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Apart from cytotoxicity, inhibitors of the COX-2 enzyme have demonstrated additional effects important for cancer treatment (such as radiosensitization of tumor cells and cell antimigratory effects); however, the relationship between the inhibition of other inflammation-related enzyme 5-LOX inhibitors and anticancer activity is still not well understood. In our study, the cytotoxicity of thirteen COX-2 and 5-LOX inhibitors previously presented by our group (1-13) was tested on three cancer cell lines (HCT 116, HT-29 and BxPC-3) and one healthy cell line (MRC-5). Compounds 3, 5, 6 and 7 showed moderate cytotoxicity, but good selectivity towards cancer cell lines. IC50 values were in the range of 22.99-51.66 µM (HCT 116 cell line), 8.63-41.20 µM (BxPC-3 cell line) and 24.78-81.60 µM (HT-29 cell line; compound 7 > 100 µM). In comparison to tested, commercially available COX-2 and 5-LOX inhibitors, both cytotoxicity and selectivity were increased. The addition of compounds 6 and 7 to irradiation treatment showed the most significant decrease in cell proliferation of the HT-29 cell line (p < 0.001). The antimigratory potential of the best dual COX-2 and 5-LOX inhibitors (compounds 1, 2, 3 and 5) was tested by a wound-healing assay using the SW620 cell line. Compounds 1 and 3 were singled out as compounds with the most potent effect (relative wound closure was 3.20% (24 h), 5,08% (48 h) for compound 1 and 3.86% (24 h), 7.68% (48 h) for compound 3). Considering all these results, compound 3 stood out as the compound with the most optimal biological activity, with the best dual COX-2 and 5-LOX inhibitory activity, good selectivity towards tested cancer cell lines, significant cell antimigratory potential and a lack of toxic effects at therapeutic doses.

Gentamicin sulfate is a potent broad spectrum aminoglycoside antibiotic which is used against Gram- positive and Gram-negative bacteria. A simple, isocratic HPLC method for separation, identification and determination of gentamicin and parabens (methylparaben and propylparaben) was developed and validated. To our knowledge there is no report about simultaneous determination of those three ana- lytes in pharmaceutical products. The optimum chromatographic conditions were achieved on CN column with a mobile phase consisting of 0.15% triethylamine in 10 mM KH 2 PO4 aqueous solution (final pH 3.0 adjusted with H 3 PO 4 ) and methanol in the ratio 70:30 (v/v), providing selective quan- tification of analytes within 5 min. The method was successfully validated according to ICH guidelines acceptance criteria in terms of selectivity, linearity, accuracy, precision and robustness. The linearity of the method was proved in defined concentrations ranges for gentamicin (0.32–1.04 mg mL1 ), meth- ylparaben (0.0072–0.0234 mg mL1 ) and propylparaben (0.0008–0.0026 mg mL1 ). Relative standard deviations calculated for all analytes in precision testing were <2% (analysis repeatability) and <3% (intermediate precision). Recovery values were between 98.87% and 101.67%. Chromatographic pa- rameters are not significantly influenced by small variations of column temperature, pH and molarity of KH2 PO 4 . Finally, the method was successfully applied for quantitative determination of gentamicin and parabens in commercially available solution for injection. Proposed HPLC method is found to be promising in terms of simplicity, analysis times and non-use of derivatization and ion-pair agents.

