119 research outputs found
Primena veze izmeÄu inflamacije i tumora u razvoju jedinjenja sa antitumorskom aktivnoÅ”Äu
It is estimated that up to 20% of cancer-related deaths are linked with inflammation
(1). Inhibition of inflammatory enzymes COX-2 and 5-LOX impacts cancer cells directly, or
indirectly via tumor microenvironment. Wider anticancer potential has been investigated for
a small group of COX-2 inhibitors (2), while there are no such data for dual COX-2 and 5-LOX
inhibitors. The main aim of the project is to select the most promising anticancer drug
candidates from a group of COX-2 and dual COX-2 and 5-LOX inhibitors (newly synthesized
and previously synthesized). New compounds will be designed using structure-based and
ligand-based in silico methods and synthesized. Cytotoxicity will be evaluated towards four
cancer cell lines by MTT assay. Wider anticancer potential of selected compounds, which
includes synergism with conventional chemotherapy and radiotherapy, inhibition of
angiogenesis and activity towards multidrug resistant cancer cells, will be investigated and
lead compounds will be identified. Mechanisms of action of lead compounds will be
proposed after bioinformatics analysis of genes expression. In vitro evaluation of passive
gastrointestinal absorption (PAMPA and BMC), binding to human serum albumin (HPLC and
electrochemistry) and metabolism (human liver microsomes) will be performed. QSPR,
QSRR and QSMARt models will be created and, together with analysis of metabolism, will be
used for the optimization of structures of lead compounds. The project will result in the
development of new anticancer drug candidates, make new and strengthen previously
established scientific collaborations and give starting point for potential clinical evaluations
of lead compounds.Procenjuje se da je do 20% smrtnih sluÄajeva koji su posledica tumora povezano sa
inflamacijom (1). Inhibicija enzima inflamacije COX-2 i 5-LOX utiÄe na tumorske Äelije
direktno ili indirektno preko tumorskog mikrookruženja. Širi antitumorski potencijal je do
sada ispitan za malu grupu COX-2 inhibitora (2), dok takva istraživanja nisu do sada vrŔena
na dualnim COX-2 i 5-LOX inhibitorima. Glavni cilj projekta je da se identifikuju najbolji
kandidati za antitumorske lekove iz grupe COX-2 i grupe dualnih inhibitora COX-2 i 5-LOX
(novosintetisana i prethodno sintetisana jedinjenja). Nova jedinjenja Äe biti dizajnirana
primenom in silico metoda koje se zasnivaju na poznavanju strukture receptora i liganda,
nakon Äega Äe biti sintetisana. CitotoksiÄnost Äe biti ispitana na Äetiri tumorske Äelijske linije
primenom MTT testa. Å iri antitumorski potencijal odabranih jedinjenja, koji podrazumeva
sinergistiÄko dejstvo sa konvencionalnom hemoterapijom i radioterapijom, inhibiciju
angiogeneze i aktivnost prema multidrug rezistentnim Äelijskim linijama, Äe biti ispitan,
nakon Äega Äe biti identifikovana vodeÄa (lead) jedinjenja. Mehanizam delovanja vodeÄih
jedinjenja Äe biti predložen nakon bioinformatiÄke analize ekspresije gena. BiÄe izvrÅ”ena in
vitro procena pasivne gastrointestinalne apsorpcije (PAMPA i BMC metodama), vezivanja za
humani serumski albumin (HPLC i elektrohemijkim metodama) i metabolizma primenom
humanih mikrozomnih enzima jetre. QSPR, QSRR i QSMARt modeli Äe biti formirani i, zajedno
sa analizom metabolizma, biÄe upotrebljeni za optimizaciju struktura vodeÄih jedinjenja.
