their specific toxicity in tested impurities 3, 4).
The suitability of the material to be used for sampling and the suitability of the sampling medium should
be determined. The ability to recover samples accurately may be affected by the choice of the sampling
material. A major complication when using rinse solutions are probably very dilute solutions since the
quantitative content of the active substance is not always possible to determine by the available
analytical methods1, 3).
Carmoisine (E 122) is a red coloured synthetic food
azo dye, a derivative of diazosulfonaphthalenes, a
crystalline substance that is soluble in water. It is
used to provide a certain color for many medicines as
for individual matters and mixed with other dyes 5).
Carmoisine
6)
Methods for determining the dyes in food and medicines by HPLC are known . But these methods are
laborious, they require a lot of time and use expensive equipment that carries great expenses to
pharmaceutical manufacturers.
1)
2)
PIC/S document PI006-3. Cleaning validation. 2007; 27 p.
U.
S.
Food
and
Drug Administration.
Guide
to
inspections
validation
of
cleaning
processes.
http://www.fda.gov/ICECI/Inspections/InspectionGuides/ucm074922.htm
3)
Shklyaev S.A. Kyiv: Pharmaceutical Journal 2012; 5, 33-37.
4)
Zavyalova I.E., Sharahova E.F. Siberia: Siberian Medical Review 2010; 63(3), 57-62.
16
5)
Smirnov E. V. Directory St. Petersburg: ed. Profession 2009; 352 p.
6)
Minioti K. S., Sakellariou C. F., Thomaidis N. S. Elsevier: Anal. Chim. Acta 2007; 583(1), 103-110.
17
CLOSE THE GATE! ANTIFUNGAL STYRYLQUINOLINES AS CDR1
SUBSTRATES
ROBERT MUSIOL1, ANNA KRASOWSKA2, JOANNA SZCZEPANIAK2, WIOLETA CIESLIK1,
EWELINA SPACZYNSKA1
1
Institute of Chemistry, University of Silesia, Katowice, Poland
2
Faculty of Biotechnology, University of Wroclaw, Poland
[email protected].
The worldwide appearance of fungal infections have grown enormously during last few decades. This,
surely increased our demand for drugs and therapeutical strategies 1,2. Until the 1940s, comparatively
few agents were available for the treatment of systemic fungal infections. The first antifungal agent from
that time was griseofulvin, isolated in 1939. The development of polyene antifungals with Amphotericin
B is first major advance in medical mycology. The continuous search for new and less toxic antifungal
drugs led finally to the discovery of the azoles. The very first drug of this class was Chlormidazole
introduced in 1958 for the treatment of topical mycosis. Although some disadvantages, it began a new
era of antifungal drugs. In this regards the late 1960s, may be considered as dawn of the azole
antimycotics3. On the other hand it was also beginning of our continuous struggle with the very
discouraging aspect of mycoses – drug resistance.
Fungal species have evolved a multitude of mechanisms to survive exposure to xenobiotic or harmful
substances. Extensive biochemical studies have highlighted a significant diversity in the mechanisms
by which fungi can became. These mechanisms involve; increased levels of the cellular target or
alteration of its molecular structure (mutations) which reduce the efficacy of a drug, reduced intracellular
accumulation of antifungal drug or alterations in ergosterol synthesis 4. Since drug resistance can
develop as a stepwise process over time; these mechanisms may combine with each other 5.
Fungal cells in plasma membranes possess multidrug efflux transporters, which are responsible for
important aspect of resistance.
Figure 1. Two main efflux pump proteins responsible for fungal resistance.
18
The two main classes of pumps are the ATP-binding cassette (ABC) and the major facilitator
superfamily (MFS)6. The CDR1 and CDR2 (Candida drug resistance 1 or 2) genes encode two
homologous transporters of the ABC family, and the MDR1 (multi-drug resistance 1) gene encodes the
MFS (Figure 1)7. The mechanisms involve the upregulation of genes encoding the efflux transporters.
The overexpression of MDR1 is responsible for specific resistance to fluconazole exclusively. By
contrast, the overexpression of CDR genes is associated with cross-resistance to different azole
derivatives, including fluconazole, itraconazole and ketoconazole. Deletion of CDR1 and CDR2 confers
hypersensitivity to azoles5,8. Other multidrug efflux transporter genes of the ABC class are CDR3,
CDR4 and CDR59, for which only partial sequences are known. CDR3 is expressed at very low levels,
but this expression does not appear to be dependent on resistance to azole derivatives. Limited data on
CDR4 and CDR5 suggest that these genes are not involved in the mechanisms of azole resistance 10.
Our team is working on antifungal compounds for roughly ten years 11-13. We have designed and
synthesized series of quinoline related structures as potential antimycotic drugs. Quinoline derivatives
are particularly important as privileged structures for antimicrobial agents14. In our approach we
highlighted the structural resemblance between quinolines and allylamine antimycotics as terbinafine or
naftifine. Then heterocyclic moiety was reinforced with bunch of substituents that have been selected
form library of active small molecular quinolines with known antifungal activity as shown on Figure 2.
N
H
R
N
R = phenyl, hetaryl
Cl
X
Cl
N
OH
R´
X
N
R
OH
Figure 2. Design of styrylquinolines with antifungal potency.
The designed compounds were synthesised according to modified procedures and our developed state
of the art microwave assisted synthesis allowing high yields in relatively short period of time. Biological
activities of those compounds were tested against broad spectrum of fungal and bacterial strains.
Several of the designed compounds appeared more active than standard drugs. For additional tests
some mutant strains of Candida albicans were chosen. These were CDR1Δ, CDR2Δ with deletion of
Cdr1p or Cdr2p exporters respectively and wild type with correct functionality for both pumps. The most
interesting compounds shown promising activity against Cdr1p strain and synergy with fluconazole.
Prolonged incubation with styrylquinolines cause also overexpression and delocalisation of the efflux
pump.
19
To conclude the styrylquinolines appeared interesting leading structures for antifungal agents. Their
activities are up to order of magnitude higher than that of fluconazole. At the same time these
compounds are less vulnerable to resistance.
Financial support of NCN grant 2013/09/B/NZ7/00423 is acknowledged.
1)
Kauffman C.A. Clin. Infect. Dis. 2004; 39, 588-590
2)
McNeil M. M., et al. Clin. Infect. Dis. 2001; 33, 641-617.
3)
Musiol R., Kowalczyk W. Curr. Med. Chem. 2012;19, 1378–1388.
4)
Sanglard D. Mycologist, 2003; 17, 74-78.
5)
Lupetti A., et al. Trends Mol. Med. 2002; 8, 76-81.
6)
Joseph-Horne T., Hollomon D. W. FEMS Microbiol. Lett. 1997; 149, 141-149.
7)
Fera M. T., et al. Expert Rev. Anti Infect. Ther. 2009; 7, 981-998.
8)
Rai V., et al. Biochemistry, 2005; 44, 6650-6661.
9)
Shukla S., et al.Biochemistry, 2007; 46, 12081-12090.
10)
Sanglard D. Curr. Opin. Microbiol., 2002; 5, 379–385.
11)
Musiol R., et al. Bioorg Med Chem. 2006; 14, 3592–3598.
12)
Jampilek J., et al. Molecules. 2009; 14, 4246–4265.
13)
Jampilek J., et al.Molecules. 2009;14: 1145–1159.
14)
Musiol R., et al. Curr Med Chem. 2010; 17, 1960–1973.
20
HPLC DETERMINATION OF HEPATOTOXIC PEPTIDES IN WATER
RESERVOIRS WITH OCCURENCE OF INVASIVE SPECIES OF
PECTINATELLA MAGNIFICA
JIŘÍ PAZOUREK* 1, KAREL ŠMEJKAL 1, PETER KOLLÁR 1, JOSEF RAJCHARD 2
1
Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences Brno, Czech Republic
2
Agriculture faculty, University of South Bohemia, České Budějovice, Czech Republic
[email protected]
Introduction
Pectinatella magnifica (PM) is an invasive bryozoan species, which makes giant colonies in fresh water.
Its occurrence in the Czech Republic was for the first time documented in 2003 in Třeboňsko area
(south Bohemia). Colonies of P. magnifica live on submerged branches of Salix spp., on submerged
woody species rests and less frequently on littoral vegetation (e.g. Typha latifolia), at stones and other
objects. The species occurs mostly in large numbers and its life activity probably has a significant
influence on the composition of the food chain (it is a filtrating species) and the values of hydrochemical
and physicochemical parameters. Not only PM itself can affect the quality of surrounding water - PM
colonies are joined with algae and cyanobacteria during its lifetime
1)
. These symbionts can also
produce toxic compounds to the water reservoirs. The goal of this contribution was to identify and
determine contingent toxins known in water with cyanobacteria (microcystins, nodularins).
Experimental methods
HPLC conditions:

DIONEX Ultimate 3000, software Chromeleon

HPLC column Ascentis Express RP-amide 2.7 um, 150x2.1 mm.

mobile phase was acetonitrile/water with 0.1% formic acid, flow rate 0.5 ml/min, gradient elution

UV detection at 240 nm
Sample preparation

samples from various water reservoirs were stored lyophilized. Prior to analysis they were
extracted to methanol/water mixture, sonicated and filtrated.
Results and discussion
An HPLC method
2)
was transferred and modified for our purpose. Total time of HPLC analysis
including an equilibration stage was 20 minutes, the analytes retention time was less than 7 minutes
(see Fig. 1).
21
Fig. 1. Typical chromatogram of standard mixture of hepatotoxic peptides.
The method was applied to samples collected in the area of South Bohemia in the seasons 2012-2013.
On a model reservoir (Veselí I) with average production of biomass, we also estimated a threshold
concentration in water that could be determined by the method. The concentrations are 0.130 ng/L, and
8.2 ng/L, resp.
Conclusions
A rapid method for determination of microcystins and nodularin was developed and applied to real
samples from South Bohemia water reservoirs. We can conclude that the invasion of PM in water
reservoirs of the Czech Republic does not represent a toxicity risk in this moment.3)
The work was supported by a project P503/12/0337 of Grant Agency of Czech Republic (GAČR).
1)
Šetlíková, I., et al. Biologia, 2013; 68(6), 1136-1141.
2)
Bláhová L., et al. Environ. Chem. Lett. 2008; 6, 223-227.
3)
Pazourek J., HPCL analysis of toxins in lyophilised Pectinatella magnifica, Lecture on International Conference
„Invasive aquatic animal species" held in České Budějovice, Agriculture Faculty, University of South Bohemia,
České Budějovice, 9.12.2014.
22
CHEMOINFORMATICS, A TOOL FOR DRUG DISCOVERY, AN
UPGRADE FOR A NEW CENTURY: PROBLEMS, CONCEPTS,
DEFINITIONS
JAROSLAW POLANSKI
Institute of Chemistry, University of Silesia, Katowice Poland; [email protected].
Chemoinformatics is a term that has been coined to describe a discipline organizing and coordinating
the application of informatics methods in chemistry. At the semantic level, two elements constructing
the designation chemoinformatics clearly relate this discipline to chemistry and informatics. The
designation originated in the 90’, when a new name was sought for the discipline that uses computers
in drug design. The basic problems, concepts and definitions will be discussed during the lecture.
Drug design is a central arena where chemoinformatics is played. With the development of the methods
we are more and more aware that we need to understand better a formal structure of this new
discipline. The problems, concepts and definitions of this discipline will be discussed during the lecture.
At the semantic level two elements constructing the designation chemoinformatics clearly relate this
discipline to chemistry and informatics. Chemistry focuses on chemical compounds. But what do we
mean by a chemical compound? Substantial definitions of chemistry were formulated by IUPAC. This
includes especially:

Chemical element

Chemical species

Chemical substance

Measurement

Mixture

Molecular entity

Molecule

Property
Quite surprisingly we are not formally defining chemical compound [1, 2]. What is the reason for that?
Chemoinformatics focuses on molecular descriptors and properties. Despite common belief real
measured properties are rare in chemistry. Thus, we are to predict them, which is especially important
in molecular design.
Some major problems focused by chemoinformatics are listed below to realize how important they are
in molecular design:
Chemical data documentation and searches: database searches and management
Property prediction: compounds’ series (FCS) are mapped from FCS into VCS; two basic versions
are available, i.e., property vs. property or structure vs. property. However, in the design step for novel
23
compounds in VCS (we can design both compounds VCS or FCS where some properties can be
registered in databases and/or literature) we are always to use a structure version (no property is
available in VCS).
Calculating
molecular descriptors: 2D
(chemical graphs) and
3D
(molecular modeling)
representations are mapped into single numbers (0D); vectors, fingerprints(1D); matrixes, surface maps
(2D); surface, atomic representations, force fields, virtual or real receptor data (3D); etc.
Molecular modeling: molecular structure 3D data generation in silico from their 2D or 1D
representations. This includes both predictions of molecular topography and molecular descriptors i.e.,
atomic annotations data by calculated (simulated) atomic properties.
Structure elucidation: (synonym: structure-spectra correlations) property to molecular structure is
mapped in FCS when we are attempting to find a structure having a certain spectra or in structure to
property version (VCS and FCS) we are simulating a spectra for a given molecule (substance).
Mapping structure to activity (SAR): a series of structures (FCS substances) is needed to study
SAR, which are usually synthesized. The real goal of SAR is usually to predict the training series for
designing new structures and substances by property predictions in a qualitative or quantitative
procedures.
Synthesis design: designing organic synthesis in product (target molecule) to reagent (synthon,
reagent equivalent) mapping.
Where are critical points of the current molecular design? How current dogma of drug development
influences the progress in pharma? Are we getting better and more innovative in pharma R&D and how
to evaluate this [3]? These are problems that will be discussed during the lecture.
1)
Polanski J., Gesteiger J. Computer Representation of Chemical Compounds, in: Handbook of Computational
Chemistry, Springer, 2015, in print.
2)
IUPAC Goldbook. http://goldbook.iupac.org.
3)
Polanski J., Bogocz J., Drug Discov. Today, 2015, in print.
24
NEW
DERIVATIVES
OF
CURCUMIN
WITH
POTENTIAL
ANTICANCER AND ANTIRADICAL ACTIVITY
NATALIA MIKLÁŠOVÁ, LUCIA RIŠIAŇOVÁ, JINDRA VALENTOVÁ, SAMUEL VARÉNYI,
FERDINAND DEVÍNSKY
Department of chemical theory of drugs, Faculty of Pharmacy, Comenius University in Bratislava, Slovakia;
[email protected]
Introduction
Curcumin (1,7-bis(4-hydroxy-3-methoxy phenyl)hepta-1,6-diene-3,5-dione) the yellow spice used for
anti-inflammatory, anticancer, antioxidant activities, AIDS and Alzheimer´s disorders 1), exhibits a poor
bioavailability. This issue might be improved by using adjuvants like piperine, liposomal curcumin,
nanoparticles, phospholipid complex and structural analogues of curcumin. 2) Different derivatives of
curcumin can coordinate with metallic ions to form complexes with various biological potential, for
example palladium complexes of curcumin describe the antitumoral and antioxidant activities on human
prostate cancer cells.3) Several Knoevenagel condensates of curcumin and their copper complexes
inhibit the TNF-α induced NF-κB activation and proliferation of human leukemic cells. Schiff bases of
such derivatives induce the cell growth inhibition in colon and pancreatic cancer cells. Therefore, these
compounds are interesting for a chemopreventive and therapeutic activity against cancer and for their
increasing bioavailability.4)
Fourteen Knoevenagel condensates of two curcuminoids were synthesized and structurally
characterized by NMR, IR, UVspectra and their antiradical activity was determined.
Knoevenagel condensates were synthesised by general procedure 5) 1 mmol of curcumin dissolved in
toluene was reacted with a corresponding aromatic aldehyde (2 mmol) in a dropwise manner with
continuous stirring, in the presence of a catalytic amount of piperidine and acetic acid. The reaction
mixture was refluxed overnight and the final product was isolated by column chromatography.
Literature studies shown that 4-arylidene curcumin analogues inhibit growth of lung cancer cells with
less concentrations than curcumin, acting as potential compounds for development against cancer and
inflammatory diseases.5) In our laboratory fourteen curcuminoids were synthesised, as Fig.1 illustrates,
with the expectation that the condensates will exhibit significant antiradical activity.
25
O
O
H
O
OH
R
R
R
140 oC
R1
OH
R=OCH3; OCH2CH3
R1= OCH3; OCH2CH3; OH; H
R
piperidine/AcOH
+
HO
O
HO
OH
R2
R1
R2
Fig 1: Synthesis of Knoevenagel condensates of curcumin and its analogue
This work was supported by the Slovak Research and Development Agency under the contract No. APVV-051612 and FaF UK/10/2015, FaF UK/11/2015.
1)
Zambre A.P., et al. Synth. React. Inorg. Met.-Org. Chem. 37(1), 2007; 19-27.
2)
Anand P., et al. Mol. pharmaceutics, 2007; 4(6), 807-818.
3)
Miklášová N., et al. Inorg. Chem. Comm., 2014; 46, 229-233.
4)
Padhye S., et al. Pharm. res., 2009; 26(8), 1874-1880.
5)
ZuO Y., et al. Eur. J. Med. Chem. 2012; 55, 346–357.
26
POTENTIAL ANTICANCER AGENT HYPERICIN AND ITS MODEL
COMPOUND EMODIN: INTERACTION WITH DNA
JANA STANIČOVÁ, VALÉRIA VEREBOVÁ, ALENA STREJČKOVÁ
Department of Chemistry, Biochemistry & Biophysics, Institute of Biophysics, The University of Veterinary
Medicine and Pharmacy, Košice, Slovakia; [email protected]
Hypericin (HYP, Fig. 1a) is a natural photosensitizing pigment occurring in plants of the genus
Hypericum. HYP under light illumination displays anti-proliferative and cytotoxic effects on many tumour
cell lines1, 2). These properties together with minimal dark toxicity, tumour selectivity and high clearance
from the host body make HYP a very promising agent in photodynamic therapy of cancer. Due to its
prospective pharmaceutical utilization, this interesting molecule has been the subject of many scientific
research groups3-6).
Our team has been dealing with the interaction of HYP and its derivatives (such as emodin, quinizarin,
danthron, alizarin) with important bio macromolecules to better understand a drug´s mechanism of
action.
The interaction of the drug with linear calf thymus DNA is the main aim of this contribution. The binding
constant and interaction mode have been determined by spectrophotometric methods. Model
compound of HYP, emodin (E, Fig. 1b), was considered to our study for its structural similarity with HYP
molecule. In addition, emodin, a plant derived anthraquinone, was found to be a photosensitizer which
possess anti-tumor, anti-bacterial, anti-viral, anti-inflammatory, and myorelaxing activities7,8). Detailed
knowledge of the interaction of HYP and E with the DNA at the molecular level is extremely important in
determination of their distribution and therapeutic effect in the body.
OH
O
OH
OH
HO
CH3
HO
CH3
O
HO
OH
CH3
O
OH
Fig. 1. Chemical
O
OH
a)
b)
structure of hypericin (a) and
emodin (b)
This work was supported by grant KEGA No. 014 UVLF – 4/2013. Authors are pleased to thank Dr. P. Miškovský
and Dr. R. Varhač for the possibility to use experimental equipment in their laboratories.
1)
Lopez-Bazzocchi I., et al. Photochem. Photobiol.1991; 54, 95-98.
27
2)
Andreoni A., et al. Photochem. Photobiol. 1994; 59, 529-533.
3)
Miškovský P. Curr. Drug Targets 2002; 3, 55-84.
4)
Dougherty T.J., et al. Natl. Cancer Inst. 1998; 90, 889–905.
5)
Kiesslich T., et al.Curr. Med. Chem. 2006; 13, 2189-20201.
6)
Karioti A., et al. Int. J. Mol. Sci. 2010; 11, 562-594.
7)
Radovic J., et al. Food chem. tox. 2012; 50, 9, 3181 – 3189.
8)
Poliaček I., et al. Phys. Res. 2003; 52, 6, 749-762.
28
SYNTHESIS AND ANTIOXIDANT ACTIVITY OF PHENYLCARBAMIC
ACID DERIVATIVES ACTING ON CARDIOVASCULAR SYSTEM
LUKÁŠ
STANZEL1,
SICHROVSKÁ1,
IVA
MATEJ
MARUNIAK1,
KAPUSTÍKOVÁ1,
IVAN
MALÍK1,
ĽUBICA
SEDLÁROVÁ1,
EVA
HAVRANOVÁ
JOZEF
CSÖLLEI2
1
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University, Bratislava, Slovak
Republic
2
Department of Chemical Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences,
Brno, Czech Republic
[email protected]
In
search
of
new
perspective
antagonists
of
β-adrenergic
receptors,
alkylesters
of
4{2hydroxy3[(4pyridin2yl)piperazine1yl]propoxy}phenylcarbamic acid monochlorides, alkyl =
methyl-buthyl, (labelled as 4a1-4d1) have been evaluated. They might act as the potential antioxidants,
therefore the aim of current study was to investigate their antioxidant profile by applying three in vitro
methods.
Following mentioned, the capability of these compounds to reduce relatively stable reference 2,2diphenyl-1-picrylhydrazyl (DPPH) radicals and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)
(ABTS) radicals in performed spectrophotometric analyses and to influence redox potential using FRAP
method was inspected. The position of alkyl side chain bonded to lipophilic aromatic fragment as well
as the electrostatic effects, which were induced by pyridin-2-yl substitution, have been considered the
essential factors, which have influenced an antioxidant potential of investigated molecules.
Relatively high lipophilicity of all tested compounds might be relevant, but any connection between
length of alkyl side chain and activity has not been revealed.
29
POTENTIAL THERAPEUTIC APLICATIONS OF PRENYLATED
PHENOLS
KAREL ŠMEJKAL1,2
1
Department of Natural Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences Brno,
Czech Republic
2
Department of Molecular biology and Pharmaceutical Biotechnology, Faculty of Pharmacy, University of
Veterinary and Pharmaceutical Sciences Brno, Czech Republic
[email protected]
Introduction.
The flavonoids are a group of secondary metabolites biosynthetically derived from shikimic acid and
polyketide pathways. They are plant pigments containing benzopyrane substituted with a phenyl ring at
position 2 or 3. Their aglycones can be lipophilic; their lipophilicity can be further enhanced by
methylating the hydroxyl groups, or by prenylation or geranylation at different positions on the skeleton.
The prenyl or geranyl moiety may be modified in different ways and enhances the interaction with
organism.
Experimental methods.
Different chromatographic methods were used for isolation of series of prenylated phenols.
Experimental in vitro and in vivo studies have revealed many biological and pharmacological activities
of flavonoids. We worked with prenylated compounds from Morus alba and Paulownia tomentosa1-5).
Results and discussion.
Inflammation is a multiple and complex response by the body to infection or injury. Prenylated
compounds show pleiotropic effects and can modulate a broad spectrum of inflammatory regulatory
nodes. Their antiphlogistic action combines many particular effects: it can be mediated by several
pathways: via anti-oxidant and pro-oxidant effects, by interacting directly with pro-inflammatory proteins,
and by interacting with signal pathways and inhibiting the expression of inflammation-related genes. In
vivo tests have confirmed all of the effects of flavonoids previously observed in in vitro experiments.
The antibacterial effect of prenylated substances was confirmed, some synergy with standard
antibiotics was observed against MRSA1-5).
Conclusion.
Prenylated phenols are good candidates for further research to discovered new therapeutics for
treatment of diseases connected with bacterial infection and inflammatory disorders.
1)
Šmejkal K., et.al. J. Nat. Prod. 2008; 71 (4), 706–709.
30
2)
Navrátilová A., et al. Phytochemistry 2013; 89, 104–113.
3)
Hošek J., et al. J. Nat. Prod. 2011, 74 (4), 614–619.
4)
Hanáková Z., et al. J. Nat. Prod. 2015, 78, 850-863.
5)
Vochyánová Z., et al. Fitoterapia 2015, 101, 201-207.
31
THE POSSIBILITIES OF HPLC WITH POST-COLUMN REACTION IN
THE
SEARCH
FOR
PLANT-DERIVED
ANTIOXIDANTS
AND
INHIBITORS OF ENZYME ACTIVITIES.
MILAN ŽEMLIČKA
Department of Natural Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences Brno,
[email protected]
Despite the high number of publications on the topic of antioxidants that have degraded this topic and,
therefore, just the antioxidant activity itself is almost impossible to publish, plant-derived antioxidants
still remain a commercially very important and scientifically interesting group of food supplements. In
the search for active constituents from plant extracts an idea occurred at the end of the 20th century:
HPLC instrumentation linked up with post-column reaction with a highly colored synthetic radical was
used 1). DPPH· and ABTS+· are the most frequently used radicals, but other oxidation-reduction or
enzymatic reactions have extended the use of this way of determination.
The basic arrangement of the instrumentation is shown in the following scheme 2):
This method is suitable for the identification of an antioxidant or other biologically active substance
without their previous isolation. It allows to determine which compounds and to what extend are
responsible for the specific activity and it rationalizes the means of isolation.
This lecture summarizes the current state of this kind of experiments and points out its benefits and
limitations.
1)
Yamaguchi T., et al. Biosci. Biotechnol. Biochem. 1998; 62(6) 1201-1204.
2)
Pazourek J., Václavík J., Žemlička M. Food Chem. 2011; 125, 785–790.
32
Posters
Nr.
Authors
Title of Poster
P1.
AMBROŻKIEWICZ W.
P2.
P9.
AMIĆ A., PAVIĆ V., MOLNAR M.
BAK A., KOZIK V., WYSZOMIRSKI M.,
MAGDZIARZ T., SMOLINSKI A.,
POLANSKI J.
BEDLOVIČOVÁ Z., UNGVARSKÁ
MAĽUČKÁ L., SALAYOVÁ A.,
HARVANOVÁ J., OČENÁŠ P.
CZAPLIŃSKA B., MROZEKWILCZKIEWICZ A., RAMS-BARON M.,
SPACZYŃSKA E., KORZEC M.,
MUSIOŁ R.
ČAVOJSKÝ T., PAŠKOVÁ L., BILKA F.,
SLOVÁK L., BAUEROVÁ K.,
PAULÍKOVÁ I.
ČERNÍKOVÁ A., JAMPÍLEK J., BOBÁĽ
P.
DAŇKOVÁ I., MALANÍK M.,
DALL´ACQUA S., GAZDOVÁ M.,
HANÁKOVÁ Z.
DOKUPILOVÁ S., VEIZEROVÁ L.,
GALBA J., MIKUŠ P.
THE USAGE OF CATALYSTS IN PRODUCTION OF THE
DRUGS
SYNTHESIS OF NEW OXADIAZOLE BASED ANTIOXIDANTS
MODELING OF DYE-FIBER AFFINITY USING RD SOM-4DQSAR APPROACH: APPLICATION TO SET OF
ANTHRAQUINONE DERIVATIVES
DETERMINATION OF BIOLOGICALLY ACTIVE
COMPOUNDS IN FUNGI OF GENUS CORDYCEPS
SINENSIS BY HPLC AND NMR
P10.
FARSA O., BRKA P.
P11.
FAZEKAS T., MICIAN J.
P12.
FORGÁCSOVÁ A., MIKUŠ P.,
HAVRÁNEK E., MELNÍK M.
P13.
GÁLUSOVÁ A., ANDRIAMAINTY F.,
MIKLÁŠ R.
P14.
GARAJ V.
P15.
GAZVODA M., POLANC S.
P3.
P4.
P5.
P6.
P7.
P8.
P16.
P17.
P18.
P19.
GONĚC T., KMEŤKO J., POSPÍŠILOVÁ
Š., KOS J., ZADRAŽILOVÁ I.,
JAMPÍLEK J.
GRUBEROVÁ L., KRATOCHVÍL B.,
SEILEROVÁ L.
HAVRANOVÁ SICHROVSKÁ L.,
STANZEL L., MALÍK I., MARUNIAK M.,
KAPUSTÍKOVÁ I., SEDLÁROVÁ E.,
CSӦLLEI J.
CHRIPKOVÁ M., ANTOLIKOVÁ N.,
ZIGO F., BUDOVSKÁ M., MOJŽIŠ J.,
TAKÁČ L., TOROPILOVÁ D.
CHARACTERIZATION OF NEW GREEN FLUOROPHORES
BASED ON QUINOLINE SCAFFOLD.
STUDY OF BUTYRYLCHOLINESTERASE WITHIN
EXPERIMENTAL ARTHRITIS
ANALOGS OF ALAPTIDE AND THEIR EFFECT ON THE
PENETRATION OF DRUGS
NEWLY IDENTIFIED CHEMICAL CONSTITUENTS OF
PARASITIC PLANT LATHRAEA SQUAMARIA L.
ANALYSIS OF FLAVONOIDS IN GRAPE LEAVES BY HPLCDAD-MS/MS
FROM AN OLD DRUG TO A NEW ONE: SYNTHESIS OF
VALPROATE FROM 5,5-DIPROPYLBARBITURIC ACID
MOBILE APPLICATIONS AND THEIR POTENTIAL FOR USE
IN PHARMACEUTICAL PRACTICE
PREPARATION AND ANALYSIS OF GALLIUM (III)
COMPLEXES WITH AMINOCARBOXYLATES AND
SULFONAMIDES
DETERMINATION OF CMC OF CATIONIC TENSIDE IN
AQUEOUS AND MIXED WATER-ALCOHOL SOLUTIONS
VIRTUAL SCREENING OF COMBINATORIAL LIBRARY OF
1,3,5-TRIAZINE MOIETY SUBSTITUTED WITH
BENZENESULFONAMIDES AND DIFFERENT AMINES
SYNTHESIS AND BIOLOGICAL ACTIVITY OF SELECTED
CINNAMIC ACID DERIVATIVES
SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF N(HALOGEN-PHENYL)-1-HYDROXYNAPHTHALENE-2CARBOXAMIDES
SOLUBILITY AND DISSOLUTION RATEIMPROVEMENT OF
CANDESARTAN CILEXETIL
ESTIMATION OF LIPOHYDROPHILIC PROPERTIES OF
MOLECULES WITH POTENTIAL Β3-AGONISTIC ACTIVITY
ANTIPROLIFERATIVE EFFECT OF 1-METHOXYBRASSININ
33
P20.
JĘDRZEJOWSKA A., MATUSSEK M.,
KOZIK V., BĄK A., ZADRAŽILOVÁ I.,
JAMPÍLEK J.
P21.
JEŽKO P., SÁRAZOVÁ Z.
P22.
KAPKOWSKI M., SŁOTA M.,
POLAŃSKI J.
P23.
KAPUSTÍKOVÁ I., MARUNIAK M.,
HAVRANOVÁ-SICHROVSKÁ L.,
STANZEL L., MALÍK I., SEDLÁROVÁ E.
P24.
KARABANOVICH G., ROH J.,
JIRKOVSKÁ-VÁVROVÁ A., VÁVROVÁ
K., ŠIMŮNEK T.
SYNTHESIS AND STUDY OF CARDIOPROTECTIVE
ACTIVITY OF NEW DEXRAZOXANE ANALOGS
P25.
KASTNER P., BURDOVÁ K.,
PILAŘOVÁ P.
1. HPLC METHOD FOR STABILITY EVALUATION OF
PHARMACEUTICAL PREPARATION CONTAINING SODIUM
PICOSULFATE
P26.
P27.
KORZEC M., RZYCKA R., SENKAŁA
S., SZPACZYŃSKA E., CZAPLIŃSKA
B., CIEŚLIK W., MROZEKWILCZKIEWICZ A., RAMS-BARON M.,
MUSIOŁ R., POLAŃSKI J.
KOS J., GONĚC T., ZADRAŽILOVÁ I.,
VALEŠOVÁ M., POSPÍŠILOVÁ Š.,
SPACZYŃSKA E., JAMPÍLEK J.
P28.
KOZAKIEWICZ D., POLAŃSKI J.
P29.
KOZIK V., BĄK A.
P30.
KRAJČIOVÁ D., MIKUŠ P., MELNÍK M.,
HAVRÁNEK E.
P31.
KRÁTKÝ M., STOLAŘÍKOVÁ J.,
VINŠOVÁ J.
P32.
KROUTIL A., CSÖLLEI J.
P33.
KRYVANYCH O., BEVZ N., HARNA N.,
BEVZ O.
P34.
KUBÍNOVÁ R., SRNÁNKOVÁ H.
P35.
LUKAČOVIČOVÁ O., HAVRÁNEK E.
P36.
MALARZ K., RAMS-BARON M.,
MROZEK-WILCZKIEWICZ A., SERDA
M., POLAŃSKI J., MUSIOŁ R.
P37.
MARÁKOVÁ K., PIEŠŤANSKÝ J.,
MIKUŠ P.
P38.
MARVANOVA P., MOKRY P., HUMPA
O., PADRTOVA T.
SYNTHESIS AND BIOLOGICAL PROPERTIES OF CHOSEN
SYMMETRICAL AMIDES AND THIOAMIDES OF
TEREPHTHALIC ACID
STUDY OF STRUCTURE AND ADMET PROPERTIES OF AT1 RECEPTOR ANTAGONISTS
CATALYTIC GAS-PHASE GLYCEROL PROCESSING OVER
SIO2-, CU-, NI- AND FE- SUPPORTED AU
NANOPARTICLES
DETERMINATION OF ACID-BASE DISSOCIATION
CONSTANTS OF NEWLY SYNTHESIZED
ARYLETHANOLAMINE DERIVATIVES USING CAPILLARY
ZONE ELECTROPHORESIS
SYNTHESIS OF QUINOLINE DERIVATIVES BY USING
NANO-PD/CU CATALYST IN THE SEARCH OF NEW
FLUOROPHORES
RING-SUBSTITUTED 1-HYDROXYNAPHTHALENE-2CARBOXANILIDES AS POTENTIAL ANTIBACTERIAL
AGENTS
SYNTHESIS OF NOVEL CARBAMATES AND
THIOSEMICARBAZONES AS POTENTIAL ANTICANCER
DRUG CANDIDATES
THE STUDY OF EPR AND UV-VIS JUICE FROM
POMEGRANATES
PREPARATION AND ANALYSIS OF COPPER (II)
COMPLEXES WITH AMINOCARBOXYLATES AND
SULFONAMIDES
SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF NOVEL
SULFONAMIDE DERIVATIVES
SYNTHESIS OF ARYLOXYAMINEPROPANOLES AND
ARYLAMINEPROPANOLES
USE OF 2,6-DICHLOROQUINONE-4-CHLORIMIDE FOR
ASSAY OF PHENYLEPHRINE HYDROCHLORIDE IN
COMBINED TABLETS WITH PARACETAMOL AND
CHLORPHENIRAMINE MALEATE
FLAVONOIDS FROM TAGETES PATULA L. AS INHIBITORS
OF ACETYL/BUTYRYLCHOLINESTERASE
RADIONUCLIDE X-RAY FLUORESCENCE
SPECTROMETRY IN ELEMENTAL ANALYSIS OF MEDICAL
PLANTS AND MEDICINES IN USE TO LOWER
CHOLESTEROL LEVELS
EFFECTS OF THIOSEMICARBAZONES ANALOGS ON
OXIDATIVE STRESS AND CELLS DEATH IN HUMAN
COLON CANCER.
ON-LINE HYPHENATION OF CAPILLARY
ELECTROPHORESIS AND MASS SPECTROMETRY USED
FOR THE ANALYSIS OF BIOGENIC AMINES IN GRAPE
LEAVES
SYNTHESIS OF NEW
ARYLCARBONYLOXYAMINOPROPANOL DERIVATIVES
CONTAINING N-PHENYLPIPERAZINE MOIETY
34
P40.
MIKLÁŠOVÁ N., RIŠIAŇOVÁ L.,
VALENTOVÁ J., VARÉNYI S.,
DEVÍNSKY F.
MIKULOVÁ M., SÝKOROVÁ M.,
HAVRÁNEK E.
P41.
MOKRÝ M., ŠRÁMKOVÁ M.
P39.
P42.
P43.
P44.
P45.
P46.
P47.
P48.
MULARSKI J., CIEŚLIK W.,
SZCZEPANIAK J., KRASOWSKA A.,
MUSIOŁ R.
MYHAL A., DOBROVA A.,
GOLOVCHENKO O., GEORGIYANTS
V.
MYZNIKOV L.V., MELNIKOVA Y.V.,
ZEVATSKII Y.E.
NALEPA P., MROZEK-WILCZKIEWICZ
A., POLAŃSKI J.
NĚMEČEK J., KARABANOVICH G.,
ROH J., VALÁŠKOVÁ L., VÁVROVÁ K.,
STOLAŘÍKOVÁ J., PÁVEK P.,
KLIMEŠOVÁ V., HRABÁLEK A.
ONDREJKOVÁ A., SÜLI J.,
KOLČÁKOVÁ L., ONDREJKA R.,
ČECHVALA P., BENKÖ Z., PROKEŠ
M.
PADRTOVÁ T., MOKRÝ P.,
ODEHNALOVÁ K., MARVANOVÁ P.,
HUMPA O.
P49.
PIEŠŤANSKÝ J., MARÁKOVÁ K.,
MIKUŠ P.
P50.
PILAŘOVÁ P, KUŽELOVÁ K.,
KASTNER P.
P51.
PÍŽOVÁ H., BOBÁĽ P.
P52.
PLANKOVÁ A., HAVRÁNEK E., MIKUŠ
P.
P53.
REJMUND M, POLAŃSKI J.
P54.
SALANCI E., ANDRIAMAINTY F.,
MALÍK I.
P55.
P56.
SEMELKOVÁ L., JANĎOUREK O.,
PATEROVÁ P., KONEČNÁ K.,
DOLEŽAL M., ZITKO J.
SPACZYŃSKA E., CIEŚLIK W.,
MROZEK-WILCZKIEWICZ A., RAMSBARON M., MALARZ K., MUSIOŁ R.
P57.
STOPKOVÁ l., BEZÁKOVÁ Ž.
P58.
SÜLI J., KOLČÁKOVÁ L.,
HARVANOVÁ J., ONDREJKOVÁ A.
P59.
ŠULEKOVÁ M., HUDÁK A.
P60.
TARHAJOVÁ M., HÉTHELYI E. B.,
BÖSZÖRMÉNYI A., SZOLGAI G.,
TÓTH J., CZIGLE S.
NEW DERIVATIVES OF CURCUMIN WITH POTENTIAL
TARGET-SPECIFIC RADIOPHARMACEUTICALS: LTRYPTOPHAN AS A RADIONUCLIDE CARRIER
HPLC ANALYSIS OF SOME NON-STEROIDAL
ANTOFLOGISTICS
ANTIFUNGAL STYRYLQUINOLINES PROBABLY ACT AS A
CELLULAR RAZOR BLADE
VALIDATION OF SPECTROPHOTOMETRIC METHOD OF
ASSAYING METRONIDAZOLE IN CAPSULES
BARGELLINI REACTION IN FUNCTIONALIZATION OF
HETEROCYCLIC COMPOUNDS
STABLE GOLD NANOPARTICLES – SYNTHESIS,
BIOCONJUGATION AND APPLICATION
NITRO GROUP-CONTAINING OXA- AND THIADIAZOLES AS
POTENTIAL ANTITUBERCULAR AGENTS
INFLUENCE OF Β-CAROTENE ON ANTIGENIC
EFFECTIVENESS OF INACTIVATED RABIES VACCINE
WITH SQUALENE ADJUVANT
SYNTHESIS AND EVALUATION OF
HETEROARYLCARBONYLOXYAMINOPROPANOLS AND
THEIR QUATERNARY AMMONIUM SALTS
HYDRODYNAMICALLY CLOSED CITP-CZE COUPLED WITH
TANDEM MASS SPECTROMETRY FOR DETERMIANTION
OF ANTIGRIPAL DRUGS IN HUMAN URINE
HPLC HODNOCENÍ PIROXIKAMU V PLAZMĚ S VYUŽITÍM
SPME A DEPROTEINACE
AN OPTIMIZED AND SCALABLE SYNTHESIS OF
PROPYLPHOSPHONIC ANHYDRIDE
STANOVENIE SELÉNU A ZINKU V KRVNEJ PLAZME
PACIENTOV S ATOPICKOU DERMATITÍDOU
MICROWAVE – ASSISTED SYNTHESIS AND
CHARACTERIZATION OF THIOSEMICARBAZONES BASED
ON 3-AMINOPYRIDINE-2-CARBOXALDEHYDE
STUDY OF THE CMC OF 1-[3-(3ALCOXYPHENYLCARBAMOYLOXY)-2-HYDROXYPROPYL]4-(4-FLUOROPHENYL)PIPERAZINIUMCHLORIDE
SYNTHESIS AND BIOLOGICAL EVALUATION OF NBENZYL-3-BENZYLAMINOPYRAZINE-2-CARBOXAMIDES
SYNTHESIS, BIOLOGICAL ACTIVITY AND FLUORESCENCE
PROPERTIES OF STYRYLQUINOLINE DERIVATIVES
SOLUBILITY OF VALSARTAN - INFLUENCE OF CATIONIC
SURFACTANTS ON THE EXTENT OF SOLUBILIZATION
ANTIOXIDATIVE PROTECTION OF INACTIVATED RABIES
VACCINE WITH SQUALENE ADJUVANT BY Β-CAROTENE
DETERMINATION OF THE COLORANTS IN VITAMIN E BY
HPLC WITH PHOTODIODE ARRAY DETECTION
ANALÝZA PRCHAVÝCH ZLOŽIEK RASTLÍN RODU
HYLOCEREUS
35
P61.
VALÁŠKOVÁ L, ROH J,
KARABANOVICH G, NĚMEČEK J,
STOLAŘÍKOVÁ J, VÁVROVÁ K,
KLIMEŠOVÁ V, HRABÁLEK A
P62.
VALENTOVÁ M., ROHÁLOVÁ J.
P63.
VOSÁTKA R., KRÁTKÝ M., VINŠOVÁ
J.
P64.
YAKOVENKO O., MASLIY Y.
NITRO GROUP-CONTAINING SUBSTITUTED
TETRAZOLES: NEW HIGHLY EFFICIENT
ANTITUBERCULAR AGENTS
BIOLOGICAL ACTIVITY OF SELECTED SPECIES OF
MACROMYCETES
SYNTHESIS OF TRICLOSAN DERIVATIVES AND THEIR
ANTIMYCOBACTERIAL EFFECT
THE DEVELOPMENT OF DENTAL DRUG IN THE FORM OF
MEDICATED CHEWING GUM
36
POTENTIAL USAGE OF NEW NANOCATALYSTS FOR ORGANIC
SYNTHESIS
WERONIKA AMBROŻKIEWICZ1, JAROSŁAW POLAŃSKI1
1
Institute of Chemistry, University of Silesia, Katowice, Poland; [email protected]
Introduction
Over recent years, nanometals are widely study due to their catalytic properties, in particular, in
combination with various carriers
1)
. Recently there has been developed an efficient method for
disperse the nanoparticles on the amorphous SiO2 surface with the ultrasound assistance 2). Further
research led to improvement of this method of transfer of nanoparticles to the target metal grains
3)
.
Metals used as carriers are also involved in the catalysis and can act as a synergist to increase the
effectiveness of the active ingredients. This type of structures can be especially useful in organic
synthesis, where a new efficient catalysts are constantly required 4).
Experimental
The preparation of amorphous silica included usage of Stöber process. Precursors of nanoparticles
was added into obtained carrier, then sonicated, dried and reduced in a hydrogen furnace. Nanometals
on intermediate carriers were transferred onto metal powder via etching by aqueous solution of sodium
hydroxide. Catalytic properties were tested in the oxidation reactions of various compounds.
There have been obtained series of nanoformations based on transition metals dispersed on
amorphous silica and a series of bimetallic nanostructures, which indicated the catalytic potential.
1)
Korzec M., et al. J. Cat. 2014, 313, 1-8
2)
Bartczak P., et al. Patent application P.405270
3)
Bartczak P., et al. Patent application P.405268
4)
Kapkowski M., et al. J. Cat., 2014, 319, 110-118
37
SYNTHESIS OF NEW OXADIAZOLE BASED ANTIOXIDANTS
ANA AMIĆ1, VALENTINA PAVIĆ1, MAJA MOLNAR2
1
Department of Biology, Josip Juraj Strossmayer University of Osijek, Croatia
2
Faculty of Food Technology, Josip Juraj Strossmayer University of Osijek, Croatia
Oxadiazoles are a class of heterocyclic compounds with a wide range of biological activities. Their
combination with coumarin core is interesting synthetic approach and could yield some new biologically
active compounds including antioxidants.
A series of new oxadiazoles was synthesized from coumarin Schiff bases in reaction with acetic
anhydride. Their structure was confirmed with various spectral techniques and their antioxidant activity
was determined as DPPH scavenging activity. A combination of coumarin core with oxadiazole ring was
found to be an excellent approach for obtaining antioxidants, since some compounds had an excellent