Drug discovery and development is a very challenging, expensive and time-consuming process. Impressive technological advances in computer sciences and molecular biology have made it possible to use computer-aided drug design (CADD) methods in various stages of the drug discovery and development pipeline. Nowadays, CADD presents an efficacious and indispensable tool, widely used in medicinal chemistry, to lead rational drug design and synthesis of novel compounds. In this article, an overview of commonly used CADD approaches from hit identification to lead optimization was presented. Moreover, different aspects of design of multitarget ligands for neuropsychiatric and anti-inflammatory diseases were summarized. Apparently, designing multi-target directed ligands for treatment of various complex diseases may offer better efficacy, and fewer side effects. Antipsychotics that act through aminergic G protein-coupled receptors (GPCRs), especially dopamine D2 and serotonin 5-HT2A receptors, are the best option for treatment of various symptoms associated with neuropsychiatric disorders. Furthermore, multi-target directed cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) inhibitors are also a successful approach to aid the discovery of new anti-inflammatory drugs with fewer side effects. Overall, employing CADD approaches in the process of rational drug design provides a great opportunity for future development, allowing rapid identification of compounds with the optimal polypharmacological profile., Proces otkrića i razvoja lekova je veoma zahtevan, skup i dugotrajan. Veliki tehnološki napredak u molekularnoj biologiji i kompjuterskim naukama je omogućio primenu metoda kompjuterski potpomognutog dizajniranja lekova (CADD) u različitim fazama procesa otkrića i razvoja lekova. Danas CADD predstavlja efikasnu i nezamenljivu alatku, koja se široko koristi u medicinskoj hemiji za racionalni dizajn i sintezu novih jedinjenja. U ovom preglednom radu biće prikazani CADD pristupi koji se najčešće koriste od procesa identifikacije hit jedinjenja do optimizacije lead jedinjenja. Pored toga, biće predstavljeni različiti aspekti u dizajnu višeciljnih liganada za neuropsihijatrijske i inflamatorne bolesti. Pokazano je da su ova jedinjenja veoma efikasna u lečenju složenih bolesti zbog veće efikasnosti i manje neželjenih efekata koje izazivaju. Antipsihotici koji deluju preko aminergičnih G-protein spregnutih receptora (GPCR), posebno preko dopaminskih D2 i serotoninskih 5-HT2A receptora, predstavljaju najbolju opcija za lečenje različitih simptoma povezanih sa neuropsihijatrijskim poremećajima. Pored toga, dizajn i sinteza dualnih inhibitora ciklooksigenaze-2 (COX-2) i 5lipoksigenaze (5-LOX) takođe predstavlja uspešan pristup u otkrivanju novih antiinflamatornih lekova sa manje neželjenih efekata. Na kraju se može zaključiti da primena CADD metoda u procesu racionalnog dizajniranja lekova pruža značajnu priliku za dalji napredak jer omogućava brzu identifikaciju jedinjenja sa optimalnim polifarmakološkim profilom.

A series of eleven 9-acridinyl amino acid derivatives were synthesized using a two-step procedure. Cytotoxicity was tested on the K562 and A549 cancer cell lines and normal diploid cell line MRC5 using the MTT assay. Compounds 6, 7, 8 and 9 were the most active, with IC50 values comparable to or lower than that of chemotherapeutic agent amsacrine. 8 and 9 were especially effective in the A549 cell line (IC50 ≈ 6 μM), which is of special interest since amsacrine is not sufficiently active in lung cancer patients. Cell cycle analysis revealed that 7 and 9 caused G2/M block, amsacrine caused arrest in the S phase, while 6 and 8 induced apoptotic cell death independently of the cell cycle regulation. In comparison to amsacrine, 6, 7, 8, and 9 showed similar inhibitory potential towards topoisomerase II, whereas only 7 showed DNA intercalation properties. In contrast to amsacrine, 6, 7, 8 and 9 showed a lack of toxicity towards unstimulated normal human leucocytes.

A simple and convenient reversed-phase high-performance liquid chromatography (RP-HPLC) method for simultaneous separation, identification, and determination of sodium metabisulfite and sodium benzoate in pharmaceutical formulation has been developed and validated. Chromatographic separation was achieved on RP column Zorbax Extend C-18 (150 x 4.6 mm i.d., 3.5 mu m particles), and mixture of 0.1% phosphoric acid and acetonitrile in the ratio 62: 38 (v/v) was used as a mobile phase. The flow rate was set at 1.0 mL/min with detection wavelength of 275 nm. The method was successfully validated according to International Conference on Harmonization (ICH) guidelines acceptance criteria. The method is selective, as no interferences were observed at retention times corresponding to these analytes. Results of regression analyses (r) and statistical insignificance of calibration curve intercepts (p) proved linearity of the method in defined concentration ranges for sodium metabisulfite and sodium benzoate (0.05-0.15 mg/mL). Relative standard deviations calculated for both analytes in precision testing were below the limits defined for active pharmaceutical ingredients (analysis repeatability: lt 2%; intermediate precision: lt 3%). Recovery values were between 98.16% and 101.94%. According to results of robustness testing, chromatographic parameters are not significantly influenced by small variation of acetonitrile content in mobile phase, column temperature, and flow rate. Finally, the method was applied for quantitative determination of investigated preservatives in real sample analysis.

