Karboanhidrazės (CA) yra metalofermentai, katalizuojantys virsmus tarp anglies dioksido ir bikarbonato. Jų slopinimas gali būti taikomas gydyti tokias skirtingas ligas kaip glaukoma, vėžys, nutukimas, epilepsija, osteoporozė ir kt. Šiuo metu yra beveik 30 mažamolekulinių junginių, kurie naudojami kaip vaistai, su padidėjusiu karboanhidrazių aktyvumu susijusioms ligoms gydyti. Darbe tirta rekombinantinių žmogaus karboanhidrazių I, II, VII, IX ir XIII sąveika su sulfonamidiniais ligandais. Įvertintas tirtų baltymų stabilumas skirtingomis eksperimentinėmis sąlygomis, nustatyta priešvėžinio taikinio CA IX oligomerinė būsena. Modeliniais baltymais naudojant karboanhidrazes, praplėstos terminio poslinkio metodo taikymo ribos. Išmatuoti 40 naujų susintetintų junginių sąveikos su karboanhidrazėmis termodinaminiai parametrai, išanalizuota CA XIII sąveikos su sulfonamidiniais slopikliais termodinamika, atskiriant tikruosius, nuo eksperimento sąlygų ir susijusių reakcijų nepriklausančius jungimosi parametrus. / Carbonic anhydrases (CAs) are metalloenzymes that catalyze the conversion between carbon dioxide and bicarbonate. Their inhibition can be applied for treatment of different diseases, such as glaucoma, cancer, obesity, epilepsy, osteoporosis, etc. There are nearly 30 small molecule ligands that are used as drugs for carbonic anhydrase related diseases. In this work interaction between recombinant human carbonic anhydrases I, II, VII, IX, XIII and sulfonamide ligands was analysed. Stability of selected carbonic anhydrases was evaluated in different experimental conditions. Oligomeric structure of anticancer target CA IX was determined. Using carbonic anhydrases as model proteins, the application range of thermal shift assay was extended. Binding parameters of 40 new compounds to human carbonic anhydrases were measured. The binding thermodynamics of sulfonamide ligands to CA XIII was analyzed and intrinsic binding parameters, independent of the experimental conditions and linked protonation reactions, were determined.

Carbonic anhydrases (CAs) are metalloenzymes that catalyze the conversion between carbon dioxide and bicarbonate. Their inhibition can be applied for treatment of different diseases, such as glaucoma, cancer, obesity, epilepsy, osteoporosis, etc. There are nearly 30 small molecule ligands that are used as drugs for carbonic anhydrase related diseases. In this work interaction between recombinant human carbonic anhydrases I, II, VII, IX, XIII and sulfonamide ligands was analysed. Stability of selected carbonic anhydrases was evaluated in different experimental conditions. Oligomeric structure of anticancer target CA IX was determined. Using carbonic anhydrases as model proteins, the application range of thermal shift assay was extended. Binding parameters of 40 new compounds to human carbonic anhydrases were measured. The binding thermodynamics of sulfonamide ligands to CA XIII was analyzed and intrinsic binding parameters, independent of the experimental conditions and linked protonation reactions, were determined. / Karboanhidrazės (CA) yra metalofermentai, katalizuojantys virsmus tarp anglies dioksido ir bikarbonato. Jų slopinimas gali būti taikomas gydyti tokias skirtingas ligas kaip glaukoma, vėžys, nutukimas, epilepsija, osteoporozė ir kt. Šiuo metu yra beveik 30 mažamolekulinių junginių, kurie naudojami kaip vaistai, su padidėjusiu karboanhidrazių aktyvumu susijusioms ligoms gydyti. Darbe tirta rekombinantinių žmogaus karboanhidrazių I, II, VII, IX ir XIII sąveika su sulfonamidiniais ligandais. Įvertintas tirtų baltymų stabilumas skirtingomis eksperimentinėmis sąlygomis, nustatyta priešvėžinio taikinio CA IX oligomerinė būsena. Modeliniais baltymais naudojant karboanhidrazes, praplėstos terminio poslinkio metodo taikymo ribos. Išmatuoti 40 naujų susintetintų junginių sąveikos su karboanhidrazėmis termodinaminiai parametrai, išanalizuota CA XIII sąveikos su sulfonamidiniais slopikliais termodinamika, atskiriant tikruosius, nuo eksperimento sąlygų ir susijusių reakcijų nepriklausančius jungimosi parametrus.