antioxidant activity.
38
MODELING OF DYE-FIBER AFFINITY USING RD SOM-4D-QSAR
APPROACH:
APPLICATION
TO
SET
OF
ANTHRAQUINONE
DERIVATIVES
ANDRZEJ BAK1, VIOLETTA KOZIK2, MIROSLAW WYSZOMIRSKI3, TOMASZ MAGDZIARZ1, ADAM
SMOLINSKI4 AND JAROSLAW POLANSKI1
1
Department of Organic Chemistry, Institute of Chemistry, University of Silesia, Katowice, Poland
2
Department of Organic Synthesis, Institute of Chemistry, University of Silesia, Katowice, Poland
3
Faculty of Materials and Environmental Sciences, University of Bielsko-Biala,Poland
4
Central Mining Institute, Department of Energy Saving and Air Protection, Central Mining Institute
Katowice,Poland
[email protected]
Introduction
A comprehensive projection of the molecular structure into the compound property space has been in
the center of interest of modern chemistry for quite some time. An expansion of rational design
techniques, particularly the quantitative activity-structure relationship (QSAR), and its variants have
been described in various studies. However, a diversity of inter/intramolecular phenomena involved in
the interaction of a molecule with the corresponding environment makes the modeling of the
environmental
response
a complicated issue.
In spite of the satisfactory QSAR assessments focused on modeling dye-cellulose affinity, it is still
controversial whether a pharmacophore concept or its extended idea of tinctophore might be directly
applied in dye chemistry to predict tinctorial properties. The arrangement of dye molecules on the
polysachaccharide surface suggested existence of binding cavities or sites of higher or lower affinities
in the crystalline region capable of incorporating a dye molecule. Despite considerable variations
between dye tinctophores and drug pharmacophores some RI mD-(Q)SAR assessments have been
conducted, which indicated that the shape-determined dyeing affinity rules as a cofactor and
significantly contributed to the electrostatic field distribution.
A major objective of the presented study is a systematic analysis of tinctorial properties of
anthraquinone dyes using a standard and neural formalism of 4D-QSAR method together with the IVEPLS procedure. The applied 4D-QSAR methods take into consideration an impact of receptor-like
cavities on a supramolecular cellulose structure during a dye-fiber interaction modeling. The dye-fiber
39
geometry is crucial to explain dyeing affinity and can provide better insight into dye-cellulose
interactions. The application of the RD 4D-QSAR and its neural counterpart, namely RD SOM-4DQSAR was presented to verify a pharmacophore and/or tinctophores concept in dye chemistry and
discuss the rules governing QSAR modeling. Additionally, we examined performance of the IVE-PLS
method employed for the variable elimination of the dye data sets with the external model validations
using SDEP and q 2 test estimators, respectively. The automated data reduction using IVE-PLS
procedure represents a filter for selecting only those descriptors, which have the highest individual
weightings to the observed dye affinity indicating areas potentially relevant for dye-fiber interactions.
Moreover, the stochastic SMV procedure to investigate the predictive ability of the method for a large
population of 4D-QSAR models was employed. The obtained findings were compared with the
previously published RI 3D/4D-QSAR models for the corresponding anthraquinone trainings sets. The
neutral (protonated) and anionic (deprotonated) forms of anthraquinone scaffold were examined in
order to deal with the uncertainty of the dye ionization state. The results are comparable to both the
neutral and anionic dye sets regardless of the occupancy and charge descriptors applied, respectively.
Conclusions
The
presented
RD
4D-QSAR
methodology
together
with
IVE-PLS
procedure
provides
a robust and predictive modeling technique, which facilitates detailed specification of the molecular
motifs significantly contributing to the fiber-dye affinity.
40
DETERMINATION OF BIOLOGICALLY ACTIVE COMPOUNDS IN
FUNGI OF GENUS CORDYCEPS SINENSIS BY HPLC AND NMR
ZDENKA BEDLOVIČOVÁ, LUCIA UNGVARSKÁ MAĽUČKÁ, ANETA SALAYOVÁ, JARMILA
HARVANOVÁ, PETER OČENÁŠ
University of Veterinary Medicine and Pharmacy, Košice, Slovakia, [email protected]
Cordyceps sinensis is the fungi parasiting larvae, pupae and imagoes of insect as well as fruiting
bodies of truffles of the genus Elaphomyces 1). The fungi is known in both the traditional Chinese
medicine and in the modern medicinal methods. It is used as dietary supplement (CORDYCEPS
MRL.®, ACAI DETOX®). The fact of Cordyceps sinensis consequence is supported by many scientific
studies, which showed its positive effects for example in anti-tumor therapy2), in treatment of HIV/AIDS,
asthma, liver diseases and it also has positive effect on female fertility etc3). This work is focused on the
basic research of identification studied biologically active compounds presented in fungi by HPLC (High
Performance Liquid Chromatography) and NMR (Nuclear Magnetic Resonance).
1)
Hobbs Ch. Medicinal mushrooms: an exploration of tradition, healing, and culture. Santa Cruz (CA): Botanica
Press 1995; 251 s.
2)
Wang B. J., et al. Food Chem. Tox. 2005; 43, 543-552.
3)
Gong Y. X. et al. Chromatogr. A 2004; 1055, 215-221.
41
CHARACTERIZATION OF NEW GREEN FLUOROPHORES BASED
ON QUINOLINE SCAFFOLD.
BARBARA CZAPLIŃSKA1, ANNA MROZEK-WILCZKIEWICZ2,3, MARZENA RAMS-BARON2,3,
EWELINA SPACZYŃSKA1, MATEUSZ KORZEC1, ROBERT MUSIOŁ1
1
2
A. Chełkowski Institute of Physics, University of Silesia, Katowice, Poland
3
Silesian Center for Education and Interdisciplinary Research, Chorzow, Poland
Introduction
Fluorescence phenomenon is widely used in chemical and life sciences. As a tool of investigation, it
allows us to gain insight into a world of biological structures and cellular mechanisms. Moreover,
fluorescent dyes are commonly used to imaging of cancer and neurological diseases, as indicators for
aminoacids and sugars or as micro- and macroelements sesnsors [1,2]. Molecule has to meet a
number of conditions to become a fluorescent dye, whose are not always easy to gain. That is why
searching for new, better fluorophores is still of interest.
This work presents synthesis and characterization of over a dozen of novel Schiff bases based on a
quinoline scaffold that exhibit fluorescence properties.
Starting from 2-methylquinoline, we obtained 2-(4-amino-trans-styryl)quinoline, which constituted a
starting material for further Shiff bases synthesis [3]. The last reaction step was carried out under
microwave conditions. The structures of newly synthesized compounds were confirmed by 1HNMR and
13
CNMR
spectroscopy.
Their
biological
activity
spectrum
was
checked
on
human
colon
adenocarcinoma cells (HCT116). The fluorescence properties were measured at room temperature with
using
of
U-2900
spectrophotometer
(Hitachi)
and
F-7000 spectrofluorimeter (Hitachi). Additonally, Gaussian09 software package was applied in
theoretical DFT and TD DFT calculations with 6-311+G(d,p) as a basis set and CAM-B3LYP as a
hybrid exchange-correlation functional.
In result we designed and synthesized thirteen hitherto undescribed compounds in general synthesis
pathway presented in Fig.1.
They exhibit good fluorescence properties with emission in green light region and quite large Stokes
shifts (around 100 nm). Moreover, they show positive and negative solvatochromism dependent on
change of dipole moment of molecule in ground and excited state. TD DFT calculations were carried
out with use of two exchange-correlation functionals – B3LYP and CAM-B3LYP, the latter seems to be
42
a better choice. Examination of biological activity did not implicated any significant results – in general,
all of them turned out to be inactive.
Fig. 1: General synthesis pathway of new fluorophores.
Conclusions
Thirteen novel fluorophores based on quinoline scaffold were synthesized and described. New
compounds exhibit fluorescence properties which were examined. Additionally, theoretical calculations
using the density functional theory (DFT) and time-dependent DFT methods were carried out and then
compared with experimental data. To gain a deeper understanding of spectroscopic nature, orbital
energy analysis was also performed. Moreover, their biological activity spectrum was also assayed. As
general conclusion the presented compounds may become useful dyes for biological applications.
1)
Orzeł Ł., Dąbrowski J., Zastosowanie pomiarów fluorescencji w biochemii i chemii bionieorganicznej, Zespół
Fizykochemii Koordynacyjnej i Bionieorganicznej,Wydział chemii UJ
2)
Tsien R., Fluorescence readouts of biochemistry in live cells and organizm, Molecular Imaging: Principles and
practice, 2008, p. 808-828
3)
Cieślik W., et al. Bioorg.& Med.Chem., 2012, 20, 6960-6968
4)
Valeur B, Molecular Fluorescence: Principles and Applications, 2001
43
STUDY OF BUTYRYLCHOLINESTERASE WITHIN EXPERIMENTAL
ARTHRITIS
TOMÁŠ ČAVOJSKÝ1, LUDMIĽA
PAŠKOVÁ1, FRANTIŠEK BILKA1, LUKÁŠ
SLOVÁK 2,
KATARÍNA BAUEROVÁ2, INGRID PAULÍKOVÁ1
1
Department of Cell and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University, Bratislava,
Slovakia
2
Institute of Experimental Pharmacology and Toxicology, Slovak academy of sciences, Bratislava, Slovakia
[email protected]
Introduction
Inflammation is a protective response whose goal is to eliminate the injury-inducing agent, prevent
tissue damage and initiate the repair process. BuChE is involved in the several functions: in lipoprotein
metabolism, hydrolysis of acetylcholine and non-choline esters1. Important function is suggested for
BuChE also in the cholinergic anti-inflammatory pathway mediated by the neurotransmitter
acetylcholine (Ach) which exerts and directs inhibition of the pro-inflammatory cytokines production2.
Increasing of BuChE activity may lead to the greater hydrolytic destruction, this resulted in a reduction
of Ach concentration, what could trigger and perpetuate systemic inflammation. Rheumatoid arthritis
(RA) is the most common inflammatory rheumatic disease, manifested by inflammation of the joints
also in other organs especially the lungs, blood vessels and heart. The reason of RA is determined by
autoimmune processes which leads to the deficit of T-suppressor function of lymphocytes 3.
Characteristic for the RA is increased levels of proinflammatory cytokines, mostly prominent are
considered Il-1β and TNFα.
Materials and methods
For simulation of RA conditions we used the model of experimental arthritis. Experimental arthritis was
induced by intradermal injection of a suspension of Mycobacterium butyricum injected to the base of the
tail.. The injection contained 100 mg of heat-killed mycobacteria suspended in incomplete Freund's
adjuvant (0.1 ml). The experiment lasted 28 days, involving 3 groups of the experimental rat animals:
the group of RA, the group of treated RA with methotrexate and the control group; each contained 10
animals. We determined BuChE activity by Ellman method and its biotransformation activity using
BCH2 as a substrate. We also determined the mRNA transcriptional level of BuChE.
The model of RA was confirmed by arthritic score, body weight change and reduced expression of CRP
and IL-1β protein level in plasma. Under the inflammatory conditions, we observed a significant
decrease of BuChE biotransformation activity, which accounted for 50% of the original activity in the rat
44
lungs and even 30% in the liver comparing to the control group. The kinetics of the enzyme hydrolysis
of BuChE was also altered significantly. In both organs of the inflammatory model we observed a
significant increase of the KM value. The expression of BuChE gene has shown interesting finding by
increased transription level of BuChE in the liver (p˂0,05), and by contrast the significant reduction in
the lungs (p˂0,0001) and spleen (p˂0,001).
Conclusions
Decreased biotransformation activity of BuChE in both tissues may eventually result in an increased
concentration of the drug in the cell with serious consequences for the treatment.
This work was supported by grants: APVV 0052-10, UK/296/2014 and VEGA 2/0045/11, 2/0044/15. This
contribution is the result of the project implementation: Comenius University in Bratislava Science Park supported
by the research and development operational programme funded by the ERDF. Grant number: ITMS
26240220086.
1)
Patocka J., Kuca K., Jun D. Acta. Med. 2004, 47, 215-28.
2)
Rosas-Ballina M., Tracey K.J. J. Intern. Med. 2009, 265, 663-79.
3)
Gravallese E.M. Ann. Rheum. Dis. 2003, 61,84-86.
45
SELECTED ALAPTIDE ANALOGUES AND THEIR EFFECT
ON THE PENETRATION OF THEOPHYLLINE
ANETA CERNIKOVA, PAVEL BOBAL, JOSEF JAMPILEK
Department of Chemical Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical
Sciences, Brno, Czech Republic; [email protected]
There are many ways of administration of drugs into the organism. Possibilities of
transdermal application of drugs are extensively studied. The composition of the outermost
horny layer of the skin (stratum corneum) protects the body against the permeation of most
xenobiotics. There are many approaches to increase the penetration of drug substances
through the skin. One of them is the use of chemical penetration modifiers (CPMs). The
exact mechanism of action of CPMs is not known, but there are several hypotheses of
probable pathways of the penetration of drugs, which can be modified by CPMs
can be administered on the skin in the form of creams, ointments, gels
4)
1–3)
. Drugs
as well as
transdermal patches (patches are commonly used for application of contraceptives, smoking
cessation, reduction of pain, etc.)
1, 2)
. CPMs modify the penetration/permeation of drugs
across the stratum corneum both is the case when local effect of the drug in the skin is
desired (e.g., corticosteroids) and in the case when the systemic effect in the organism is
required (e.g., hormonal contraceptives).
Our research team began to study alaptide as a potential penetration/permeation modifier
several years ago
5–9)
. Alaptide (8-methyl-6,9-diazaspiro[4.5]decane-7,10-dione) is a cyclic
dipeptide that is capable to interact with components of the skin. It is an original Czech
compound prepared in the 80s of the 20th century. Alaptide showed significant skin curative
effect, while any toxic, teratogenic or embryotoxic effects were not observed
10–12)
. The aim of
this study is to investigate how the application of selected alaptide analogues (AAs)
9)
affects
the permeation of theophylline from various media, including propylene glycol (mixed with
water, 1:1) and physiological buffer (pH = 7.4), through full-thickness pig ear skin using static
Franz cells.
This study was supported by the IGA VFU Brno 302/2015/FaF.
1)
Jampilek J., Brychtova K. Med. Res. Rev. 2012; 32, 907–947.
2)
Jampilek J. J. Bioequiv. Availab. 2013; 5, 233–235.
3)
Cernikova A., Jampilek J. Chem. Listy 2014; 108, 7–16.
4)
Jampilek J., et al. Patent WO/2013/020527 A1, 2013.
5)
Opatrilova R., et al. Sci. World J. 2013; 2013, Article ID 787283 (8 pages).
6)
Opatrilova R., Jampilek J. ADMET 2014; 2, 56–62.
7)
Cernikova A., et al. Mil. Med. Sci. Lett. 2014; 83, 34–39.
8)
Jampilek J., et al. Czech Patent Application PV 2014-416, 2014.
9)
Cernikova A., et al. ADMET 2014; 2, 248–253.
10)
Kasafirek E., et al. Belg. 897843, 1984 & CS 231227, 1986.
11)
Radl S., Kasafirek E., Krejci I. Drugs Fut. 1990; 15, 445–447.
12)
Jampilek J., et al. Patent WO/2014/019556 A1, 2014.
NEWLY
IDENTIFIED
CHEMICAL
CONSTITUENTS
OF
PARASITIC PLANT LATHRAEA SQUAMARIA L.
IVANA DAŇKOVÁ1, MILAN MALANÍK1, STEFANO DALL´ACQUA2, MARKÉTA
GAZDOVÁ1, ZUZANA HANÁKOVÁ1
1
Department of Natural Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical
Sciences Brno, Czech Republic
2
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Italy
Lathraea squamaria L. (Orobanchaceae) is a non-chlorophyllous root parasitic plant.
According to the prior phytochemical investigations
1,2)
, the vegetative parts of the plant
contain mainly iridoid glycosides (e.g. aucubin, melampyroside) and phenolic acids.
In previous research work carried out on our department phenylpropanoid glycosides
and benzoic acid were identified, these compounds were described in this plant species
for the first time. Moreover, we have found out that other important chemical constituents
of the plant with potential biological activity have not been determined yet. The aim of the
present study was to bring a new knowledge about other chemical constituents of the plant.
The extract of the underground part of the plant was prepared by ultrasound-assisted
maceration in methanol and analysed by means of HPLC on reverse phase. We focused
on several polar substances which have not been identified in this plant so far. Subsequently,
the methanolic extract was subjected to liquid-liquid fractionation with chloroform,
ethylacetate and water successively; the ethylacetate aliquot was used for separation. Four
fractions of unknown compounds were isolated by means of the preparative liquid
chromatography and analysed by using available spectral methods (UV, IR, HRMS, 1H NMR,
13
C NMR). Based on the results of mass spectrometry and nuclear magnetic resonance
and according to the comparison with the data from literature 3), the
iridoid
scyphiphin B1.
The
presence
of
this
O
O
substance from the fraction 4 was identified as non-glycosidic
H
type
of compounds has been determined in Lathraea squamaria L. for
the first time. Identification of the remaining three compounds from
isolated
fractions
will
be
continued.
We
suppose
that they could have similar chemical structure.
1)
Swiatek, L., Dombrowicz, E. Pol. J. Pharmacol. Pharm. 1976; 28, 105-109.
2)
Grabias, B., et al. Phytochemistry. 1993; 32, 1489-1491.
3)
Zeng, Y. B., et al. Chin. Chem. Lett. 2007; 18, 1509-1511.
O
HO
H
OH
CH3
ANALYSIS OF FLAVONOIDS IN GRAPE LEAVES BY HPLCDAD-MS/MS
SVETLANA DOKUPILOVÁ, LUCIA VEIZEROVÁ, JAROSLAV GALBA, PETER MIKUŠ
Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy, Comenius
University, Bratislava, Slovakia, [email protected]
Introduction
Flavonoids are water soluble polyphenolic molecules, that belong to a group of plant
metabolites thought to provide health benefits through cell signalling pathways and
antioxidant effects. flavonoids are the most abundant and widely studied, and have enjoyed
greater attention among grape researchers1.
Grape leaves of various grape varietes obtained from Research Institute of Viticulture and
Enology in Slovakia were analysed. Samples were collected in September 2013. Leaves
were dried and frozen (-20°C) until treated prior analysis. Samples were homogenized and
2,5g of each sample was extracted with 40 ml mixture of methanol/water 80:20 (v/v) for
120min and 2x20 ml dichlórmethene was used for sample clean-up. 2,5 ml of watermethanolic part was dried and dissloved in mobile phase. Analysis was performed on LC
Agilent 1200 Infinity System (Agilent Technologies, USA) with Agilent 1260 Infinity Diode
Array Detector and
Agilent 6520 Accurate-Mass Quadrupole Time-of-Flight. Separation
column was Kromasil C18 (4,0×150 mm, 3,5 µm). Mobile phase consisted of acetonitrile and
formate buffer (ammonium formate with concentration 10 mmol/l and pH adjusted to 3,1 by
formic acid). Gradient elution 10% -95% of acetonitrile in 62min. was used.
MS Detection conditions were: negative mode (ESI-), capillary voltage: 3,5 kV, drying gas
temperature: 300 °C, drying gas flow rate: 10 l/min, nebulizer pressure: 40 psi, fragmentor
voltage: 140 V, collision energy: 20 eV.
Identification of the separated compounds was carried out using a UV, MS and MS/MS
spectra. 7 quercetin and 4 kaempferol glycoside flavonoids (quercetin-3-O-rutinoside,
quercetin-glucuronide,
quercetin-3-O-galactoside,
quercetin-3-O-glucoside,
quercetin-
pentoside 1, kaempferol-3-O-rutinoside, kaempferol-hexoside 1, quercetin-pentoside 2,
quercetin-3-O-rhamnoside,
kaempferol-hexoside
2,
kaempferol-deoxyhexoside)
were
identified. DAD detector and external calibration curves of quercetin and kaempferol (in the
range 5-100 µg/ml) were used for quantification. 9 of the 13 identified compounds could be
quantified (see Table 1).
Table 1. content of glycoside flavonoids in grape leaves:
Compound
Range of content (μg/g)
quercetin-3-O-rutinoside
41,79 - 1576,58
quercetin-glucuronide
1385,49 - 7284,86
quercetin-3-O-galactoside
95,49 - 1152,52
quercetin-3-O-glucoside
83,51 - 4665,55
quercetin-pentoside 1
<LOD - 1022,68
kaempferol-3-O-rutinoside
<LOQ - 913,51
kaempferol-hexoside 1
<LOQ - 1348,04
quercetin-3-O-rhamnoside
<LOQ - 1307,75
kaempferol-hexoside 2
<LOQ - 2380,98
It was found, that the highest contained flavonoid was: quercetin-glucuronide, the content of
which ranged from 1385 to 7284 μg/g. The amount of other components varied from 41,79
μg/g to maximum value of 4665 μg/g. The method validation confirmed sufficient linearity,
precision, accuracy and separation efficiency. Low limits of detection and limit of
quantification (LOD = 1,12 μg/ml, LOQ = 3,73 μg/ml for quercetin and LOD = 2,35 μg/ml,
LOQ = 7,84 μg/ml for kaempferol) were detected.
Conclusions
It was confirmed that the proposed method is suitable for the comprehensive analysis of
complex plant matrices, such as grape leaves. The results of research will contribute to the
creation of profiles of pharmaceutically active substances contained in the leaves of nearly
30 Slovak vine varieties that could be used in the future to prepare nutraceuticals.
Research was supported by grant: APVV-0550-11
Liu, C. et al. Food chem., 2013; 136, 643-649.
FROM AN OLD DRUG TO A NEW ONE: SYNTHESIS OF
VALPROATE FROM 5,5-DIPROPYLBARBITURIC ACID
OLDŘICH FARSA, PAVOL BRKA
Sciences Brno, Czech Republic;
Introduction
Barbiturates are old-fashioned and mainly obsolete sedatives, hypnotics and antiepileptics.
On the contrary, valproic acid and its salts are modern and popular therapeutics used not
only as antiepileptics. That is why we have proposed usage of our old department reserves
of 5,5-dipropylbarbituric acid as a as the strarting material for new experiment in our
Medicinal Chemistry practical courses in which valproic acid will be prepared.
Experiments involved optimization of two synthetic steps: hydrolysis of 5,5-dipropylbarbituric
acid to 2,2-dipropylmalonic acid and decarboxylation of the latter to 2-propylpentanoic, eg.
valproic acid.
The two-stage synthetic procedure for synthesis of valproic acid has been developed. The
first stage is the basic hydrolysis of 5,5-dipropylbarbituric acid to 2,2-dipropylmalonic acid in
larger amount1). This step will be performed by a laboratory technician to avoid the contact of
students with the barbiturate. The second stage is the thermal decarboxylation of 2,2dipropylmalonic acid to valproic acid 2). This step will be performed by students either in a
microwave reactor or by a conventional heating. After the reserve 5,5-dipropylbarbituric acid
will be consumed the first stage will be replaced with a conventional synthesis of 2,2dipropylmalonic acid from diethyl malonate.
Conclusions
A simple two-step synthetic procedure which enables the synthesis of valproic acid from 5,5dipropylbarbituric acid suitable for practical courses has been developed.
1)
Aspelund H., Skoglund L. Acta Acad. Aboensis Math. et Phys. 1937; 10, 14-36.
2)
Helavi V. B., et al. J. Chem. Research (S), 2003; 174-175.
MOBILE APPLICATIONS AND THEIR POTENTIAL FOR USE
IN PHARMACEUTICAL PRACTICE
TOMÁŠ FAZEKAŠ, JURAJ MIČIAN
Dept. of Physical Chemistry of Drugs, Faculty of Pharmacy, Comenius University in Bratislava,
Slovakia, [email protected]
Mobile health (m-health) applications installed on smartphones appear as a good possibility
for improvement of specific interactions between and within healthcare providers and
patients. Smartphones, integrating phone, browser, e-mail, media player, camera, GPS,
voice recognition, are currently experiencing perhaps a greater boom than in his time internet
had.
Thus m-health exhibits a growing potential for use in pharmaceutical theory and
practice.
Only to little information exists about patients m-health preferences in Slovak
Republic, mainly because the development of this technology is relatively new and rapid.
The aim of the presented work was to identify interest in using m-health related applications
among Slovak patients. Demographic data, experiences, interest, preferences and
empowerment were collected and processed with statistical software PSPP. A questionnaire
with 4 dimensions was designed, pretested and validated by factor analysis and Cronbach
alpha (>0.86). The questionnaire was administered to 200 persons with 97% response rate.
The majority of identified users were younger and belonged to the age group 18-29 years
old. The expected impact of higher education and / or professional qualification on more
intensive use of electronic services not was not been proven. M-health applications are still
not been used by the majority of Slovak patients, because of poor awareness and language
barriers. Initiative in this field expected from the state regulator respectively from a
professional organization. Development and implementation of such applications should be
supervised by pharmacists, pharmacies, importers and distributors or customers themselves.
Expected functionalities of m-health applications are mainly these: product information and
