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Vanilin je spoj iz skupine aromatskih benzaldehida. Dobro je topljiv u organskim otapalima, a metoksilna skupina čini ga topljivim u polarnim otapalima. Primarnu primjenu nalazi u prehrambenoj industriji, a također se koristi i u farmaceutskoj industriji za sintezu farmaceutskih proizvoda i u industriji parfema zahvaljujući svom ugodnom mirisu. Prirodni izvor vanilina je biljka vanilije, a prvi su se vanilijom koristili narodi Srednje Amerike još u 12. stoljeću. Prvi je put izoliran 1858. godine, sintetiziran 1874. iz koniferina, a krajem 19. stoljeća postaje komercijalno dostupan. Danas postoji više načina sinteze: biosinteza, sinteza iz lignina te kemijska sinteza iz jednostavnijih preteča poput 4-hidroksibenzaldehida s natrijevim hidroksidom. Cilj ovoga rada je usporedba rezultata sinteze vanilina iz 4-hidroksibenzaldehida provedene u studentskom laboratoriju s rezultatima dostupnim u literaturi. Vanillin is a compound belonging to the aromatic benzaldehyde group. It is mostly soluble in organic solvents, and the methoxy group makes it soluble in polar solvents. It is primarily used in the food industry but it is also used in the pharmaceutical to make pharmaceuticals and by the perfumes industry thanks to its pleasant scent. The vanilla plant is the natural source of vanillin, and, in the 12th century, the people of Central America were the first to use it. Vanillin was first isolated in 1858, synthesized in 1874 from coniferin, and became commercially available at the end of the 19th century. Today there are several ways to synthesize vanillin: biosynthesis, synthesis from lignin and chemical synthesis from simpler precursors such as 4-hydroxibenzaldehyde with sodium methoxide. The aim of this study is to compare the results of vanillin synthesis from 4-hydroxybenzaldehyde done in a student laboratory with those available in literature.

Male molekule koje se preferencijalno vežu za hibridne i višelančane strukture DNA i RNA nekovalentnim interakcijama od velikog su znanstvenog interesa, te mogu naći primjenu kao novi i bolji dijagnostički i terapeutski alati. Iako te strukture imaju važnu biološku funkciju unutar stanice mali je broj liganada koji se na njih preferencijalno vežu. Cilj ovog rada je pronaći nove strukturne motive/spojeve koji se preferencijalno vežu na hibridne i višelančane u odnosu na regularne DNA i RNA strukture. Ispitano je nekoliko grupa spojeva: benzotiazolni derivati i cijaninske boje koje su dobivene u suradnji s drugim istraživačkim grupama. Također, istražene su interakcije novosintetiziranih fenantridinskih derivata s DNA. Spojevi s preferencijalnim vezanjem na hibridne i višelančane strukture DNA i RNA odabrani su pomoću brzih metoda probira-kompeticijske dijalize i temperaturnog mekšanja smjesa te RNaza H testa. Daljnjim bioanalitičkim metodama (spektroskopija UV/Vis, fluorescencijska spektroskopija i cirkularni dikroizam) detaljno su okarakterizirane interakcije vezanja malih molekula s odabranim metama. Pokazali smo da se benzotiazolski i cijaninski derivati preferencijalno vežu na hibridne i višelančane strukture DNA i RNA u odnosu na regularne što bi se moglo iskoristiti u antitumorskoj terapiji i za detekciju struktura nukleinskih kiselina. Small molecules able to preferentially bind to specific hybrid and multistranded structures of DNA and RNA by non-covalent interactions are of great scientific interest when it comes to obtaining new and better diagnostic and therapeutic tools. Although these structures have an important biological function inside the cell, there is only a few known ligands which preferentially bind to them. The goal of this thesis is to find new structural motifs/compounds that preferentially bind to hybrid and multistranded structures compared to regular DNA and RNA structures. A small library of several groups of compounds was formed: benzothiazole derivatives and cyanine dyes obtained in collaboration with other research groups. Also, interactions of newly synthesized phenanthridine derivatives with DNA were investigated. Compounds with preferential binding to hybrid and multistranded structures of DNA and RNA were selected using competitive dialysis as a rapid screening method, as well as with melting of mixtures and RNase H assay. Further bioanalytical methods (flourescence and UV/Vis spectroscopy and circular dichroism) were used to characterize their binding interactions with selected targets in detail. The obtained results indicate that certain derivatives from the mentioned classes of compounds preferentially bind to hybrid and multistranded structures of DNA and RNA compared to regular ones, which opens the possibility of their application in antitumor therapy and for the detection of nucleic acid structures.

U sklopu ovog rada istražena su fizikalno-kemijska svojstva nanočestica titanijeva dioksida (TiO2), suspendiranih u vodenim otopinama elektrolita i polielektrolita. Dinamičkim raspršenjem svijetlosti i mikroelektroforezom određeni su hidrodinamički promjer i elektrokinetički potencijal čestica kao pokazatelji stabilnosti suspenzija. Istraživanje se fokusiralo na utjecaj promjene pH–vrijednosti, ionske jakosti, vrste i koncentracije soli te polielektrolita na stabilnost nanočestica (nanopločica i nanosfera) u suspenziji. Ispitano je kako modifikacija površinskog naboja pozitivno ili negativno nabijenim polielektrolitom utječe na stabilnost suspenzije uz dodatak soli jednovalentnih kationa. Procijenjena je učinkovitost polielektrolita kao potencijalnih stabilizatora nanočestica TiO2 u kontekstu kinetike agregacije čestica. Pokazano je za obje morfologije da uslijed prenabijanja površine nanočestica polielektrolitom, priređeni sustavi pokazuju izuzetnu stabilnost te da ne dolazi do agregacije u području izoelektrične točke. Također, ustanovljeno je da je potrebna viša koncentracija soli za agregaciju nanočestica ukoliko je polielektrolit adsorbiran na površinu čestica. In this study the physicochemical properties of titanium dioxide (TiO2) nanoparticles suspended in electrolyte and polyelectrolyte aqueous solutions were investigated The hydrodynamic diameter and the electrokinetic potential of the particles, as indicators of suspension stability, were measured using dynamic light scattering and microelectrophoresis. The research focused on the influence of changes in pH, ionic strength, type/concentration of salts and polyelectrolytes on the stability of nanoparticles (nanoplates and nanospheres) in suspension. It was examined how surface modification with either positively or negatively charged polyelectrolyte affects suspension stability in the presence of monovalent salts. The efficacy of polyelectrolytes as potential stabilizers for nanoparticles, in the context of particle aggregation kinetics, was assessed. It was shown that due to the surface overcharging by the polyelectrolytes, the prepared systems showed exceptional stability, with no aggregation near the isoelectric point. Additionally, it was found that a higher salt concentration is required for nanoparticles aggregation if the polyelectrolyte is adsorbed on the particle surface.

U ovom diplomskom radu sintetizirani su koordinacijski polimeri bakrovog(II) klorida i bakrovog(II) bromida s 4-cianopiridinom (4-CNpy), [CuCl2(4-CNpy)2]n (1) i [CuBr2(4- CNpy)2]n (2). Nastali produkti karakterizirani su infracrvenom spektroskopijom (IR), termičkom analizom (TGA/DSC) te difrakcijom rendgenskog zračenja u polikristalnom uzorku (PXRD). Kristali koordinacijskih polimera priređeni su metodom difuzije preko tekuće faze. Priređenim igličastim kristalima ispitana je mehanička fleksibilnost uslijed primjene vanjske mehaničke sile. Mehanički odziv kristala istražen je modificiranom metodom savijanja u tri točke. Kristali spojeva 1 i 2 pokazali su mehanički potaknutu elastičnosti te je određen stupanj elastičnosti. Ustanovljena mehanička fleksibilnost kristala spojeva 1 i 2 korelirana je s međumolekulskim interakcijama ostvarenim u kristalnom pakiranju priređenih spojeva. This diploma thesis presents synthesis of coordination polymers of copper(II) chloride and copper(II) bromide with 4-cyanopyridine (4-CNpy), [CuCl2(4-CNpy)2]n (1) and [CuBr2(4- CNpy)2]n (2). The obtained compounds were characterized by infrared spectroscopy (IR), thermal methods (TGA/DSC) and powder X-ray diffraction (PXRD). Crystals of the coordination polymers were prepared using liquid-liquid diffusion method. The mechanical flexibility of crystals was examined due to the application of an external mechanical force. The mechanical responsiveness of crystals was investigated via modified three-point bending experiments. Mechanically induced elasticity of crystals was observed and the extent of elasticity was calculated. Mechanical flexibility observed on crystals of compounds 1 and 2 was correlated with intermolecular interactions in crystal packings of prepared compounds.

Polielektrolitni kompleksi i višeslojevi poli(alilamin-hidroklorida) i poli(akrilne kiseline) pripravljeni su u vodenim otopinama NaF, NaCl, NaClO4, LiCl i (CH3)4NCl. Nastanak polielektrolitnih kompleksa po prvi puta istražen je visoko-razlučivom ultrazvučnom spektroskopijom. Ovom metodom ustanovljeno je da interpolielektrolitna neutralizacija rezultira ion-specifičnom agregacijom nabijenih koloidnih kompleksa. Analogan utjecaj iona određen je u slučaju odgovarajućih višeslojeva. Elipsometrom i mikroskopom atomskih sila utvrđeno je da debljina i hrapavost filma raste u nizu F − < Cl− < ClO4 − za anione odnosno (CH3)4N + < Na+ < Li+ za katione. Nadalje, ustanovljeno je da hidrofobnost višesloja također slijedi opisani trend za anione kada je polikation završni sloj filma. Zagrijavanjem polielektrolitnih višesloja tijekom njihove priprave potisnut je specifičan utjecaj iona na površinsku hrapavost i močivost zbog reorijentacije i migracije polimernih lanaca na međupovršini višesloj/zrak. Naposlijetku, provedeno istraživanje ukazuje na značajnu korelaciju između procesa interpolielektrolitne neutralizacije u otopini i na površini. Polyelectrolyte complexes and multilayers of poly(allylamine hydrochloride) and poly(acrylic acid) were prepared in aqueous solutions of NaF, NaCl, NaClO4, LiCl, and (CH3)4NCl. The formation of polyelectrolyte complexes was investigated for the first time by high-resolution ultrasonic spectroscopy. Using this method, it was established that interpolyelectrolyte neutralization results in ion-specific aggregation of charged colloidal complexes. Analogous influence of ions was observed in the case of corresponding multilayers. Using ellipsometer and atomic force microscope, it was determined that the thickness and roughness of the film increases in the sequence F− < Cl− < ClO4 − for anions and (CH3)4N + < Na+ < Li+ for cations. Furthermore, it was found that the hydrophobicity of the multilayer also follows the described trend for anions when a polycation is terminating layer of the film. By heating polyelectrolyte multilayers during their preparation, the specific influence of ions on surface roughness and wettability was suppressed due to the reorientation and migration of polymer chains at the multilayer/air interface. Finally, the conducted research indicates a significant correlation between the process of interpolyelectrolyte neutralization in solution and on the surface.

Porast rezistencije bakterija na antibiotike koji su trenutno na tržištu potaknuo je potrebu za otkrivanjem novih. Makrolidni antibiotik azitromicin koristi se za liječenje bolesti uzrokovanih Gram-pozitivnim i Gram-negativnim bakterijama. Makrozoni su konjugati azitromicina i tiosemikarbazona pripremljeni za liječenje infekcija uzrokovanih rezistentnim bakterijama, s potencijalom zaobilaženja razvijenih mehanizama rezistencije. Kako bi se dodatno povećala biološka aktivnost i farmakološka svojstva, napravljeni su kompleksi tiosemikarbazona s metalnim ionima. U ovom radu prvo je pripravljen tiosemikarbazon izveden iz 4- aminobenzojeve kiseline i salicilaldehida. Nakon toga, dobiveni tiosemikarbazon je vezan na 4''-aminopropilni derivat azitromicina dajući pripadajući makrozon. Istražena je mogućnost kompleksiranja s ionima bakra(II) i nikla(II). Sintetizirani spojevi karakterizirani su spektroskopijom NMR. The increase in bacterial resistance to currently marketed antibiotics has prompted the need to discover new ones. The macrolide antibiotic azithromycin is used to treat diseases caused by Gram-positive and Gram-negative bacteria. Macrozones are conjugates of azithromycin and thiosemicarbazones prepared to treat infections caused by resistant bacteria, with potential of bypassing the developed resistance mechanisms. In order to further increase biological activity and pharmacological properties thiosemicarbazone complexes with metal ions have been made. In this work, a thiosemicarbazone derived from 4-aminobenzoic acid and salicylaldehyde was prepared first. Subsequently, the resulting thiosemicarbazone was bound to the 4''-aminopropyl derivative of azithromycin. The possibility of complexation with copper (II) and nickel (II) ions was explored. The synthesized compounds were characterized by NMR spectroscopy.

Primjenom niza modernih računalnih metoda dobiven je detaljan uvid u interakcije koje antagonist istradefilin ostvaruje s adenozinskim A2A receptorom. Temeljem identifikacije strukturnih elemenata koji najviše doprinose vezanju, dizajnirani su derivati većeg afiniteta. Osim konvencionalnog pristupa koji uključuje uvođenje novih funkcionalnih skupina, primijenjena je i modifikacija deuteriranjem čime je postignuto poboljšano vezanje istradefilina u A2A receptor. U ovom radu izotopna supstitucija predstavlja originalan koncept moduliranja farmakodinamičkih svojstava, a dobiveni rezultati ukazuju na veću učinkovitost istradefilina obogaćenog deuterijem što bi se moglo iskoristiti za poboljšanje njegovog farmakoterapijskog profila u liječenju Parkinsonove bolesti. By utilising a series of modern computational methods, a detailed insight into the interactions of the antagonist istradefylline with the adenosine A2A receptor was obtained. Based on the identification of structural elements that contribute the most to binding, higher affinity derivatives were designed. In addition to the conventional approach that includes the introduction of new functional groups, modification by deuteration was also applied, which achieved improved binding of istradefylline to the A2A receptor. In this work, isotopic substitution represents an original concept of modulating pharmacodynamic properties, and the obtained results indicate a greater efficiency of deuterium-enriched istradefylline, which could be used to improve its pharmacotherapeutic profile in the treatment of Parkinson's disease.