<p>U disertacije je sprovedeno teorijsko istraživanje mikroskopskih i makroskopskih (termodinamičkih) osobina tankoslojnih i niskodimenzionih kristalnih struktura u okviru modela fonona akustičkog tipa. Upoređivanjem fononskih spektara i termodinamičkih svojstava u neograničenim i posmatranim nanostrukturama ustanovljeno je da se oni znatno razlikuju usled postojanja granica kod niskodimenzionih sistema. Takođe je primećeno da u fononskim spektrima niskodimenzionih struktura postoji energijski procep, &scaron;to znači da je energija pobuđivanja fonona u niskodimenzionim kristalnim strukturama veća od one u balku. Ovo uzrokuje da toplotna kapacitivnost u niskotemperaturskoj oblasti ima i znatno niže vrednosti u ograničenim strukturama nego u masovnim uzorcima.</p> / <p>In ths work, a theoretical study of microscopic and thermodynamic properties of thin-layered and low-dimensional crystalline structures &nbsp;of the model type of acoustic phonons was carried out. Comparison of the phonon spectra and &nbsp;thermodynamic properties of the infinite with the observed &nbsp;nanostructures revealed that they differ greatly because of the existence of borders in the low-dimensional systems. It is also noted that in the phonon spectra of nanostructures an energy gap exists, which means that the energy od excitation of phonons in the nanostructured crystal systems is higher than those of the bulk. This indicates that the heat capacitance in the low-temperature range ha a significantly lower values in bounded structures than in massive samples.</p>

<p>Uloga micela u razvoju farmaceutskih oblika zasniva se na njihovom potencijalu da kao transportni sistemi povećaju bioraspoloživost lekovite supstance i unaprede njen farmakokinetski profil. Micele mogu da modifikuju propustljivost biolo&scaron;kuh membrana, omoguće kontrolisano oslobađanje lekovite supstance iz farmaceutskog oblika, stabilizuju lekovitu supstancu, itd. Kombinovanje različitih surfaktanata kao gradivih jedinica omogućava konstruisanje micela sa poželjnim fizičko-hemijskim karakteristikama. Takođe, me&scaron;ovite micele između čijih gradivnih jedinica postoje sinergističke interakcije se formiraju na nižim koncentracijama surfaktanata u poređenju sa monokomponentnim micelama. U doktorskoj disertaciji je ispitana termodinamička stabilnost binarnih me&scaron;ovitih micela Brij S10 i Brij S20 surfaktanata sa polisorbatom 20, polisorbatom 60 i polisorbatom 80 kao kosurfaktantima. Analiziran je uticaj građe odabranih nejonskih surfaktanata na fizičko-hemijske parametre i stabilnost njihovih me&scaron;ovitih micela. Vrednosti kritične micelarne koncentracije su dobijene spektrofluorimetrijskim merenjima. Da bi se analizirao uticaj temperature na termodinamičku stabilnost micela, merenja su vr&scaron;ena na sledećim temperaturama: 273.15 K, 283.15 K, 293.15 K, 303.15 K i 313.15 K. Rezultati su tumačeni sa aspekta teorije regularnih rastvora (regular solution theory &ndash; RST), uz primenu Porterove jednačine. Na osnovu rezultata istraživanja, predloženi su modeli monokomponentnih i binarnih me&scaron;ovitih micela. Utvrđeno je da u svim analiziranim binarnim me&scaron;ovitim sistemima postoji dodatna entropija. Može se doneti zaključak da pretpostavka RST o isključivo entalpijskoj prirodi dodatne Gibsove energije nije primenljiva na ispitivane binarne sisteme. Utvrđeno je da postojanje razlike u dužini hidrofobnih segmenata monomera dodatno stabilizuje me&scaron;ovitu micelu. Sa porastom temperature, ovaj stabilizacioni efekat entropijskog porekla postaje izraženiji. Prisustvo dvostruke veze u ugljovodoničnom lancu polisorbata 80 doprinosi povećanoj rigidnosti njegovih molekula. Rezultat je smanjena termodinamička stabilnost me&scaron;ovitih micela koje sadrže polisorbat 80 u poređenju sa me&scaron;ovitim micelama koje sadrže njegov zasićeni homolog, polisorbat 60. Utvrđeno je da razlika u dužini polarnih segmenata monomera koji grade me&scaron;ovitu micelu utiče na stepen hidratacije micele, a time i na njenu stabilnost.