Rezultat projekta Äe biti novi kandidati za antitumorske lekove, uspostavljanje novih i
jaÄanje postojeÄih nauÄno-istraživaÄkih saradnji i postavljanje polazne taÄke za potencijalna
kliniÄka ispitivanja vodeÄih jedinjenja.VIII Kongres farmaceuta Srbije sa meÄunarodnim uÄeÅ”Äem, 12-15.10.2022. Beogra
Sinteza i fiziÄkoāhemijska karakterizacija tri novosintetisana derivata sulfhidroksamske kiseline kao potencijalnih dualnih inhibitora enzima ciklooksigenazeā2 i 5ālipooksigenaze
Inflammatory mediators derived from arachidonic acid by the enzymes
cyclooxygenase (COX) and lipoxygenase (LOX) are involved in the pathogenesis of various
inflammatory diseases. Inhibition of the COX pathway is thought to lead to potentiation of
the LOX pathway, and inhibition of both pathways represents a rational approach to the
design and development of more effective and safer anti-inflammatory drugs (1). The aim of
this study was the synthesis and physico-chemical characterization of 1f, 1g and 1h
derivatives derived from a previously conducted 3D-QSAR study and molecular docking (2).
The sulfhydroxamic acid derivative 1f was synthesized in a two-step process. Sulfonyl
chloride was synthesized from commercially available sulfonic acid and thionyl chloride in
the presence of a catalytic amount of DMF. Sulfhydroxamic acid was further obtained from
previously synthesized sulfonyl chloride and hydroxylamine hydrochloride in the presence
of 10% NaHCO3 solution. Sulfhydroxamic acid derivatives, 1g and 1h were synthesized from
commercially available sulfonyl chlorides and hydroxylamine hydrochloride in the presence
of 10% NaHCO3 solution. The reaction mixtures were purified by liquid-liquid extraction and
preparative TLC to give derivatives 1f, 1g, 1h in yields: 26%, 53% and 63%. The structure
and purity of the synthesized compounds were confirmed by determination of the melting
points and spectroscopic techniques (ATR-FTIR, 1H-NMR, 13C-NMR, MS/MS). Based on a
previously conducted in silico study, the voluminous sulfhydroxamic group is responsible for
iron chelation within 5-LOX active center and for COX-2 selectivity, as COX-2 has wider side
pocket, so potent inhibition of COX-2 and 5-LOX enzymes is expected.Inflamatorni medijatori koji nastaju iz arahidonske kiseline posredstvom enzima
ciklooksigenaze (COX) i lipooksigenaze (LOX) uÄestvuju u patogenezi brojnih inflamatornih
oboljenja. Smatra se da inhibicija COX puta dovodi do potenciranja LOX puta, te inhibicija oba
puta predstavlja racionalan pristup dizajniranja i razvoja novih, efikasnijih i bezbednijih
antiinflamatornih lekova (1). Cilj rada je bila sinteza i fiziÄko-hemijska karakterizacija
derivata 1f, 1g i 1h proisteklih iz prethodno sprovedene 3D-QSAR studije i molekulskog
docking-a (2). Derivat sulfhidroksamske kiseline 1f je sintetisan u dvostepenom postupku pri
Äemu se najpre iz komercijalno dostupne sulfonske kiseline i tionil hlorida u prisustvu
katalitiÄke koliÄine DMF dobija odgovarajuÄi sulfonil hlorid. OdgovarajuÄa sulfhidroksamska
kiselina (1f) se dobija u drugom koraku iz prethodno sintetisanog sulfonil hlorida i
hidroksilamin-hidrohlorida u prisustvu 10% rastvora NaHCO3 . Derivati sulfhidroksamske
kiseline, 1g i 1h su sintetisani jednostepenim postupkom iz komercijalno dostupnih sulfonil
hlorida i hidroksilamin-hidrohlorida u prisustvu 10% rastvora NaHCO3 . Nakon postupka
sinteze jedinjenja su preÄiÅ”Äena teÄno-teÄnom ekstrakcijom i preparativnom TLC pri Äemu se
dobijaju derivati 1f, 1g, 1h u prinosima 26%, 53% i 63%. Struktura i ÄistoÄa sintetisanih
jedinjenja je potvrÄena odreÄivanjem temperature topljenja i spektroskopskim (ATR-FTIR,
1 H-NMR, 13 C-NMR, MS/MS) tehnikama. Na osnovu prethodno sprovedene in silico studije,
voluminozni centar i sulfhidroksamska grupa sintetisanih derivata su odgovorni za COX-2
selektivnost zbog prisustva Ŕireg vezivnog mesta unutar COX-2 enzima, dok
sulfhidroksamska grupa unutar 5-LOX enzima helira gvožÄe aktivnog centra i inhibira enzim,