The oxytocin receptor (OTR) and the vasopressin 1a receptor (V1aR) are two G protein-coupled receptors involved in mediating various physiological processes and social and sexual behaviors. Producing specific antibodies to study these receptors is challenging due to their sequence homology and structure similarity, along with two additional closely related vasopressin receptors. Thus, this thesis aimed to validate ten antibodies to OTR and seven antibodies to V1aR produced against distinctive receptor peptide epitopes, using western blot and immunoprecipitation followed by mass spectrometry. The validation process pointed towards one V1aR antibody (1G1), which showed promising results in both western blot and immunoprecipitation experiments, and needs to be further tested. The best OTR antibody identified in immunoprecipitation experiments (1B2) did not perform well in western blots. Oksitocinski receptor (OTR) i vazopresinski 1a receptor (V1aR) dva su receptora spregnuta sa G-proteinom uključena u posredovanje raznih fizioloških procesa te društvenog i seksualnog ponašanja. Proizvodnja specifičnih antitijela za proučavanje ovih receptora predstavlja izazov zbog sličnosti njihovog slijeda i strukture, kao i sličnosti dva dodatna blisko povezana vazopresinska receptora. Stoga, cilj je ovog rada bio validirati deset antitijela za OTR i sedam antitijela za V1aR proizvedenih prema karakterističnim peptidnim epitopima receptora, koristeći western blot i imunoprecipitaciju povezanu s masenom spektrometrijom. Jedno antitijelo za V1aR (1G1) u procesu validacije pokazalo je obećavajuće rezultate u eksperimentima western blota i imunoprecipitacije. Najbolje od OTR antitijela identificiranih u eksperimentima imunoprecipitacije (1B2) nije se pokazalo uspješnim u western blotovima.

U ovom su radu istražene mogućnosti primjene metode LC-SPE/krio NMR za praćenje kemijskih reakcija i određivanje strukture sporednih produkata i/ili razgradnih produkata on-line koji nastaju u reakciji, s naglaskom na bioaktivne molekule i lijekove. Za razvoj metodologije odabrana su tri modelna sustava. Za svaki modelni sustav istraženi su uvjeti kromatografskog odjeljivanja komponenata reakcijskih smjesâ, zatim parametri za učinkovitu ekstrakciju odabranih analita na SPE-sorbense te utjecaj parametara ekstrakcije na osjetljivost krio-probe NMR. Za identifikaciju i strukturnu karakterizaciju izoliranih spojeva korištene su spektroskopija NMR i tehnika LC-MS. Prvi modelni sustav bili su karbohidrazidi i njihovi derivati koji imaju kelirajuća svojstva te antioksidativno i antiepileptičko djelovanje. Tijekom analize karbohidrazida istražena je osjetljivost sustava LC-SPE/krio NMR. Drugi modelni sustav bio je etodolak koji pripada skupini nesteroidnih protuupalnih lijekova. Analizom 7-etiltriptofola, polazne tvari u sintezi etodolaka, identificirano je i strukturno okarakterizirano 17 onečišćenja. Treći modelni sustav bili su makrozoni, novi amidni konjugati azitromicina i derivata tiosemikarbazida, koji imaju zadovoljavajuće antibakterijsko djelovanje na odabrane susceptibilne i rezistentne bakterijske sojeve. Analizirano je ukupno 6 reakcijskih smjesâ iz triju različitih klasa novosintetiziranih makrozona. Identificirana onečišćenja bila su sporedni produkti i razgradni produkti makrolidne i tiosemikarbazonske strukture, te dijastereomeri makrozona. Konfiguracije dijastereomerâ predložene su na temelju analize spektara NOESY i računskih simulacija molekulske dinamike. Provedeno istraživanje rezultiralo je razvojem pouzdane, brže i učinkovitije metode za analizu složenih smjesa spojeva koristeći pritom znatno manje količine uzorka i organskih otapala u usporedbi s postojećim metodama off-line. Metoda LC-SPE/krio NMR može imati širu primjenu u ekstrakciji, pročišćavanju i karakterizaciji novosintetiziranih spojeva i/ili onečišćenja iz reakcijskih smjesâ. In this doctoral thesis, the possibilities of applying the LC-SPE/cryo NMR method for monitoring chemical reactions and determining the structure of by-products and/or degradation products on-line, with an emphasis on bioactive molecules and drugs, were explored. Three different model systems were explored for methodology development. For each model system, the parameters for chromatographic separation of the reaction mixtures components, as well as parameters for the efficient extraction of the selected analytes on SPE-sorbents, were optimized. Hence, the influence of the SPE-parameters on the NMR cryoprobe sensitivity was also evaluated. NMR spectroscopy and LC-MS were used for identification and structural characterization of the extracted mixture components. The first model system were carbohydrazides and their derivatives, which are known as chelating reagents and also show antioxidative and antiepileptic effects. Sensitivity of LC-SPE/cryo NMR system was determined by analyzing carbohydrazides. Etodolac, a nonsteroidal anti-inflammatory drug, was chosen as the second model system. During the analysis of 7-ethyltryptophol, a starting material in the synthesis of etodolac, 17 impurities were identified and structurally characterized. The third model system were macrozones, new amide conjugates of azitromycin and thiosemicarbazide derivatives which exhibit good antibacterial activity against susceptible and resistant bacterial strains. Six reaction mixtures from three different classes of newly synthesized macrozones were analysed. Identified impurities were classified as macrolide and non-macrolide related by-products and degradation products, as well as macrozone diastereomers. Configurations of diastereomers were proposed on the basis of NOESY NMR spectroscopy data and molecular dynamic simulations. This research resulted in development of a reliable, faster and more efficient methodology for complex mixture analysis using significantly smaller amount of sample and organic solvents in comparison with standard off-line methods. LC-SPE/cryo NMR method can be applied for the extraction, purification and characterization of newly synthesized compounds and/or impurities from reaction mixtures.

Ciklički peptidi predstavljaju posebno zanimljivu klasu supramolekulskih anionskih receptora zbog svoje raznolikosti i vrlo dobrih kompleksacijskih svojstava. Prvi korak sinteze cikličkih peptida je priprava njihovih linearnih analoga, a zatim se provodi reakcija ciklizacije. Najvažnija komponenta uspješne ciklizacije je konformacijska preorganizacija linearnog peptida u kojoj dolazi do približavanja N i C reaktivnih krajeva. Preorganizacija se može pospješiti korištenjem templatnih iona. U ovom radu istraženo je kompleksiranje metilnih estera linearnog tetra i heksafenilalanina s halogenidnim i oksoanionima u acetonitrilu. Konstante stabilnosti nastalih kompleksa određene su metodama 1H NMR spektroskopije, spektrofluorimetrije i spektroskopije cirkularnog dikroizma. Strukturne karakteristike anionskih kompleksa oba receptora istražene su simulacijama molekulske dinamike. Rezultati upućuju na to da su anioni koordinirani pomoću amidnih i amino protona peptida. Većina nastalih kompleksa je stehiometrije 1:1, osim u slučaju dihidrogenfosfata kod kojeg je uočena i stehiometrija 1:2 (receptor:anion) Cyclic peptides represent a particularly interesting class of supramolecular anion receptors due to their diversity and good complexation properties. The first step in the synthesis of cyclic peptides is the preparation of their linear analogues after which the cyclization is carried out. The most important factor for a successful cyclization is the conformational reorganization of the linear peptide, in which the N and C reactive ends approach each other. Reorganization can be enhanced using templating ions. In this work, the complexation of methyl esters of linear tetra and hexaphenylalanine with halide and oxoanions in acetonitrile was investigated. The stability constants of corresponding complexes were determined by 1H NMR spectroscopy, spectrofluorimetry and circular dichroism spectroscopy. The structural characteristics of the anionic complexes of both receptors were investigated by molecular dynamics simulations as well. The results indicate that the anions are coordinated by the amide and amino protons of the peptide. Most of the resulting complexes are of 1:1 stoichiometry, except in the case of dihydrogen phosphate, where a 1:2 stoichiometry (receptor:anion) was also observed.

The development of new approaches in the treatment of Parkinson's disease presents a challenge due to the presence of the blood-brain barrier, which hinders effective drug delivery to the brain. Due to their unique physicochemical properties, nanoparticles have the potential to improve conventional therapeutic approaches and enable targeted drug delivery across the blood-brain barrier. The main goal of this doctoral thesis was to synthesize and characterize differently functionalized gold and selenium nanoparticles and to examine their interaction with catechols (levodopa and dopamine) in order to design more effective and safer therapeutic systems for Parkinson's disease. The process of drug binding to the nano-surface was investigated using various spectroscopic, laser and microscopic methods, while the bloodbrain barrier permeability of prepared nano-systems was tested under in vitro settings. The obtained results have provided a better understanding of the processes that occur during the binding of catechol drugs to gold and selenium nanoparticles. This will potentially enable the development of a nano-formulation with targeted drug delivery to the brain with the aim of improving the therapy of Parkinson's disease. Razvoj novih pristupa u liječenju Parkinsonove bolesti predstavlja izazov zbog prisutnosti krvno-moždane barijere koja otežava učinkovitu dostavu lijekova u mozak. Zahvaljujući jedinstvenim fizikalno-kemijskim svojstvima, nanočestice imaju potencijal poboljšati konvencionalne terapijske pristupe i omogućiti ciljanu isporuku lijekova preko krvnomoždane barijere. Glavni cilj ovog doktorskog rada bio je sintetizirati i karakterizirati različito funkcionalizirane nanočestice zlata i selena te ispitati njihovo međudjelovanje s kateholima (levodopa i dopamin) u svrhu dizajniranja učinkovitijih i sigurnijih terapijskih sustava za liječenje Parkinsonove bolesti. Proces vezanja lijekova na nano-površinu ispitivao se pomoću različitih spektroskopskih, laserskih i mikroskopskih metoda, a zatim su se provela istraživanja propusnosti krvno-moždane barijere za novopripravljene nanosustave u in vitro uvjetima. Dobiveni rezultati omogućili su bolje razumijevanje procesa koji se događaju prilikom vezanja lijekova kateholne strukture i novih nanočestica zlata i selena. Na taj način potiče se razvoj nano-formulacija s ciljanom isporukom lijekova u mozak s ciljem poboljšanja terapije za Parkinsonovu bolest.

U okviru ovog rada pripravljene su i u čvrstom stanju istražene tetraklorokupratne(II), tetrabromokupratne(II) i tetrakloromanganatne(II) soli izvedene iz monoamina, diamina i triamina različitih geometrijskih karakteristika i kemijskih funkcionalnosti. Poseban naglasak pri istraživnju posvećen je strukturnim karakteristikama priređenih spojeva, njihovim termičkim svojstvima, te odabranim magnetskim i električnim svojstvima. Iz difrakcijske strukturne analize priređenih spojeva, te simetrijske i statističke analize literaturno opisanih primjera, postavljena su geometrijska ograničenja i očekivanja za ovaj tip spojeva, koja su omogućila predlaganje smislenih strukturnih modela iz podataka difrakcije na prahu. Predložena je prava simetrija feroelektrične faze etilamonijevog tetraklorokuprata(II) iz podataka neutronske difrakcije, određene su strukture triju aperiodičnih kristala. Izučene su promjene u strukturi metilamonijevih halogenokuprata(II) uslijed supstitucijskog miješanja na položaju halogenidnog iona. Analizirana su magnetska svojstva slojevitih hibrida u ovisnosti o odabranim gradivnim ionima i dimenzionalnosti anorganske okosnice, te su kvantificirane modulacije u magnetskim svojstvima u supstitucijski miješanim kristalima. In this work, tetrachlorocuprate(II), tetrabromocuprate(II) and tetrachloromanganate(II) salts derived from monoamines, diamines and triamines with different geometric characteristics and chemical functionalities were prepared and investigated in the solid state. An emphasis was given to the structural characteristics, thermal properties, and selected magnetic and electrical properties of prepared compounds. Geometric limitations and expectations were derived for this type of materials from the diffraction structural analysis of prepared compounds, and the symmetry and statistical analysis of those described in the literature, which enabled the proposal of meaningful structural models from powder diffraction data. The true symmetry of the ferroelectric phase of ethylammonium tetrachlorocuprate was proposed from neutron diffraction data. Structures of three aperiodic crystals were determined. Changes in the structure of methylammonium tetrahalocuprates(II) due to substitutional mixing of halide ions were quantified. The magnetic properties of the layered hybrids were analyzed in relation to the selection of building ions and the dimensionality of the inorganic framework. Modulations of the magnetic properties in substitutionally mixed crystals were quantified.

This thesis investigates the technical feasibility of bovine muscle progenitor stem cells as a source for in vitro meat cultivation. The cells were isolated using enzymatic digestion. Bovine muscle progenitor stem cells were identified using the paired box protein, Pax7 using immunocytochemical assays and fluorescence microscopy. The cells were treated with SB203580, a common inhibitor for the p38 mitogen activated protein kinase pathway to examine its influence on the cell’s proliferation and differentiation capabilities. However, it didn’t yield the expected outcome. Collagen type 1, gelatine and laminin were used as coatings and compared to see their influence on the selectivity of the target cell type, of which laminin was the most promising. Lastly, cells were sorted via fluorescence activated cell sorting using CD29, CD56 as positive, and CD31 and CD45 as negative cell surface markers, which proved to be an auspicious method for the purification of the cell culture. Ova disertacija istražuje tehničku izvedivost korištenja matičnih stanica prekursora mišićnih stanica iz goveda za uzgoj mesa in vitro. Stanice su izolirane primjenom enzimatske digestije. Matične stanice prekursori mišićnih stanica iz goveda identificirane su pomoću transkripcijskog faktora Pax7, uporabom imunocitokemijskih testova i fluorescentne mikroskopije. Stanice su tretirane SB203580, inhibitorom metaboličkog puta p38 mitogen-aktivirane proteinske kinaze, kako bi se ispitao njezin utjecaj na sposobnosti proliferacije i diferencijacije stanica. Međutim, korištenje istog nije dalo očekivani rezultat. Stanično posuđe tretirano je raznovrsnim premazima: kolagen tip 1, želatina i laminin, kako bi se promatrao njihov utjecaj na proliferaciju i selektivnost prema stanicama od interesa. Laminin se pokazao kao najobećavajući. Konačno, stanice su sortirane putem fluorescentno aktiviranog sortiranja stanica primjenom CD29, CD56 kao pozitivnih i CD31 i CD45 kao negativnih površinskih staničnih markera, što se pokazalo izvrsnom metodom za pročišćavanje kulture stanica.