</p> / <p>The role of the micelles in pharmaceutical formulation lies in their ability to, when used as drug delivery systems, increase the bioavailability of the drug and enhance its pharmacokinetic profile. Micelles may modify the permeability of biomembranes, enable controlled release from drug delivery systems, stabilize the drug, etc. By combining different surfactants as building units it is possible to engineer micelles with favorable physicochemical characteristics. Also, the mixed micelles between whose building units synergistic interactions exist are formed on lower concentrations of surfactants in comparison to single-component micelles. In the doctoral dissertation the thermodynamic stability of binary mixed micelles built of Brij S10 and Brij S20 with polysorbate 20, polysorbate 60 and polysorbate 80 as co-surfactants is examined. The influence of the structure of selected nonionic surfactants on physicochemical parameters and the stability of their mixed micelles is analyzed. Critical micelle concentration values were obtained by spectrofluorimetric measurements. In order to analyze the influence of the temperature on the micelles&rsquo; thermodynamic stability, measurements were conducted on following temperatures: 273.15 K, 283.15 K, 293.15 K, 303.15 K and 313.15 K. Obtained results were studied using regular solution theory (RST) and Porter&rsquo;s equation. Based on the research results, models of both single-component and binary mixed micelles are introduced. It is deduced that in all analyzed binary mixed systems the excess entropy exists. A conclusion can be made that the assumption of RST regarding solely enthalpic nature of the excess Gibbs energy is not applicable to investigated binary systems. The difference in the length of the hydrophobic segments of monomers additionally stabilizes the mixed micelle. With the increase in the temperature, this stabilizing effect of the entropic nature is more noticeable. The existence of the double bond in polysorbate 80 hydrocarbon chain contributes to the increased rigidity of its molecules. As a result, the thermodynamic stability of the mixed micelles containing polysorbate 80 is lesser than the stability of the mixed micelles containing its saturated homologue, polysorbate 60. It is determined that the difference in length of polar segments of mixed micelle monomers affects the micelle hydration and therefore, its stability.</p>

<p>Binarne me&scaron;ovite micele su značajne u farmaceutsko-farmakolo&scaron;kim primenama, u kojima lek kao povr&scaron;inska aktivna supstanca može da bude gradivna jedinica binarne micelle (promena bioraspoloživosti, mehanizam resorpcije itd.). Takođe su važne i me&scaron;ovite micele kod kojih su gradivne jedinice biosurfaktanti (biokompatibilni solubilizatori lekova). U kozmetičkoj, prehrambenoj i industriji detergenata me&scaron;ovite micele su uveliko zastupljene. Važno je nalaženje binarnih sistema surfaktanata koji u me&scaron;ovitim micelama poseduju sinergističko međudelovanje (negativna vrednost interakcionog parametra). Takva binarna sme&scaron;a surfaktanata ima nižu vrednost kritične micelarne koncentracije od hidrofobnije komponente sme&scaron;e. U prvom delu rada je ispitivan uticaj dužine hidrofobnog segmenta homolognih Tween-ova iz me&scaron;ovitih micela sa Triton-om X100 na interakcioni parametra ( i, j &beta; ) i dodatnu Gibbs-ovu energiju (GE ), takođe ispitivao se i uticaj temperature na stabilnost proučavanih me&scaron;ovitih micela (273,15-328,115 K). Kritične micelarne koncentracije čistih surfaktanata i njihovih binarnih sme&scaron;a su određeni spektrofluorometrijski uz primenu pirena kao probnog molekula. Nađeno je da na svakoj ispitivanoj temperaturi povećanje dužine zasićenog alifatičnog niza polisorbata povećava stabilnost me&scaron;ovite micele ( , 0 i j &beta; &lt; i GE &lt; 0 ) u odnosu