te se oÄekuje potentna dualna COX-2 i 5-LOX inhibitorna aktivnost sintetisanih derivata.VIII Kongres farmaceuta Srbije sa meÄunarodnim uÄeÅ”Äem, 12-15.10.2022. Beogra
Primena hromatografskih tehnika u optimizaciji procesa preÄiÅ”Äavanja amida kortienske kiseline iz hidrokortizona i etil estra L-glicina
Soft ('antedrug') glucocorticoids are pharmacologically active compounds which are biotransformed in a predictable and controllable way to inactive and non-toxic metabolites. Amides of cortienic acids (17(-carboxamide derivatives of glucocorticoids) are potential soft drugs with fewer side effects than traditional glucocorticoids. The purification of 17(- carboxamide derivatives of hydrocortisone was explained using the amide of hydrocortisonederived cortienic acid and ethyl ester of L-glycine as an example and performed by use of column chromatography and preparative thin-layer chromatography (TLC). The optimization of purification process was performed employing analytical TLC and reversed-phase highperformance liquid chromatography (RP-HPLC). The mobile phase that enables best chromatographic separation of the amide from impurities on TLC plate (chloroform-methanol (95:5 V/V)) was selected and modified (reduction of polarity and addition of glacial acetic acid) to be used for the column chromatography and preparative TLC purification. It was confirmed by use of RP-HPLC that purification procedures applied in this study resulted in pure (96.2 %) amide.Soft ('antedrug') glukokortikoidi su farmakoloÅ”ki aktivna jedinjenja koja se biotransformiÅ”u na predvidiv i kontrolisan naÄin do neaktivnih i netoksiÄnih metabolita. Amidi kortienskih kiselina (17(-karboksamidni derivati glukokortikoida) su potencijalni soft lekovi sa manje neželjenih efekata u odnosu na konvencionalne glukokortikoide. PreÄiÅ”Äavanje 17(- karboksamidnih derivata hidrokortizona je prikazano na primeru amida kortienske kiseline iz hidrokortizona i etil estra L-glicina i vrÅ”i se primenom hromatografije na koloni i preparativne tankoslojne hromatografije (TLC). Optimizacija procesa preÄiÅ”Äavanja je izvrÅ”ena primenom analitiÄke TLC i reverzno-fazne teÄne hromatografije pod visokim pritiskom (RP-HPLC). Mobilna faza koja omoguÄuje najbolje hromatografsko razdvajanje amida od neÄistoÄa na TLC ploÄici (hloroform-metanol (95:5 V/V) je odabrana i izvrÅ”ena je njena modifikacija (smanjenje polarnosti, odnosno dodatak glacijalne sirÄetne kliseline) u cilju preÄiÅ”Äavanja hromatografijom na koloni, odnosno preparativnom TLC. Primenom RP-HPLC je potvrÄeno da su navedeni postupci preÄiÅ”Äavanja omoguÄili dobijanje amida stepena ÄistoÄe 96,2 %
Molecular Docking Analysis of Novel Thiourea Derivatives of Naproxen with Potential Antitumor Activity
Naproxen, as a propionic acid derivative, causes serious gastrointestinal side effects due to the presence of free carboxylic group. In that sense, masking of carboxylic group with other pharmacophores may be a promising strategy to decrease gastrointestinal toxicity. Thiourea derivatives have been intensively investigated as potential antitumor drugs, whereby their activity is based on potential inhibition of protein kinases, topoisomerases, carbonic anhydrase and sirtuins. In addition, it was shown that inhibition of certain protein kinases might reverse resistance to chemotherapeutic drugs by enhancing the cell death in the presence of low concentrations of drug. Twenty new thiourea derivatives of naproxen were designed and their binding to four selected protein kinases involved in tumor multidrug resistance (AKT2, mTOR, EGFR and VEGFR1) was estimated using two molecular docking programs (AutoDock Vina and OEDocking). According to OEDocking, the highest potential to inhibit AKT2 and mTor has derivative 1, while derivative 20 demonstrates the highest potential towards EGFR and VEGFR1. According to AutoDock Vina, the highest potential for inhibition of EGFR, AKT2 and VEGFR1 have derivatives 16 and 17. Therefore, derivatives 1, 16, 17 and 20 are potentially the most potent protein kinase inhibitors that could be further synthesized and tested for anticancer activity