U sklopu diplomskog rada ispitani su mehanokemijski postupci sinteze koordinacijskih spojeva bakra(II) i aminokiselina s alifatskim bočnim ograncima. Koristeći LAG metodu, odnosno otapalom potpomognuto mljevenje, uspješno su pripravljeni koordinacijski spojevi Cu(L-Ala)2, Cu(L-Leu)2, Cu(L-Ile)2, Cu(L-Val)2, Cu(L-Pro)2 i ternarni spojevi Cu(L-Ala)(L-Ile) i Cu(L-Ala)( L-Val). Prilikom sinteza ternarnih spojeva, u 8 od 10 sinteza su dobiveni koordinacijski spojevi bakra s istovrsnim aminokiselinama, što ukazuje na selektivnost Cu(II) iona prema aminokiselinama, koja prati sljedeći trend: L-Leu ≈ L-Pro > L-Ile ≈ L-Val > L-Ala. Produkti su karakterizirani koristeći spektroskopiju IR (tehnika ATR) i rendgensku difrakciju u polikristalnom uzorku. Produkti trans-Cu(L-Val)2 i cis-Cu(L-Ile)2(H2O) su karakterizirani i rendgenskom difrakcijom na monokristalnom uzorku te su određene molekulske i kristalne strukture. U slučaju trans-bis(L-valinato)bakra(II), metalni centar ima oktaedarsku koordinaciju uz deformaciju geometrije što je posljedica Jahn-Tellerovog efekta. Kod cisbis(L-izoleucinato)bakra(II) je metalni centar pentakoordiniran te je geometrija kvadratno piramidalna. This diploma thesis presents an investigation of mechanochemical synthesis of coordination compounds of copper(II) and amino acids containing aliphatic side chains. Using the LAG method (liquid assisted grinding), the following compounds were successfully obtained: Cu(L-Ala)2, Cu(L-Leu)2, Cu(L-Ile)2, Cu(L-Val)2, Cu(L-Pro)2, Cu(L-Ala)( L-Ile) and Cu(L-Ala)(L-Val). When trying the synthesis of ternary compounds, coordination compounds of copper with identical amino acids were obtained in 8 out of 10 syntheses, which indicates the selectivity of Cu(II) ions towards amino acids, which follows the trend: L-Leu ≈ L-Pro > L-Ile ≈ L-Val > L-Ala. Obtained complexes were identified using IR spectroscopy (ATR technique) and powder X-ray diffraction. Molecular and crystal structures of trans-Cu(L-Val)2 and cis-Cu(L-Ile)2(H2O) were deermined using single crystal X-ray diffraction. Jahn-Teller effect was noticed as the geometry of copper ion in trans-Cu(L-Val)2 is distorted octahedral, whereas in the case of cis-Cu(L-Ile)2(H2O) the copper ion is pentacoordinated and the geometry is square pyramidal.

ADP-ribozilacija je kovalentna modifikacija koja kontrolira brojne važne stanične procese od bakterija do čovjeka. ADP-ribozilacijski sustav čine ADP-ribozil-transferaze koje prenose ADPribozu na ciljnu (makro)molekulu i ADP-ribozil-hidrolaze koje osiguravaju reverzibilnost procesa. Glavni predstavnici ADP-ribozil-hidrolaza su makrodomenski proteini zastupljeni u sve tri domene života uključujući i viruse. U ovom doktorskom radu istraživana su dva makrodomenska proteina, SCO6735 iz bakterije Streptomyces coelicolor i Mac1 iz virusa SARS-CoV-2. Pokazano je da SCO6735 uklanja ADP-ribozilaciju s jednolančane DNA modificirane na timinu. Na temelju računalnih i eksperimentalnih rezultata predložen je novi katalitički mehanizam hidrolize ADPriboze s DNA karakterističan za skupinu proteina sličnih SCO6735. Makrodomena Mac1 inhibira imunološki odgovor domaćina čime potiče virusnu replikaciju. Stoga ima terapeutski potencijal za liječenje bolesti uzrokovane virusom SARS-CoV-2. U tu svrhu provedena je virtualna pretraga baze malih molekula koja je rezultirala testiranjem 27 spojeva metodom izotermalne titracijske kalorimetrije. Izmjereni afiniteti vezanja potvrdili su dobro vezanje nekoliko spojeva, uključujući FDA-odobrene lijekove Dasatinib i Metotreksat. ADP-ribosylation is a covalent modification that controls essential cellular processes, conserved from bacteria to humans. The ADP-ribosylation system consists of ADP-ribosyl transferases that catalyse the transfer of ADP-ribose to a target (macro)molecule, while ADP-ribosyl hydrolases ensure reversibility. Macrodomain proteins are ADP-ribosyl hydrolases present in all three domains of life and viruses. In this dissertation, two macrodomain proteins were investigated: SCO6735 from Streptomyces coelicolor and Mac1 encoded by the virus SARS-CoV-2. SCO6735 was experimentally shown to act antagonistically to a bacterial toxin known to impair replication by ADP -ribosylation of thymidines in single-stranded DNA. Furthermore, a novel catalytic mechanism of DNA-(ADP-ribose) hydrolysis characteristic of the subgroup of SCO6735-related proteins was proposed. The Mac1 macrodomain is a good therapeutic target, because it inhibits the host immune response, thus allowing virus replication. In search of potential inhibitors, a virtual screening of a compound library was performed. The screening resulted in testing of 27 molecules with the isothermal titration calorimetry. Determined binding affinities suggested good binding of several molecules, including the FDA-approved drugs Dasatinib and Methotrexate.

Makrolidni antibiotici pripadaju skupni makrolida koji se široko primjenjuju za liječenje infekcija gornjih i donjih dišnih puteva. Mehanizam djelovanja makrolida temelji se na vezanju za 23S rRNA u peptidil-transferaznom centru bakterijskog ribosoma ili u njegovoj blizini, što sprječava izlaz novosintetiziranih peptida i inhibira sintezu proteina. Pojava višestruke bakterijske rezistencije predstavlja ozbiljnu prijetnju za ljudsko zdravlje, što je potaknulo sintezu novih derivata makrolida koji će pokazivati bolju bioaktivnost. Vezanjem poznatih makrolida i bioaktivnih tiosemikarbazona dobiveni su novi konjugati, makrozoni, koji uspješno djeluju protiv nekih rezistentnih bakterijskih sojeva. U ovom radu okarakterizirane su interakcije nekoliko 4''-aril-makrozona s biološkim metama, poput albumina iz goveđeg seruma te ribosoma iz bakterije Escherichia coli. Istraživanja su provedena kombinacijom metoda spektrofluorimetrije, spektrofotometrije i cirkularnog dikroizma. Kao kompetitor u fluorimetrijskim mjerenjima korišten je flourescirajući derivat azitromicina, 9a-NBD-azitromicin. Podaci su zatim obrađeni metodama multivarijatne analize kako bi se odredile konstante vezanja. Due to their anti-infective activity, macrolide antibiotics (macrolides) are widely used for the treatment of upper and lower respiratory tract infections. Their mechanism of action is based on reversible binding to 23S rRNA at or near the peptidyl-transferase center of the bacterial ribosome, which prevents the release of newly synthesized peptides and inhibits the protein synthesis. The emergence of bacterial resistance represents a serious threat to human health, which prompted the synthesis of new macrolide derivatives with a better bioactivity. Linking known macrolides to bioactive thiosemicarbazones resulted with new conjugates, macrozones, active against some resistant bacterial strains. In this work, the interactions of several 4''-aryl-macrozones with biological targets, such as bovine serum albumin and Escherichia coli ribosomes, were characterized. The studies were carried out by combining spectrofluorimetric, spectrophotometric and circular dichroism methods. Competition fluorimetric experiments were conducted using a fluorescent derivative of azithromycin, 9a-NBD-azithromycin. The data were further processed with multivariate analysis methods to determine the binding constants.

Na zeolitima strukturnih tipova FAU i MFI (ZSM-5) napravljene su modifikacije tijekom ili nakon sinteze kao bi se poboljšala njihova katalitička aktivnost. Pripremljeni uzorci su modificirani dodatkom mezoporoznog templata N,N,N-trimetilheksadekan-1-amonijeva bromida (CTAB) ili jetkani vodenom otopinom natrijeva hidroksida, kako bi se povećao broj mezopora u kristalima i olakšala difuzija reaktanata do aktivnog mjesta. Jakost aktivnog mjesta ugađana je kationskom zamjenom s ionima Mg2+ , Co2+, Ni2+ i Zn2+ (u vodenoj otopini i mehanokemijski). Utjecaj pojedinog metalnog kationa na broj i jakost Lewisovih i Brønstedovih kiselih mjesta u materijalu praćen je infracrvenom spektroskopijom u vakuumu pomoću adsorpcije probne molekule (deuterirani acetonitril). Broj pojedinih kiselih mjesta dobiven je integriranjem pojedinih krivulja nakon dekonvolucije spektra vezanog D3-acetonitrila, anjihova jakost određena je iz količine nakon zagrijavanja na 150, 200 i 300 °C. Na odabranim uzorcima napravljeni su katalitički testovi konverzije metanola u olefine. Modifications during and after sythesis were made on FAU and MFI (ZSM-5) zeolite type samples in order to enhance their catalytic activity. Prepared samples were modified with mesoporosity template N,N,N-Trimethylhexadecan-1-aminium bromide (CTAB) or etched with water solution of sodium hydroxide to increase the volume of mesopores in crystals and faciliate diffusion of reactnats to the active site. The strength of active sites was modified by cationic exchange with Mg2+ , Co2+ , Ni2+ and Zn2+ ions (in water solution and mechanochemicaly). Effect of metal cation on number and strength of Lewis and Brønsted acid sites was studied by infrared spectroscopy in vacuum with the adsorption of probe molecule (deuterated acetonitrile). The number of various types of acid sites was calculated from the integrated curves after deconvolution of spectra of adsorbed D3-acetonitrile, while their strength is determined after heating at 150, 200 and 300 °C. Catalytic tests of methanol to olefine conversion (MTO) were preformed on selected samples.

Kolagen je najprisutniji protein u čovjekovu tijelu. Ima strukturu trostruke zavojnice i određeni sadržaj aminokiselina. Stanicama daje čvrstoću i potporu zbog stabiliziranosti unutarmolekulskim interakcijama. Sintetiziraju ga fibroblasti. Fibroblasti sintetiziraju i druge komponente izvanstaničnog matriksa. Tijekom godina, zbog nakupljanja oštećenih i pogrešno strukturiranih proteina, koža gubi svoja svojstva te se različitim metodama pokušava taj proces usporiti. Najznačajnija je primjena peptida kolagena koji nastaju djelovanjem enzima na denaturirani kolagen koji zbog svojih svojstava potiču stvaranje kolagenih vlakana, potiču sintezu hijaluronske kiseline i diobu stanica fibroblasta., Collagen is the most abundant protein in the human body. It has a structure of triple helix and a specific content of amino acids. It gives strength and support to cells as it is stabilized by intramolecular interactions. It is synthesized by fibroblasts. Fibroblasts also synthesize other components of the extracellular matrix. Over the years, the skin loses its properties due to the accumulation of damaged and misstructured proteins, and various methods are used to slow down this process. The most important method is the application of collagen peptides, which are produced by the action of enzymes on denatured collagen and due to their properties stimulate the formation of collagen fibers, the synthesis of hylauronic acid and the division of fibroblast cells.

Uporaba tekućih goriva je svakodnevnica suvremenog čovjeka. Dizel, benzin, kerozin, tekući naftni plin su među prvima na koje se misli kada se spominje pojam gorivo. Zbog svih svojstava pogodan su izbor za motore s unutrašnjim sagorijevanjem pri čemu se, njihovim izgaranjem, toplinska energija pretvara u mehaničku. Ugljikovodični lanci, ponajprije alkani, čine sastav tekućih goriva. Dodatkom aditiva uklanjaju se nepoželjna svojstva goriva i poboljšavaju karakteristike motora. Pogodna metoda za analizu tekućih goriva je plinska kromatografija koja zbog velikog broja teorijskih tavana dobro razdvaja komponente bliskih svojstava. Reprezentativni uzorci dizela i benzina su korišteni za analizu sastava goriva plinskom kromatografijom., The use of liquid fuels is part of people's everyday lives today. Diesel, gasoline, kerosene, liquid gas are among the first that come to mind when the term fuel is mentioned. Due to their composition, they are suitable for internal combustion engines, in which the ignition of the fuel converts thermal energy into mechanical energy. The composition of liquid fuels consists of hydrocarbon chains, mainly alkanes. Additives help to remove undesirable compounds from the fuels and improve the properties of the engine. A suitable method for the analysis of liquid fuels is gas chromatography, which separates components with similar properties well due to the large number of theoretical plates. Representative samples of diesel and gasoline fuels were used for the gas chromatographic analysis.