comparison, generics checker, product availability checker, pricing, ability to evaluate
medicines and pharmacies, drug reminder, pill identifier, drug interaction checker. The lowest
interest by respondents was given to the opportunity to contact the pharmacist trough an
application, so customers still prefer personal consultations with a pharmacist.
Based on results of our research we think that a typical Slovak customer is ready to utilize
electronic pharmacy services. His requirements are exactly defined but he expects that these
will be provided to him free of charge. It will be interesting to follow up, how these customer
needs will be reflected by the domestic market in the near future.1,2
1)
BAILEY, Stacy Cooper, et al. JAMIA, 2014, 21.3: 542-546
2)
POWER, Jessica MH, et al. Health Pol. Tech., 2014, 3.4: 296-305.
PREPARATION
AND
COMPLEXES
WITH
ANALYSIS
OF
GALLIUM
AMINOCARBOXYLATES
(III)
AND
SULFONAMIDES
1
ANDREA FORGÁCSOVÁ, 1,2PETER MIKUŠ, 1EMIL HAVRÁNEK AND 1,3MILAN MELNÍK
1
Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy of Comenius
University in Bratislava, Slovakia
2
Toxicological and Antidoping Center, Faculty of Pharmacy, Comenius University in Bratislava,
Slovakia
3
Department of Inorganic Chemistry, Slovak Technical University, Bratislava, Slovakia
[email protected];
Gallium complexes has shown efficacy in the treatment of several apparently diverse
disorders. Gallium has some medically useful radionuclides which have made extensive
contribution in both the diagnosis and therapy of some diseases especially with usage in
oncology, for example
67
Ga,
68
Ga. Besides platinum, gallium is also very promising for the
treatment of some cancer diseases. Specificity of the Ga pharmaceuticals toward the target
is usually applied as a complex. Complexation with organic ligands has been recognized as
a promising strategy for creating new diagnostic and tumour-inhibiting compounds with a
number of advantages. Chelating agents like macrocyclic chelators (DOTA, NOTA) are used
to form more stabile complexes. Radiolabeled amino acids and peptides are of increasing
interest in nuclear medicine with special emphasis to peptides labelled with positron emitters.
Our research was aimed at the development of ternary Ga complexes with selected amino
carboxylates and sulphonamides as potential diagnostic and/or therapeutic agents in nuclear
medicine and oncology.
Acknowledgement: This work was supported by the projects VEGA 1/0873/15, KEGA 022UK4/2015, FaFUK/14/2015 and carried out in the Toxicological and Antidoping Center and at the
University in Bratislava.
DETERMINATION OF CMC OF CATIONIC TENSIDE IN
AQUEOUS AND MIXED WATER-ALCOHOL SOLUTIONS
ANDREA GÁLUSOVÁ, FILS ANDRIAMAINTY, ROMAN MIKLÁŠ
Dept. of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University in Bratislava, Slovakia
The aim of this paper was to determine influence of short-chain alcohols on CMC of cationic
tenside
which
belongs
to
the
group
of
quaternary
ammonium
compounds
(1182-RM-12-14). CMC was determined in aqueous and mixed water-alcohol solutions methanol, ethanol and propanol with concentration 0,1M, 0,2M and 0,5M by measuring
absorbance of pyrene in tenside solution. The dependence of absorbance vs. surfactant
concentration for unmasked pyrene peaks has exhibited typical sigmoidal plot. The
Sigmoidal-Boltzmann equation was used to determine the CMC of studied compound in
solutions with different alcohol concentrations. In studied concentration range the CMC was
decreased by increasing concentration of alcohol, the most by methanol, less by ethanol and
the least by propanol.
The work was supported by the grant No. UK/265/2015.
1)
Tanford C. The Hydrophobic Effect: Formulation of Micelles and Biological Membranes. 2nd edition.
New York: Wiley-Interscience Publication 1980; 233 p.
2)
Huang J., Mao M., Zhu B. Colloids surf. A 1999; 155, 339-348.
3)
Bielawska M., et al. Colloids Surf. A 2013; 424, 81-88.
4)
Basu Ray G., Chakraborty I., Moulik S. J. Colloid Interface Sci. 2006; 294, 248-254.
5)
Kalyansundaram K., Thomas J. K. J. Am. Chem. Soc. 1977; 99, 2039-2044.
6)
Khan Z. H., Hhanna B. N. J. Chem. Phys. 1973; 59, 3015.
7)
Aguiar J., et al. J. Colloid Interface Sci. 2003; 258, 116-122.
VIRTUAL SCREENING OF COMBINATORIAL LIBRARY OF
1,3,5-TRIAZINE
MOIETY
SUBSTITUTED
WITH
BENZENESULFONAMIDES AND DIFFERENT AMINES
VLADIMÍR GARAJ
Department of Pharmaceutical Chemistry, Faculty of Pharmacy UK, Bratislava, Slovakia
[email protected]
Carbonic anhydrase (CA) IX is one of the proteins that are involved in cancerogenesis in
hypoxic breast tumors. Indeed, it is present in high amounts in hypoxic tumor cells, where it
plays a crucial role in metabolic reprogramming due to the scarcity of oxygen, triggered by
the hypoxia inducible factor 1 transcription factor. In such tumors, CA IX is involved, along
with other proteins, in tumor pH regulation and its survival in a harsh environment (including
hypoxia and acidity). CA IX is also validated as an imaging and treatment target for hypoxic
tumors and metastasis1).
A virtual library of novel benzenesulfonamides was formed by substitution of Cl on
cyanurchloride with 18 different amines similarly to previous experimental works 2). The library
of virtually assembled compounds was computationally screened against human CA IX and
CA II using Glide and quantum polarized ligand docking workflow from Schrödinger. ADMET
properties were predicted for compounds with best score from docking to CA IX and best
ratio of CA IX / CA II score using QuickProp.
This research was financed by grant VEGA No. 1/0743/13.
1)
Supuran C. T., Winum J. Y. Expert. Opin. Drug Discov. 2015; 10, 591–597.
2)
Carta F., et al. Bioorg. Med. Chem. 2011; 19, 3105–3119.
SYNTHESIS AND BIOLOGICAL ACTIVITY OF SELECTED
CINNAMIC ACID DERIVATIVES
MARTIN GAZVODA, SLOVENKO POLANC
Faculty
of
Chemistry
and
Chemical
Technology,
University
of
Ljubljana,
Slovenia;
[email protected]
The enzymes AKR1C1‒AKR1C4, members of the aldo-keto reductase superfamily, catalyse
the interconversions of 3-, 17- and 20-ketosteroids with the corresponding 3α/β-, 17β-, 20αhydroxysteroids to varying extents, using NADPH as a cofactor 1). In this way they can control
the ligand occupancy and trans-activation of androgen, estrogen and progesterone receptors
by modulating the concentrations of the active steroids. The AKR1C enzymes are also
involved in the prostaglandin and neurosteroid production and inactivation, and in the
metabolism of xenobiotics. Among them, AKR1C3 preferentially acts as a 17-ketosteroid
reductase and converts a weak androgen 4-androstene-3,17-dione to a potent androgen
testosterone, and estrone to a potent estrogen 17β-estradiol. AKR1C3 also catalyses the
reduction of prostaglandin H2 (PGH2) into PGF2α, and PGD2 into 11β-PGF2, thereby
diverting the biosynthesis of prostanoids away from the antiproliferative J-series. Catalysing
these reactions AKR1C3 represents an important target enzyme for the development of
potential drugs for a treatment of the hormone dependent and hormone independent forms of
cancer.
1)
Penning T. M., Byrns, M. C. Ann. N. Y. Acad. Sci. 2009, 1155, 33‒42 and references therein.
2)
Byrns M. C., et al. J. Steroid Biochem. Mol. Biol. 2010, 118, 177‒187.
SYNTHESIS
AND
ANTIMICROBIAL
ACTIVITY
OF
N-
(HALOGEN-PHENYL)-1-HYDROXYNAPHTHALENE-2CARBOXAMIDES
TOMÁŠ GONĚC, JURAJ KMEŤKO, JIŘÍ KOS, ŠÁRKA POSPÍŠILOVÁ, IVETA
ZADRAŽILOVÁ, JOSEF JAMPÍLEK
Sciences Brno, Czech Republic, [email protected]
Introduction
Recent studies have shown that, despite antibacterial therapy, methicillin-resistant
Staphylococcus aureus (MRSA) infections are still associated with serious clinical
consequences, especially treatment failure, higher morbidity and mortality, prolonged
hospitalization, increased health care costs, etc. Activity against MRSA is of a great
importance in the new generation of antibacterial agents because of the worldwide
increasing prevalence of this pathogen, more frequent antibiotic resistance to available antiMRSA drugs, their toxicity and general lack of oral agents 1). Some recently published
derivatives with halogen substitution showed promising activities. 2)
The aim of recent work was to synthetize series of N-(phenyl)-1-hydroxynaphthalene-2carboxamides with more halogen atoms on anilide fragment (Fig. 1). 39 compounds with di-,
tri-, tetra- and penta- halogen-substituted anilide ring were prepared according to well
approved microwave-assisted synthetic method2). The identities of compounds were
confirmed by
1
H and
13
C NMR, HRMS and IR spectroscopy. Preliminary results of
antibacterial activity against four strains of Staphylococcus aureus (among them 3 are
methicillin-resistant) showed that 10 compounds have comparable or even higher MIC than
standards ciprofloxacin and ampicillin.
NH
R
OH
O
Fig. 1. Studied compounds. R: di-, tri-, tetra-, penta-,-F, -Cl, -Br
Project was supported by IGA 320/2015/FAF
1)
Goněc T., et al. Molecules 2015, 20, 9767-9787.
2)
Goněc T., et al. Bioorg. Med. Chem. 2013, 21, 6531-6541
SOLUBILITY AND DISSOLUTION RATE IMPROVEMENT OF
CANDESARTAN CILEXETIL
LUCIE GRUBEROVÁ1, BOHUMIL KRATOCHVÍL1, LENKA SEILEROVÁ1
1
Department of Solid State Chemistry, University of Chemistry and Technology Prague, Czech
Republic; [email protected]
Candesartan cilexetil (CC) is a antagonist of angiotensin II receptor, commonly used to treat
hypertension. Based on solubility and absorption characteristics, CC is classified as a class II
drug in the Biopharmaceutics Classification System. Candesartan cilexetil is highly lipophilic
compound and is practically insoluble in water. Low solubility of CC across the physiological
pH range resulted in an incomplete absorption from the gastrointestinal tract (GIT). Solubility
and rate of dissolution are the most important determinants of candesartan cilexetil
absorption in the GIT. The aim of this investigation is to improve solubility and dissolution
rate using solid dispersion of candesartan cilexetil prepared by hot-melt extrusion.
Creation of sink conditions for poorly soluble compounds is limited by a large volume of
medium. Volume of the medium can be decreased by addition of surfactants which can
significantly increase drug solubility. The first objective of the study was to investigate the
solubility of candesartan cilexetil in a phosphate buffer with different ionic strength, pH value,
type and amount of surfactant (Tween 20 or 80). Tween 20 has significantly increased the
solubility of CC, with a bigger effect observed at higher concentration (0,70 %, w/w). Higher
concentration of Tween 80 resulted in slightly increase of CC solubility. It was observed that
the increased ionic strength of phosphate salts allows to increase solubility of CC. The
solubilization of candesartan cilexetil is more effective at higher pH. Results demonstrate that
at least 240 ml of 0,05 mol/l of phosphate buffer at pH 6.5 with 0,35 % of Tween 20 is
required for 8 mg content of CC in tablets in order to achieve minimal sink conditions in
dissolution studies (according to the US Pharmacopeia).
The second aim of this study was to characterize the solid dispersion of candesartan cilexetil
prepared by hot-melt extrusion (HME). HME is currently generating more and more interest,
as the percentage of newly produced poorly soluble drugs is constantly increasing. Hot-melt
extrusion is still an emerging technology and its potential in pharmaceutical technology has
not yet been fully explored. In this study, extruder with single screw was used.
Polyvinylpyrrolidone (PVP 25) and meglumine as carrier material, CC and selected
excipients were used to create different batches for extrusion. The extruded material was
pulverized and then sifted through sieve to obtain uniform size particles. Subsequently, the
powder was compressed to tablets (0,5-2,0 t) or filled into gelatin capsules (4 or 8 mg CC).
The physicochemical properties of solid dispersions were determined by XRDP, DSC and
FTIR. All prepared tablets were evaluated for hardness, friability and disintegration time.
Dissolution studies were carried out on samples with suitable characteristics. For these
samples was proved enhanced dissolution profile in comparison to physical mixture and pure
drug. The highest amount of candesartan cilexetil dissolved after 120 minute was 89 %. The
faster dissolution rate and higher solubility of solid dispersions can be attributed to entrapped
CC into the hydrophilic coat of carriers and excipients and its crystallinity changes.
Financial support fromspecific university research (MSMT No 20/2015).
ESTIMATION OF LIPOHYDROPHILIC PROPERTIES OF
MOLECULES WITH POTENTIAL β3-AGONISTIC ACTIVITY
ĽUBICA HAVRANOVÁ SICHROVSKÁ1, LUKÁŠ STANZEL1, IVAN MALÍK1, MATEJ
MARUNIAK1, IVA KAPUSTÍKOVÁ1, EVA SEDLÁROVÁ1, JOZEF CSӦLLEI2
1
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University Bratislava,
Slovak Republic
2
Sciences, Brno, Czech Republic
[email protected]
The paper present the lipophilicity study of four newly syntethized substances potentially
active as β3-adrenergic receptor agonists labelled as BL-14S2-BL-44S2. Partition coefficient
(log P) was estimated in three systems consisted of lipophilic and hydrophilic medium.
Generally accepted shake-flask method for obtaining the log P values in these mediums was
used. The non-polar phase was composed of octan-1-ol, cyclohexane, heptane, whereby the
aqueous phase was always represented by phosphate buffer
with pH = 7,4. Into the
lipophilic enviroment of octan-1-ol, all of the substances were able to penetrate and the
values of log PO were from 0,90 to 1,70. With the elongation of alkoxylcarbonylamino
fragment, the log PO values increases constantly. As cyclohexane and heptane are highly
lipophile solvents, only the substance BL-44S2 with butoxylcarbonylamino moiety was able
to penetrate into them. Partition coefficient in medium cyclohexane/phosphate buffer (log PC)
system was of value 0,22 and was higher than partition coefficient estimated in
heptane/phosphate buffer, which was of value 0,17, in consequence of different structural
character of molecules of the solvents metioned above.
3-{4-[(alkoxycarbonyl)amino]phenoxy}-N-{2-[4-(aminosulfonyl)phenyl]ethyl}-2-hydroxypropan-1-ammonium
chlorides
BL-14S2: R = CH3
BL-34S2: R = C3H7
BL-24S2: R = C2H5
BL-44S2: R = C4H9
Figure 1. Structure of evaluated molecules BL-14S2-BL-44S2
ANTIPROLIFERATIVE EFFECT OF 1-METHOXYBRASSININ
1
MARTINA CHRIPKOVÁ, 1NATALIA ANTOLIKOVÁ, 2FRANTIŠEK ZIGO, 3MARIANA
BUDOVSKÁ, 4JÁN MOJŽIŠ, 5LADISLAV TAKÁČ, 6DENISA TOROPILOVÁ*
1
Department of Human and Clinical Pharmacology, University of Veterinary Medicine and Pharmacy,
Kosice, Slovakia
2
Department of Animal husbandry, University of Veterinary Medicine and Pharmacy, Kosice, Slovakia
3
Department of Organic Chemistry, Institute of Chemical Sciences, Faculty of Science, Pavol Jozef
Safarik University, Kosice, Slovakia
4
Department of Pharmacology, Faculty of Medicine, Pavol Jozef Safarik University, Kosice, Slovakia
5
Department of the Environment, Veterinary legislation and Economy, University of Veterinary
Medicine and Pharmacy, Kosice, Slovakia
6
Department of Biology, Zoology and Radiobiology, University of Veterinary Medicine and Pharmacy,
Kosice, Slovakia
[email protected]
Several epidemiologic studies suggest that consumption of cruciferous vegetables may be
particularly effective in reducing cancer risk at several organ sites 1), 2). Indole phytoalexins
represent a specific group of phytoalexins synthesized by plants of the family Cruciferae.
Phytoalexins are anti-microbial secondary metabolites of low molecular weight produced by
plants "de novo" after exposure to biological, physical or chemical stress 3). These substances
are usually produced in small quantities. Chemical synthesis can provide access to
reasonable amounts of phytoalexins that are necessary to evaluate their biological activities.
Indole phytoalexins have been reported to exhibit several biological activities, including
chemopreventive4) antiproliferative,
antifungal5),
antiprotozoal6)
and
anticarcinogenic7)
activities. The unique structural feature of the majority of indole phytoalexins is the presence
of an indole ring and side chain or another heterocycle, containing a nitrogen atom and one
or two sulpfur atoms8). Until now 44 indole phytoalexins, i.e. metabolites have been isolated
and their structure elucidated9). 1-methoxybrassinin, brassinin and cyclobrassinin were the
first cruciferous phytoalexins, isolated from the Chinese cabbage after infection with the
bacterium Pseudomonas cichorii
10)
. These natural substances have been suggested as a
potential anti-tumor agents but little is known about their inhibitory mechanism on the growth
of cancer cells.
The present study was conducted to examine the effects of 1-methoxybrassinin on cell
proliferation in the different human cancer cell lines.
This work was supported by VEGA 1/0322/14 and KEGA 008UVLF-4/2014
1)
Talalay P., Fahey J. W. J Nutr. 2001; 131, 3027-3033.
2)
Tse G., Eslick G. D. Nutr. Cancer. 2014; 66, 128-139.
3)
Dixon R. A., Lamb, C. J. Annu Rev Plant Biol. 1990; 41, 339-367.
4)
Mehta R. G., et al. Carcinogenesis. 1995; 16, 399-404.
5)
Pedras M. S., et al. J Org Chem. 2004; 69, 4471-4476.
6)
Mezencev R., et al. Exp Parasitol. 2009; 122, 66–69.
7)
Izutani Y., et al. In.t J. Oncol. 2012; 40, 816-824.
8)
Pedras M. S., et al. Phytochemistry. 2000; 53, 161-176.
9)
Pedras M. S., Yaya E. E. Phytochemistry. 2010; 71, 1191–1197.
10)
Takasugi M., et al. J. Chem. Soc., Chem. Commun.1986; 14, 1077–1078.
SYNTHESIS AND BIOLOGICAL PROPERTIES OF CHOSEN
SYMMETRICAL
AMIDES
AND
THIOAMIDES
OF
TEREPHTHALIC ACID
AGNIESZKA
JĘDRZEJOWSKA1,
MAREK
MATUSSEK1,
VIOLETTA
KOZIK1,
ANDRZEJ BĄK1, IVETA ZADRAŽILOVÁ2, JOSEPH JAMPÍLEK2
1
University of Silesia, Institute of Chemistry, Katowice, Poland
2
University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic
[email protected]
The modern Organic Chemistry is a research area, which connects not only the synthesis of
new chemical compounds which have interesting properties, but also to design and predict of
attractive structures and properties. The new compounds are widely used in various
industries such as pharmaceutical, electronics, chemical and others.
Diamides of terephthalic acid obtained in the reactions with amino acids have interesting
properties. Functionalization using bioactive compounds is attractive in terms of synthetic, as
is possible in this way to get new active analogs.
Marek Matussek is co-financed by the European Social Fund, the project DoktoRIS.
STUDY OF STRUCTURE AND ADMET PROPERTIES OF AT 1
RECEPTOR ANTAGONISTS
PAVOL JEŽKO, ZUZANA SÁRAZOVÁ
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University in Bratislava,
Slovakia; [email protected]
The renin-angiotensin-aldosterone system plays a key role in the regulation of blood
pressure. This system is involved in the pathogenesis of hypertension, congestive heart
failure, and chronic renal failure. The main effector peptide, the angiotensin II, interacts with
AT1 and AT2 angiotensin receptors. AT1 receptors antagonists are useful in the treatment of
hypertension, heart failure, and renal disease.
For the study of the structure was used program Jaguar. Lipinsky rules of five, polar surface
area and ADME properties were studied in QikProp program. Toxicity of studied drugs was
calculated in program VirtualToxLab.
After geometry optimalization, by method HF/6-31G(d), were found the most stable
conformers. The ortho position of the acid group is responsible for the non-coplanar
conformation of the biphenyl part of AT1 receptors antagonists. Some drugs are absorbed in
lower amount. Drugs which contain two acidic groups and have higher polar surface area are
used as prodrugs. Low toxicity of AT1 receptor antagonists is in good agreement with low
affinity to hERG K+ channel. Agonistic effect on PPARγ receptor was shown in some AT 1
receptor antagonists (irbesartan, telmisartan a valsartan).
A COMPARISON OF SIO2-, CU-, AND NI-SUPPORTED AU
NANOPARTICLES
FOR
SELECTIVE
GLYCEROL
OXIDATION TO ACETIC ACID
MACIEJ KAPKOWSKI, MONIKA SŁOTA, JAROSŁAW POLAŃSKI
University of
Silesia in Katowice, Institute of Chemistry, University of Silesia, Katowice, Poland
[email protected]
We tested for the first time the efficiency of SiO 2-, Cu-, and Ni-supported Au in deep glycerol
oxidation in a diluted and viscous H2O2/H2O liquid phase. Acetic acid (AA), the C 2 oxidate,
was preferentially formed in such a system. High conversion (100%) and AA yields (90%)
were observed for the sol-gel SiO2-suppported Au in diluted solutions. Although with the
increase of glycerol concentration in the viscous liquid phase these values decreased to ca.
40% (conversion) and 20% (AA yield), the addition of acetonitrile improved the AA yield to
ca. 40%, while the surfactants were found to be capable of a many-fold enhancement of the
catalyst activity at the room temperature highly-viscous liquid phase. High performances
were also observed for the bimetallic Au/Cu and Au/Ni catalysts obtained by nano-Au
transfer; however, these catalysts were destroyed during the reaction by the Cu or Ni
leaching effect.
The research was co-financed by the National Research and Development Center (NCBiR) under
Grant ORGANOMET No: PBS2/A5/40/2014. Author Maciej Kapkowski expresses his appreciation for
the support of the DoktoRIS - Scholarship program for innovative Silesia, which is co-financed by the
European Union within the framework of the ESF.
DETERMINATION
CONSTANTS
OF
OF
ACID-BASE
DISSOCIATION
NEWLY
SYNTHESIZED
ARYLETHANOLAMINE DERIVATIVES USING CAPILLARY
ZONE ELECTROPHORESIS
IVA KAPUSTÍKOVÁ, MATEJ MARUNIAK, ĽUBICA HAVRANOVÁ-SICHROVSKÁ,
LUKÁŠ STANZEL, IVAN MALÍK, EVA SEDLÁROVÁ
Dep. of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University in Bratislava, Slovakia;
[email protected]
The acid-base dissociation constant (pKa) is an important physico-chemical parameter which
affects drug pharmacokinetics and toxicity (ADMET: absorption, distribution, metabolism,
excretion, toxicity). The early-obtained information about these drug properties may decrease
costly development process of a compound with poor pharmaceutical prospect.
Determination of pKa using capillary zone electrophoresis was based on measurement of the
effective electrophoretic mobility of studied substances as a function of separation electrolyte
pH. This relationship was then fitted by a sigmoidal curve, where pKa is the value of pH in the
sigmoid inflection point. pKa values obtained in buffer solution were recalculated
to thermodynamic p𝐾aT witch correspond to water medium.
Dissociation
constants
of
eight
newly
synthesized
arylethanolamine
derivatives,
with potential α and β-adrenolytic properties, were determined. As was assumed previously
from their chemical structure, pKa correspond to protonated piperazine nitrogen proximal
to the arylethanolamine chain. The pKa values of compounds does not refer about evident
influence of chemical structure (substituents pyridine-2-yl or pyrimidine-2-yl; methyl–butyl)
on acidity of the molecule.
SYNTHESIS
AND
STUDY
OF
CARDIOPROTECTIVE
ACTIVITY OF NEW DEXRAZOXANE ANALOGS
GALINA KARABANOVICH, JAROSLAV ROH, ANNA JIRKOVSKÁ-VÁVROVÁ,
KATEŘINA VÁVROVÁ, TOMÁŠ ŠIMŮNEK
Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Czech Republic;
[email protected].
Anthracycline antibiotics (doxorubicin, daunorubicin) are widely used anti-neoplastic agents.
However, they have the serious drawback – cardiotoxicity, which may be caused by inhibition
and/or poisoning of topoisomerase-IIβ in cardiomyocytes, the iron-mediated generation of
reactive oxygen species or accumulation of metabolic products of the anthracyclines in the
heart.
The only substance, which is effective against anthracycline induced cardiotoxicity is
dexrazoxane (DEX). Nowadays, there are two theory about of its mechanism of action:
protection of cardiomyocytes via its metal-chelating hydrolysis product ADR-925 or through
the interaction with topoisomerase IIβ (TOP2B).
As the mechanism of DEX action is not still clearly known, the intensive structure –
cardioprotective activity study is needed.
Figure. 1 Dexrazoxane (DEX) and its metabolite ADR-925
Our work is focused on the study of the effects of changes in linker between two piperazine2,6-dionecycles of razoxane (racemic form of DEX), replacement of one or both piperazine2,6-diones for other cycles and the influence of terminal alkylation of razoxane on
cardioprotective activity.
The study was supported by the Czech Science Foundation project 13-15008S and Charles University
in Prague (SVV 260 183).
HPLC
METHOD
FOR
STABILITY
EVALUATION
OF
PHARMACEUTICAL PREPARATION CONTAINING SODIUM
PICOSULFATE
PETR KASTNER, KATEŘINA BURDOVÁ, PAVLA PILAŘOVÁ
Department of Pharmaceutical Chemistry and Drug Control, Faculty of Pharmacy in Hradec Králové,
Charles University, Czech Republic; [email protected]
Introduction Nowadays, HPLC is the most spread and powerful analytical tool in drug
control. It is the first choice method for the solution of all problems connected with evaluation
of related substances and assay of the active substance as well as preservative. This
separation method was thus used for development of analytical method which enables
evaluation of oral liquid drug preparation containing sodium picosulfate as favorite laxative
drug.
Experimental method HPLC separation of components was achieved with LiChroCART®,