Acetylsalicylic acid belongs to nonsteroidal anti-inflammatory drugs with antiinflammatory, analgesic and antipyretic properties. The aim of this work was development and validation of HPLC method for qualitative and quantitative analysis of acetylsalicylic acid and its major degradation product, salicylic acid, in dosage forms. The optimal separation was achived using Zorbax Eclipse XDB-C18 Analytical column (4,6x150 mm, particle size 5 μm) thermostated at 35°C. Mobile phase consisted of eluents A and B in ratio 65:35 (V/V). As the eluent A, water of HPLC purity and 85% phosphoric acid in ratio 80:0.5 (V/V) were used, while acetonitrile was used as the eluent B. The flow rate was 1.0 mL/min and UV detection was performed at 240 nm. The method was validated in terms of selectivity, linearity, precision, accuracy and robustness for both analytes, as well as limits of detection and quantification for salicylic acid. The obtained results meet the requirements of analytical procedures validation. The proposed HPLC method was applied in qualitative and quantitative analysis of acetylsalicylic and salicylic acids in six different forms of drugs. All obtained results meet the requirements of manufacturer specifications. The established HPLC method was found to be rapid, simple, accurate and selective for simultaneous determination of acetylsalicylic and salicylic acids in dosage forms., Acetilsalicilna kiselina pripada grupi nesteroidnih antiinflamatornih lekova koji ispoljavaju antiinflamatorno, analgetičko i antipiretičko delovanje. Cilj ovog rada je bio razvoj i validacija HPLC metode za kvalitativnu i kvantitativnu analizu acetilsalicilne kiseline i njenog degradacionog proizvoda, salicilne kiseline, u doziranim oblicima. Optimalno razdvajanje ispitivanih analita postignuto je na koloni Zorbax Eclipse XDB-C18 Analytical (4,6x150 mm, veličina čestica 5 μm) na temperaturi od 35°C. Mobilnu fazu čine smeša A i B u odnosu 65:35 (V/V). Smeša A je voda HPLC čistoće i 85% fosforna kiselina u odnosu 80:0,5 (V/V), a B je acetonitril. Protok je bio 1,0 mL/min, a talasna dužina detekcije 240 nm. Metoda je validirana i ispitani su sledeći parametri validacije: selektivnost, linearnost, preciznost, tačnost i robusnost za oba analita, kao i limiti detekcije i kvantifikacije za salicilnu kiselinu. Dobijene vrednosti su u skladu sa definisanim kriterijumima za validaciju metode. Predložena HPLC metoda je primenjena za kvalitativnu i kvantitativnu analizu acetilsalicilne i salicilne kiseline u šest različitih komercijalnih preparata. Svi rezultati ispitivanja su u dozvoljenim granicama specifikacija proizvođača. Predložena HPLC metoda pod datim eksperimentalnim uslovima predstavlja brz, jednostavan, tačan i selektivan postupak za istovremeno određivanje acetilsalicilne i salicilne kiseline u doziranim oblicima.

The biological activity of three previously synthesized 17 beta-carboxamide glucocorticoids (BG, BEG, and MPEA) was tested in vitro on mitogen stimulated and non-stimulated peripheral blood mononuclear cells (MNCs) and granulocytes from human healthy donors, and the results were compared to the conventional glucocorticoid dexamethasone. The tested 17 beta-carboxamide glucocorticoids did not induce decreases in MNC viability and proliferation, while modulation of reactive oxygen species (ROS) synthesis in granulocytes was dependent on the cell donor. The obtained results indicate the possibility of avoidance of strong lymphocyte suppression, which is generally recognized during administration of conventional glucocorticoids. Furthermore, the metabolism of the tested derivatives was predicted in silico. The predicted metabolites were synthesized and the in silico results were confirmed by in vitro evaluation of the metabolism of BG, BEG, and MPEA in human serum and in cultures of peripheral blood MNCs. The results of the biological activity and metabolism evaluation and of previous in vivo evaluations of biological activity indicate the soft drug nature of BG, BEG, and MPEA. In order to be fully considered as soft glucocorticoids, further investigations on the toxicity and activity of the formed metabolites are required.

Acridine ring can be found in molecules used in many different spheres, including industry and medicine. Nowadays, even acridines with antibacterial activity are of research interest due to increasing bacterial resistance. Some acridine derivatives showed antimalarial or antiviral activity. Acridine derivatives were also investigated for antitumor activity due to the interaction with topoisomerase II and DNA base pairs. Considering these possible uses of acridine derivatives, this work was made as overview of all significant structure characteristics for specific action of these compounds.