Proantocijanidini iz sjemenki grožđa su polimerni fenolni spojevi koji imaju blagotvoran učinak na čovjekovo zdravlje. Najpoznatiji su po svom antioksidativnom svojstvu, a štite od bolesti tako što smanjuju količine slobodnih radikala u organizmu koji oštećuju stanice. Njihova svojstva su u mnogome određena su stupnjem polimerizacije odnosno brojem monomernih jedinica od kojih su sastavljeni. Ispitana je metoda određivanja stupnja polimerizacije proantocijanidina iz sjemenki grožđa floroglucinolom umjesto benzil merkaptana, koji je izrazito neugodnog mirisa i nezgodan za korištenje. Postupak s floroglucinolom ne zahtjeva rad u digestoru niti posebnu opremu. Uzorci su bile sjemenke autohtonih hrvatskih sorti grožđa Plavca malog, Crljenka i Babića. Uzorci su analizirani korištenjem tekućinske kromatografije visoke djelotvornosti te je iz tih podataka izračunat stupanj polimerizacije proantocijanidina. Uzorci su analizirani korištenjem tekućinske kromatografije visoke djelotvornosti., Proanthocyanidins from grape seeds are polymeric phenolic compounds that have a beneficial effect on human health. They are best known for their antioxidant properties, protecting against disease by reducing the amount of free radicals in the body that damage cells. Their properties are largely determined by the degree of polymerization, that is, by the number of monomer units of which they are composed. The method for determinination of the polymerisation degree of proantocyanidins from grape seeds with phloroglucinol instead of benzyl mercaptan, which has an extremely unpleasant smell and is inconvenient to use, was examined. The procedure with phloroglucinol does not require a digester or special equipment. The samples were seeds of indigenous Croatian grape varieties Plavac mali, Crljenak and Babić. The samples were analyzed using high performance liquid chromatography and the degree of polymerization of proanthocyanidin was calculated from these data.

Rod Veronica pripada porodici Plantaginaceae te ima veliki broj vrsta rasprostranjenih na brojnim staništima, što ukazuje na njihovu veliku adaptivnost. Nepovoljni životni uvjeti utječu na rast, razvoj i reprodukciju biljaka. Kako bi se zaštitile, biljke su razvile mehanizme izlučivanja specijaliziranih metabolita. Tim metabolitima pripadaju eterična ulja, mješavine slobodnih hlapljivih spojeva dobivenih sekundarnim metaboličkim putevima. Kao nusprodukti izolacije eteričnih ulja nastaju hidrolati, nekoć dosta zanemarivani i smatrani otpadnim produktom, danas predstavljaju temu od velikog interesa. Predmet istraživanja ovog rada su hidrolati vrste roda Veronica umjerenog staništa. Tehnike korištene za izolaciju su mikrovalna ekstrakcija i hidrodestilacija po Clevenger-u., The genus Veronica belongs to the Plantaginaceae family and it has a large number of species distributed in numerous habitats, which indicates their great adaptability. Unfavorable living conditions affect the growth, development and reproduction of plants. In order to protect themselves they have developed the mechanisms for secretion of specialized metabolites. These metabolites include essential oils, mixtures of free volatile compounds obtained by secondary metabolic pathways. Hydrosols are formed as byproducts of the isolation of essential oils. Once largely neglected and considered a waste product, today they represent a topic of great interest. The subject of this research are hydrosols of the species from genus Veronica from moderate habitats. Techniques used for isolation are microwave extraction and Clevenger hydrodistillation.

U sklopu ovog diplomskog rada razvijen je sintetski put priprave amidnih, disupstituiranih ferocenskih derivata s desmuramil-peptidnom (DMP) i manoznom podjedinicom. Derivati se međusobno razlikuju u broju metilenskih skupina (n = 0-4) na poveznici između ferocenskog i šećernog dijela molekule. U ovom je radu pripravljena ciljna molekula 11, s poveznicom od četiri metilenske skupine i to vezanjem sljedećih strukturnih motiva: O-benzil zaštićene manoze s glikolnom poveznicom, ferocenkarboksilne kiseline koja je esterificirana, a potom i modificirana anhidridom glutarne kiseline u disusptituiranu ferocensku kiselinu 6, te benzilom zaštićenog derivata DMP-a 7. Amidne veze su ostvarene pomoću pouzdane i uhodane EDC/HOBt metode koja se često koristi u tu svrhu u peptidnoj kemiji. Sintetski put priprave ciljne molekule završava korakom uklanjanja benzilne zaštite hidrogenolizom. Provedena je detaljna strukturna karakterizacija svih sintetiziranih prekursora i konačnog spoja ( 1H, DEPTQ NMR i IR spektroskopija, spektrometrija masa ESI-MS). Ovako optimizirana metoda poslužit će za sintezu preostalih spojeva ove serije u budućim istraživanjima čiji je konačni cilj ispitivanje njihove adjuvantske aktivnosti. In this Thesis the synthetic route for the preparation of amide, disubstituted ferrocene derivatives with desmuramyl peptide (DMP) and mannose subunit was optimized. The structures of final compounds vary in the number of methylene groups (n = 0-4) between the ferrocene and the sugar part of the molecule. Target molecule 11, which has four methylene groups, was prepared in this work by combining following structural motifs: O-benzyl protected mannose with glycolic linker, ferrocene carboxylic acid which was converted to ester first and subsequently modified with glutaric anhydride to disubstituted ferrocene acid 6, and benzyl protected DMP derivative 7. Amide bonds are formed by reliable and wellestablished EDC/HOBt method which is often used in peptide synthesis for this purpose. The synthetic route to target molecule ends with benzyl protection removal by hydrogenolysis. Structural characterization of all precursors as well as target compound was performed by 1H, DEPTQ NMR and IR spectroscopies as well as mass spectrometry (ESI-MS). Optimized synthetic route presented here will serve for the preparation of remaining compounds in this series. The final aim is the evaluation of their adjuvant activity.

Kinetika otapanja (100), (110) i (111) plohe fluorita u vodi praćena je potenciometrijski mjerenjem koncentracije Ca2+, F– i H+ iona u otopini ion-selektivnim elektrodama tijekom 84 dana otapanja. Tenziometrijom i mikroskopom atomskih sila praćen je utjecaj otapanja na promjenu močivosti površine i topografiju (100), (110) i (111) ploha fluorita mjerenjem kontaktnog kuta i faktora hrapavosti prije i nakon otapanja. Površinski potencijal (100), (110) i (111) plohe monokristala fluorita tijekom 24 sata otapanja mjeren je fluoritnim monokristalnim elektrodama, a elektrokinetički potencijal nanočestica fluorita tijekom 37 dana otapanja određen je mjerenjem elektroforetske pokretljivosti nanočestica fluorita. Istražen je i utjecaj H+ , OH– , F– i Ca2+ iona na elektrokinetički potencijal nanočestica fluorita. Metodama statičkog i dinamičkog raspršenja svjetlosti određen je hidrodinamički promjer i promjer giracije nanočestica fluorita. Određeni su koeficijenti brzine difuzije iona u otopinu za (100), (110) i (111) plohu te iznose: 0,005 s–1 ; 0,045 s–1 i 0,03 s–1 . Ion selective electrodes were used to study dissolution kinetics of (100), (110) and (111) fluorite crystallographic planes by measuring Ca2+, F– and H+ ion concentration in the solution. Tensiometry and atomic force microscopy were employed for measuring dissolution impact on contact angle and surface roughness of specified fluorite crystallographic planes. Surface potential change of mentioned fluorite crystallographic planes during 24 hours of dissolution was investigated using a single crystal electrode. The electrokinetic potential of fluorite nanoparticles during dissolution was determined by measuring electrophoretic mobility. The influence of H+ , OH– , F– i Ca2+ ions on the electrokinetic potential of fluorite nanoparticles was also investigated. The hydrodynamic diameter and diameter of gyration of fluorite nanoparticles were determined using static and dynamic light scattering methods. The ion diffusion rate coefficients in the solution for (100), (110) and (111) planes were obtained: 0.005 s–1 , 0.045 s–1 and 0.03 s–1 respectively.

U ovom radu istražene su mogućnosti korištenja Calvetovog kalorimetra za izravno izotermno kalorimetrijsko određivanje reakcijskih entalpija procesa u čvrstom stanju. Reakcije karboksilnih kiselina i 4-N,N-dimetilaminopiridina provedene su uz značajan suvišak kiselina te su potpomognute dodatkom male količine metanola pri čemu dolazi do nastajanja kokristala soli pridinijevog karboksilata s karboksilnom kiselinom. Određene su odgovarajuće reakcijske entalpije, a izmjerene su i reakcije stvaranja kokristala iz prethodno dobivenih soli. Dobivene vrijednosti reakcijskih entalpija i reaktivnost istraživanih sustava diskutirani su s obzirom na strukturu korištenih kiselina, njihove kiselosti i mogućnosti ostvarivanja vodikovih veza. Neposrednom izotermnom kalorimetrijom određene su i reakcijske entalpije taljenja ekvimolarne smjese benzofenona i difenilamina pri 25 °C te nastajanja kokristala iz metastabilne tekućine uslijed dodatka klice kokristala. Produkti svih istraženih reakcija identificirani su na temelju difraktograma praškastih uzoraka. Reakcijske entalpije izmjerene neposredno izotermnom kalorimetrijom uspoređene su s onima određenim na temelju termodinamičkih ciklusa. U tu svrhu kalorimetrijski su određene entalpije otapanja krutih reaktanata i produkata te entalpije odgovarajućih reakcija u otopini. Rezultati dobiveni pomoću obje metode vrlo dobro se slažu čime je potvrđena pouzdanost razvijene metode izravnog izotermnog kalorimetrijskog određivanja entalpija reakcija u čvrstom stanju. The potential of using a Calvet calorimeter for direct determination of reaction enthalpies for processes between solid reactants at isothermal conditions was investigated in this work. The reactions of carboxylic acids with 4-N,N-dimethylaminopyridine were carried out using a significant excess of the acid and assisted by addition of methanol. Co-crystals of protonated carboxylic acids with the corresponding salts were the favoured products at the conditions used. The reaction enthalpies related to formation of the co-crystals from either free base or previously prepared salts were determined. The obtained results were discussed with respect to the molecular structures and pKa values of the studied acids and their potential to form hydrogen bonds. The enthalpy changes related to the melting od benzophenon and diphenylamine mixed in equimolar ratio and subsequent co-crystal formation from the metastable melt by addition of a crystalline seed were measured. The reaction products were identified by powder X-ray diffraction which also enabled the determination of reaction extent. The results of direct calorimetric measurements were validated by correlation with those based on Hess's law. This yielded an excellent data agreement, confirming the reliability of the herein developed methodology.

Morski plodovi čine važnu sastavnicu uravnotežene ljudske prehrane, ali njihovom konzumacijom može doći do intoksikacije ukoliko su kontaminirani fitoplanktonskim toksinima. Proizvode ih fitoplanktonske alge, a nakupljaju se u morskim organizmima koji se hrane fitoplanktonom. Fitoplanktonski toksini su skupina kompleksnih organskih spojeva, a podijeljeni su u šest skupina s obzirom na simptome trovanja koje izazivaju. Pored negativnih utjecaja na ljudsko zdravlje kao bioaktivne molekule imaju i potencijala u razvoju novih lijekova. Zabrinjavajuća činjenica je što s porastom temperatura mora i oceana dolazi do povećane produkcije ovih toksina koji se pojavljuju i na mjestima gdje ih do sad nije bilo. Analiza uzorka školjkaša provedena je s ciljem detekcije i identifikacije poznatih i novih vrsta toksina na području Jadranskog mora. Kao metoda se koristila tekućinska kromatografija uparena s masenom spektrometrijom visoke rezolucije., Seafood is an important component of a balanced diet, but their consumption can lead to intoxication if they are contaminated with phytoplankton toxins. They are produced by phytoplankton algae and accumulate in marine organisms that feed on phytoplankton. Phytoplankton toxins are a group of complex organic compounds divided into six groups according to the symptoms they cause. In addition to negative effects on human health, as bioactive molecules they also have potential in the development of new drugs. The worrying fact is that the rise in sea and ocean temperatures leads to an increased production of these toxins, which appear in places where they were not detected before. The analysis of the shellfish sample was carried out with the aim of detecting and identifying known and new types of toxins in the Adriatic Sea. The method used was liquid chromatography coupled with high-resolution mass spectrometry.

Mjerena su površinska svojstva i otapanje čestica amorfne silike veličine 200 nm. Metodom potenciometrijske masene titracije određen je interval u kojem se nalazi točka nul-naboja. Metodom elektroforeze određena je izoelektrična točka čestica silike u vodenoj otopini pri dvije ionske jakosti u prisutnosti i odsutnosti otopljene silikatne kiseline. Proces otapanja koloidnih čestica silike u vodenim otopinama različite pH vrijednosti praćen je metodama dinamičkog i statičkog raspršenja svjetlosti te spektrometrijom masa uz induktivno spregnutu plazmu. Silika se otapa otpuštanjem SiO(OH)+ i OH– iona s površine pri čemu nastaju nabijene praznine koje mijenjaju površinski naboj. Površinski naboj određuje brzinu otapanja silike. Nastanak pozitivno nabijenih praznina ubrzava otpuštanje SiO(OH)+ , a nastajanje negativno nabijenih praznina ubrzava otpuštanje OH– iona. Brzina otapanja amorfne silike povećava se od rc  10– 16 mol s–1 cm–2 (pri pH = 3) do rc  10–14 mol s–1 cm–2 (pri pH = 11). Vrijednosti brzine otapanja uspoređene su s empirijskom jednadžbom za otapanje amorfne silike prethodno predloženoj u literaturi. Surface properties and dissolution of 200 nm amorphous silica particles were measured. The interval of point of zero charge of silica particles in aqeous solutions was determined by means of potentiometric mass titration. The isoelectric point was determined electrophoreticaly at two ionic strenghts, and in the presence and the absence of dissolved silicic acid. Dissolution process of silica particles in aqueous solutions with different pH values was monitored by means of dynamic and static light scattering, and inductively coupled plasma - mass spectrometry. Silica dissolution occurs via removal of SiO(OH)+ , and OH– from the silica surface. Surface charge regulates the rate of silica dissolution. Creation of positively charged vacancies accelerates the removal of SiO(OH)+ , while creation of negatively charged vacancies accelerates the removal of OH– ions. The rate of dissolution i.e. the rate of cation removal from silica surface, increases from rc  10–16 mol s–1 cm–2 (at pH = 3) to rc  10–14 mol s–1 cm–2 (at pH = 11). The values of dissolution rate were compared with previously proposed empirical equation for amorphous silica dissolution.