250x4,0, Purospher® STAR, RP – 18C, 5μm column using UV detector at 263 nm. The
mobile phase consisted of a buffer, acetonitrile and isopropylalcohol in the ratio of 55:43:2
(v/v/v).
The
buffer
contained
sodium
dihydrogen
phosphate,
water
R
and
cetyltrimethylammonium bromide. The pH value was adjusted by phosphoric acid to 7.0. The
temperature of the column was 40 °C, injection volume 4 μl, the flow rate was adjusted to 1.0
ml/min.
Results and discussion The developed method was based on valid pharmacopoeial one 1),
it was optimized for and then it was validated
2)
as sufficiently selective, precise, accurate,
linear and sensitive. Robustness was tested by means of Plackett-Burman design
3)
and it
was found, that ratio of the acetonitrile in the mobile phase has the biggest influence on
separation from all proved parameters. On the other hand, flow, buffer concentration and
ratio of propan-2-ol in the mobile phase little affect the separation.
Fig. 1. A typical chromatogram of pharmaceutical preparation containing sodium picosulfate as active
ingredient, sodium benzoate as preservative and impurity A, which is the main hydrolytical degradation
productof the aktive substance.
Conclusions The developed method enables simultaneous evaluation of the content of
active ingredient, its related substances and preservative. It was sufficiently validated from
the point of selectivity, linearity, precision, accuracy, sensitivity and robustness and can be
used for release control of the pharmaceutical preparation and for its stability studies, too. This work was supported by a research project SVV 260062
1)
European pharmacopoeia. 8th ed. Strasbourg: Council Of Europe, 2013. ISBN 978-92-871-7525-0
2)
ICH [online]. 2013 [cit. 2013-12-11]. Available from:
http://www.ich.org/products/guidelines/quality/quality-single/article/validation-of-analytical-procedurestext-and-methodology.html
3)
Holík M. Chem. listy. 98, 2004.
SYNTHESIS OF QUINOLINE DERIVATIVES BY USING
NANO-PD/CU CATALYST IN THE SEARCH OF NEW
FLUOROPHORES.
MATEUSZ KORZEC1, ROKSANA RZYCKA1, SANDRA SENKAŁA1, EWELINA
SZPACZYŃSKA1,
BARBARA
MROZEK-WILCZKIEWICZ2,
CZAPLIŃSKA1,
MARZENA
WIOLETTA
RAMS-BARON2,
CIEŚLIK1,
ROBERT
ANNA
MUSIOŁ1,
JAROSŁAW POLAŃSKI1
1
2
Silesian Center for Education and Interdisciplinary Research, Chorzow, Poland
[email protected]
The application of fluorescence spectroscopy and imaging in biological systems has
expanded tremendously over the past decades. Fluorescence spectroscopy and timeresolved fluorescence are considered to be primarily research tools in biochemistry and
biophysics. Fluorescence is now a dominant methodology used in many fields of science i.e.,
biotechnology, flow cytometry, medical diagnostics, DNA sequencing, forensics, genetic
analysis and other 1). Imaging of biological structures by fluorescence microscopy acquires
special importance in the diagnosis of cancer, for example, in photodynamic diagnosis (PDD)
in urology
2, 3)
or brain imaging
4, 5)
. Recently, fluorescent properties of styrylochinolines have
been discovered and used, for example, for quantification of zinc in urban runoff
combined therapeutic and diagnostic in protein misfolding diseases in brain cells
6)
5)
, in
, as
fluorescent sensor for Fe2+ 7), demonstrates multicolor fluorescence upon addition of different
metal cations
8)
. Fluorescent properties of dye can be dependent on perturbation of their
emission by proximity to conducting particles or surfaces
DNA
11, 12)
9, 10)
and the cyanine determination oligonucleotides
interactions with fluorescent dyes have also been investigated
, as shown in the studies of
13, 14)
. Recently, metal ions
15, 16, 17, 18, 19)
A series of quinoline derivatives were designed based on the styrylquinoline system (Fig. 1).
The results indicated that representative compounds are biologically inactive (cell culture,
MTS assay) but have promising physicochemical properties (Stokes shift, quantum yield)
and preferentially incorporated into the plasma membrane or any other intracellular
organelles 20).
The financial support of the National Center for Science NCN grants 2014/13/D/NZ7/00322 (A.M.W.),
2012/07/N/NZ7/02110 (W.C.), 2013/09/B/NZ700423 (R.M.) is greatly appreciated. E.S. and M.K.
appreciate the DoktoRIS studentship and W.C appreciate the CITTRFUŚ fellowship. Nano-Pd/Cu was
prepared under National Research and Development Center (NCBiR) Grant ORGANOMET no:
PBS2/A5/40/2014.
1)
Lakowicz J. R. Principles of Fluorescence Spectroscopy. 3rd edition Boston: Springer, 2006.
2)
Pytel A., Schmeller N. Adult Urology. 2002; 59, 216-219.
3)
Jichlinski P., Jacqmin D. Eur. ur. suppl. 2008; 7, 529–535.
4)
Gomer Ch. J. Photodynamic Therapy . In: Kostron H. Photodynamic diagnosis and therapy and the
brain. New York: Springer 2010. http://link.springer.com/book/10.1007/978-1-60761-697-9
5)
Staderini M., et al. ACS Med. Chem. Lett. 2013; 4, 225-229.
6)
Hafuka A. et al. Water Res. 2014; 54, 12-20.
7)
Praveen L., et al. Tetrahedron Letters. 2010; 51, 6626-6629.
8)
Shiraishi Y. et al. Cem. Eur. J. 2011; 17, 8324-8332.
9)
Green B. Eur. J. Biochem. 1970; 14, 567-574.
10)
Lakowicz J. R., et al. Anal Biochem. 2003; 320, 13-20.
11)
Malicka J., et al. Biochem Biophys Res Commun. 2003; 306, 213-218.
12)
Malicka J. et al. Anal Biochem. 2003; 315, 160-169.
13)
14)
15)
Sankaran N. B., et al. Proc. Indian Acad. Sci. 2002; 114, 539-545.
16)
Mastiholi B. M., et al. Optik. 2013; 124, 261-264.
17)
Sutter J. U., et al. Methods Appl. Fluoresc. 2014; 2, 1-8.
18)
Elrobyab S. A. K., et al. Molecular Simulation. 2011; 37, 940-952.
19)
Asselin J., et al. Advances in Chemistry. 2014, 1-16.
20)
Rams-Baron M., et al. Plos One. 2015; 10, 1-17.
RING-SUBSTITUTED
CARBOXANILIDES
1-HYDROXYNAPHTHALENE-2AS
POTENTIAL
ANTIBACTERIAL
AGENTS
JIŘÍ KOS1,*, TOMÁŠ GONĚC1, IVETA ZADRAŽILOVÁ1, MIROSLAVA VALEŠOVÁ1,
ŠÁRKA POSPÍŠILOVÁ1, EWELINA SPACZYŃSKA1, JOSEF JAMPÍLEK1
1
Sciences, Palackeho 1/3, 612 42 Brno, Czech Republic
[email protected]
The antibiotic resistance of important Gram-positive pathogen, Staphylococcus aureus, has
become one of the most challenging and persistent worldwide health problems. Because of
the changing features of MRSA, it is one of the most difficult bacteria for clinicians to treat.
The emergence of resistance to currently available drugs, their toxicity and general lack of
oral agents justify an urgent need for new anti-MRSA agents 1,2.
In this study, a series of twenty-six ring-substituted 1-hydroxynaphthalene-2-carboxanilides
was prepared and characterized. In vitro antibacterial activity of the discussed compounds
was evaluated against three clinical isolates of methicillin-resistant Staphylococcus aureus
(MRSA 63718, MRSA SA 630 and MRSA SA 3202). S. aureus ATCC 29213 was used as
reference and quality control strain. Some of the tested compounds (for example, N-[4chloro-3-(trifluoromethyl)phenyl]-1-hydroxynaphthalene-2-carboxamide,
(trifluoromethyl)phenyl]-1-hydroxynaphthalene-2-carboxamide
(trifluoromethyl)phenyl]-1-hydroxynaphthalene-2-carboxamide)
and
showed
activity comparable with or higher than that of the standard ampicillin.
This study was supported by the IGA VFU Brno 322/2015/FaF
1, Kallen, A.J.; et al. JAMA 2010, 304, 641–647.
2, Liu, C.; et al. Clin. Infect. Dis. 2011, 52, 285–292.
N-[3-fluoro-4N-[4-bromo-3the
antibacterial
SYNTHESIS
OF
NOVEL
CARBAMATES
AND
THIOSEMICARBAZONES AS POTENTIAL ANTICANCERS
DARIUSZ KOZAKIEWICZ, JAROSŁAW POLAŃSKI
University of Silesia, Institute of Chemistry, Department of Organic Chemistry, Katowice, Poland
Carbamate and thiosemicarbazone moieties are one of the privileged structural
arrangements, widely deployed within pharmaceutical and crop protection industries. Their
broad scope of utility, especially as anticancers or antimicrobial drugs, rendered them as an
object of great interest for many years
and
recent
20
years
of
1-3
. Thus, their area of utility was broadly explored,
research
has
proven
that
junction
of
carbamate
or
thiosemicarbazone moieties with aromatic ketones and phenylethylamines yield a series of
structures that display a wide scope of bioactivity
4-6
. Our research objective is a synthesis of
carbamates and thiosemicarbazones based on the 1-phenyl-2-amine-1-ketone moieties as
shown on scheme below.
O
O
R1
Cl
HN
O
R1
O
N
O
H2 N
NH
H2 N
R2
R1
NH
S
R2
NH 2
S
H2 N
O
NH
R2
NH
N
NH 2
S
S
N
HN
NH
NH 2
R1
O
O
Scheme 1 | Synthesis of carbamates and thiosemicarbazones
Such a structure has its origins in the nature. Extensive studies displayed [7], that
functionalization of naturally occurring products by means of diverse substituents would lead
to novel compounds with diverse bioactivity profile. An example of such approach is the lead
structure of natural chrysanthemic acid, a molecular fragment used as a skeleton of many
novel insecticides. Additionally derivatives of cathinone, substance derived from Khat plant,
are convenient starting materials in synthesis of tetrahydroisochinolines. Cathinones itself
have a good synthetic availability. We have synthesized a series of the above mentioned
compounds as potential anticancer drug candidates. At the moment the obtained compounds
are tested for their biological activity.
1. Morgan, L. R. et al. Cancer Chemother Pharmacol 2009, 64, 829–835
2. Janganati, V. et al. Bioorg.Med. Chem. Lett. 2014, 24, 3499–3502
3. Cravatt, B. et al. WO 2012/058115
4. Turan-Zitouni, G. Eur. J. Med. Chem. 2008, 43, 981-985
5. Fatondji, H. R. Med Chem Res, 2013, 22, 2151–2162
6. Du, X. J. Med. Chem. 2002, 45, 2695-2707
7. Deng, Li, et al. J. Integr. Agricult. 2014, 13, 1320-1330
STUDY
OF
EPR
AND
UV-VIS
JUICES
FROM
POMEGRANATE
VIOLETTA KOZIK1, ANDRZEJ BAK2
1
Department of Organic Synthesis, Institute of Chemistry, University of Silesia, Katowice, Poland
2
[email protected]
Pomegranate products show great differences in the contents and quality of polyphenols.
This concerns both juices and juice extracts 1). Main active ingredients include fatty acids
(saturated, 20%; monounsaturated, 10%; punicic acid, 70%; phytosterols, 0.40.6%, with
beta-sitosterol, stigmasterol and campesterol prevailing), phytoestrogens (genistein,
daidzein, coumestans), steride estrogens (17-alpha estradiol, 0.3%, and estrone), gammatocopherol and polyphenols, 0.015%. Both pomegranate juice and pomegranate seed oil
show strong antioxidant properties, comparable with those of green tea and superior to red
wine2).
1)
Fischer-Zorn, M., Ara, V. Flüssiges Obst., 2007, 08, 386-393
2)
Gil, M.I., et al. J. Agric. Food Chem. 2000, 48, 4581-4589
PREPARATION
COMPLEXES
AND
WITH
ANALYSIS
OF
COPPER
AMINOCARBOXYLATES
(II)
AND
SULFONAMIDES
1
DOMINIKA KRAJČIOVÁ, 1,2PETER MIKUŠ, 1EMIL HAVRÁNEK, 1,3MILAN MELNÍK
1
Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy of Comenius
University in Bratislava, Slovakia
2
Toxicological and Antidoping Center, Faculty of Pharmacy, Comenius University in Bratislava,
Slovakia
3
Department of Inorganic Chemistry, Slovak Technical University, Bratislava, Slovakia
[email protected]
There have been some notable developments in several areas of organometallic
pharmaceuticals that have potentially far-reaching importance for future medical applications
and a research as well. One of the significant developments in the field of oncology and
hematology is the application of copper complexes. The chemistry of copper compounds has
been extensively investigated, and the relationship between structure and reactivity is of a
major importance. Copper creates stable complexes with a wide variety of organic molecules
(in our review work they are amino carboxylates and sulfonamides in the presence of
macrocyclic chelators). The aim of our experimental investigation is designing and
preparation of ternary copper (II) complexes with selected amino carboxylates and
sulfonamides. Although such ternary complexes have not been prepared and characterized
so far, one can expect of their inhibition activity toward carbonic anhydrase IX.
This work was supported by the projects VEGA 1/0873/15, KEGA 022UK-4/2015, FaFUK/20/2015 and
carried out in the Toxicological and Antidoping Center and at the Department of Pharmaceutical
Analysis and Nuclear Pharmacy, Faculty of Pharmacy, Comenius University in Bratislava.
SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF NOVEL
SULFONAMIDE DERIVATIVES
MARTIN KRÁTKÝ1, JIŘINA STOLAŘÍKOVÁ2, JARMILA VINŠOVÁ1
1
Department of Inorganic and Organic Chemistry, Faculty of Pharmacy, Charles University, Hradec
Králové, Czech Republic
2
Laboratory for Mycobacterial Diagnostics and Tuberculosis, Regional Institute of Public Health in
Ostrava, Czech Republic
[email protected]
The alarming progression of drug resistance among human pathogens justifies the
development of novel antimicrobial agents. Research studies should provide new
antimicrobial molecules active against, e.g., Mycobacterium tuberculosis, nontuberculous
mycobacteria, methicillin-resistant Staphylococcus aureus (MRSA) and polyresistant Gramnegative species 1).
The modification of known drugs represents an effective approach in drug design.
Sulfonamides have been widely used for the therapy of various bacterial infections. However,
they share some disadvantages. More recently, sulfonamides were considered useless for
the treatment of tuberculosis but concomitantly useful for the therapy of some
nontuberculous mycobacterial infections. Based on the findings that M. tuberculosis strains
are susceptible in vitro to clinically achievable concentrations of sulfamethoxazole 1, this
molecule has been “resurrected” and novel sulfonamides have recently been reported as
potential antimycobacterial agents
1), 2)
. Additionally, a wide range of urea derivatives were
found to display antimycobacterial activity 3).
That´s why sulfamethoxazole-based ureas 2 and their cyclic analogues imidazolidine-2,4,5triones 3 have been designed, synthesised, and evaluated as potential antimicrobial agents.
In our previous paper 1), we found that N-heptyl sulfamethoxazole-based urea displayed a
significant activity against nontuberculous mycobacteria. Based on this finding, we involved
also N-alkyl derivatives with a various length of alkyl chain.
The work was financially supported by the Research Project IGA NT 13346 (2012).
1)
Krátký M., et al. Chem. Pap. 2015; 69, 1108–1117.
2)
Krátký M., et al. Eur. J. Med. Chem. 2012; 50, 433–440.
3)
Brown J. R., et al. Bioorg. Med. Chem. 2011; 19, 5585–5595.
SYNTHESIS
OF
ARYLOXYAMINEPROPANOLES
AND
ARYLAMINEPROPANOLES
ALEŠ KROUTIL, JOSEF CSÖLLEI
Science Brno, Czech Republic
Introduction:
Aryloxyaminepropanole and arylaminepropanole dervatives, containing a
carbamate group with potential antidysrythmic activity were synthesised.
Experimental methods: The starting material for aryloxyaminepropanole derivatives was 4aminophenole, which gives the carbamate byl reaction with alkylchloroformiate. From this
intermediate were prepared a aryloxyaminopropanole derivatives by two step reaction with
epichlorhydrine and consequently with an secondary amine.
In case of arylaminopropanoles was the starting material 4-aminoacetophenone. By reaction
with alkylchloroformiate were obtained apropriate carbamates. The next step was the
Mannich reaction, provided with paraformaldehyde and a secondary amine. These
compounds were isolated and consequently reduced by NaBH 4 to final products.
From all mentioned final products were prepared salts with hydrochloric acid or fumaric acid.
OH
2
N
O
O
R
R
R
3
1
O
NH
Fig. 1 – Aryloxyaminopropanoles structure
OH
R
O
R
N
1
O
2
NH
R
3
Fig. 2 – Arylaminopropanoles structure
Results and discussion: The final compounds ant their salts were characterised by
common analytical methods (IR, NMR). For the Mannich reactions were used different
solvents and conditions with various yields. The hydrochlorides or fumarates prepared were
crystalline solids.
THE USE OF 2,6-DICHLOROQUINONE-4-CHLORIMIDE FOR
QUANTITATIVE DETERMINATION OF PHENYLEPHRINE
HYDROCHLORIDE
IN
COMBINED
TABLETS
WITH
PARACETAMOL AND CHLORPHENIRAMINE MALEATE
OLEKSANDR KRYVANYCH, NATALIIA BEVZ, NATALIIA HARNA, OLENA BEVZ
National Univerzity of Pharmacy, Kharkiv, Ukraine; [email protected]
This work is devoted to development and study of the validation characteristics of the method
for quantitative determination of phenylephrine hydrochloride in such dosage forms as tablets
in combination with other active ingredients based on the reaction with Gibbs reagent.
It has been found that unlike paracetamol, phenylephrine hydrochloride interacts with 2,6dichloroquinone-4-chlorimide in the alkaline medium forming a coloured compound with the
absorption maximum at the wavelength of 615 nm.
It has been experimentally proven that other active pharmaceutical ingredients and
excipients of tablets do not impede the quantitative determination of phenylephrine
hydrochloride. When studying the validation characteristics of the method the following
results have been obtained: linearity a=|-0.4251|<7.7, b=1.0054, precision, the correlation
coefficient r=0.9995. The results allow to recommend this method for analysis of
phenylephrine hydrochloride in other laboratories.
FLAVONOIDS FROM TAGETES PATULA L. AS INHIBITORS
OF ACETYL/BUTYRYLCHOLINESTERASE
RENATA KUBÍNOVÁ, HANA SRNÁNKOVÁ
Sciences, Brno, Czech Republic; [email protected]
Introduction: Tagetes patula L. (Asteraceae), popularly known as dwarf marigold or French
marigold, is an annual plant with 20–30 cm height, native to North America and widely
disseminated worldwide. The extract from plant has antibacterial and antifungal activity and
acts as insecticide. The phytochemical study of T. patula has resulted in the isolation of the
chemical constituents, such as flavonoids, benzofurans and thiophenes.
1)
Alzheimer's disease (AD), a very common form of dementia among the elderly, is in relation
to brain deficits in acetylcholine. Acetylcholinesterase (AChE) predominates in healthy brain
and with butyrylcholinesterase (BuChE) considered to play role in regulating brain
acetylcholine levels. Both enzymes therefore represent legitimate therapeutic targets for
symptomatic treatment of AD.2)
Experimental methods: Aerial section of T. patula was extracted with methanol at room
temperature for 30 minutes in an ultrasonic bath. The extract was concentrated in vacuo to a
dark gum and lyophilized. The lyophilized extract was column chromatographed on silica gel
by eluting with mobile phase chloroform-methanol with increasing polarity. Biological
activities were measured by Microplate Reader (inhibition AChE/BuChE using Ellman
method, antioxidant activity using method of DPPH radical).
Results and discussion: Fraction 13 yielded patuletin (2 mg), fraction 26 its hexoside
probably patulitrin (4 mg). These compounds were identified by HPLC-DAD-MS and FT-IR
and by comparison with literature data. Isolated flavonoids acted particularly as inhibitors of
BuChE, their activity was better than standard galantamine. Antioxidant activity of isolated
flavonoids was lower than standard quercetin.
Conclusions:
In consideration of fact that BuChE activity is progressively increased in
patients with AD, it may not be an advantage for a cholinesterase inhibitor to be selective for
AChE.2) Good balance in AChE/BuChE inhibition of flavonoids from T. patula may result in
higher efficacy. Antioxidant activity of compounds can contribute to a better beneficial effect
in the treatment of AD.
1)
Maresa D., et al. Microbiol Res 2004; 159; 295–304.
2)
Katalinic M., et al. Europ J Med Chem 2010; 45; 186-192.
RADIONUCLIDE X-RAY FLUORESCENCE SPECTROMETRY
IN ELEMENTAL ANALYSIS OF MEDICAL PLANTS AND
MEDICINES IN USE TO LOWER CHOLESTEROL LEVELS
OĽGA LUKAČOVIČOVÁ1, EMIL HAVRÁNEK1
1
University in Bratislava.Slovakia; [email protected]
Man receives portion of total cholesterol from food, or is formed directly in the body. When
cholesterol levels in the blood exceeds a certain amount, can occur in the body harmful and
very serious changes. Patients in addition to medication often used phytotherapy. In order to
rapidly achieve the desired effect, they drink tea in large volumes. Tea, although they are
medicinal plants, may contain not desirable and dangerous elements. It is therefore
necessary to select an appropriate analytical method for quality control of all the preparations
that a person is taking. For elemental analysis (Ti, Cr, Mn, Fe, Ni, Cu, Zn, Pb) of commercial
tea on cholesterol, plant Veronica officinalis, L. and Radix taraxaci and drug ATORIS® was
used Radionuclide X-ray Fluorescence Spectrometry. For analysis the excitation radiation
source was used
238
Pu, semiconductor detector Si/Li for detection and for signal processing
was used multichannel analyzer ORTEC® (the arrangement of the sample-source-detector:
reflective side geometry). Samples for analysis were compressed into tablets of defined
shape and weight. In the measured spectra were presented energy maxima (peaks),
corresponding to specific elements. The areas of evaluated peaks in samples were
compared with areas in prepared standards. They were calculated detection limits from
prepared standards/calibration curves on plants (Cr-1.31/1.40 μg/g; Mn-1.20/1.31 μg/g; Fe1.34/1.22 μg/g; Ni-1.37/1.45μg/g; Cu-1.72/1.80 μg/g; Zn-1.59/1.65 μg/g; Pb-1.89/1.96 μg/g),
also from calibration curves on ATORIS® (Cr-1.62 μg/g; Mn-1.05 μg/g; Fe-1.99 μg/g; Ni-1.57
μg/g; Cu-1.77 μg/g; Zn-1.03 μg/g; Pb-1.93μg/g). The content of elements in biological
samples was evaluated from results of prepared standard addition of selected elements on
plant matrix and by parameters of calibration curves. ATORIS ® was evaluated by similar
standard and calibration curves, but for its distinct character (organic matrix) was for their
preparation benzoic acid selected as a matrix, except the own matrix. Because of the high
content of titanium were prepared special standards, using solid Ti2O3 mixed in benzoic acid
and constructed calibration curve for the high content of this element. The content of titanium
in the tablet of drug was 2.13 mg in 1 g of tablet.
Financial support this study was provided by the Slovak Grant Agency for Science under the project
VEGA No.1/0873/15 and UK/195/2015.
EFFECTS OF
THIOSEMICARBAZONES
ANALOGS ON
OXIDATIVE STRESS AND CELLS DEATH IN HUMAN
COLON CANCER.
KATARZYNA
MALARZ1,
MARZENA
RAMS-BARON2,3,
ANNA
MROZEK-
WILCZKIEWICZ2,3, MACIEJ SERDA1, JAROSŁAW POLAŃSKI1, ROBERT MUSIOŁ1
1
2
Department of Solid State Physics, A. Chełkowski Institute of Physics, University of Silesia, Katowice,
Poland
3
Silesian Center for Education and Interdisciplinary Research, Chorzów, Poland
[email protected]
Introduction
Iron plays an important role in the life cycle of the cell. This essential element is required for
several key processes such as DNA synthesis, mitochondrial electron transport, synthesis of
heme, and as a co-factor for many redox enzymes1). Furthermore, iron is involved in the
Fenton and the Haber-Weiss reactions, whose lead to generation of reactive oxygen species
(ROS). ROS together with antioxidants may also play a key role in anti-cancer therapies that
exploit oxidative stress. Accumulation of ROS in response to external stimuli may affect the
regulation and expression of certain genes. This in consequence modulate the cellular
response through cell cycle arrest, activation of transcription factors and the triggering of
apoptosis2). Currently, most of chemotherapeutic agents causes cancer cells death by
stimulating ROS generation as at least one part of the mechanism. Induction of oxidative
stress by drugs accompanied reduction of the level of oxidative enzymes and their ability to
restore homeostasis. Glutathione (GSH) is one of the most prevalent reducing agents thiols
in living cells, making it one of the most important cellular antioxidants. During oxidative
stress reduced form of GSH is oxidized to glutathione disulfide (GSSG). Decreases in
cellular GSH levels triggers the unfolding protein response, peroxidation lipids and
apoptosis3).
Thiosemicarbazones are compounds, which are characterized by a wide spectrum of
biological properties including the antiproliferative activities. The most possible mechanism of
action includes generation of reactive oxygen species, iron chelation or inhibition of the
ribonucleotide reductase (RR) - enzyme necessary in the synthesis of DNA4).
Obtained series of novel derivatives thiosemicarbazones have been tested for cytotoxic
activity on two cell lines of human colon carcinoma: HCT116 wild type and HCT116 with
knockout TP53 gene. The antiproliferative activities were determined using the MTS assay.
Then, selected derivatives thiosemicarbazones showing the highest antiproliferative activity
have been tested for generation reactive oxygen species, and the subsequent impact on
levels of mitochondrial GSH. Formation of reactive oxygen species was evaluated using the
oxidation-sensitive dye CellROX® Green Reagent. A qualitative analysis carried out using
fluorescence microscopy techniques and a qualitative analysis were performed using
multiplate reader (Synergy4, Biotek). The changes of intracellular GSH levels were
determined using a commercial GSH-Glo™ Glutathione Assay. The next step was to
determine of caspases 3/7 activation and their participation on the apoptosis pathway.
To understand the overall mechanism of action of these compounds we focused on their
ability to induce oxidative stress, DNA damages and apoptosis. The series of novel