na idealnu me&scaron;ovitu micelu. Prisustvo C9 dvostruke veze u ostatku oleinske kiseline Tween-a 80 umanjuje sinergistički efekat ovog polisorbata u odnosu na Tween 60, koji ima hidrofobni segment sa istim brojem C atoma, ali bez olefinske veze. Dodatna slobodna entalpija ispod temperature 298.15 K određena je entropijskim doprinosem, a iznad 298.15 K entalpijskim doprinosom. U drugom delu rada ispitivane su binarne sme&scaron;e surfaktanata Tween 85/Triton X100 i Tween 85/Triton X165 u vodenom rastvoru u intervalu temperature od 273.15 K do 323.15 K. Pri temperaturama od 273.15 K i 283.15 K između različitih gradivnih jedinica binarnih me&scaron;ovitih micela postoje sinergističke interakcije na čiju veličinu ne utiče razlika u dužini polioksietlienskih nizova Triton-a X100 i Triton-a X165. Na većim vrednostima temperature duži polioksietilenski niz Triton-a X165 pokazuje stabilizirajuči efekat u me&scaron;ovitoj miceli sa Tween-om 85 u odnosu na me&scaron;ovite micele Tween 85/Triton X100. Kod ispitivanih binarnim me&scaron;ovitim micela ne može se prihvatiti aproksimacija regularnih sme&scaron;a o nula vrednosti dodatne entropije formiranja binarne me&scaron;ovite micele.</p> / <p>Binary micelles are important in pharmaceutical-pharmacological applications, in which drug as surface active substance can be building unit of binary micelle (influencing bioavailability change, change of resorption mechanism etc.). Mixed micelles in which building units are biosurfactants (biocompatible drug solubilizers) are also important. In cosmetics, food and detergent industry mixed micelles are widely present. It is important to find binary surfactant systems having synergistic interactions (negative value of interaction parameter). That binary surfactant mixture has lower critical micelle concentration value than more hydrophobic component of the mixture. In the first parth of this work, influence of hydrophobic segment length of homologues Tweens in mixed micelles with Triton X-100 on the interaction parameter value ( i, j &beta; ) and an excess Gibbs energy (GE ) was investigated. Also, influence of temperature on the stability of investigated mixed micelles has been investigated (273.15-328.15 K). Critical micelle concentration values of pure surfactants and their binary mixtures were determined spectrofluorimetrically using pyrene as a probe molecule. It was found for every investigated temperature that increase in length of saturated aliphatic side chain of a polisorbate increases the stability of investigated mixed micelles (&beta; &lt; and GE &lt; 0 ) compared to ideal mixed micelles. Presence of the C9 double bond in oleic acid residue of Tween 80 reduces synergistic effect of this polysorbate compared to Tween 60 which contains hydrophobic segment with the same number of C atoms but without olefin bond. Excess free enthalpy below the 298.15 K is determined with entropic contribution, but above 298.15 K with enthalpic contribution. In the second parth of this work, binary surfactant mixtures of Tween 85/Triton X100 and Tween 85/Triton X165 have been investigated in water solution in the temperature range from 273.15 K to 323.15 K. At the temperatures of 273.15 K and 283.15 K between different building units of binary mixed micelles synergistic interactions exist, which size is not affected by difference in length of the polyoxyethylene chains. At higher temperatures, longer polyoxyethylene chain of Triton X165 shows more stabilizing effect in mixed micelle with Tween 85 than in mixed micelles of Tween 85/Triton X100. For investigated binary mixed micelles, regular solution theory approximation about zero value of excess entropy of binary mixed micelle formation cannot be accepted.</p>