Ovaj diplomski rad sastoji se od istraživačkoga i metodičkog dijela. U istraživačkom dijelu opisana je priprava O-metiloksima kinuklidin-3-ona klasičnom sintezom te primjenom mehanokemije i mikrovalnog zračenja. Spoj je pripravljen reakcijom hidrokloridnih soli kinuklidin-3-ona i metoksiamina bez dodatka baze. Kvaternizacijom kinuklidinskog dušikova atoma pripravljenog Ometiloksima s metil-jodidom, benzil-bromidom te različito supstituiranim benzil-bromidima (nitro skupine i atomi halogena u meta i para položaju) pripravljena je serija od 10 novih kvaternih derivata kinuklidin-3-on-O-metiloksima. Reakcije su praćene spektroskopijom ATR. Pripravljenim spojevima koji do sada nisu opisani u literaturi, određeno je talište te su strukture dobivenih spojeva potvrđene spektrometrijom masa, infracrvenom spektroskopijom, te 1D i 2D spektroskopijom NMR. Cilj metodičkog dijela rada je potaknuti nastavnike kemije na kreativnost u nastavi temeljenoj na učenju otkrivanjem unutar koncepta Enzimi. Navedena tema je izborni sadržaj u novom kurikulumu Kemije četvrtoga razreda gimnazije. Osmišljen je pokus na temelju kojega su oblikovani metodičkodidaktički materijali za jedan 90-minutni nastavni sat. Pri tome su postavljeni odgojno-obrazovni ciljevi temeljeni na prethodnim učeničkim znanjima, vještinama i sposobnostima. This thesis consists of a research and methodological part. The research part describes the preparation of O-methyloxime of quinuclidin-3-one by classical synthesis and the application of mechanochemistry and microwave radiation. The compound was prepared by the reaction of hydrochloride salts of quinuclidin-3-one and methoxyamine without the addition of a base. By quaternization of the quinuclidine nitrogen atom of prepared O-methyloxime with methyl iodide, benzyl bromide and differently substituted benzyl bromides (nitro groups and halogen atoms in the meta and para positions), a series of 10 new quaternary derivatives of quinuclidine-3-one-O-methyloxime was prepared. The reactions were monitored by ATR spectroscopy. The melting points of the prepared compounds, which have not been described in the literature so far, was determined, and the structures of the obtained compounds were confirmed by mass spectrometry, infrared spectroscopy, and 1D and 2D NMR spectroscopy. The goal of the methodological part of thesis is to encourage chemistry teachers to be creative in teaching based on learning through discovery within the concept of Enzymes. The mentioned topic is an optional content in the new chemistry curriculum of the fourth grade of high school. An experiment was designed on the basis of which methodological and didactic materials were designed for one 90-minute lesson. In doing so, educational goals were set based on previous student knowledge, skills and abilities.

Rod Veronica obuhvaća velik broj vrsta koje su zbog svojih osobina prilagođene životu na raznim staništima. Opisanih pet vrsta iz ovog rada žive pretežito na obalama rijeka, potoka i jezera. Izolacijom eteričnih ulja kao nusprodukti nastaju hidrolati, cvjetne vodice obogaćene spojevima iz ulja. Postoje mnoge klasične i nekonvencionalne metode izolacije eteričnih ulja. Za određivanje masene koncentracije hidrolata korištene su metode vodene destilacije u aparaturi po Clevengeru i mikrovalna ekstrakcija., The genus Veronica includes a large number of species that, due to their characteristics, are adapted to life in various habitats. The five species described in this thesis live mainly on the banks of rivers, streams and lakes. Isolation of essential oils produces hydrosols as by-products, floral waters enriched with compounds from oils. There are many classical and unconventional methods of isolating essential oils. The methods of water distillation in a Clevenger apparatus and microwave extraction were used to determine the mass concentrations of the hydrosols.

DNA-ovisna metaloproteaza Spartan (SPRTN) održava genomski integritet sudjelujući u popravku križnih veza DNA i proteina (engl. DNA-protein crosslink, DPC), čestog tipa oštećenja DNA. Citotoksičnost DPC-ova pripisuje se ometanju staničnih procesa koji uključuju DNA (replikacija, transkripcija i popravak) te predstavljaju prepreku u održavanju genomske stabilnosti. Pokazano je da SPRTN sudjeluje u vezanju AAA+ ATP-aze p97, čija je najpoznatija uloga odmotavanje ubikvitinitanih proteina. Rješavanje kristalne strukture kompleksa SPRTN:p97 i njegova strukturna karakterizacija omogućila bi dublje razumijevanje uloge p97 u popravku DPC oštećenja. Stoga su humani SPRTN i p97 te njihovi skraćeni konstrukti, prekomjerno eksprimirani i pročišćeni kromatografskim metodama. Metodom povlačenja proteina okarakterizirana je interakcija navedenog kompleksa. Kako bi se pokušalo odrediti stehiometriju nastalog kompleksa napravljena je elektroforeza na poliakrilamidnom gelu u nativnim uvjetima. Kompleks proteina SPRTN i p97 kristaliziran je metodom difuzije para u svrhu rješavanja njihove kristalne strukture. Inicijalni kristalizacijski uvjeti su detaljno optimizirani kako bi se pripremili monokristali zadovoljavajuće veličine i kvalitete, što bi u konačnici omogučilo rješavanje kristalne strukture kompleksa SPRTN:p97. The DNA-dependent metalloprotease Spartan (SPRTN) functions in maintaining genomic integrity by participating in the repair of DNA-protein crosslinks (DPCs), a common type of DNA damage. The cytotoxicity of DPCs is attributed to the disruption of cellular processes involving DNA (e.g. replication, transcription and repair) and poses a challenge in the maintenance of genomic stability. It was shown that SPRTN participates in the binding of the AAA+ ATPase p97, well known for its function in unwinding of ubiquitinated proteins. Solving the crystal structure of the SPRTN:p97 complex and its functional characterization would allow a deeper understanding of the p97 role in DPC damage repair. Therefore, human SPRTN and p97, as well as several of their deletion mutants were overexpressed and purified by standard chromatography techniques. Subsequently, complex formation between SPRTN and p97 was tested and characterized via pull-down assays. To estimate the stoichiometry of the complex, native polyacrylamide gel electrophoresis was applied. SPRTN and p97 complex was subjected to crystallization by vapor diffusion method. Initial crystallization conditions were further optimized with the aim to obtain larger and higher quality diffracting crystals. Taken together, this set the stage for obtaining the structure of the SPRTN:p97 complex.

Cilj ovog rada je pregled teorije vitamina C te izrada infracrvenog spektra za pet različitih uzoraka koji sadrže vitamin C. Prikazana su strukturna, fizikalna i kemijska svojstva ovog vitamina. Sprječava bolest skorbut pa se naziva askorbinska kiselina. Vitamin je L-enantiomer molekule. Gotovo je planarni peteročlani laktonski prsten s postraničnim lancem. Ima mogućnost reverzibilne oksidacije do dehidroaskorbinske kiseline i stvaranja relativno stabilnih slobodnih radikala te može djelovati kao sakupljač slobodnih radikala u reakcijama koje uključuju reaktivne oksidacijske vrste (ROS). Nasuprot tome, u nekim slučajevima može djelovati i prooksidativno. Albert Szent-Györgyi je 1928. godine izolirao molekulu vitamina C, a stvarnu strukturu je 1933. godine razradio Norman Haworth, nakon čega je Tadeus Reichstein objavio i patentirao komercijalno izvedivu metodu sinteze iz glukoze. Navedene su brojne funkcije i važnost kod biljaka, životinja i ljudi. Sudjeluje u zaštiti drugih antioksidansa, pomaže tijelu da apsorbira željezo, može inaktivirati i neutralizirati histamin te ima ulogu u imunološkom sustavu. Povezuje se sa zaštitom lipida, DNK i proteina od oksidansa te djeluje u mnogim mono- i dioksigenazama za održavanje metala u reduciranom stanju. Neophodan je za ispravno odvijanje fotosinteze te može kontrolirati diobu, produljenje i diferencijaciju stanica, kao i programiranu staničnu smrt. Navedene su količine vitamina C u prehrambenim izvorima te dnevne potrebe kod ljudi za sve uzraste. Kod većih doza ne dolazi do potpune apsorpcije, a velik dio apsorbirane doze se ne metabolizira i izlučuje urinom. Glavni metaboliti askorbinske kiseline u organizmu su dehidroaskorbinska kiselina, 2,3-diketo-gulonska kiselina i oksalna kiselina. Dostupne su brojne analitičke metode za opis vitamina C od kojih je u ovom radu korištena infracrvena spektroskopija s Fourierovom transformacijom. Vitamin C ima mnoge primjene u medicini te prehrambenoj, kemijskoj i kozmetičkoj industriji. The aim of this paper is to review the theory of vitamin C and to create an infrared spectrum for five different samples containing vitamin C. The structural, physical and chemical properties of this vitamin are presented. It prevents the disease scurvy, which is why it got the name ascorbic acid. Vitamin is the L-enantiomer of the molecule. It is an almost planar five-membered lactone ring with a side chain. In addition to the possibility of reversible oxidation to dehydroascorbic acid, it can form relatively stable free radicals and can act as a scavenger of free radicals in reactions involving reactive oxidizing species (ROS). On the other hand, in some cases it can also act as a pro-oxidant. Albert Szent-Györgyi isolated the vitamin C molecule in 1928, and the actual structure was worked out by Norman Haworth in 1933, after which Tadeus Reichstein published and patented a commercially viable method of synthesis from glucose. Numerous functions and importance in plants, animals and humans are listed. It participates in the protection of other antioxidants, helps the body to absorb iron, can inactivate and neutralize histamine and plays a role in the immune system. Further, it is associated with the protection of lipids, DNA and proteins from oxidants and acts in many mono- and dioxygenases to maintain metals in a reduced state. It is necessary for the proper development of photosynthesis and can control the division, elongation and differentiation of cells, as well as programmed cell death. Amounts of vitamin C in food sources and daily requirements for people of all ages are listed. With larger doses, complete absorption does not occur, and a large part of the absorbed dose is not metabolized and is excreted in the urine. The main metabolites of ascorbic acid in the body are dehydroascorbic acid, 2,3-diketo-gulonic acid and oxalic acid. Numerous analytical methods are available for the description of vitamin C, of which infrared spectroscopy with Fourier transformation was used in this work. Vitamin C has many applications in medicine as well as in the food, chemical and cosmetic industries.

U posljednje je vrijeme pojava mikroplastičnih čestica pod sve većim povećalom javnosti. Mikroplastika se nikada neće u potpunosti razgraditi, a negativno utječe na okoliš, ljudsko zdravlje te zdravlje drugih živih bića. Naime probavni sustav organizma ne može probaviti mikroplastiku što predstavlja rizik za ekosustav. Mikroplastika na sebe može vezati kemikalije iz okoliša, kao i bakterije i mikrobe koji potom mogu stvoriti biofilm. Upravo bi se zbog toga problem mikroplastike trebao ozbiljno shvatiti jer dođe li mikroplastika do ljudskog organizma ondje može otpustiti toksične spojeve kao i mikrobe koji su u njoj vezani, oštećujući time i ljudsko zdravlje. Kako bi se mikroplastika uklonila postoji nekoliko rješenja od kojih je u ovom radu predstavljena ekstrakcija mikroplastike ferofluidima. Ferofluidi predstavljaju vrlo snažnu magnetizirajuću tekućinu sastavljenu od tekućeg medija koji sadrži čvrste magnetske čestice vrlo male veličine, koloidno raspršene u tekućini nosača. Uklanjanje mikroplastike ferofluidima vrlo je efikasno jer se čak i najmanje čestice mikroplastike mogu vezati na ferofluid te se potom pomoću magneta mogu odstraniti. U ovom je radu korišten ferofluid na bazi sintetskog ulja te šest otopina mikroplastičnih čestica (polistirena (PS) i polivinil klorida (PVC)) u rasponu veličine od nešto manje od 300 μm do 500 μm. Dokazano je da je tehnika uklanjanja mikroplastike ferofluidom učinkovita i brza. Čestice polistirena imale su općenito bolju učinkovitost uklanjanja od čestica polivil klorida. Također je ustvrđeno da se veće čestice od 355-500 μm bolje ekstrahiraju od manjih čestica manje od 300 μm. Recently, the appearance of microplastic particles has come under increasing public scrutiny. Microplastics cannot be completely decomposed, and they have a negative impact on the environment, human health, and the health of other living beings. Namely, the digestive system of the organism cannot digest microplastics, which represents a health risk for the ecosystem. Microplastics can bind chemicals from the environment, as well as bacteria and microbes that then create a biofilm. It is precisely for this reason that the problem of microplastics should be taken seriously, because if microplastics reach the human body, they can release toxic compounds as well as the microbes bound in it, damaging human health. In order to remove microplastics, there are several solutions, of which the extraction of microplastics with ferrofluids is presented in this paper. Ferrofluids represent a very strong magnetizing liquid composed of a liquid medium, containing solid magnetic particles of very small size, colloidally dispersed in a liquid carrier. The removal of microplastics with ferrofluids is very effective because even the smallest particles of microplastics can bind to the ferrofluid and be removed using a magnet. In this work, ferrofluid based on synthetic oil and six solutions of microplastic particles (polystyrene (PS) and polyvinyl chloride (PVC)) in the size range from slightly less than 300 μm to 500 μm were used. The microplastic removal technique with ferrofluid has been proven to be effective and fast. Polystyrene particles had generally better removal efficiency than polyvinyl chloride particles. It was also found that larger particles of 355-500 μm are easier to extracted than smaller particles of less than 300 μm.