thiosemicarbazone derivatives, that have been tested, exhibited significant ability to inhibit
proliferation of cancer cells. In particular, di-2-pyridylketone, and 8-hydroxyquinoline moieties
induce a high antiproliferative activity on nanomolar level. The analysis of quantitative and
qualitative confirmed the generation of reactive oxygen species by selected compounds. The
increase of ROS above baseline we demonstrated after 3 or 6 hours of application of the
compounds. Generation of ROS was correlated with significant decreased the levels
intracellular GSH. This result suggests that the cell death induced by derivatives
thiosemicarbazones was not associated with caspase-3/7 activation.
Conclusions
In
summary,
the
present
data
reveal
that
selected
investigated
derivatives
thiosemicarbazones affect the ability to induce oxidative stress in human colon carcinoma
cells. Thiosemicarbazones induced reduction in intracellular GSH levels, without significantly
increasing ROS concentration. ROS-mediated oxidative damage and significant reduction in
the level of intracellular GSH may lead to mitochondrial dysfunction and cell death by
apoptosis, which is the desired effect of the cancer therapy.
The reported studies are financial supported by the Polish National Center for Science (NCN, grant no
2014/13/D/NZ7/00322).
1)
Rao V. A. Antioxidants & Redox Signaling, 2013; 18(8), 930–55.
2)
Manda G.,et al. Curr Chem. Biol. 2009; 3: 342-366.
3)
Circu M. L., Aw T. Y. Free Rad. Bio.&Med., 2010; 48: 749–762.
4)
Richardson D. R., Kalinowski D. S. et. al. Biochim. et Biophys. Acta, 2009; 1790: 702-17.
ON-LINE
HYPHENATION
OF
CAPILLARY
ELECTROPHORESIS AND MASS SPECTROMETRY USED
FOR THE ANALYSIS OF BIOGENIC AMINES IN GRAPE
LEAVES
KATARÍNA MARÁKOVÁ, JURAJ PIEŠŤANSKÝ, PETER MIKUŠ
University in Bratislava, Slovak Republic
[email protected]
The quality and safety of food products have a great impact on human health. The grape and
its products contain a wide range of substances. Biogenic amines (BA) are nitrogencontaining low molecular weight organic bases with aliphatic (putrescine, cadaverine),
aromatic (phenylethylamine, tyramine) or heterocyclic (histamine) structure, which are
commonly found in various foodstuff. BAs can be formed in food by decarboxylation of
corresponding amino acids by microbial enzymes. Under normal conditions, exogenous
amines ingested as part of the diet are absorbed from the food and quickly detoxified in the
organism via amine oxidases or by conjugation. However, if normal catabolic routes of
amines are inhibited or if the amount uptaken is large, this can result in several physiological
effects (migraines, headaches, nausea, hypo- or hypertension, cardiac palpitations and
anaphylactic shock)1. Histamine and tyramine are known to be the main cause of food
intoxication, although other amines such as putrescine, cadaverine and phanylethylamine
may intensify the undesirable effect of histamine. The content of biogenic amines in food
should be monitored for their potential toxicity and the fact that the quantity of biogenic
amines can be used as the food quality marker, while it is difficult to degrade BAs by high
temperature treatment. Due to the current importance of food BAs in quality control and
consumer safety, there is still a challenge to develop new methods for their fast, reliable
analysis in samples of different foods 1,2.
The research was supported by the projects APVV-0550-11, VEGA No. 1/0873/15, KEGA No. 022UK4/2015 and carried out in the Toxicological and Antidoping Center (TAC) of Faculty of Pharmacy,
Comenius University.
1
Simo, C., Moreno-rribas, M.V., Cifuentes, A. J. Chromatogr. A 1195 (2008) 150-156.
2
Beneduce, L., Romano, A., Capozzi V., Lucas, P. Ann. Microbiol. 60 (2010) 573-578.
SYNTHESIS
OF
NEW
ARYLCARBONYLOXYAMINOPROPANOL DERIVATIVES AS
POTENTIAL CARDIOVASCULAR DRUGS
PAVLÍNA MARVANOVÁ, PETR MOKRÝ, TEREZA PADRTOVÁ, OTAKAR HUMPA
1
Department of Chemical drugs, University of Veterinary and Pharmaceutical sciences Brno, Czech
republic
2
Josef Dadok National NMR Centre, CEITEC, Masaryk University Brno, Czech republic
[email protected]
Antagonists of β-adrenergic receptors are widely used in therapy of many cardiovascular
indications because of their positive effects on cardiovascular system. Moreover, newly
synthesized compounds have also other beneficial effects such as vasodilatory activity,
antioxidant activity and ultrashort effect. These features can improve haemodynamic and
metabolic profile, prevent atherosclerosis complication or ischaemia-reperfusion injury and
can be used for treatment of urgent cases. 1
The aim of this study was synthesis and evaluation of 4 hydrochlorides of 2-hydroxy-3-(4phenylpiperazin-1-yl)
propyl
4-alkoxybenzoates
(Fig.
1),
new
(arylcarbonyloxy)aminopropanol derivatives as potential β-blockers for treatment of
cardiovascular diseases and their complications.
OH
O
O
H+
N
Cl
N
R: -CH 3 to -CH 2CH2CH2CH3
Fig.1
O
R
This study was supported by Open access project, ID number LM2011020, funded by the Ministry of
Education, Youth and Sports of the Czech Republic.
1
Tengler, J. et al. Cent. Eur. J. Chem. 2013, 11, 1757-1767
NEW DERIVATIVES OF CURCUMIN WITH POTENTIAL
NATALIA MIKLÁŠOVÁ, LUCIA RIŠIAŇOVÁ, JINDRA VALENTOVÁ, SAMUEL
VARÉNYI, FERDINAND DEVÍNSKY
Department of Chemical Theory of Drugs, Faculty of Pharmacy, Comenius University, Bratislava,
Introduction
Curcumin (1,7-bis(4-hydroxy-3-methoxy phenyl)hepta-1,6-diene-3,5-dione) the yellow spice
used for anti-inflammatory, anticancer, antioxidant activities, AIDS and Alzheimer´s
disorders1), exhibits a poor bioavailability. This issue might be improved by using adjuvants
like piperine, liposomal curcumin, nanoparticles, phospholipid complex and structural
analogues of curcumin.2) Different derivatives of curcumin can coordinate with metallic ions
to form complexes with various biological potential, for example palladium complexes of
curcumin describe the antitumoral and antioxidant activities on human prostate cancer cells.3)
Several Knoevenagel condensates of curcumin and their copper complexes inhibit the TNF-α
induced NF-κB activation and proliferation of human leukemic cells. Schiff bases of such
derivatives induce the cell growth inhibition in colon and pancreatic cancer cells. Therefore,
these compounds are interesting for a chemopreventive and therapeutic activity against
cancer and for their increasing bioavailability.4)
Fourteen Knoevenagel condensates of two curcuminoids were synthesized and structurally
characterized by NMR, IR, UVspectra and their antiradical activity was determined.
Knoevenagel condensates were synthesised by general procedure 5)1 mmol of curcumin
dissolved in toluene was reacted with a corresponding aromatic aldehyde (2 mmol) in a
dropwise manner with continuous stirring, in the presence of a catalytic amount of piperidine
and acetic acid. The reaction mixture was refluxed overnight and the final product was
isolated by column chromatography.
Literature studies shown that 4-arylidene curcumin analogues inhibit growth of lung cancer
cells with less concentrations than curcumin, acting as
potential compounds for
development against cancer and inflammatory diseases.5) In our laboratory fourteen
curcuminoids were synthesised, as Fig.1 illustrates, with the expectation that the
condensates will exhibit significant antiradical activity.
Fig 1: Synthesis of Knoevenagel condensates of curcumin and its analogue
O
O
H
O
OH
R
R
R
140 oC
R1
OH
R=OCH3; OCH2CH3
R
piperidine/AcOH
+
HO
O
HO
OH
R2
R1
R2
We are grateful to Faculty of Pharmacy, Comenius University in Bratislava and Department of
Chemical Theory of Drugs for support.
This work was supported by the Slovak Research and Development Agency under the contract No.
APVV-0516-12 and FaF UK/10/2015, FaF UK/11/2015.
1)
Zambre A.P., et al. Synth. React. Inorg., Met.-Org., Nano-Met. Chem. 37(1), 2007; 19-27.
2)
Anand P., et al. Mol. pharm. 2007; 4(6), 807-818.
3)
Miklášová N., et al. Inorg. Chem. Commun., 2014; 46, 229-233.
4)
Padhye S., et al. Pharm. research, 2009; 26(8), 1874-1880.
5)
ZuO Y., et al. Eur. J. Med. Chem. 2012; 55, 346–357.
TARGET-SPECIFIC RADIOPHARMACEUTICALS:
L-TRYPTOPHAN AS A RADIONUCLIDE CARRIER
MÁRIA MIKULOVÁ, MIROSLAVA SÝKOROVÁ, EMIL HAVRÁNEK
University in Bratislava, Slovakia; [email protected]
In recent years a lot of attention has been paid to development and evaluation of
radiopharmaceuticals in the field of nuclear medicine and pharmacy. Radiopharmaceuticals
based on peptides have come to the foreground because of their high affinity with
transporters or receptors overexpressed in many tumours. Many amino acids, short peptides
and polypeptides are labeled with radionuclides for use in a diagnosis and therapy of various
diseases. The most frequently used radiolabeled target-specific peptides are analogues
related to hormone somatostatine. This work is focused on an initial amino acid L-tryptophan,
the significant serotonine and melatonine precursor. Radiolabeling procedure of L-tryptophan
with technetium-99m by direct method, using stannous chloride as reducing agent, is
described in this work. The aim of this work is to observe convenient reaction conditions for
the complex preparation and labeling efficiency control of resulting complex. Analytical
separation methods as paper chromatography, thin-layer chromatography and paper
electrophoresis with radiometric detection were used for radiolabeling efficiency control.
Complex preparation was carried out under various pH conditions, where the pH 6,83 was
proved to be the most appropriate, using the acetate buffer. The formation of two complexes
of
99m
Tc-L-tryptophan was observed. Since radiolabeled tryptophan and its metabolites are
accumulated in the brain and various tumour cells, it can be used for brain activity imaging,
diagnosis of serious diseases and futher peptide structures studies.
This work was supported by the grant UK/195/2015 and project VEGA 1/0873/15.
Dijkgraaf I., Boerman O.C. et al. Med. Chem. 2007; 7, 543-551.
Jager P.L. et al. J. Nucl. Med. 2001; 42, 432-445.
HPLC
ANALYSIS
OF
SOME
NON-STEROIDAL
ANTIFLOGISTICS
MILAN MOKRÝ, MARKÉTA ŠRÁMKOVÁ
Department of Pharmaceutical Chemistry and Drug Analysis, Faculty of Pharmacy in Hradec Králové,
Introduction A chromatographic method was developed for the detection and quatification of
four nonsteroidal antiflogistic drugs – ketorolac, tiaprofenic acid, suprofen and ketoprofen.
The HPLC analysis is carried out using a C18 column and UV detection at 314 nm.
Experimental methods The aim of this work was to elaborate chromatographic conditions
for simultaneous analytical evaluation of mixture of these drugs. Standard stock solutions at
a concentration of 1 mg/ml were prepared by dissolving the appropriate amount of individual
drug in 10 ml of methanol and stored at 4 °C, working solutions by dilution of standard stock
solutions with mobile phase. The concentrations of the standard stock solutions were 0.05
mg/ml. Separations of ketorolac, tiaprofenic acid, suprofen and ketoprofen were performed
on a Discovery HS C18 (150 x 4.6 mm) column (Supelco, Bellefonte, USA). The mobile
phase was acetonitrile and 0.01 M potassium dihydrogen phosphate buffer adjusted to pH
3.0 by addition of phosphoric acid (40/60, v/v) and was pumped with a flow rate of 1 ml/min.
Results and discussion A typical chromatogram of resolved components is shown in Fig. 1.
Elaborated chromatographic condition, ambient temperature for column and HPLC system
inclusive were found to best for analysis. The described method has been validated for
specifity, linearity, system suitability, accuracy and intermediate precision.
Fig. 1. A typical chromatogram of mixture of antiflogistics (all 50 μg/ml). Sequence of
peaks:ketorolac, suprofen, tiaprofenic acid, ketoprofen.
Conclusions A simple HPLC method with UV detection has been succesfully developed to
determine ketorolac, tiaprofenic acid, suprofen and ketoprofen. The method has linear
response in working range and is accurate and precise.
This work was supported by a research project SVV 260062
1)
Maboundou C., et al. J. Chromatogr. B. 1994; 657, 173-183.
2)
Kulo A., et al. Chromatographia 2014; 77, 803-812.
ANTIFUNGAL STYRYLQUINOLINES PROBABLY ACT AS A
CELLULAR RAZOR BLADE
JACEK
MULARSKI,
WIOLETA
CIEŚLIK,
JOANNA
SZCZEPANIAK*,
ANNA
KRASOWSKA*, ROBERT MUSIOŁ
* Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland
Ergosterol, a fungal specific cellular building block, is most abundand in the plasma
membrane, but has been found within membranes of many organelles as well. Its ubiquity
allows to design ergosterol specific drug therapies. Azoles are the most widely deployed
antifungals in clinics and were found to inhibit the biosynthesis of ergosterol. Nonetheless,
multi drug resistance has arisen from their excessive use. On the other hand polyenes such
as amphotericin (AmB) primarily kill yeast by simply binding plasma membrane ergosterol
[1]. Such agregates form ion channels what leads to cell damage. Unfortunatelly, some AmB
affinity for cholesterol has been observed and intrinsic high toxicity correlates with
exceptional activity profile, limiting its use only to fight systemic fungal infections as the last
line of defense. Despite azoles are in treatment more than two decades, our understanding
of
the
processes
specific
cellular
disrupted
ergosterol
by
deprivation
HO
evaluating
we
have
O
been
styrylquinolines
(SQ) bioactivity profile [2]. It
Cl
O
OH
OH
OH
OH
O
O
OH
Cl
O
remains insufficient.
Recently,
OH
HO
HO
N
OH
O
HO
O
OH
NH 2
amphotericin B
[AmB]
styrylquinoline
[SQ]
appears that these small molecules reveal excellent antifungal activity. The ongoing research
has already exposed some mechanistic diversity. For instance, resembling some AmB
structural relationships, such as conjugated molecular π orbitals, which are known for
creating a rigid molecular surface. Therefore, it can be assumed that SQ may act as
ergosterol-selective cellular razor blade. There is also evidence of intracellular ergosterol
complexation. As an assistance to those observations and development of further lead
optimization we've designed NMR – based experiment. 1H NMR has been chosen for
preliminary studies. This sensitive technique enables maintenance of solutes at marginal
desired level. Chemical shifts as well through-space coupling constants of SQ – ergosterol
supramolecular complexes should be a measure of their antifungal activity and SQ –
cholesterol as a toxicity coefficient. The experiment can also illustrate qualitatively how the
drug interacts with its particular sterol ligand, and quantitatively, determining its complexation
ratio at particular concentration.
Acknowledgement: National science centre grant no 2013/09/B/NZ7/00423
1
Proc. Natl. Acad. Sci. USA 109 (2012) 11156-11159
2
Bioorg. Med. Chem. (2012) 6960-6968
VALIDATION OF SPECTROPHOTOMETRIC METHOD OF
ASSAYING METRONIDAZOLE IN CAPSULES
ARTEM
MYHAL,
ANNA
DOBROVA,
OLGA
GOLOVCHENKO,
VICTORIYA
GEORGIYANTS
The National University of Pharmacy, Ukraine, Kharkiv; [email protected]
Introduction
The international community imposes stringent requirements for the quality and safety of
products. Nevertheless, counterfeiting of medicines is a special social danger and an urgent
problem. Therefore, the suggested methods of quality control must comply with regulatory
requirements and fully confirm the quality of goods.
Validation of quality control methods is recommended by the State Pharmacopoeia of
Ukraine (SPhU) and the world's leading Pharmacopoeias during the registration of
medicines. This procedure aims to experimentally verify the correctness and accuracy of the
above methods1-3, 6, 7, 10-13.
The guarantee of the safety and effectiveness of drugs is also their stability and absence of
side effects of interaction that can take place in the organism, with different active
ingredients. This applies to simultaneously appointed drugs and components of food, drinks,
mineral water, food additives that people use independently in their daily diet. Patients rarely
adhere to the recommended diet, without changing their eating habits. Therefore, the study
of influence of the most common cases of interaction on bioavailability and pharmacological
activity of prescribed drugs is relevant.
In order to experimentally prove the expediency or inadmissibility of the combined use of
metronidazole as a helicobacter drug with others drugs, food, mineral water and other
beverages rich in metal cations, we carried out validation of analytical method of assaying
metronidazole in capsules by the standard method within the UV-spectrophotometric method
according to the SPhU requirements, which is planned for use in studying the bioavailability
of products of metronidazole interaction with metal salts.
1)
Bagirov V.L., et al. Pharm. Ind. 2007; 58 p.
2)
Bezugly P.O., et al. ed. NUPh, Golden pages 2013; 552 p.
3)
Chung Ch. Ch. et al. Analytical method validation and instrument performance verification. John
Wiley & Sons, Inc 2004, 11–51.
6)
Grizodub O.I., et al. Pharmakom 2004; 3, 3-17.
7)
Grizodub O.I., et al. Pharmakom 2004; 2, 20-34.
10)
State Pharmacopoeia of Ukraine. State Enterprise "Scientific and expert pharmacopoeia center".
1st ed. Kharkiv: RIREH 2001; 556 p.
11)
App. 1., 1st ed. Kharkiv: RIREH 2004; 494 p.
12)
13)
BARGELLINI
REACTION
IN
FUNCTIONALIZATION
OF
HETEROCYCLIC COMPOUNDS
L.V. MYZNIKOV, Y.V. MELNIKOVA, Y.E. ZEVATSKII
Saint-Petersburg State University of Technology and Design, Russia
The progress in chemistry of heterocyclic compounds is impossible without the use of
innovative approaches to their functionalization. One of such approaches is application
multicomponent reactions for synthesis and functionalization of heterocyclic compounds.
Multicomponent reactions very attractive because they allow several simple molecules
assemble to complex multifunctional structures which difficult to prepare with other methods.
One of such multicomponent reactions - Bargellini reaction - was discovered for more than
100 years ago1. Bargellini reaction up to date was not applied for fuctionalization of
heterocyclic compounds.
We have found that heterocyclic thiols and NH heterocycles may act as nucleophiles in
Bargellini reaction:
O
CHCl3
+
+
Het-SH
Het
S
+
OH
O
O
CHCl3
O
+
Het(NH)
Het
OH
Bargellini reaction can be used for functionalization series structure divergent heterocyclic
thioles and NH heterocycles. The reaction is carried out without solvent with excess of a
ketone and chloroform at 0°C. At this temperature reaction is completed in 12 h, the yields of
carboxylic acids 64-95%. Substances which can be prepared by the presented procedure are
interesting in consequence of their biological activity. It was shown that such compounds are
useful for treatment of glaucoma2, diabetes3 and metabolic diseases4.
1
Bargellini, G. Gazz. Chim. Ital. 1906, 36, 329.
2
Bartels, S.P. Pat. 20080058373 USA.
3
Bigge, C.F., et al. Pat. 20030171377 USA.
4
Jomard, A., et al. Pat. 20070065471 USA.
STABLE
GOLD
NANOPARTICLES
–
SYNTHESIS,
BIOCONJUGATION AND APPLICATION
PAWEŁ NALEPA1, ANNA MROZEK-WILCZKIEWICZ2, JAROSŁAW POLAŃSKI1
1
Institute of Chemistry, Faculty of Mathematics, Physics and Chemistry, University of Silesia in
Katowice, Poland
2
Silesian Center for Education and Interdisciplinary Research, Chorzów, Poland
[email protected]
Gold nanoparticles (GNPs) are good biocompatible materials due to their special physical
and chemical properties1. There is more and more research where GNPs are used in the
medical imaging and in the cancer therapy. Nanotechniques, the usage and production of
nanomaterials has been developed in our team for many years 2,3,4. Furthermore, we
synthesise various compounds which have anticancer activity and can be developed into
novel anticancer treatment5,6. Therefore, it is not surprising that we decided to combine these
two paths of our studies.
Indeed, there are many examples described in the literature of chemically 9 and biologically10
functionalized GNPs; however, these conjugates are very often instable. Although GNPs are
rather stable in period of many months, modified nanoparticles – conjugates GNPs to drug,
dye or antibody are often instable in a long period, particularly in the presence of high salts
and proteins, which is the essence of human body11.
There exist some reports where glycerol is used as an efficient reducing agent 13,14. Glycerol
is an environmental friendly and completely biocompatible reagent. Thus we decided to use
glycerol and synthesise GNPs with various concentration of cross-linked polyvinylpyrrolidone
(PVP) as a stabilizing agent. We also wanted to determine size, shape, dispersity and
stability of the obtained GNPs. Our aim was to obtain stable GNPs with diameter less than
60 nm because these GNPs have the most appropriate physical and biological properties 1.
The next step will be coupling nanoparticles with our potential drugs. These bioconjugates
will be examinated in the in vivo biological studies.
The experiments were conducted under National Research and Development Center (NCBiR) Grant
ORGANOMET no: PBS2/A5/40/2014 (J.P.).
The financial support of the National Center for Science NCN grants 2014/13/D/NZ7/00322 (A.M.W.)
is also greatly appreciated.
1) Dreaden E.C., et al. Chem. Soc. Rev. 2011; 40, 3391-3404
2)
Kapkowski M., et al. J. Catal. 2014; 319, 110–118.
3)
Korzec M., et al. J. Catal. 2014; 313, 1–8.
4) Bujak P., et al. J. of Catal. 2012; 295, 15–21.
5)
Mrozek-Wilczkiewicz A., et al. ACS Med. Chem. Lett. 2014; 5, 336–339.
6) Serda M., et al. PLOS One 2014; 9, e110291
9) Katz E., Shipway A.N., Willner I. Chemically functionalized metal nanoparticles. In: Liz-Marzán,
L.M., Kamat, P.V. eds. Nanoscale Materials, Springer US 2003
10) Sperling R.A., Parak W.J. Adv. Nanopart. 2010; 368, 1333-1383
11) Liu S., Han M. Adv. Funct. Mater. 2005; 15, 961-967
13) Díaz-Álvarez A.E. Appl. Sci. 2013; 3, 55-69
14) Kouz J., Varma R.S. Chem. Commun. 2013; 49, 692-694
NITRO GROUP-CONTAINING OXA- AND THIADIAZOLES AS
POTENTIAL ANTITUBERCULAR AGENTS
JAN
NĚMEČEK,1
GALINA
KARABANOVICH,1
JAROSLAV
ROH,1
LENKA
VALÁŠKOVÁ,1 KATEŘINA VÁVROVÁ,1 JIŘINA STOLAŘÍKOVÁ,2 PETR PÁVEK,1
VĚRA KLIMEŠOVÁ,1 ALEXANDR HRABÁLEK1
1
Department of Inorganic and Organic Chemistry, Faculty of Pharmacy in Hradec Králové, Charles
University in Prague, Czech Republic
2
Department of Bacteriology and Mycology, Regional Institute of Public Health, Ostrava, Czech
Republic
[email protected]
Tuberculosis (TB) occurs in every part of the world and in 2013, 1.5 million people died from
TB. Although TB is curable disease, increasing amount of multidrug-resistant and extensively
drug-resistant strains, which respond to only few available medicines, underline the need for
new antiTB agents with different modes of action.
Based on our lead compounds, 5-(dinitrobenzylsulfanyl)tetrazoles, we prepared a series of
1,3,4-oxadiazoles and 1,3,4-thiadiazoles and studied the effect of the individual structural
fragments of these compounds on the antimycobacterial activity, selectivity towards
mycobacteria and toxicity.
Antimycobacterial activity of all prepared compounds were evaluated against four
mycobacterial strains, compounds with the most promising activities were further evaluated
against six MDR/XDR-TB strains. To study the selectivity of these compounds, their IC 50 on
human cell lines and MIC for selected bacterial and fungal strains were evaluated.
Furthermore, acute oral toxicity and 14-days repeated dose toxicity experiments were
performed. Selected compounds showed excellent antimycobacterial activity with MIC values
reaching 0.03 µM against drug susceptible and MDR/XDR-TB strains. Furthermore, these
compounds showed high selectivity and low in vitro and in vivo toxicity.
This work was supported by the Czech Science Foundation project (14-08423S), “Pre-seed activities
of CU out of Prague“, reg. n. CZ.1.05/3.1.00/13.0284 and "Pre-seed activities of CU out of Prague II",
reg. n. CZ.1.05/3.1.00/14.0299
INFLUENCE
OF
Β-CAROTENE
ON
ANTIGENIC
EFFECTIVENESS OF INACTIVATED RABIES VACCINE
WITH SQUALENE ADJUVANT
ONDREJKOVÁ A. 1, SÜLI J.2, KOLČÁKOVÁ L.2, ONDREJKA R.1, ČECHVALA P.1,
BENKÖ Z.1, PROKEŠ M.1
1
Department of Epizootology and Parasitology
2
Department of Chemistry, Biochemistry and Biophysics
University of Veterinary Medicine and Pharmacy in Košice, Slovakia
[email protected]
Squalene adjuvants are successfully used for potentiating the efficacy of inactivated vaccines
for human and veterinary use. Commercially produced squalene adjuvant formulations are
added to vaccines either liquid or lyophilized 1). The experimental squalene adjuvant is a
formulation that allows the preparation of adjuvanted vaccine in one step – aqueous phase of
adjuvant is formed directly by inactivated viral suspension 2;3). This adjuvant has proved to be
effective in the application for the target species of domestic animals4;5).
In view of the increased sensitivity to oxidation, α-tocopherol – an effective in vivo antioxidant for the lipids is added to some of the squalene adjuvant formulations. Several types
of squalene emulsions with α-tocopherol are developed for the preparation of vaccines, e.g.
SE (stable emulsion) or SB62, which is suitable for human influenza vaccines 6;7). However, in
in vitro systems α-tocopherol can cause prooxidation, while the β-carotene effectively