<p>Da bi se dobio sistem surfaktanata željenih osobina moguće je hemijski modifikovati već postojeće molekule povr&scaron;inski aktivnih supstanci, a druga mogućnost je konstrukcija binarnih sme&scaron;a surfaktanata. U farmaceutskoj i prehrambenoj industriji uveliko se primenjuju binarne sme&scaron;e povr&scaron;inski aktivnih molekula. Ukoliko je binarna me&scaron;ovita micela termodinamički stabilnija od hipotetičke idealne binarne me&scaron;ovite micele, onda je kritična micelarna koncentracija binarne sme&scaron;e surfaktanata niža čak i od hidrofobnije gradivne jedinice me&scaron;ovite micele, &scaron;to znači da je za isti efekat povr&scaron;inske aktivnosti potrebna manja količina binarne sme&scaron;e nego čistog surfaktanta. Različite gradivne jedinice binarne micele u micelarnoj pseudofazi mogu formirati specifične regije koje mogu vezivati lekove određenih strukturnih karakteristika. Pogodno je da jedna gradivna jedinica bude krute konformacije, npr. soli žučnih kiselina, dok je druga gradivna jedinica konformaciono pokretljiva (ugljovodonični nizovi iznad C10). Na taj način se povećava zapremina hidrofobne micelarne faze u odnosu na zapreminu hidrofobne micelarne faze monokomponentne micelle konformaciono krutog surfaktanta, &scaron;to povećava solubilizacioni kapacitet me&scaron;ovite micele u odnosu na monokomponentnu micelu krutog surfaktanta. Povećanjem dužine ugljovodoničnog niza konformaciono pokretnog surfaktanta povećava se stepen unutra&scaron;nje pokretljivosti u hidrofobnom domenu me&scaron;ovite micele, &scaron;to takođe povećava verovatnoću prihvatanja molekula gosta. Micelarni sistemi, kako monokomponentnih micela tako i binarnih me&scaron;ovitih micela dodatno se mogu termodinamički stabilizovati povećanjem jonske jačine rastvora. Za hidrataciju katjona tro&scaron;e se molekuli vode iz sistema, &scaron;to povećava efekat desolvatizacije hidrofobne povr&scaron;ine surfaktanata, pa se zbog toga pospe&scaron;uje samoasocijacija.</p> / <p>To obtain the surfactant system with the desired properties it is possible to chemically modify existing molecules of surface active agents. The other possibility is the construction of binary mixtures of surfactants. Binary mixtures of surface active molecules are widely used In the pharmaceutical and food industry. If the binary mixture micelle is more thermodynamically stable than the hypothetical ideal binary mixed micelle, then the critical micellar concentration (CMC) of the binary mixture of surfactants is even lower than the CMC of the more hydrophobic building block of the binary mixture. That means that for the same effect of surface activity less the amount of the binary mixture than the pure surfactants is required. The different building blocks of binary micelles in micelar pseudophase can form specific regions that can bind drugs of certain structural characteristics. It is suitable that one building block is of a rigid conformation, i.e. bile acid salts, while the second building block is of a flexible conformation (above C10 hydrocarbon arrays). In this way the volume of the hydrophobic micellar phase is increased in relation to the volume of the hydrophobic micellar phase of the monocomponent micelles of conformationally rigid surfactant, which increases the capacity of solubilisation of the mixed micelles, compared to the mono-component surfactant micelle of the rigid conformation. By increasing the length of the hydrocarbon array of the the conformational flexible surfactant, the degree of internal mobility in the hydrophobic domain of mixed micelles is also increased, which also increases the likelihood of acceptance of guest molecules. Micellar systems, of both monocomponent micelles and mixed micelles can be additionally thermodynamically stabilized by increasing the ionic strength of the solution. The hydration of cations uses the molecules of water from the system, which increases the effect of desolvatisation of the hydrophobic surface of the surfactants, and therefore promotes self-association.</p>