Rak je jedna od najrasprostranjenijih bolesti današnjice i često se javlja uslijed mutacija ili promjena u ekspresiji gena koji kodiraju za proteine uključene u kontrolu rasta i razvoja stanica. RRAS2 je primjer takvog proteina te pripada skupini malih GTPaza. Često je prekomjerno eksprimiran u različitim tipovima raka kod ljudi. Homolog proteina RRAS2 pronađen je i kod evolucijski puno starijih i jednostavnijih životinja – spužvi. Spužve predstavljaju dobar modelni organizam jer imaju jednostavnu morfologiju, a kompleksan genom, te većina gena iz spužvi pokazuje značajnu očuvanost s homolozima kod kralješnjaka. Kako bismo dobili bolji uvid u biokemijske karakteristike homologa RRAS2 iz spužve Eunapius subterraneus, klonirana je spužvina cDNA za RRAS2 te unesena u odgovarajući ekspresijski vektor. Napravljena je prekomjerna ekspresija rekombinantnog proteina u bakteriji Escherichia coli. Analizirana je i potvrđena intrinzična GTPazna aktivnost spužvinog proteina RRAS2, što ukazuje na očuvanost biokemijskih svojstava ovog onkogena. U metodičkom dijelu ovog rada dan je prijedlog nastavnog sata temeljen na istraživačkoj nastavi, za odabranu temu (Enzimi). Pregledana je stručna i udžbenička literatura, oblikovani su prikladni didaktičko-metodički materijali te su navedeni mogući problemi koji se mogu javiti tijekom izvedbe nastavnog sata istraživačkog učenja. Cancer is one of the most widespread diseases nowadays and often occurs due to mutations or changes in the expression levels of genes that control cell growth and development. RRAS2 is one of such proteins and a small GTPase. The homologue of the human RRAS2 was found in evolutionarily much older and simpler animals – sponges. Despite their simple morphology, sponges have a complex genome with many genes very similar to their vertebrate homologues. To get a better insight into the biochemical characteristics of the RRAS2 homologue from the sponge Eunapius subterraneus, the cDNA for RRAS2 was cloned and inserted into an appropriate expression vector. E. coli cells were chemically transformed for the expression of the RRAS2 recombinant protein. The GTPase activity of the sponge RRAS2 was analyzed and confirmed, which indicates that the biochemical properties of this oncogene are evolutionary conserved. In the methodological part, a proposal of inquiry-based lesson, for the selected topic (Enzymes) is given. Professional and textbook literature was reviewed, appropriate didactic-methodical materials were designed, and possible problems that may arise during the implementation of the inquiry-based learning were highlighted.

Polielektroliti su makromolekule koje se sastoje od ponavljajućih podjedinica koje se mogu ionizirati u vodenoj otopini. Uzastopnom naizmjeničnom adsorpcijom polielektrolita različitih naboja na supstrat, pomoću „sloj po sloj“ metode, nastaje polielektrolitni višesloj. Polielektrolitni višeslojevi se mogu koristiti kao antibakterijski filmovi. Na adheziju bakterija utječu hrapavost, naboj i hidrofilnost površine. U ovom radu istražena su svojstva polielektrolitnog višesloja koji se sastoji od kitozana i poli(akrilne kiseline), te višeslojeva na koje je adsorbiran lizozim. Rezultati mjerenja zeta potencijala i potencijala strujanja pokazali su da je površina istraženih filmova negativno nabijena pri fiziološkom pH. Utvrđeno je da je površina pripravljenih višeslojeva hidrofilna i približno jednake hrapavosti koja odgovara glatkim površinama. Adhezija bakterija Pseudomonas aeruginosa i Staphylococcus aureus istražena je pomoću pretražnog elektronskog mikroskopa te je ustanovljeno da su antibakterijska svojstva filmova izraženija kada je lizozim zadnji sloj. Polyelectrolytes are macromolecules comprised of repeating units that could be ionized in an aqueous solution. Polyelectrolyte multilayers are prepared by alternating adsorption on a substrate using „layer by layer“ method. Polyelectrolyte multilayers can be used as antibacterial films. Bacterial adhesion depends on properties such as roughness, charge and hydrophilicity. In this thesis, these properties of a polyelectrolyte multilayer consisting of chitosan and poly(acrylic acid), as well as those with an added layer of lysozyme were studied. Zeta and streaming potential results showed that the examined surfaces were negatively charged at physiological pH. The surfaces of all prepared multilayers were hydrophylic and had similar roughness which corresponds to smooth surfaces. Adhesion of bacteria Pseudomonas aeruginosa and Staphylococcus aureus was studied using scanning electron microscopy and it was shown that antibacterial properties were more pronounced in the case with lysozyme as the terminating layer.

Nesteroidni protuupalni predlijek nabumeton se u jetri metabolizira u svoj farmakološki akivan oblik, 6-metoksi-2-naftiloctenu kiselinu, koja djeluje kao selektivan inhibitor izoenzima ciklooksigenaza-2. Prema biofarmaceutskom sustavu klasifikacije djelatnih tvari, nabumeton pripada drugoj skupini koju karakteriziraju slaba topljivost i visoka permeabilnost. Njegova se topljivost može poboljšati nastajanjem inkluzijskog kompleksa s cikličkim oligosaharidima, ciklodekstrinima. Provođenjem studija prisilne razgradnje dobivaju se informacije o stabilnosti lijeka i njegovim razgradnim produktima koji utječu na njegovu učinkovitost i sigurnost. U sklopu ovog diplomskog rada korišten je spregnuti sustav tekućinske kromatografije ultravisoke djelotvornosti i spektrometrije masa visokog razlučivanja za analizu razgradnih produkata nabumetona nastalih u kiselom, lužnatom, i oksidirajućem mediju, pri sobnoj ili povišenoj temperaturi, uz ili bez prisutnosti β-ciklodekstrina. Strukture i sheme fragmentiranja razgradnih produkata nabumetona pretpostavljene su na temelju rezultata analize tandemnom spektrometrijom masa. Nonsteroidal anti-inflammatory prodrug nabumetone is metabolized in the liver into its pharmacologically active metabolite, 6-methoxy-2-naphthylacetic acid, which acts as a selective cyclooxygenase-2 inhibitor. According to the Biopharmaceutical Classification System of drug substances, nabumetone is classified in the second group which is characterized by low solubility and high permeability. Its solubility can be increased by the formation of inclusion complexes with cyclic oligosaccharides cyclodextrins. Forced degradation studies provide information about drug stability and its degradation products which affect its effectiveness and safety. In this work, ultra performance liquid chromatography coupled with high-resolution mass spectrometry was used for the analysis of nabumetone degradation products formed in acidic, alkaline, and oxidizing media, at room or elevated temperature, with or without the presence of β-cyclodextrin. Structures and fragmentation schemes of nabumetone degradants were proposed according to the results of tandem mass spectrometry analysis.

U ovom radu ispitana je mogućnost kokristalizacije soli bis(1,10- fenantrolin)kloridobakrova(II) klorida s perhalogeniranim donorima halogenske veze. Pokusima kokristalizacije mehanokemijskom sintezom pristupljeno je na dva načina. Prvi način uključivao je mljevenje soli koordinacijskog spoja bakra(II) i donora halogenske veze u različitim stehiometrijskim omjerima, dok je drugi način bila višekomponentna mehanokemijska sinteza u jednom koraku, odnosno mljevenje bakrova(II) klorida dihidrata, 1,10-fenantrolina monohidrata i donora halogenske veze u različitim stehiometrijskim omjerima. Mehanokemijskom sintezom pripravljeno je 8 novih kristalnih produkata koji su okarakterizirani difrakcijom rentgenskog zračenja na praškastom uzorku, dok je sintezom iz otopine pripravljeno 6 jediničnih kristala. Kokristalima pripravljenim iz otopine difrakcijom rentgenskog zračenja na jediničnom kristalu određena je kristalna i molekulska struktura, a termička stabilnost ispitana je termogravimetrijskom analizom. Strukturnom analizom utvrđeno je da u svim pripravljenim kokristalima nekoordinirani i koordinirani kloridi na atomu bakra sudjeluju u halogenskoj vezi Cl···I s donorima halogenske veze, a konačni ishod reakcije ne ovisi o načinu mehanokemijske sinteze. Within this work, cocrystallization of bis(1,10-phenanthroline)chloridocopper(II) chloride with perhalogenated halogen bond donors was studied. Cocrystallization experiments were approached in two ways. Firstly, grinding a complex salt of copper(II) and a halogen bond donor, and secondly, grinding copper(II) chloride dihydrate, 1,10-phenanthroline monohydrate and a halogen bond donor in different stoichiometric ratios. Eight new crystalline products were prepared by mechanochemical synthesis and characterized by powder X-ray diffraction, while six single crystals were prepared by crystallization from solution. The crystal and molecular structure of the cocrystals prepared from the solution was determined by single crystal X-ray diffraction. Thermal stability was tested by thermogravimetric analysis. Structural analysis revealed that in all prepared cocrystals, both coordinated and uncoordinated chloride participate in a halogen bond with the iodine atom of the halogen bond donor, and the final outcome of the reaction does not depend on the approach to mechanochemical synthesis.

Porozni organski polimeri pokazali su se prikladnima za selektivnu adsorpciju ugljikova dioksida (CO2). U ovom istraživanju metodama računalne kemije proučeno je šest organskih poroznih sustava organiziranih u kovalentne organske mreže. Istražen je utjecaj dviju građevnih jedinki (fenilnim skupinama trisuptituirani amin i piridin) i triju dušik-dušik poveznica (azo, azoksi i azodioksidna veza) te utjecaj različitih konfiguracija slaganja susjednih dvodimenzijskih slojeva na adsorpcijska svojstva materijala. Periodičkim metodama teorije funkcionala gustoće (DFT) optimizirane su strukture kovalentnih organskih mreža, podvrgnute Monte Carlo simulacijama te su izračunate adsorpcijske izoterme i prikazane raspodjele gustoće adsorbiranih molekula CO2 i N2. Rezultati su uspoređeni s podacima o vezanju molekula dobivenim na temelju izračunatih vrijednosti elektrostatskog potencijala. Porozni sustavi izgrađeni od aminskih građevnih jedinki povezanih azodioksidnim vezama pokazali su najbolju adsorpciju CO2. Porous organic polymers have proven to be suitable for the selective carbon dioxide (CO2) adsorption. In this research, six organic porous systems organized into covalent organic frameworks were examined using computational chemistry methods. The influence of two building blocks (triphenyl substituted amine and pyridine) and three nitrogen-nitrogen linkages (azo, azoxy and azodioxy) and the influence of different neighbouring twodimensional layer stacking configurations on the adsorption properties of the material were examined. Using periodic density functional theory (DFT) methods, covalent organic framework structures were optimized and subjected to Monte Carlo simulations to compute adsorption isotherms along with density plots of adsorbed CO2 and N2 molecules. The results were compared with other data obtained from the calculated electrostatic potential values. Porous systems constructed from amine building blocks linked by azodioxy bonds have shown the best CO2 adsorption.

U okviru ovog diplomskog rada pripravljene su halogenidne soli alifatskih triamina dietilentriamina ((H3DETA)Cl3), N-(2-aminoetil)-1,3-propandiamina ((H3N-1,3-PDA)Cl3 i (H3N-1,3-PDA)Br3) kao i kloridna sol trietilentetraaamina ((H4TETA)Cl4). Klasičnom otopinskom sintezom pripravljena je serija hibridno organsko-anorganskih halogenokuprata(II): (H3DETA)[CuCl5], (H3DETA)[CuBr5], (H3N-1,3-PDA)[CuCl5], (H3N1,3-PDA)[CuBr5], (H4TETA)[CuCl6] i (H4PEEA)[CuBr6]·H2O. Anorganske graĎevne jedinke kod svih prireĎenih halogenometalata su stehiometrije CuX4, te se većinom udružuju u slojevite okosnice. Kod hibridnih halogenometalata sa slojevitom strukturom dolazi do ugraĎivanja organskih kationa izmeĎu anorganskih slojeva. Organski kationi povezuju se putem protoniranih terminalnih amino skupina za halogenokupratne(II) jedinke, a meĎusobno se udružuju putem N‒H···X···H‒N vodikovih veza. Pripravljenim hibridnim halogenometalatima odreĎene su kristalne strukture pomoću difrakcije rendgenskog zračenja na jediničnom kristalu, a spojevi su dodatno okarakterizirani pomoću infracrvene spektroskopije, difrakcije rendgenskog zračenja na polikristalnom uzorku te metodama termičke analize. In this work, halide salts of aliphatic triamines diethylenetriamine, ((H3DETA)Cl3), N-(2- aminoethyl)-1,3-propanediamine, (H3N-1,3-PDA)Cl3 and (H3N-1,3 -PDA)Br3, and the chloride salt of triethylenetetraamine, (H4TETA)Cl4, were prepared. A series of hybrid organic-inorganic halocuprates(II) were prepared by solution crystallization: (H3DETA)[CuCl5], (H3DETA)[CuBr5], (H3N-1,3-PDA)[CuCl5], (H3N-1,3-PDA)[CuBr5], (H4TETA)[CuCl6] and (H4PEEA)[CuBr6]·H2O. The inorganic building blocks of all prepared halogenometalates have stoichiometry CuX4, and they are mostly combined into layered frameworks. In the case of hybrid halometallates with a layered structure, organic cations are incorporated between the inorganic layers. Organic cations are connected via protonated terminal amino groups to halocuprate(II) units, and interlinked to each other via N‒ H···X···H‒N hydrogen bonds. The crystal structures of the prepared hybrid halogenometalates were determined using single-crystal X-ray diffraction, and the compounds were additionally characterized using infrared spectroscopy, powder X-ray diffraction and thermal analyses.

Alosterija je jedna od najvažnijih pojava u regulaciji metaboličke i genetske aktivnosti u organizmu. Identifikacija alosteričkih signalnih puteva pomoću računalnih tehnika predstavlja moćan alat u proučavanju ponašanja proteina uslijed vezanja modulatora što može služiti u otkrivanju novih lijekova.