protects against oxidation of squalene8-10). Various inactivated adjuvant vaccines with αtocopherol induced faster and higher production of antibodies in tested animals compared to
control vaccines11;12). However, we have no data about vaccines containing β-carotene.
Antioxidant protection of squalene by β-carotene is satisfactory9;10), but it is also important to
know, whether the addition of the antioxidant alter the effectiveness of vaccines.
In this work we examined how is the antigenic activity of inactivated rabies vaccine with
squalene adjuvants affected by β-carotene.
This work was financed by scientific project APVV-0605-12 (Slovak Republic).
1)
Fox C.B. Molecules 2009; 14, 3286–3312.
2)
Süli J., et al. Vaccine 2004; 22, 3464–3469.
3)
Beníšek Z., et al. Vaccine 2004; 22, 3470–3474.
4)
Süli J., et al. A new adjuvant inactivated rabies vaccine. In: Dodet, B., et al. eds. Rabies in Europe.
Dev. Biol. Basel: Basel Karger 2006; 125, 313.
5)
Süli J., et al. Acta Vet. Beograd 2010; 60, 597–603.
6)
Leesman G. D. US Patent 6630161 B1, 2003. http://www.google.com/patents/US6630161.
7)
Reed S. G., et al. Trends Immunol. 2009; 30, 23–32.
8)
Mueller L., Boehm V. Molecules 2011; 16, 1055–1069.
9)
Süli J., et al. Folia Vet. 2012; 56, Suppl. II, 56–58.
10)
Süli J., et al. Influence of β-carotene on antigenic effectiveness of inactivated rabies vaccine with
squalene adjuvant. In: Sborník 44. Konference Syntéza a analýza liečiv. Brno: 2015.
11)
Tengerdy R. P. Vaccine 1991; 9, 273–276.
12)
Franchini A. et al. Poultry Sci. 1995; 74, 666–671.
SYNTHESIS
AND
EVALUATION
OF
HETEROARYLCARBONYLOXYAMINOPROPANOLS
AND
THEIR QUATERNARY AMMONIUM SALTS
TEREZA
PADRTOVÁ1,
MARKÉTA
DALECKÁ1,
PETR
MOKRÝ1,
KLÁRA
ODEHNALOVÁ1, PAVLÍNA MARVANOVÁ1, OTAKAR HUMPA2
1
Sciences, Brno, Czech Republic
2
Josef Dadok National NMR Centre, CEITEC, Masaryk University, Brno, Czech Republic
[email protected]
Introduction
Recent WHO document shows that about 33.3% of the world population suffers from
diseases related to the cardiovascular system.1 Beta blockers are among the drugs of first
choice in the therapy of these diseases, but unfortunately they have a wide range of adverse
effects. Synthesis of new derivatives with ultrashort effect aims to reduce the adverse effects
of these substances and their improved pharmacological profile.
Preparation of the final compounds was carried out by microwave iradiation and the method
was compared with conventional techniques. The structure of the compounds was
determined by measuring the 1D and 2D-NMR, IR and MS. Physicochemical parameters of
the resulting compounds were also determined: lipophilicity parameter (LogP) by HPLC, and
acid-base dissociation constant (pKa) by capillary electrophoresis. Quaternary amines was
determined by surface activity.
HO
HO
+
O
Z
-
-
R
3
N
X
X
N
R
4
O
Z
2
R
+
N
2
R
3
R
O
O
1
1
R
R
Fig. 1: structure of final compounds
This study was supported by the IGA VFU Brno 315/2015/FaF and Open access project, ID number
LM2011020, funded by the Ministry of Education, Youth and Sports of the Czech Republic.
1)
World health organization, World health statistics 2014, Geneva.
http://www.who.int/gho/publications/world_health_statistics/2014/en/
HYDRODYNAMICALLY CLOSED CITP-CZE COUPLED WITH
TANDEM MASS SPECTROMETRY FOR DETERMIANTION
OF ANTIGRIPAL DRUGS IN HUMAN URINE
JURAJ PIEŠŤANSKÝ, KATARÍNA MARÁKOVÁ, PETER MIKUŠ
University, Bratislava, Slovakia;[email protected]
Introduction
Paracetamol, pheniramine, and phenylephrine are typical drugs presented in over-thecounter (OTC) medicines against cough, cold and flu. These three drugs with different
pharmacotherapeutic activity can be described as antigripal drugs. Paracetamol (PCM) is an
analgesic and antipyretic, pheniramine (PHM) is a H1-antihistamine, and phenylephrine
(PHE) is a α1-adrenergic receptor agonist, that is used as a nasal decongestant. Theraflu ® is
a typical commercial OTC medicine with presence of these drugs.
A modular capillary electrophoresis analyzer EA-102 (Villa Labeco, Spišská Nová Ves,
Slovakia), assembled in the column-coupling configuration (hydrodynamically closed) of the
separation unit, was used in this work for performing the CITP-CZE runs (cationic regime of
the separation). The samples were injected by a 10 µL Hamilton syringe via a rubber septum
into the injection block of the analyzer. An CITP column was provided with an 800 µm I.D.
polytetrafluorethylene (PTFE) capillary tube of a 90 mm length and a contactless conductivity
detector. A CZE column was the same as the CITP one except for a 300 µm I.D. and a 160
mm length. The CZE column was provided with an on-column conductivity detector (Villa
Labeco). Win ACES software, ver. 1.4 (Faculty of Natural Sciences Comenius University,
Bratislava, Slovakia) was used for controlling the analytical protocol. The experiments were
performed in constant current mode at 20ºC. The driving currents applied were 300 µA (ITP)
and 40 µA (CZE).
Identification and quantification of the analytes were performed on a triple quadrupole mass
spectrometer (QqQ) Agilent 6410 Series Triple Quadrupole (Agilent Technologies, Santa
Clara, CA, USA) coupled to the CE analyzer through an ESI interface and an elution block
(coupling CE with ESI) proposed by Foret et al.1). The mass spectrometer operated in the
positive-ion mode. The MS detection was carried out in the multiple reaction monitoring
(MRM) mode.
CITP-CZE-ESI-QqQ system was optimized in order to obtain sufficient sample clean-up and
good performance parameters so the method could be used in the direct analysis of
antigripal drugs in human urine. The optimum CITP-CZE conditions were: i) CITP – LE = 10
mM NH4Ac + 20 mM HAc (pH = 4,5), TE = 10 mM HAc (pH = 3,1), driving current 300 µA; ii)
CZE – BGE = 10 mM HAc (pH = 3,1), driving current 40 µA. PCM did not migrate properly in
the CE system without electroosmotic flow (EOF) and it could not be detected in
hydrodynamically closed CE separation system under given CITP conditions. Therefore,
PCM was not considered further in this study. The optimum composition of the spray liquid
was methanol/water (50/50, v/v) with 0,1% (v/v) HAc and its flow rate was 2 µL.min -1. For the
ESI-QQQ mass spectrometer, optimum parameters were as follows: i) nebulizer pressure (15
psi); ii) drying gas temperature (300°C); iii) drying gas flow rate (5 L.min -1); iv) capillary
voltage (5000 V); v) fragmentor voltage (100 V) and vi) collision energy (10 eV – PHM, 8 eV
– PHE). Mass spectrometry measurements were carried in MRM mode with the following
transitions for pheniramine: m/z 241.2 → 196.1 (quantifier) and 241.2 → 168.1 (identifier)
and for phenylephrine: m/z 168.1 → 150.3 (quantifier) and 168.1 → 135.1 (identifier).
For validation of the proposed method, a model human matrix spiked with PHM and PHE
standards was used. Calculated performance parameters confirmed good linearity (r 2 ˃
0,999), accuracy (R ˃ 98.0%) and precision (RSD tm ˂ 1.50% and RSDarea ˂ 8.00%) of the
proposed method. Finally, the CITP-CZE-MS/MS method was applied to monitor the level of
excreted PHM and PHE in human urine after oral administration of one dose of Theraflu ® (20
mg pheniramine maleate, 10 mg phenylephrine hydrochloride per one dose – sachet) to the
volunteers. Samples were collected in various time intervals (0-24h). The concentration
profiles of excreted pheniramine and phenylephrine in human urine were successfully
obtained by means of the studied method. Pheniramine metabolite, monodesmethylpheniramine, was also identified in the real urine samples (identifier – m/z 227.2 → 118.2
and quantifier – m/z 227.2 → 196.1).
Conclusions
The presented CITP-CZE-ESI-MS/MS combination represent a powerful tool for the analysis
of relative unpretreated (only diluted) biological samples containing different trace and/or
ultratrace concentration levels of cationic drugs and their metabolic products.
This work was supported by the projects VEGA 1/0873/15, KEGA 022UK-4/2015 and by the grant
from the Faculty of Pharmacy Comenius University (FP CU), namely FaF UK/3/2015 and carried out in
the Toxicological and Antidoping Center (TAC) FP CU.
1)
Tomáš R., et al. J. Chromatogr. A 2010; 1217, 4144–4149.
HPLC EVALUATION OF PIROXICAM IN PLASMA USING
SPME AND PRECIPITATION
PAVLA PILAŘOVÁ, KRISTÝNA KUŽELOVÁ, PETR KASTNER
Department of Pharmaceutical Chemistry and Drug Control, Faculty of Pharmacy in Hradec Králové,
Introduction Work is focused on the development, validation and comparison of two
bioanalytical methods for the determination of piroxicam in plasma. SPME and protein
precipitation was tested. Evaluation isolation procedures were performed using HPLC with
UV detection.
Experimental method Piroxicam was isolated from spiked plasma using SPME and protein
precipitation. Plasma was always adjusted to pH 2,5. SPME was composed of 20 minutes
sorption on PDMS/DVB fiber and 20 minutes desorption into 200 ul of methanol. After
desorption to 200 μl of methanol was added 50 μl of acid purified water and this solution was
injected on the column. In the second method the sample of plasma was precipitated with
acetonitril, 30 seconds was shaken and then 5 minutes centrifuged at 5 000 G. Then the
supernatant was removed and it was analyzed with HPLC. HPLC separation of piroxicam
and IS was carry out on the column with reversed phase C 18 Nova-Pak (4μm) 150 x 3,9
mm, Waters. The mobile phase consisted of water and acetonitrile (60:40 v/v), pH of solution
was adjusted to 2,5 using the formic acid. The flow rate was 1 ml/min, the injection volume
was 20 μl and the temperature was set at 40°C. For detection was used 333 nm.
Results and discussion SPME and protein precipitation were validated according to FDA.
Specificity, accuracy, precision, recovery, linearity, limit of detection a quantification and
stability were monitored.
uV
11000
10000
9000
8000
7000
IS
6000
5000
4000
3000
2000
1000
0
-1000
-2000
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5
8.0
8.5
9.0
9.5
min
Fig. 1. A typical chromatogram of piroxicam (5 μg/ml) and IS (5 μg/ml) after precipitation of acetonitrile.
Conclusions After comparison, validation parameters, it was found that the isolation of
piroxicam using protein precipitation was more precise, more sensitive, it was provided more
accurate results. This method gave higher recovery. However, SPME can also use for
evaluation of piroxicam in plasma.
This work was supported by a research project SVV 260062
1)
Lord, H., Pawliszyn, J. J. of Chrom.A. 2000, 885, 153–193
2)
Juntinga, L., Peng, Ch., Suzukib, O. Forensic Sci. Int.. 1998, 97, 93–100
3)
Guidance for Industry, Bioanalytical Method Validation. U.S. Department of Health and Human
Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for
Veterinary Medicine (CVM). May 2001. BP
AN
OPTIMIZED
AND
SCALABLE
SYNTHESIS
OF
PROPYLPHOSPHONIC ANHYDRIDE
HANA PÍŽOVÁ, PAVEL BOBÁĽ
Sciences Brno, Czech Republic; [email protected]
Introduction
Propylphosphonic anhydride (T3P) is a powerful water scavenger and coupling reagent,
which has been used for a large number of chemical transformations. A convenient four
steps synthesis of propylphosphonic anhydride (T3P) is described, in an overall yield of 51%.
All of the synthesized compounds were characterized by 1H,
13
C and
31
P NMR, LC-MS and
GC-MS.
Commercially available diethyl phosphonate was transformed by Michaelis-Becker reaction
to diethyl propylphosphonate, followed by acidic hydrolysis to propylphosphonic acid. The
transformation into T3P was carried out in refluxing acetic anhydride with subsequent
distillation of oligomeric phosphonic acid anhydride intermediate.
Although propylphosphonic anhydride is commercially available as 50 wt % solution the
solvent range is quite limited (DMF, Ethyl acetate). For many applications, these two solvents
are not appropriate. Therefore we investigate the synthesis of T3P in neat form with further
application in a solvent of choice.
Conclusions
We have developed an optimized method for the synthesis of cyclic propylphosphonic
anhydride (T3P), which can be conveniently applied to a wide variety of chemical
transformations. This process represents a superior alternative method to produce this
important green reagent in comparison with literature reports 1.
1)
Pizova H., Bobal P. Tetrahedron Lett. 2015; 56, 2014-2017.
DETERMINATION OF SELENIUM AND ZINC IN CLINICAL
PLASMA
SAMPLES
OF
PATIENTS
WITH
ATOPIC
DERMATITIS
PLANKOVÁ, A., HAVRÁNEK, E. , MIKUŠ, P.
University, Bratislava, Slovakia; [email protected]
Galvanostatic stripping chronopotentiometry (GSC) was developed and applied for the
determination of selenium (Se) and zinc (Zn) in human plasma. In this work GSC based on
composite carbon electrode coated by a gold layer was optimized concerning various
electrochemical parameters. Along with this, the sample preparation was optimized with
respect to mineralization conditions. The human plasma samples mineralized in an autoclave
under the optimized conditions (160◦C, 100 min, 22 mol/l HNO3) were appropriately diluted by
background electrolyte solution (0.100 mol/l H2SO4 + 0.001 mol/l HCl). The proposed method
was characterized by excellent performance parameters, the limit of detection was 0.2 ng/ml,
accuracy < 5 %, reproducibility < 4 %.
The proposed method was applied for the
investigation of the relationship between atopic dermatitis and Se and Zn concentration in
human plasma. Here, patients suffering from atopic dermatitis were monitored during their
treatment with a pharmaceutical preparation containing inorganic Se and Zn (Zinkosel®). At
the beginning of the experiment, the concentration level of Se in plasma of 100 dermatitic
patients was in the interval 25.20–56.56 ng/ml (average - 40.28 ng/ml) and of Zn of 20
dermatitic patients was in the interval 571.20-726.20 ng/ml (average - 651.30 ng/ml). After 24
months therapy increased levels of Se in plasma 47.39–75.17 ng/ml (average - 63.15 ng/ml)
and of Zn 581.30–777.10 ng/ml (average - 712.90 ng/ml) were detected in 76% of the
patients with an improvement of the clinical state in 65% of the patients (dermatitic symptoms
were suppressed). Great advantage of the proposed method is its extremely high sensitivity
(sub ng/ml concentrations). It allows performing not only ultratrace but also microscale
analyses that could be essential when limited amounts of sample are available. Simple
analytical procedure and relatively short analysis time provide good conditions for a routine
use of the proposed method.
This work was supported by grants VEGA MŠVVaŠ 1/0664/12.
Planková A., et al. Pharmazie 5, 2010; 327 - 330.
Haid J., et al. Slov. Lekár 14, 2004; 242 - 243.
MICROWAVE
–
ASSISTED
SYNTHESIS
AND
CHARACTERIZATION OF THIOSEMICARBAZONES BASED
ON 3-AMINOPYRIDINE-2-CARBOXALDEHYDE
MARTA REJMUND, JAROSŁAW POLAŃSKI
Department of Organic Chemistry, Institute of Chemistry, University of Silesia, Katowice, Poland;
[email protected]
Introduction
Thiosemicarbazones (TSC) are an important class of organic compounds with great
pharmaceutical value. TSC exhibit anticancer, antifungal and antibacterial activity
1)
. They
are a versatile ligands due to possess potential donor atoms – especially sulfur and nitrogen
and they could chelating transition metal ions. The presence of multiple donor atoms within
the same ligand multiplying coordination modes and affects the properties of ligands and
complexes. Considering all of the properties it is important to be able to synthesize new
series of thiosemicarbazones which shows high biological activity 2).
Triapine is already used in medical practice, but several clinical phase II studies proved that
Triapine is inactive against solid tumors like advanced adenocarcinoma of the pancreas or
non-small-cell lung cancer. It is the reason why scientists still investigate a new series of
thiosemicarbazones which should shows biological activity without any side effects
3,4)
.
In synthesis of thiosemicarbazones we used three steps method. In the first step we have
synthesized the derivative of (1,1’-thiocarbonyl)bis-1-H-imidazole and in the next step we
added hydrazine hydrate. In the third and final step we condensed the obtained
thiosemicarbazides with corresponding aldehyde.
General method for the synthesis of thiosemicarbazides:
Into a glass tube provided with magnetic stirrer we inserted a suitable derivative piperazine,
morpholine or thiomorpholine and (1,1'-thiocarbonyl) bis-1H-imidazole. A tub was sealed with
a septum capped and stirred for 24 h at room temperature. The obtained intermediate was
extracted three times with distilled water. The collected organic phase was dried and
evaporated on a rotary evaporator.
Obtained derivative of thioketone was heated 2 h under reflux of the solvent with hydrazine
hydrate in ethanol. The final thiosemicarbazide was allowed to stayed overnight in
temperature -20 ºC and after 24 h was crystallized from methanol.
General method for the synthesis of thiosemicarbazones:
Into a glass tube provided with magnetic stirrer, we inserted equimolar amounts of 3aminopyridine-2-carboxaldehyde and the corresponding thiosemicarbazide. We used ethanol
as a solvent and two drops of concentrated acetic acid as a catalyst. The glass tube was
sealed and placed in to a microwave reactor at 83 ºC for 20 minutes. The obtained
thiosemicarbazones was crystallized from methanol.
In result of our research twelve thiosemicarbazides were prepared by using a reflux method
(2h in ethanol). Twelve thiosemicarbazones were synthesized by using a microwave –
assisted methodology, all of them are novel compounds. The method using a microwave
irradiation permits to obtain products in high purity in a short time with satisfactory yields. The
obtained thiosemicarbazides and thiosemicarbazones were fully characterized by 1H- and
13
C-NMR spectroscopy. Thiosemicarbazones were characterized also by two-dimensional
nuclear magnetic resonance spectroscopy (HMQC and COSY ). The structures of the
received TSCs were confirmed by using Mass Spectrometry (MS) and the purity were
confirmed by using TLC and HPLC technique. The examination for the biological of these
compounds is in progress.
Conclusions
In summary we presented twelve never described before in the literature thiosemicarbazones
obtained from piperazine, morpholine and thiomorpholine derivatives with 3-aminopyridine-2carboxaldehyde. The structures of the synthesized compounds (based on the same
aldehyde as Triapine) suggests a potentially high biological activity both in relation to the
tumor and fungal cells, as confirmed by studies performed initially.
1)
Moretto dos Reis C., et al. Molecules. 2011; 16, 10668-10684.
2)
Cunha S., Silva T. Tetrahedron Lett. 2009; 50, 2090-2093.
3)
Kalinowski D.S., Richardson D.R. Pharm. Rev. 2005; 57, 575-577.
4)
Heffeter P., et al. Bio. Pharm. 2012; 83,1623-1633.
STUDY
OF
THE
CMC
OF
1-[3-(3-
ALCOXYPHENYLCARBAMOYLOXY)-2-HYDROXYPROPYL]4-(4-FLUOROPHENYL)PIPERAZINIUMCHLORIDE
EVA SALANCI, FILS ANDRIAMAINTY, IVAN MALÍK
Comenius University in Bratislava, Faculty of Pharmacy, Department of Pharmaceutical Chemistry,
Bratislava; Slovakia
Introduction: Self-association of surfactants into micelles plays an essential role in various
fields of biological applications and industrial processes. Micelles are unstable units created
by noncovalent aggregation of individual monomers of surfactant. Several possible methods,
for example, surface tension, light scattering, NMR, conductometry and calorimetry can be
used for the determination of critical micelle concentration (CMC). Another approach for the
study are spectral methods like UV/VIS and fluorescence spectroscopy, which are using
different probes for the evaluation of CMC.
Aim: Our study was focused on the micellization of 1-[3-(3- alcoxyphenylcarbamoyloxy)-2hydroxypropyl]-4-(4-fluorophenyl)piperaziniumchloride in aqueous solutions and calculation
of micellization parameters.
Methods:
The
value
of
the
critical
micelle
concentration
of
alcoxyphenylcarbamoyloxy)-2-hydroxypropyl]-4-(4-fluorophenyl)piperaziniumchloride
1-[3-(3in
aqueous solution was determined by the method of pyrene absorption and the calculation of
pyrene I1/I3 ratio. The absorption spectroscopy in ultraviolet region of spectrum was based
on analysis of changes in characteristic absorption spectrum of pyrene in the presence of
surfactant molecule. The value of CMC will be determined as a dependence of the plots of
the sum of absorbances of visible pyrene peaks from the surfactant concentration.
The emission fluorescence spectroscopy using the probe pyrene was applied for
determination of the CMC from the detection of the pyrene I1/I3 ratio as a function of the
concentration surfactant molecules
1, 2
.
Conclusion: The typical absorbtion spectrum of pyrene was described by increasing
Boltzmann- type sigmoid. The typical emission spectrum of pyrene I1/I3 ratio was described
by decreasing sigmoid, again by Boltzmann- type. The values of CMC determined by both
methods were comparable. We would also like to use the method of optical density for the
future analysis.
The work was supported by the grant No. UK/182/2015.
1
Aguiar J., et al. J. Colloid Interface Sci. 2003; 258, 116-122.
2
Basu Ray G., et al. J. Colloid Interface Sci. 2006; 294, 248-254.
SYNTHESIS
AND
BIOLOGICAL
EVALUATION
OF
N-
BENZYL-3-BENZYLAMINOPYRAZINE-2-CARBOXAMIDES
LUCIA SEMELKOVÁ1, ONDŘEJ JANĎOUREK1, PAVLA PATEROVÁ2, KLÁRA
KONEČNÁ1, MARTIN DOLEŽAL1, JAN ZITKO1
1
Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Czech Republic
2
Department of Clinical Microbiology, University Hospital, Hradec Králové, Czech Republic
[email protected]
Introduction
Tuberculosis (TB) is a serious infectious disease, which has been one of the most common
causes of death around the world for a long time. There were 9.0 million new TB cases and
1.5 million TB deaths in 2013.1) Although TB is slowly declining each year and it is estimated
that 37 million lives were saved between 2000 and 2013 through effective diagnosis and
treatment, there is a danger of resistant forms development and co-infection with HIV.1)
Pyrazine ring was chosen as a basic part for drug design because of his presence in many
clinically used substances, especially in pyrazinamide (PZA) – first-line antituberculotic drug
used in TB treatment. PZA has a multi-target effect: acidification of cytoplasm 2) and its active
form pyrazinoic acid inhibits trans-translation3), inhibits Fatty Acid Synthase I4) and Aspartate
decarboxylase5). Synthesis of novel PZA derivatives is one of the perspective ways for the
new drugs development. Series of 13 new substituted N-benzyl-3-chloropyrazine-2carboxamides
and
11
new
N-benzyl-3-(benzylamino)pyrazine-2-carboxamides
were
prepared and evaluated for in vitro anti-infective activity.
Starting nitrile compound (I) was hydrolysed to corresponding acid (II), which was converted
to precursors 1-13 via aminolysis of its chloride. Final compounds 14-24 were prepared by
using microwave-assisted synthesis.
Antimycobacterial assays were performed by microdilution panel method. Compounds were
tested against Mycobacterium tuberculosis H37Rv, M. kansasii, M. avium and M. smegmatis.
Isoniazid was used as a standard. Antibacterial and antifungal evaluation was performed
against eight bacterial or fungal strains by microdilution panel method.
The most active compounds against M. tuberculosis were N-(2-methylbenzyl)-3-[(2methylbenzyl)amino]pyrazine-2-carboxamide
and
N-(3,4-dichlorobenzyl)-3-[(3,4-
dichlorobenzyl)amino]pyrazine-2-carboxamide with MIC = 12.5 µg/mL. From the antibacterial
assays
the
most
effective
compounds
were
3-chloro-N-(2-chlorobenzyl)pyrazine-2-
carboxamide with MIC = 7.81 µmol/L against Staphylococcus aureus and MIC =
15.31 µmol/L
against
Staphylococcus
epidermidis
and
N-(4-methoxybenzyl)-3-[
(4-
methoxybenzyl)amino]pyrazine-2-carboxamide with MIC = 62.5 µmol/L against MRSA.
Conclusions
24 novel compounds were prepared. All compounds were characterized by NMR and IR
spectra, melting point, calculated lipophilic parameters (log P, Clog P) and elemental
analysis. The most active compounds showed the same antimycobacterial activity as
pyrazinamide.
This study was supported by the Grant Agency of Charles University, project B-CH/1594214, SVV 260
183 and by the European Social Fund and the state budget of the Czech Republic, project TEAB no.
CZ.1.07/2.3.00/20.0235.
1)
World Health Organization. Global Tuberculosis Report 2014. WHO/HTM/TB/2014.08.
2)
Singh, P., et al. J. of Communicable Diseases. 2006; 38, 288.
3)
Shi, W., et al. Science. 2011; 333, 1630-1632.
4)
Sayahi, H., et al., Chemistry & Biodiversity, 2012; 9, 2582-2596.
5)
Shi, W., et al., Emerging Microbes & Infections, 2014; 3.
SYNTHESIS, BIOLOGICAL ACTIVITY AND FLUORESCENCE
PROPERTIES OF STYRYLQUINOLINE DERIVATIVES
EWELINA SPACZYŃSKA, WIOLETA CIEŚLIK, ANNA MROZEK-WILCZKIEWICZ,
MARZENA RAMS-BARON, KATARZYNA MALARZ, ROBERT MUSIOŁ
University of Silesia, Katowice, Poland; [email protected]
Introduction
Compounds bearing a quinoline moiety are well known due to their broad biological activities.
A number of them have been widely investigated and clinically used as antifungal or
antibacterial agents1,2). Styrylquinoline derivatives have gained strong attention recently due
to their activity as perspective HIV integrase inhibitors3). There are many reports on the
synthesis and applications of styryl dyes build on quinoline 4,5,6). Recently, some quinolinebased compounds have been synthesized and reported as potent antitumor agents 7). With