<p>U ovoj doktorskoj disertaciji proučavano je građenje&nbsp;kompleksa kobalta(II) sa hloridnim i &nbsp;bromidnim jonima u&nbsp;rastopima koji sadrže jednu neorgansku so i jedno organsko&nbsp;jedinjenje i/ili vodu pri različitom sastavu sistema i na različitim&nbsp;temperaturama. Jedan od sistema je amonijum-nitrat &ndash;&nbsp;acetamid &ndash; voda sastava NH₄NO₃∙(2,61-z)CH₃CONH₂∙zH₂O (z =&nbsp;0,0 1,61 i 2,61), a drugi sistem je amonijum&ndash;nitrat &ndash; dimetilsulfoksid&nbsp;(DMSO) sastava NH₄NO₃∙zDMSO (z = 1 &ndash; 6), &nbsp;na&nbsp;različitim temperaturama (35, 45, 55 i 65 ^oC).&nbsp;Cilj ovog istraživanja je bio određivanje stabilnosti&nbsp;kompleksa kobalta koji se grade sa sa halogenidnim jonima i&nbsp;komponentama rastvarača, pronalaženje kvalitativne i kvantitativne&nbsp;zavisnosti konstanti stabilnosti kompleksa kobalta(II) sa&nbsp;halogenidnim jonima od temperature i molskog odnosa&nbsp;komponenti sistema NH₄NO₃∙(2,61-z)CH₃CONH₂∙zH₂O odnosno &nbsp;NH₄NO₃∙zDMSO, određivanje termodinamičkih parametara koji&nbsp;karakteri&scaron;u reakcije asocijacije u ovim sistemima.</p> / <p>In this dissertation the complex formation between cobalt(II) and halide ions in the melts consisting of one inorganic salt, an organic compound and/or water has been studied at different temperatures. Two molten salt systems have been investigated: 1) &nbsp;ammonium nitrate-acetamide-water system NH₄NO₃∙(2,61-z)CH₃CONH₂∙zH₂O (z = 0.0 1.61 2.61) and 2) ammonium nitratedimethyl sulfoxide NH₄NO₃∙zDMSO (z = 1 &ndash; 6), at four different temperatures: 35, 45, 55 and 65 ^oC. The purpose of this work was to determine stability of cobalt(II) complexes formed with the halide ions and the components of the solvents, to discribe the qualitative and quantitative relationships between the stability of the complexes and the melt composition, as well as the changes in the cobalt(II) coordination due to a complex formation. Thermodynamic parameters for cobalt(II) &ndash; halide association process in these melts were alsodetermined.</p>

<p>U ovoj doktorskoj disertaciji proučavano je građenje kompleksa&nbsp;kobalta(II) sa hloridnim i &nbsp;bromidnim jonima u vodenim&nbsp;rastopima tipa&nbsp;xCa(NO3)2&middot;zH₂O&nbsp;&ndash; (1-x)NH₄NO_3&nbsp;pri različitom sastavu soli (x= 0,3 &ndash; 0,9), različitom sadržaju vode (z= 2,67 &ndash; 6,67) i na različitim temperaturama (45, 55 i 65^oC).</p><p>Istraživanja su imala za cilj proučavanje reakcije građenja&nbsp;kompleksa kobalta(II) sa hloridnim i bromidnim jonima u&nbsp;vodenim rastopima xCa(NO₃)₂&middot;zH₂O &ndash; (1&ndash;x)NH₄NO₃, &nbsp;uticaja&nbsp;temperature, sastava elektrolita i sadržaja vode na proces&nbsp;kompleksiranja, kao i određivanje termodinamičkih parametara&nbsp;koji karakteri&scaron;u reakcije asocijacije u ovim sistemima. Ovakvi&nbsp;rastopi soli su posebno interesantni zbog svoje niske tačke&nbsp;topljenja i visoke latentne toplote topljenja pa se mogu koristiti&nbsp;kao fazno-promenljivi materijali za skladi&scaron;tenje toplotne&nbsp;energije.</p> / <p>In this thesis absorption spectra of cobalt(II) chloride and&nbsp;bromide in calcium nitrate &ndash; ammonium nitrate &ndash; water&nbsp;system of the composition&nbsp; xCa(NO₃)₂&middot;zH₂O &ndash; (1&ndash;x)NH₄NO_3&nbsp;(x= 0.30 - 0.90 and z= 2.67 &ndash; 6.67) have been&nbsp;investigated in the wavelength range 400-800 nm at three&nbsp;different temperatures: 45, 55 and 65^oC.</p><p>Temparature, composition of the melt and water&nbsp;content influence on complex formation reactions between&nbsp;cobalt(II) and halide ions in aqueous xCa(NO₃)₂&middot;zH₂O &ndash;<br />(1&ndash;x)NH₄NO₃ melts have been studied. Thermodynamic&nbsp;parameters for cobalt(II) &ndash; halide association process in&nbsp;different solvents also were determined.</p><p>Investigated systems are interesting because of&nbsp;their high values of latent heat of fusion and low melting&nbsp;points. Due to these reasons, some melts are proposed for&nbsp;<br />heat energy storage materials, usually known as phase&nbsp;change materials (PCM).</p>