Uređajem za statičko raspršenje svjetlosti Mastersizer3000 određivane su brzine agregacije čestica SiO2 (S-1000, S-200 i S-80) i TiO2 (T-250, T-50-70 i T-15). Mjerenja su provedena za dvije pH vrijednosti pH (pH = 3 i pH = 7) i pri dvije ionske jakosti (Ic = 0,1 mol dm–3 i Ic = 0,001 mol dm–3 ). Ispitano je kako pH i ionska jakost utječu na brzinu agregacije navedenih čestica. Iz raspodjele veličine čestica po volumenu izračunate su ovisnosti brojčane koncentracije čestica o vremenu i određene konstante brzine agregacije. Za čestice SiO2 (S200) izmjereno je statičko raspršenje svjetlosti pri pH = 4,5 za niz ionskih jakosti te je određena kritična koagulacijska koncentracija koja iznosi cccc = 0,0326 mol dm–3 . Uređajem ZetasizerPro metodom dinamičkog raspršenja svjetlosti za čestice SiO2 (S-1000, S-200 i S-80) i TiO2 (T250, T-50-70 i T-15) određene su ovisnosti veličine čestica o pH, a metodom elektroforeze određene su izoelektrične točke. Aggregation rates of SiO2 (S-1000, S-200 and S-80) and TiO2 (T-250, T-50-70 and T-15) particles were determined using static light scattrering device Mastersizer3000. Measurements were performed for two pH values (pH = 3 and pH = 7) and at two ionic strengths ((Ic = 0,1 mol dm–3 i Ic = 0,001 mol dm–3 ). It was investigated how pH and ionic strength affect the speed of aggregation of the mentioned particles. The numerical concentration of particles in time are calculated from the prarticle size distribution by volume and used to determine aggregation rate constant. Static light scattering was measured for SiO2 (S-200) particles at pH = 4,5 for a range of ionic strengths and the critical coagulation concentration was determined, which is cccc = 0,0326 mol dm–3 . The ZetasizerPro device was used to detemine the dependence of particle size on pH using using the dynamic light scattering method and isoelectric points using the electrophoresis method for SiO2 (S-1000, S-200 and S-80) and TiO2 (T-250, T-50-70 and T15) particles.

U ovom diplomskom radu sintetizirana su tri prekursora Ugijeve reakcije koji sadrže oksimsku i karboksilnu skupinu: 6-(2-hidroksiiminometilimidazol-1-il)heksanska kiselina, 2-(4- hidroksiiminometil-pirazol-1-il)heksanska kiselina i 7-(5-metil-3-(hidroksiimino)-2- oksoindolin-1-il)heksanska kiselina. Dobiveni 5-metilisatinski prekursor poslužio je kao reaktant za Ugijevu reakciju s benzil-aminom, tert-butil-izocijanidom i formaldehidom pri čemu se usporedila mogućnost izvođenja sinteze u otopini, mikrovalnom reaktoru te mehanokemijski. Pripravljeni spojevi pročišćeni su dostupnim kromatografskim tehnikama i karakterizirani dostupnim analitičkim metodama (UPLC-UV/MS, 1D, 2D NMR). U računalnom dijelu rada uspoređeno je vezanje produkta Ugijeve reakcije i literaturno opisanih reaktivatora u aktivnom mjestu sarinom inhibiranoga enzima acetilkolinesteraze pomoću molekulskog uklapanja. In this diploma thesis new Ugi reaction's precursors containing oxime and carboxyl groups were synthesized: 6-(2-hydroxyiminomethylimidazol-1-yl)hexanoic acid, 2-(4- hydroxyiminomethyl-pyrazol-1-yl)hexanoic acid and 7-(5-methyl-3-(hydroxyimino)-2- oxoindolin-1-yl)hexanoic acid. The 5-methylisatin precursor was used as a reactant for the Ugi reaction with benzyl-amine, tert-butyl-isocyanide and formaldehyde. Besides the classical syntheses in solution, the Ugi reaction was performed in a microwave reactor as well as mechanochemically. The compounds were purified by available chromatographic techniques and characterized by available analytical methods (UPLC-UV/MS, 1D, 2D NMR). The binding of the Ugi reaction's products and acetylcholinesterase reactivators in the active site of the sarininhibited enzyme acetylcholinesterase were compared by molecular docking.

U ovom diplomskom radu sintetizirana su tri koordinacijska spoja kobalta(II) korištena kao akceptori halogenske veze – dikloro-, dibromo- i dijodo-(6,6´´-dibrom-2,2´:6´,2´´- terpiridin)kobalt(II). Koordinacijski spojevi kokristalizirani su s odabranim perfluoriranim jodbenzenskim spojevima kao donorima halogenske veze s ciljem usporedbe potencijala halogenog liganda kao akceptora halogenske veze Co–X∙∙∙I (X = Cl, Br, I) kao i izučavanja utjecaja bromovih atoma na periferiji terpiridinskog liganda na ostvarivanje halogenskih veza, kao i njihov akceptorski potencijal. Sinteza koordinacijskih spojeva i kokristala provedena je mehanokemijski, dok su jedinični kristali dobiveni kristalizacijom iz otopine. Mehanokemijskom sintezom ukupno je dobiveno osam novih kristalnih produkata koji su okarakterizirani difrakcijom rendgenskog zračenja na praškastom uzorku. Za one produkte čiji su jedinični kristali uspješno priređeni dodatno su okarakterizirani difrakcijom rendgenskog zračenja u jediničnom kristalu i termogravimetrijskom analizom. Strukturnom analizom kokristala utvrđeno je kako je dominantna međumolekulska interakcija Co–X∙∙∙I (X = Cl, Br, I). Atomi broma na periferiji liganda sudjeluju u kontaktima Br∙∙∙H, dok samo u jednom kokristalu ostvaruju halogenske veze Co–Br∙∙∙Br i Br∙∙∙I u kojima sudjeluju kao donori i kao akceptori. In this diploma thesis three cobalt(II) coordination compounds were synthesized and used as halogen bond acceptors – dichloro-, dibromo-, and diiodo-(6,6"-dibromo-2,2':6',2'- terpyridine)cobalt(II). Coordination compounds were cocrystallized with selected perfluorinated iodobenzene compounds as halogen bond donors in order to compare the potential for halogen bonding of the halogen atoms located peripherally on the terpyridine ligands, as well as their acceptor potential. Mechanochemical methods were used to synthesize bulk coordination compounds and cocrystals, whereas crystallization from solution was used to obtain single crystals. Mechanochemical synthesis resulted in eight new crystalline phases which were then identified using X-ray powder diffraction. The products were also characterized by thermogravimetric analysis and single crystal X-ray diffraction in cases where single crystals were successfully obtained. Structural analysis of the compounds revealed that the dominant intermolecular interaction is Co–X∙∙∙I (X = halogen). The bromine atoms on the ligand's periphery participate in Br∙∙∙H contacts, but in one cocrystal they participate in the halogen bonds Co–Br∙∙∙Br and Br∙∙∙I, where they act as both acceptors and donors.

Rak dojke, s dominantnim podtipom HR+ HER2−, najčešći je rak u svijetu. Jedan od načina liječenja HR+ HER2− raka dojke je kombinacijom CDK4/6 inhibitora i inhibitora aromataze, često uz lijekove ribociklib i letrozol. Kapilarna elektroforeza metoda je koja se iz dana u dan sve više razvija te bi se s odgovarajućom reproducibilnošću i osjetljivošću mogla koristiti u svrhu terapijskog praćenja lijekova. U ovom je radu razvijena i validirana metoda za određivanje ribocikliba, letrozola i ibuprofena kapilarnom elektroforezom uz ketoprofen kao unutarnji standard. Separacija je postignuta uz 75 mmol dm−3 fosfat, 8,5 mmol dm−3 sulfobutileter-β-ciklodekstrin (SBE-β-CD) i 5 % izopropanola u radnom puferu pri naponu 15 kV i temperaturi 25 °C. Uzorak je injektiran hidrodinamički pri tlaku −50 mbar u trajanju 5 sekundi, nakon injektiranja ultračiste vode 6 sekundi. Metoda je precizna, linearna u rasponu masene koncentracije od 1 do 75 µg cm−3 , specifična, robusna za promjenu temperature, koncentracije SBE-β-CD te napona. Breast cancer is the most common type of cancer in the world, with subtype HR+ HER2− being most prevalent. HR+ HER2− breast cancer can be treated with hormone therapy, with ribociclib (CDK 4/6 inhibitor) and letrozole (aromatase inhibitor) being a commonly used medicine combination. Capillary electrophoresis is a rapidly developing method which could be used for therapeutic drug monitoring purposes after providing acceptable reproducibility and sensitivity. A method for analysis of ribociclib, letrozole and ibuprofen by capillary electrophoresis using ketoprofene as an internal standard was developed and validated. The separation was obtained with 75 mmol dm−3 phosphate, 8,5 mmol dm−3 sulphobutylether-β-cyclodextrin (SBE-β-CD) and 5% isopropanol. The hydrodynamic injection was performed at −50 mbar pressure for 5 seconds, after the injection of ultrapure water for 6 seconds. The method is precise, linear in a range of mass concentration of 1 to 75 µg cm−3 , specific and robust for the change of temperature, SBE-β-CD concentration and voltage.

Derivati benzotiazola s amidinskim funkcionalnim skupinama pokazuju značajnu biološku aktivnost. Četiri novosintetizirana derivata bis-benzotiazola supstituirana amidinom spektroskopski su okarakterizirana te im je određena temperaturna stabilnost. Ispitane su interakcije derivata bis-benzotiazola supstituiranih amidinom s DNA, RNA i hibridne DNA:RNA polinukleotide. Kompeticijskom dijalizom probrani su polinukleotidi prema kojima spojevi pokazuju najveći afinitet i selektivnost, te su oni korišteni u daljnjem istraživanju. Eksperimenti temperature mekšanja pokazali su da benzotiazoli stabiliziraju dvolančane, odnosno trolančane uzvojnice polinukleotida. Dodatak polinukleotida ispitivanim spojevima uzrokovao je gašenje fluorescencije te su, na temelju fluorescencijskih titracija, izračunate ili procijenjene konstante asocijacije. Spektri titracija dobiveni metodom cirkularnog dikroizma ukazuju na mješovite načine vezanja; interkalaciju te vezanje u mali utor i agregiranje spoja duž fosfatne okosnice. Dobiveni rezultati ukazuju na selektivnost spojeva prema određenim sekvencama i strukturama nukleinskih kiselina. Amidino-substituted benzothiazole derivates showed significant biological activity. Four newly synthesized derivates of amidino-substituted bis-benzothiazoles were spectroscopically characterized and their temperature stability was determined. Interactions of amidinosubstituted bis-benzothiazoles with DNA, RNA and hybrid DNA:RNA polynucleotides were examined. Interaction of an array of polynucleotides with four benzothiazole derivates was screened by competition dialysis. According to the screening results (affinities and selectivity) polynucleotides were selected for detailed characterization. The thermal melting experiment indicated that benzothiazoles stabilised double-stranded and triple-stranded polynucleotide helixes. The addition of polynucleotide to the compound solution caused fluorescence quenching. Association constants were calculated or estimated based on fluorimetric titrations. Circular dichroism spectroscopy indicated mixed binding modes: intercalation, minor groove binding and aggregation along the phosphate backbone. The obtained results indicated benzothiazoles selectivity for specific polynucleotide sequences and structures.

U ovome radu istražena je stabilnost, biodostupnost i dijalizibilnost polifenola pinocembrina, galangina, krizina, kavene kiseline i feniletil estera kavene kiseline u uvjetima simulirane probave in vitro (oralna i želučana faza te faza probave u tankom crijevu s dva modela: centrifugiranjem i dijalizom kroz polupropusnu membranu) na dva uzorka kontinentalnog propolisa i smjesi standarda odabranih polifenola. Različitim ekstrakcijskim postupcima i korištenjem određenih nosača istraženi su ekstrakti prikladni za oralnu primjenu te analizirani validiranom metodom tekućinske kromatografije visoke djelotvornosti uz primjenu UVdetektora s nizom fotodioda (eng. High Performance Liquid Chromatography - Photodiode Array Detection, HPLC-PDA). Primjenom 2-hidroksipropil-β-ciklodekstrina (HP-β-CD) i postupkom liofilizacije dobiven je u vodi topljivi kompleks propolis-HP-β-CD s najvećim udjelom polifenola. Rezultati analiza RP-HPLC-PDA oba uzorka kontinentalnog propolisa pokazali su stabilnost odabranih polifenola tijekom svih faza simulirane probave. Biodostupnost svih polifenola u uvjetima in vitro nakon dijalize kroz polupropusnu membranu bila je vrlo niska u odnosu na model s centrifugiranjem. Nadalje, rezultati ukazuju da u modelu s centrifugiranjem nema razlike u biodostupnosti polifenola između dva uzorka kontinetalnog tipa propolisa. Vodena suspenzija liofilizata kompleksa propolis-HP-β-CD pokazala se kao potencijalna djelatna tvar u farmaceutskim proizvodima za oralnu primjenu. In this work, the stability, bioaccessibility and dialysability of polyphenols pinocembrin, galangin, chrysin, caffeic acid and phenylethyl ester of caffeic acid were investigated after in vitro simulated digestion (oral and gastric phase and small intestine phase with two models: centrifugation and dialysis through a semi-permeable membrane) on two samples of continental propolis and standard mixture of selected polyphenols. Extracts suitable for oral administration prepared by various extraction procedures and the use of certain carriers have been investigated and analyzed by validated RP-HPLC-PDA method. Using 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) carbohydrate and by lyophilisation procedure, a propolis water-soluble complex with the highest polyphenol content was prepared. The results of RP-HPLC-PDA analysis of both propolis samples showed the stability of selected polyphenols during all phases of simulated digestion. The bioaccessibility of all polyphenols in in vitro conditions after dialysis through a semi-permeable membrane was very low compared to the centrifugation model. Furthermore, the results indicate that in the model with centrifugation there is no difference in the bioaccessibility of polyphenols between the two samples of the continental type of propolis. The aqueous suspension of the lyophilisate of the propolis-HP-β-CD complex proved to be a potential active substance in pharmaceutical products for oral administration.