these in mind we focused our attention on biological activity quinoline analogues and
fluorescent compounds capable of staining cancer cells.
Novel styrylquinoline was designed by combining molecular fragments from known antiproliferative agents on the diazanaphtalene moiety. Scheme 1 depicts the synthesis of new
compounds based on styrylquinoline scaffolds. Series of quinoline derivatives were obtained
with the use of microwave-assisted synthesis.
Scheme 1. Synthesis of the studied compounds: aldehyde, Ac2O, reflux then Py/H2O or K2CO3/MeOH;
or aldehyde, microwave irradiation.
The lipophilicity of the compounds was measured by RP-HPLC. All studied compounds were
tested for their in vitro antitumor activity. Cellular proliferation was determined using the MTS
assay against the human colon carcinoma (HCT 116) cell lines with wild type p53 (p53+/+)
and with a p53 deletion (p53-/-). The most active compounds were also tested for their
cytotoxicity againts normal cells – human fibroblasts (GM 07492). Compounds localisation in
living cell cultures was studied using fluorescence microscopy. Caspase activation (-3 and 7) was measured using the luminescent Caspase-Glo 3/7 assay in HCT116 cells.
Additionally we investigated fluorescence properties of the compounds. The absorption and
fluorescence spectra were measured at room temperature with a U-2900 spectrophotometer
and an F-7000 spectrofluorimeter. The fluorescence quantum yield was determined by the
comparative method.
A series of thirty six styrylquinoline derivatives were synthesized. All compounds were used
for biological assays. In general the compounds have moderate lipophilicity suitable for good
penetration through the cell wall. Several styrylquinoline analogues were found to have
markedly greater antiproliferative activity than doxorubicin. The most interesting compounds
were 2-3 times more active against HCT116 p53-/- cells than the wild type. The subcellular
localization of compounds into human colon carcinoma cells (HCT116) suggest the quinoline
derivatives accumulate in mitochondria. Subcellular localisation of the compounds and
activating the caspase cascade insinuate the mechanism of action through mitochondrial
pathway to apoptosis in p53 independent manner. Moreover, spectroscopic characterization
for the quinoline was performed. Several quinoline derivatives were found to have interesting
fluorescent properties.
Conclusions
Styrylquinoline derivatives inhibit the proliferation of tumor cells and thus can be used as
anticancer drugs. The most active compound makes it promising for further development. A
group of quinoline analogues showing good potential as candidates for imaging agents.
Several quinoline derivatives can be successfully used as fluorescent dyes.
Ewelina Spaczyńska appreciates the support of the DoktoRIS studentship.
1)
Roth H.J., Fenner H. Arzneistoffe, 3rd Ed.; Deutscher Apotheker Verlag: Stuttgart, Germany.
2)
Harris C.R., Thorarensen A.
Curr. Med. Chem. 2004; 11, 2213–2243.
3)
Polanski J., et al. J. Med. Chem. 2002; 45, 4647-4654.
4)
Li Q., et al. ChemBioChem. 2007; 8, 1679–1687.
5)
Staderini M., et al. ACS Med. Chem. Lett. 2012; 4, 225-229.
6)
Rams-Baron M., et al. PLoS One. 2015; 10, e0131210.
7)
Musiol R., et al. Bioorg. Med. Chem. 2007; 15, 1280-1288.
Abbreviations:
HIV… Human immunodeficiency virus
RP-HPLC… Reversed-Phase High-Performance Liquid Chromatography
HCT116…Human colon carcinoma
SOLUBILITY OF VALSARTAN – INFLUENCE OF CATIONIC
SURFACTANTS ON THE EXTENT OF SOLUBILIZATION
LENKA STOPKOVÁ AND ŽELMÍRA BEZÁKOVÁ
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Comenius University in Bratislava,
Introduction: Valsartan belongs to the sartan drugs, which act at the AT1 angiotensin II
receptors as selective antagonists. Valsartan as a highly selective antihypertensive agent,
contains in its molecular structure two weak-acidic active groups which play an important role
in binding to the AT1 receptor, and a carboxylic group with a value of pKa 3.60 and a tetrazol
ring with pKa 4.70. The ionization profile of valsartan shows that at pH<1.60 exist almost
exclusively in the undissociated form at pH>6.70 as dianion. Valsartan is a drug poorly
soluble in water (0.18 g/l), where the solubility depends on the pH of the medium; it is readily
soluble in methanol, ethanol and acetonitrile. According to BCS, the drug belongs to the
class III with low permeability, poor metabolism and high solubility1),2).
Aim: The aim of the work was to study the solubility of valsartan in micellar solutions of
cationic surfactants and calculation of solubilization parameters.
Methodics: We studied the solubility of valsartan at pH 7.4 influenced by cationic surfactants
(ODTMAB, HDTMAB, TDTMAB) at room temperature 25 °C depending on time, on excess of
the drug and concentration of the surfactants (c=1-80 mM). We choose phosphate buffer
(c=3 mM) for all types of used surfactants. The samples were evaluated by
spectrophotometry and from obtained absorbance values, we calculated the valsartan
solubility – S (μg/ml). We calculated the solubilization parameters (χ, K, K M, ∆G°S) for
valsartan from the cmc (critical micellar concentration) values of solutions and the solubility
(S) of valsartan.
Conclusion: Our results showed increased solubility of valsartan in micellar solutions
compared to the solubility of drug in water. Based on our calculations, we can say, that the
values of solubility valsartan (S) in the presence of surfactants does not increase depending
on the number of carbon atoms on the hydrophobic part of the surfactants molecules. The
solubility increased in the following order HDTMAB>TDTMAB>ODTMAB. We also confirmed
the parameters of solubilization for valsartan, where with increasing values of χ, K, K M the
solubility of valsartan increased in the following order HDTMAB>TDTMAB>ODTMAB. The
values of ∆G°S were negative, indicating that the solubilization of valsartan was spontaneous.
The work was supported by the grant No. FaF UK 18/2015.
1)
Saydam M., Takka, S. J. Pharm. Sci. 2007; 32, 185˗196.
2)
Tosco, C., et al. Helvetica Chimica Acta. 2008; 91, 468˗482.
ANTIOXIDATIVE PROTECTION OF INACTIVATED RABIES
VACCINE
WITH
SQUALENE
ADJUVANT
BY
Β-CAROTENE
SÜLI J.1, KOLČÁKOVÁ L.1, HARVANOVÁ J.1, ONDREJKOVÁ A.2
1
Department of Chemistry, Biochemistry and Biophysics
2
Department of epizootiology and parasitology
University of Veterinary Medicine and Pharmacy in Košice, Slovakia; [email protected]
Introduction
When applying the modern inactivated vaccines, it is necessary to use an adjuvant for
induction of sufficient immune response. The oldest and still most widely used adjuvants are
aluminium compounds. Alum is the principal vaccine adjuvant for clinical applications but it is
a poor inducer of cellular immunity and is not an optimal adjuvant for vaccines where Th1
responses are required for protection 1). The security of alum adjuvants2) is questionable. The
Freund´s complete and incomplete adjuvants used in the past were on the present replaced
by more effective squalene emulsions, which are also easier to handle. Squalene adjuvants
are characterized by high adjuvant effect and due to their biocompatibility they are safe 3).
Squalene emulsions are generally relatively stable, but the squalene, an effective antioxidant
alone can be easily oxidized. The oxidation processes may disturb the stability of the
adjuvant emulsion and also affect its efficacy. The greatest risk of squalene oxidation arises
in time of the preparation, particularly for homogenization and during the prolonged storage
of emulsion. The appropriate solution to eliminate this problem is suggested addition of
antioxidants to squalene adjuvant. To some commercially produced adjuvant formulations is
added α-tocopherol in order to prevent oxidation of squalene. However, according to our
findings, α-tocopherol appeared under in vitro conditions as a pro-oxidant and squalene was
effectively protected by β-carotene4).
Aim of this study was to find whether β-carotene affects the particle size of the emulsion
(emulsion stability indicator) and how affects the antioxidant protection of squalene adjuvant
rabies vaccine.
This work was financed by scientific project APVV-0605-12 (Slovak Republic).
1)
Mori A., et al. Eur. J. Immunol. 2012; 42, 2709–2719.
2)
Tomljenovic L., Shaw C. A. Curr. Med. Chem. 2011; 18, 2630–2637.
3)
Fox C.B. Molecules 2009; 14, 3286–3312.
4)
Süli J., et al. Folia Vet. 2012; 56, Suppl. II, 56–58.
DETERMINATION OF THE COLORANTS IN VITAMIN E BY
HPLC WITH PHOTODIODE ARRAY DETECTION
MONIKA ŠULEKOVÁ, ALEXANDER HUDÁK
Department of chemistry, biochemistry and biophysics, Institute of pharmaceutical chemistry,
University of veterinary medicine and pharmacy in Košice, Slovakia; [email protected]
Colorants are added to drugs and nutritional supplements for commercial, psychological and
practical reasons. According to European Union regulations the content of dyes in
pharmaceutical products and food must be observed. Two colorants (Ponceau 4R, E 124
and carmine, E 120) were determined by high-performance liquid chromatography with
isocratic elution and diode-array detector. A Polaris C18-A column (250 mm x 4.6 mm), 5 μm
particle size was used as stationary phase. Mobile phase contained an Acetonitrile and
mixture CH3COONa:CH3OH (85:15 v/v) in the ratio of 20:80, v/v. The samples of vitamin E
from five different manufacturers were analyzed. In one sample of vitamin E was provided
79,5 ± 0,01 g/capsule Ponceau 4R. The other two samples contained 495,0 ± 0,02
μg/capsule, respectively 314,8 ± 0,02 μg/capsule of carmine. The obtained results for the
determination of Ponceau 4R and carmine on real samples of vitamin E demonstrated that
the described method is rather fast with a very good detection limit and quantification limit.
PLANTS OF THE GENUS HYLOCEREUS – VOLATILE
COMPOUNDS ANALYSIS
1
MIROSLAVA TARHAJOVÁ,
2
ÉVA HÉTHELYI B.,
2
ANDREA BÖSZÖRMÉNYI,
1
GÁBOR SZOLGAI, 1JAROSLAV TÓTH, 1SZILVIA CZIGLE
1
Department of Pharmacognosy and Botany, Faculty of Pharmacy, Comenius University in Bratislava,
Slovakia
2
Institute of Pharmacognosy, Faculty of Pharmacy, Semmelweiss University, Budapest, Hungary
[email protected]
Introduction: Cacti, e.g. Hylocereus (Berger) Britt., Cactaceae, are known mostly as
ornamental plants. The dragon fruits – pitaya (Hylocerei fructus) are commonly eaten in
some regions of the Americas. The taste has been described as being very bland, with a mild
sweetness, resembling a watermelon. The active compounds include betaxanthins, phenolic
compounds and terpenes.
The aim of this work was the analysis of Hylocerei fructus (Hylocereus sp. Mill., Cactaceae)
volatile constituents.
Materials: Fruits of two Hylocereus taxa: H. costaricensis (red-fleshed pitaya) and H.
undatus (white-fleshed pitaya) were analysed; collected in the Botanical Garden in Bratislava
(October, 2012).
Methods: The volatile constituents were qualitatively and quantitatively evaluated using
SPME GC-MS (Agilent 6890/5973N).
Results and discussion: We have identified 13 constituents of red-fleshed fruits, as well as
other 12 compounds in white-fleshed fruits. The following constituents were identified in all of
the red-fleshed fruits in largest percentage: cetylalcohol (62.4 %), hexadecanol (30.3 %),
and nonadecane (8.0 %). The following constituents were identified in all of the white-fleshed
fruits in largest percentage: cetylalcohol (60.5 %), hexadecanol (26.9 %), and octadecanol
(6.4 %).
Conclusions: This pilot study identified some lipophilic constituents of the fruits. Further
analyses will be carried out identify nitrogen-containing as well as flavour-bearing
compounds to compare with other cacti fruits.
This work was supported by the Slovak Grant Agency VEGA Project No. 2/0044/15 and 1/0646/14.
NITRO GROUP-CONTAINING SUBSTITUTED TETRAZOLES:
NEW HIGHLY EFFICIENT ANTITUBERCULAR AGENTS
VALÁŠKOVÁ L.1, ROH J.1, KARABANOVICH G.1, NĚMEČEK J.1, STOLAŘÍKOVÁ
J.2, VÁVROVÁ K.1, KLIMEŠOVÁ V.1, HRABÁLEK A.1
1
Department of inorganic and organic chemistry, Faculty of Pharmacy in Hradec Kralove, Charles
University in Prague, Czech Republic
2
Laboratory for dg. of mycobacteries, Departement of Bacteriology and Mycology, Regional Institute
of Public Heatlh, Ostrava, Czech Republic
[email protected]
Tuberculosis (TB) is becoming worldwide health and economic problem. Bacterial strains
causing this disease have developed resistance against approved antiTB drugs which have
been presented several decades ago and since then there was no compound introduced to
common clinical practice.
Recently,
our
group
have
developed
heteroaromatic
compounds
containing
5-(dinitrobenzylsulfanyl)tetrazole moiety with high and selective antimycobacterial activity.
Prepared compounds showed high activity against drug-susceptible M. tuberculosis with MIC
values comparable with those of the first-line antiTB drugs. Furthermore, studied compounds
possess significant activity against multidrug resistant strains, with MIC values as low as 0.25
µM and with no cross resistance with first or second line antiTB drugs. The antimycobacterial
effect is highly specific, since the compounds are inactive against bacteria or fungi and
possess low cellular toxicity and mutagenicity.
This work was supported by the Czech Science Foundation project (14-08423S), “Pre-seed activities
of CU out of Prague“, reg. n. CZ.1.05/3.1.00/13.0284 and "Pre-seed activities of CU out of Prague II",
reg. n. CZ.1.05/3.1.00/14.0299
BIOLOGICAL
ACTIVITY
OF
SELECTED
SPECIES
OF
MACROMYCETES
MARIE VALENTOVÁ, JANA ROHÁLOVÁ
Sciences Brno, Czech Republic; [email protected]
Introduction: Many species of macromycetes have been used for many years in nutrition
and in traditional medicine1) and in many of them the pharmacological activity has already
been proved.2) We focused on analysis of selected macromycetes from Fomitopsidaceae
(Polyporales): Climacocystis borealis, Daedalea quercina, Fomitopsis pinicola, Laetiporus
sulphureus,
Piptoporus
betulinus;
Polyporaceae
(Polyporales):
Fomes
fomentarius,
Trametes versicolor, Polyporus squamosus; and Tricholomataceae (Agaricales): Lepista
nuda. We assessed their biological activity and analysed extracts using HPLC-DAD and LCMS.
Experimental methods: Material was air-dried at below 35°C. Extracts in ethylacetate and
petroleum ether were prepared using microwave extractor. HPLC analysis was performed
using Agilent 1100 HPLC system with DAD, LC-MS analysis was performed on Agilent
HP1100 LC/MSD Trap VL Series, ESI. Biological activity was measured using Microplate
Reader; methods of scavenging of DPPH radical, scavenging of hydroxyl-radical, scavenging
of peroxynitrite and determination of total phenolic content were used.
Results and discussion: In preparation of extracts, as more effective solvent was found
ethyl acetate due to extraction of higher number of substances. Only a part of constituents
absorbed in the wavelenghts 280 and 350 nm; in future also other wavelenghts should be
used in the analysis. In testing the biological activity of the extracts, significant differences
between used methods were found, however, as significantly strongest antioxidant acted
Fomes fomentarius in all used methods.
Conclusions: Fungi are widely used in traditional medicine of Asian countries. However, in
many of them only little scientific information is known. In this work, we aimed to find suitable
method for extraction and analysis of fungi, which can be used in future research on higher
range of fungi with potential biological activity.
1)
Aida F.M.N.A, et al. Trends Food Sci Tech. 2009; 20; 567-575.
2)
Zhu F., et al. J. Food Compost. Anal. 2015; 41; 165-173.
SYNTHESIS OF TRICLOSAN DERIVATIVES AND THEIR
ANTIMYCOBACTERIAL EFFECT
RUDOLF VOSÁTKA, MARTIN KRÁTKÝ, JARMILA VINŠOVÁ
Department of Inorganic and Organic Chemistry, Faculty of Pharmacy, Charles University, Hradec
Králové, Czech Republic; [email protected]
Introduction
Tuberculosis (TB) represents one of the leading causes of morbidity and mortality worldwide.
Development of new potential drugs is essential because of the existence of latent TB and
development of drug-resistant TB forms (multidrug-resistant TB, extensively drug-resistant
TB and recently reported totally drug-resistant TB).1,2) Triclosan (irgasan) is a broad spectrum
antibacterial agent used in household products. Triclosan has been shown to inhibit InhA, an
essential enoyl acyl carrier protein which leads to the lysis of Mycobacterium tuberculosis.3)
Esterification of triclosan to form its prodrugs can produce compounds with improved
properties – enhanced bioavailability or absorption, higher activity and/or lower toxicity.
We used two synthetic procedures to obtain these esters. The first pathway consists in the
reaction of triclosan (1 eq.) with various acyl chlorides (1,3 eq.) in presence of triethylamine
(1,5 eq.). The second approach of the preparation triclosan esters is the Steglich
esterification. Common yields were around 70 %. Synthesized derivatives were evaluated for
their in vitro antimycobacterial activity against Mycobacterium tuberculosis H37Rv, M. avium
and two strains of M. kansasii.
It was prepared 28 triclosan esters based on various aliphatic, cycloaliphatic, aromatic and
heteroaromatic acids. 5-Chloro-2-(2,4-dichlorophenoxy)phenyl 4-bromobenzoate (TRC-B4Br) showed the best in vitro activity with minimum inhibitory concentrations (MIC) 16 µmol/L
against Mycobacterium tuberculosis H37Rv. Against next strains had the best activity 5chloro-2-(2,4-dichlorophenoxy)phenyl isonicotinate (TRC-ISO). The MIC of TRC-ISO was
similar for M. kansasii 6509/96 and better for M. avium and M. kansasii 235/80 with
comparison of INH.
Conclusions
The in vitro evaluation of 28 triclosan-based esters showed promising antimycobacterial
activity. The further research of the most active analogues will continue, particularly with
regard to cytotoxicity.
Grant dedications: IGA NT 13346 (2012)
1)
World
Health
Organization.
Global
tuberculosis
http://apps.who.int/iris/bitstream/10665/137094/1/9789241564809_eng.pdf?ua=1
2)
Krátký M., Vinšová, J. Chem. Listy. 2010; 104, 998-1005
3)
Stec J., Vilchéz C., et al. ChemMedChem. 2014; 9, 2528-2537
report
2014.
(30.6.2015)
THE DEVELOPMENT OF DENTAL DRUG IN THE FORM OF
MEDICATED CHEWING GUM
OLEKSII YAKOVENKO, JULIA MASLIY
National University of Pharmacy, Kharkiv, Ukraine; [email protected]
Every year the number of people with diseases of the mouth cavity is growing rapidly.
Therefore, the necessity for dosage forms, which would effectively contribute to the treatment
of these diseases, is an urgent problem today. Medicated chewing gum (MCG), which carry
out a role of oral drug delivery, is the optimal formulation for the prevention and treatment of
dental diseases, due to their advantages – fast local effect, no need to swallow, the
possibility of applying in any convenient place for the patient, as it does not need water,
reduced the first-pass metabolism, as well as a better perception of the patient and a more
pleasant mode of administration. Besides that, MCGs increase of secretion of saliva, which
helps digestion, cleaning and remineralization of teeth, effective in the prevention and
treatment of xerostomia, the process of mastication has massaging effect on the gingiva,
which leads to greater blood flow and microcirculation of periodontal tissues. Moreover,
properly selected active pharmaceutical ingredients (APIs) in the composition of the dosage
form provide the appropriate comprehensive treatment and preventive effect on hard tissues
of teeth, periodontal tissues and mucous. That is why the aim of our work was to develop a
dental drug of comprehensive action in the form of medicated chewing gum.
Participants
Surname, Name, Title
1.
2.
3.
AMBROŻKIEWICZ Weronika
BĄK Andrzej
Dr. hab
BEDLOVIČOVÁ Zdenka
RNDr., Ph.D.
4.
BEVZ Olena
5.
BEVZ Nataliya
Ph.D.
6.
BOBÁĽ Pavel
Ing., CSc.
7.
CAHLÍKOVÁ Lucie
Ing.
8.
CSÖLLEI Jozef
Prof. RNDr., CSc.
9.
10.
CZAPLIŃSKA Barbara
Mgr.
ČAVOJSKÝ Tomáš
PharmDr.
11.
ČERNÍKOVÁ Aneta
PharmDr.
12.
DAŇKOVÁ Ivana
PharmDr.
13.
DOKUPILOVÁ Svetlana
RNDr., Ph.D.
14.
DOLEŽAL Martin
prof. PharmDr., Ph.D.
15.
FARSA Oldřich
doc. PharmDr., Ph.D.
16.
17.
18.
19.
FAZEKAŠ Tomáš
RNDr., Ph.D.
FORGÁCSOVÁ Andrea
Mgr.
GÁLUSOVÁ Andrea
PharmDr.
GARAJ Vladimír
PharmDr., Ph.D.
Organization
University of Silesia in Katowice,
Institute of chemistry, Poland
University of Veterinary Medicine and
Pharmacy in Košice, Slovakia
National university of Pharmacy,
Kharkiv, Ukraine
Kharkiv, Ukraine
University of Veterinary and
Pharmaceutical Sciences Brno, Faculty
of Pharmacy, Czech Republic
Charles University in Prague, Faculty of
Pharmacy in Hradec Kralove, Czech
Republic
Comenius University in Bratislava,
Faculty of Pharmacy, Slovakia
Republic
Lecture/Poster Nr.
P01
P03, P20, P29
P04
P33
P33
P07, P59
L01
P18, P32
P05, P26
P06
P07
P08
P09
P55
P10
P11
P12
P13
P14
20.
GAZVODA Martin
Dr.
21.
GEORGIYANTS Victoriya
Prof.
22.
GONĚC Tomáš
PharmDr., Ph.D.
23.
GRUBEROVÁ Lucie
Ing.
24.
25.
26.
27.
28.
29.
30.
31.
32.
HAIDER Norbert
Prof. Dr.
HAIDER-SALABERGER Ingrid
Dr.
HARNA Svitlana
Dr.
HARNA Nataliia
Ph.D.
HAVRANOVÁ SICHROVSKÁ
Ľubica PharmDr.
CHRIPKOVÁ Martina
MVDr.
JAMPÍLEK Josef
Doc. PharmDr., Ph.D.
JEŽKO Pavol
PharmDr., Ph.D.
KAPKOWSKI Maciej
MSc.
33.
KAPUSTÍKOVÁ Iva
PharmDr., Ph.D.
34.
KARABANOVICH
Galina, Ing. Ph.D.
35.
KASTNER Petr
PharmDr., Ph.D.
36.
KORZEC Mateusz
MSc.
37.
KOS Jiří
Mgr.
38.
KOSMRLJ Janez
Prof.
39.
40.
KOZAKIEWICZ Dariusz
MSc.
KOZIK Violetta
Dr.
University of Ljubljana, Faculty of
Chemistry and Chemical Technology,
Slovenia
Kharkiv, Ukraine
University of chemistry and technology,
Prague, Faculty of Chemical Technology,
Czech Republic
University of Vienna, Faculty of Life
Sciences, Austria
Kharkiv, Ukraine
Kharkiv, Ukraine
Republic
Republic
University of Ljubljana, Faculty of
Chemistry and Chemical Technology,
Slovenia
P15
L02, P43
P16, P27
P17
L03
P33
P18, P23
P19
P07, P16, P20, P27
P21
P22
P18, P23
P24, P46, P60
P25, P50
P05, P26
P16, P27
L04
P28
P03, P20, P29
41.
KRAJČIOVÁ Dominika
Mgr.
42.
KRÁTKÝ Martin
PharmDr. Mgr., Ph.D.
43.
KRIUKOVA Anna Dr.
44.
KROUTIL Aleš
Mgr.
45.
KRYVANYCH Oleksandr
Ph.D.
46.
KUBÍNOVÁ Renata
47.
48.
49.
50.
51.
52.
53.
54.
55.
LUKAČOVIČOVÁ Oľga
Ing., Ph.D.
MALARZ Katarzyna
MSc.
MALEČEK Jiří
Ph.D.
MARÁKOVÁ Katarína
PharmDr., Ph.D.
MARUNIAK Matej
PharmDr., Ph.D.
MARVANOVÁ Pavlína
PharmDr.
MATERIIENKO Anna
MIKULOVÁ Mária
Mgr.
MISHCHENKO Volodymyr
Dr.
56.
MOKRÝ Milan
RNDr., CSc.
57.
MOKRÝ Petr
Mgr., Ph.D.
58.
59.
60.
61.
62.
MOLNAR Maja
MULARSKI Jacek
MSc.
MUSIOL Robert
Dr. hab.
MYHAL Artem
MYZNIKOV Leonid
Dr.
Republic
Kharkiv, Ukraine
Kharkiv, Ukraine
Tekro, s.r.o.
Kharkiv, Ukraine
Kharkiv, Ukraine
Republic
Josip Juraj Strossmayer University of
Osijek, Faculty of food technology,
Croatia
Kharkiv, Ukraine
Saint-Petersburg State University of
Technology and Design, Russia
P30
P31, P62
P32
P33
P34
P35
P36
P37, P49
P18, P23
P38, P48
L05
P40
P41
P38, P48
P02
P42
L06, P05, P26, P36,
P42
P43
P44
63.
NALEPA Paweł
MSc.
64.
NĚMEČEK Jan
Mgr.
65.
ODEHNALOVÁ Klára
Ing., Ph.D.
66.
67.
68.
69.
70.
71.
72.
ONDREJKOVÁ Anna
Doc., MVDr., Ph.D.
OPATŘILOVÁ Radka
Doc. PharmDr. Ing., Ph.D.
MBA
PADRTOVÁ Tereza
PharmDr.
PAVIĆ Valentina
PAZOUREK Jiří
Doc. RNDr. Bc., Ph.D.
PIEŠŤANSKÝ Juraj
PharmDr.
PIETRZYŃSKA Monika
Dr.
73.
PILAŘOVÁ Pavla
PharmDr., Ph.D.
74.
PÍŽOVÁ Hana
Mgr., Ph.D.
75.
76.
77.
78.
79.
80.
81.
82.
PLANKOVÁ Alexandra
RNDr., Ph.D.
POLANSKI Jaroslaw
Prof.
PROKOPENKO Yuliya
Dr.
REJMUND Marta
RIŠIAŇOVÁ Lucia
Mgr.
ROH Jaroslav
Mgr.
SALANCI Eva
PharmDr.
SAVCHENKO Lesia
Dr.
83.
SEMELKOVÁ Lucia
Mgr.
84.
SPACZYŃSKA Ewelina
Republic
Josip Juraj Strossmayer University of
Osijek, Department of biology, Croatia
Poznań University of Technology,
Faculty of Chemical Technology, Poland
Republic
Kharkiv, Ukraine
University of Silesia in Katowice, Poland
Republic
Kharkiv, Ukraine
Republic
P45
P46, P60
P48
P47, P57
P38, P48
P02
L07
P37, P49
L08
P25, P50
P51
P52
L09, P03, P22, P26,
P28, P36, P45, P53
P53
L10, P39
P24, P46, P60
P54
P55
P05, P26, P27
85.
86.
87.
88.
89.
90.
91.
Mgr.
STANIČOVÁ Jana
Doc. RNDr., Ph.D.
STANZEL Lukáš
PharmDr.
STOPKOVÁ Lenka
PharmDr.
SÜLI Judit
RNDr., Ph.D.
ŠMEJKAL Karel
ŠULEKOVÁ Monika
RNDr., Ph.D.
TARHAJOVÁ Miroslava
PharmDr.
92.
VALÁŠKOVÁ Lenka
Mgr.
93.
VALENTOVÁ Marie
PharmDr., Ph.D.
94.
VLADYMYROVA Inna
Dr.
95.
VOSÁTKA Rudolf
Mgr.
96.
YAKOVENKO Olexii
97.
ZITKO Jan
PharmDr., Ph.D.
98.
ŽEMLIČKA Milan
Doc. RNDr., CSc.
Republic
Republic
Kharkiv, Ukraine
Republic
Kharkiv, Ukraine
Republic
L11
L12, P18, P23
P56
P47, P57
L13
P58
P59
P46, P60
P61
P62
P63
P55
L14
Elektronický sborník 44. Konference Syntéza a analýza léčiv
2. - 4. září 2015
Brno
Vydala Veterinární a Farmaceutická univerzita Brno
ISBN: 978-80-7305-760-2
Abstrakty příspěvků byly redakčně kráceny a formátovány.
Zaslané celé články vyjdou v časopise Česká a Slovenská Farmacie.
Yearbook of 44th Conference Drug Synthesis and Analysis
2th – 4th September 2015
Brno
Published by University of Veterinary and Pharmaceutical sciences Brno
ISBN: 978-80-7305-760-2
Abstracts were shortened and edited by editors.
Articles will be published in Journal Česká a Slovenská Farmacie.

Book of Abstracts - Veterinární a farmaceutická univerzita Brno

Transcription

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