Octanol-water partition coefficients (log Po/w) of fifteen previously synthesized 17 beta-carboxamide glucocorticoid derivatives and prednisolone were determined using shake-flask method. The retention behavior of selected compounds was tested in five reversed-phased thin-layer chromatography (RP-TLC) systems, consisting of water and organic solvent (acetonitrile, acetone, ethanol 96%, methanol, or tetrahydrofuran), and chromatography parameters (R-M(0), S, and C-0) were calculated. Simple linear regression (SLR) analysis was performed, and the correlation between log Po/w and chromatography parameters was tested. The most appropriate RP-TLC system for log Po/w prediction was that consisting of water and ethanol 96% as the mobile phase. Statisitical evaluation of the quality of models created using this system proved their reliability for log Po/w prediction of new 17 beta-carboxamide glucocorticoid derivatives, and the model with the most favorable statistical parameters was underlined.

Type 2 diabetes is a metabolic syndrome characterized by relative insulin deficiency, insulin resistance and increased hepatic glucose output. Medications used to treat the disease (hypoglycemic agents) are divided in seven distinct classes. One of those classes, thiazolidindiones ('glitazones'), are known as insulin sensitizers and act mainly by improving peripheral uptake and utilization of glucose in muscle and fat, finally decreasing liver glucose production. These drugs activate one or more peroxisome proliferator-activated receptors (PPARs), which regulate gene expression in response to ligand binding. The thiazolidindiones differ by the nature of the groups attached to the 2,4-thiazolidindione nucleus. These agents are extensively metabolized with all metabolic changes occurring on thiazolidine ring or adjacent aryl group. Considerable interest in the metabolism of troglitazone exists, because hepatic toxicity may be associated with a quinone-type metabolite of troglitazone. Thiazolidindiones (rosiglitazone and pioglitazone) have been approved for monotherapy and combination therapy with metformin, sulfonylureas or insulin. Unfortunately, some studies and metaanalyses suggested that both drugs improve a number of cardiovascular risk factors and enhance incidence of heart failure., Dijabetes tipa 2 je metaboličko oboljenje koje karakteriše relativni nedostatak insulina, rezistencija na insulin i povećano oslobađanje glukoze iz jetre. Lekovi koji se koriste u terapiji ovog oboljenja (hipoglikemici) mogu se klasifikovati u sedam različitih grupa. Tiazolidindioni (glitazoni) povećavaju osetljivost na insulin, preuzimanje i potrošnju glukoze u mišićima i masnom tkivu, a sve to utiče na smanjeno oslobađanje glukoze iz jetre. Ovi lekovi su agonisti PPAR receptora (receptori aktivirani proliferatorom peroksizoma) i dovode do transkripcije gena uključenih u metabolizam glukoze i lipida. Tiazolidindioni se međusobno razlikuju po strukturi supstituenata na 2,4-tiazolidindionu. Podležu intenzivnom metabolizmu u jetri, sa svim metaboličkim promenama na tiazolidinskom prstenu ili susednoj aril grupi. Postoji veliko interesovanje za metabolizam troglitazona, jer se njegova hepatotoksičnost povezuje sa metabolitom troglitazona tipa hinona. Tiazolidindioni (rosigltazon i pioglitazon) su odobreni kao monoterapija ili u kombinaciji sa metforminom, derivatima sulfonilureje ili insulinom. Nažalost, novija istraživanja i meta-analize ukazale su na to da oba leka povećavaju faktore kardiovaskularnog rizika i učestalost srčane insuficijencije.

Biopartitioning micellar chromatography (BMC), which is useful to mimic the drug partitioning process in biological systems, was used as a method for determination of partition coefficients micelle/water (P-m/w) of a set of structurally diverse drugs. BMC is a chromatographic system consisting of polyoxyethylene (23) lauryl ether (Brij 35) micellar mobile phase prepared in physiological conditions and C-18 reversed stationary phase. The retention factors were determined for all selected and studied compounds and a correlation between P-m/w and oral drug absorption values were obtained. In order to study the similarity between the BMC system and other natural systems of biomembranes, the retention data on BMC were compared with permeability data obtained in Parallel artificial membrane permeation assays (PAMPA lipid model). Molecular descriptors that are believed to influence trans-cellular transport and oral drug absorption have been calculated in order to obtain a predictive and reliable model for drug permeability. Results presented in this paper indicate that a proposed descriptor for hydrophobicity P-m/w, provided, from a statistical point of view, is a better model than other hydrophobicity descriptors.