U sklopu ovog diplomskog rada pripravljeni su imini izvedeni iz 2-nitrobenzaldehida i aromatskih amina (4-, 3- i 2-halogenanilini, halogen = Cl, Br, I; te 3-nitroanilin). Mehanokemijskom sintezom i kristalizacijom iz otopine izučavana je mogućnost nastajanja kokristala pripravljenih imina s odabranim perhalogeniranim donorima halogenske veze s ciljem da se istraži potencijal različitih funkcijskih skupina imina koje se mogu ponašati kao akceptori halogenske veze. Dobiveni produkti okarakterizirani su difrakcijom rentgenskog zračenja na polikristalnom uzorku, metodama termičke analize i računalnim metodama. Za devet kokristala metodom difrakcije rentgenskog zračenja na jediničnom kristalu određene su molekulske i kristalne strukture. Strukturnom analizom utvrđeno je da je u većini kokristala dominantna supramolekulska interakcija halogenska veza N⸱⸱⸱I koja se ostvaruje između atoma joda perhalogeniranog donora halogenske veze i atoma dušika iminske skupine. U pet kokristala molekule imina se dodatno povezuju pnikogenskom vezom N⸱⸱⸱O koja se ostvaruje između atoma dušika i atoma kisika nitro skupina susjednih imina. As a part of this thesis, imines were prepared from 2-nitrobenzaldehyde and aromatic amines (4-, 3- i 2-halogenanilines, halogen = Cl, Br, I; and 3-nitroaniline). The investigation of cocrystal synthesis from prepared imines and selected halogen bond donors was carried out using mechanochemical synthesis and crystallization from solution, in order to research the possibilities of different functional groups to act as halogen bond acceptors. The obtained products were characterized by X-ray diffraction methods, by thermal analysis methods and computational methods. For nine cocrystals, crystal and molecular structures were determined using single-crystal X-ray diffraction. Structural analysis has shown N⸱⸱⸱I halogen bond between the perhalogenated halogen bond donor's iodine atom and imine nitrogen atom as the dominant supramolecular interaction in most cocrystals. In five cocrystals imine molecules are additionally connected via pnicogen bond N⸱⸱⸱O between nitro group nitrogen and oxygen atoms of neighbouring imines.

U ovom radu pripravljene su jodidne soli gvanidina u svrhu ispitivanja njihove kokatalitičke aktivnosti u reakciji aldolne kondenzacije cikloheksanona i 4-klorbenzaldehida uz katalizator L-prolin. Odabrani aciklički i ciklički gvanidinski derivati razlikuju se prema usmjerenosti NH veza, steričkim utjecajima i supstituentima. Dobivenim su spojevima ispitana kiselobazna svojstva u acetonitrilu uz referentni spoj poznate bazičnosti UV titracijom i NMR spektroskopijom. Spektrometrijom masa provedeno je kinetičko mjerenje relativnih afiniteta vezanja jodida fragmentacijom jodidom premoštenog dimera ispitivanog i referentnog gvanidina. Omjer dijastereomera produkata aldolne kondenzacije nastalog kao rezultat kokatalitičke aktivnosti gvanidina praćen je HPLC i NMR tehnikom. Pokazano je da dijastereomerni omjer produkata ovisi o kiselo-baznim svojstvima gvanidina, usmjerenosti NH veza i stvaranju međumolekulske i unutarmolekulske vodikove veze. Navedeni čimbenici utječu na stabilnost katalitičkog kompleksa među gvanidinom, supstratom i L-prolinom te time imaju izravan utjecaj na konačan ishod reakcije. In this master thesis, iodide salts of guanidine were prepared for the purpose of testing their cocatalytic activity in the aldol condensation of cyclohexanone and 4-chlorobenzaldehyde with L-proline as catalyst. Acyclic and cyclic guanidine derivatives differ by the orientation of NH bonds, steric effect and substituents. Acid-base properties of the obtained compounds were determined in acetonitrile with a reference compound of known basicity using UV titration and NMR spectroscopy. Mass spectrometry was used to measure the relative affinity of iodide binding by fragmentation of the iodide-bound dimer of the tested and reference guanidine. Diastereomeric ratio of the aldol condensation products, as a result of cocatalytic activity of guanidines, was monitored by HPLC and NMR techniques. It was shown that the diastereomeric ratio of the products depends on the acid-base properties, orientation of NH bonds in guanidine and formation of intramolecular and intermolecular hydrogen bonds. The mentioned factors influence the stability of the catalytic complex between guanidine, substrate and L-proline and thus have a direct effect on the final outcome of the reaction.

U ovom diplomskom radu ispitana je mogućnost kokristalizacije koordinacijskih spojeva vrste CoX2L (gdje je X = Cl, Br ili I, a L = 6,6'-dimetil-2,2'-bipiridin) i perhalogeniranih donora halogenske veze, s ciljem izučavanja mogućnosti nastajanja kokristala s metaloorganskom podjedinicom te uspoređivanja akceptorskog potencijala halogenidnih iona koordiniranih na metalni centar za ostvarivanje halogenske veze. Mehanokemijskom sintezom i kristalizacijom iz otopine priređeno je sedam novih kokristala koji su okarakterizirani difrakcijom rendgenskoga zračenja na praškastom uzorku, difrakcijom rendgenskoga zračenja u jediničnom kristalu i termogravimetrijskom analizom. U svim strukturno okarakteriziranim kokristalima utvrđeno je kako je halogenska veza Co−X···I (X = Cl, Br ili I) dominantna međumolekulska interakcija koja se ostvaruje između atoma joda perhalogeniranog spoja kao donora halogenske veze i halogenidnog liganda koordinacijskog spoja kobalta(II) kao akceptora halogenske veze. Pokazano je i da se korištenjem perhalogeniranih donora različite topičnosti i geometrije može ugađati supramolekulska arhitektura i termička stabilnost dobivenih produkata. Within this master thesis, the possibility of cocrystallization of coordination compound CoX2L (X = Cl, Br or I, L = 6,6'-dimethyl-2,2'-bipyridine) and perhalogenated halogen bond donors was examined, with the aim of studying the plausibility of cocrystal formation with a metalorganic subunit and comparing the ability of the halide ions coordinated to the Co(II) metal center to act as a halogen bond acceptor. Seven new cocrystals were synthesized by mechanochemical synthesis and crystallization from solution. The obtained products were characterized by powder and single crystal X-ray diffraction, and by thermogravimetric analysis. Structural analysis of the cocrystals revealed that the dominant supramolecular interactions are Co−X···I halogen bonds (X = Cl, Br or I) formed between the iodine atom of the halogen bond donor and the halogen ligand of the cobalt(II) coordination compound as a halogen bond acceptor. The results of this study also demonstrate the possibility of altering supramolecular architectures and thermal stabilities of the products by utilizing donors featuring different geometries and topicities.

Proces fosforilacije jedan je od najčešćih metoda regulacije ciljnih proteina koja omogućuje normalnu staničnu komunikaciju te doprinosi održavanju homeostaze. Odvija se pomoću proteinskih kinaza koje izvršavaju bitne uloge u gotovo svim staničnim signalnim putevima, uključujući staničnu proliferaciju, diferencijaciju, migraciju, apoptozu te metabolizam. Promijenjena aktivnost većine proteinskih kinaza, s naglaskom na proteinske tirozin-kinaze, uočena je kod mnogih ljudskih bolesti, osobito tumorskih. Upravo zato, ova vrsta enzima postala je središnja meta farmaceutske industrije za razvoj selektivnih inhibitora koji bi se mogli koristiti u liječenju takvih oboljenja. U ovom radu obrađeno je djelovanje proteinskih kinaza, s naglaskom na tirozin-kinaze, njihova signalizacija i uloga u zdravim i malignim stanicama. Također su predstavljeni pojedini inhibitori tirozin-kinaza te njihova uloga u liječenju raznih bolesti., The process of phosphorylation is one of the most common methods of regulating target proteins, as it allows normal cellular communication and contributes to the maintenance of homeostasis. It occurs with the help of protein kinases, which play important roles in most cellular signaling pathways, including cellular proliferation, differentiation, migration, apoptosis and metabolism. Altered activity of most protein kinases, with an emphasis on the protein tyrosine kinases, is observed in many human diseases, especially tumors. That is why this type of enzyme has become the main target of the pharmaceutical industry for the process of developing selective inhibitors, which could be used as treatment against such diseases. This thesis examines the impact of protein kinases, with the emphasis on the tyrosine kinases, their signaling and their role in both healthy and malignant cells. Individual inhibitors of tyrosine kinases and their role in the treatment of various diseases are also presented.

U okviru rada je iz fumarne kiseline u 4 koraka sintetizirana ciklobutan-1,2,3,4-tetrakarboksilna kiselina (cBTA) na koju su zatim amidnom vezom vezane po 4 molekule različitih aminokiselina: L-alanina, L-fenilalanina i L-prolina. Dobiveni homokiralni ligandi korišteni su za pripravu metaloorganskih mreža (MOF-ova) u solvotermalnim i mehanokemijskim reakcijama sa spojevima cirkonija(IV). Separacijska svojstva MOF-a izvedenog iz L-alanina ispitana su pomoću tri kiralna organska spoja. Katalitička svojstva istog MOF-a ispitana su na aldolnoj reakciji cikloheksanona i 4-nitrobenzaldehida. Separacija je praćena tekućinskom kromatografijom visoke djelotvornosti s kiralnom stacionarnom fazom. Sintetizirani ligandi analizirani su spektroskopijom NMR, masenom spektrometrijom i IR spektroskopijom. MOFovi su analizirani difrakcijom rentgenskog zračenja na polikristalnom uzorku, termogravimetrijom, IR spektroskopijom te spektroskopijom NMR u čvrstom stanju. Within this thesis cyclobutane-1,2,3,4-tetracarboxylic acid (cBTA) was synthesised in 4 steps starting from fumaric acid. Amide bonds were formed between cBTA and 4 molecules of different amino acids: L-alanine, L-phenylalanine and L-proline. Obtained homochiral ligands were used to prepare metal-organic frameworks (MOFs) through solvothermal and mechanochemical reactions with zirconium(IV) compounds. Separation properties of the MOF derived from L-alanine were investigated using different chiral organic compounds followed by high performance liquid chromatography with a chiral stationary phase. Catalytic properties of the MOF were investigated through the aldol reaction of cyclohexanone and 4-nitrobenzaldehyde. Synthesised ligands were analysed using NMR spectroscopy, mass spectrometry and IR spectroscopy. MOFs were analysed using powder x-ray diffraction, thermogravimetry, IR and solid-state NMR spectroscopy.

U današnje vrijeme upotreba ksenobiotika sve je češća, a samim time se i sve veće količine nekontrolirano ispuštaju u okoliš. Pesticidi, antiparazitici te ostali ksenobiotici i njihovi metaboliti, razgradni i transformacijski produkti dospijevaju u tlo i vodu čineći štetu biljnom i životinjskom svijetu. Kako su ksenobiotici spojevi složene kemijske strukture, vrlo ih je teško u potpunosti ukloniti iz vode, a često je potrebno korištenje složenih i skupih metoda obrade otpadnih voda. Metode obrade aktivnim muljem najzastupljenije su metode u obradi otpadnih voda. Cilj ovoga rada je ispitati mogućnosti uklanjanja ksenobiotika iz otpadnih voda biorazgradnjom pomoću aktivnog mulja. Određivana je mogućnost biorazgradnje antiparazitika febantela, albendazola i mebendazola te pesticida iz skupine insekticida tiakloprida, acetamiprida i klotianidina u vremenu od 48 sati tekućinskom kromatografijom visoke djelotvornosti uz praćenje pH-vrijednosti, temperature, gustoće, ukupne količine ugljika te utjecaja na mikroorganizme. Proces biorazgradnje odvijao se u pet reaktora koji su bili zaštićeni aluminijskom folijom kako bi se spriječio utjecaj Sunčeve svjetlosti na razgradnju ksenobiotika. Uklanjanje ksenobiotika određivano je tekućinskom kromatografijom visoke djelotvornosti uz detektor s nizom dioda, a uzorci su se uzimali u točno određenim vremenskim intervalima. Uz to, uzorci su se analizirali i mjerenjem ukupne količine ugljika, a pratili smo i gustoću aktivnog mulja prije procesa biorazgradnje te gustoću nakon biorazgradnje i to za reaktor s muljem, mikroplastikom i ksenobiotikom te za reaktor s muljem i ksenobiotikom. Također je za svaki ksenobiotik provedena i mikrobiološka analiza kako bi se utvrdio utjecaj pojedinog ksenobiotika na mikroorganizme koji sudjeluju u procesu biorazgradnje. Stopostotno uklanjanje postignuto je za sve antiparazitike, dok je kod pesticida uklanjanje bilo nešto lošije. Uz sam proces uklanjanja uzimala se u obzir i adsorpcija i desorpcija komponenata na površinu mikroplastike i pahuljice aktivnog mulja, a pomoću dobivenih kromatograma detektirali su se i razgradni/transformacijski produkti koji nastaju tijekom procesa biorazgradnje. Nowadays, the use of xenobiotics is becoming more and more common, and thus increasing amounts are being released into the environment in an uncontrolled manner. Pesticides, antiparasitics and other xenobiotics and their metabolites, degradation and transformation products reach the soil and water causing damage to the plant and animal life. Since xenobiotics are compounds with a complex chemical structure, it is very difficult to completely remove them from water, and it is often necessary to use complex and expensive wastewater treatment methods. Activated sludge treatment methods are the most common methods in wastewater treatment. The aim of this work is to examine the possibilities of removing xenobiotics from wastewater by biodegradation using activated sludge. The possibility of biodegradation of the antiparasitics febantel, albendazole and mebendazole and pesticides from the group of insecticides thiacloprid, acetamiprid and clothianidin was determined within 48 hours by high-performance liquid chromatography with monitoring of pH-value, temperature, density, total carbon and impact on microorganisms. The biodegradation process took place in five reactors that were protected by aluminum foil to prevent the influence of sunlight on the degradation of xenobiotics. The removal of xenobiotics was determined by high-performance liquid chromatography with a diode array detector, and samples were taken at precisely determined time intervals. In addition, the samples were analyzed by measuring the total amount of carbon, and we monitored the density of activated sludge before the biodegradation process and the density after biodegradation for the reactor with sludge, microplastics and xenobiotics and for the reactor with sludge and xenobiotics. A microbiological analysis was also carried out for each xenobiotic in order to determine the influence of each xenobiotic on the microorganisms that participate in the biodegradation process. One hundred percent removal was achieved for all antiparasitics, while the removal was slightly worse for pesticides. In addition to the removal process itself, the adsorption and desorption of components on the surface of microplastics and activated sludge flakes were also taken into account, and the resulting chromatograms were used to detect the degradation/transformation products that arise during the biodegradation process.