Synthesis and RP-TLC lipophilicity evaluation of a novel fluocinolon acetonide soft drug derivative
Cortienic acid was obtained by periodic acid oxidation of fluocinolone acetonide, whereas corresponding amide was synthesized from the cortienic acid and ethyl ester of b-alanine by dicyclohexylcarbodiimide - hydroxybenzotriazole coupling procedure. Lipophilicity of the amide was evaluated by using reversed-phase thin-layer chromatography systems, consisting of ethanol and water in various ratios, and was higher in comparison to fluocinolone acetonide and cortienic acid
Development and Validation of a New Isocratic RP-HPLC Method for Simultaneous Determination of Sodium Metabisulfite and Sodium Benzoate in Pharmaceutical Formulation
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
Razvoj i validacija metode teÄne hromatografije za odreÄivanje acetilsalicilne i salicilne kiseline u doziranim oblicima
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 Ī¼m) thermostated at 35Ā°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, analgetiÄko i antipiretiÄko 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, veliÄina Äestica 5 Ī¼m) na temperaturi od 35Ā°C. Mobilnu fazu Äine smeÅ”a A i B u odnosu 65:35 (V/V). SmeÅ”a A je voda HPLC ÄistoÄe 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 sledeÄi parametri validacije: selektivnost, linearnost, preciznost, taÄnost 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 Å”est razliÄitih komercijalnih preparata. Svi rezultati ispitivanja su u dozvoljenim granicama specifikacija proizvoÄaÄa. Predložena HPLC metoda pod datim eksperimentalnim uslovima predstavlja brz, jednostavan, taÄan i selektivan postupak za istovremeno odreÄivanje acetilsalicilne i salicilne kiseline u doziranim oblicima
Influence of Solid Drug Delivery System Formulation on Poorly Water-Soluble Drug Dissolution and Permeability
The majority of drugs have a low dissolution rate, which is a limiting step for their absorption. In this manuscript, solid dispersions (SD), solid self-microemulsifying drug delivery systems (S-SMEDDS) and solid self-nanoemulsifying drug delivery systems (S-SNEDDS) were evaluated as potential formulation strategies to increase the dissolution rate of carbamazepine. Influence of increased dissolution rate on permeability of carbamazepine was evaluated using PAMPA test. In S-SMEDDS and S-SNEDDS formulations, the ratio of liquid SMEDDS/SNEDDS and solid carrier (Neusilin((R)) UFL2) was varied, and carbamazepine content was constant. In SD formulations, the ratio of carbamazepine and Neusilin((R)) UFL2, was varied. Formulations that showed the best dissolution rate of carbamazepine (SD_1:6, SMEDDS_1:1, SNEDDS_1:6) were mutually compared, characterization of these formulations was performed by DSC, PXRD and FT-IR analyses, and a PAMPA test was done. All formulations have shown a significant increase in dissolution rate compared to pure carbamazepine and immediate-release carbamazepine tablets. Formulation S-SMEDDS_1:1 showed the fastest release rate and permeability of carbamazepine. DSC, PXRD and FT-IR analyses confirmed that in S-SMEDDS and S-SNEDDS carbamazepine remained in polymorph form III, and that it was converted to an amorphous state in SD formulations. All formulations showed increased permeability of carbamazepine, compared to pure carbamazepine
The use of PAMPA for skin permeability and retention evaluation of metilprednisolone - derived cortienic acid derivatives as potential soft glucocorticoids
Amidi ili estri kortienske kiseline metilprednizolona predstavljaju potencijalne
soft glukokortikoide sa manje izraženim neželjenim efektima u odnosu na
konvencionalne glukokortikoide. Retencija i permeabilnost su važne osobine soft
glukokortikoida za primenu na kožu koje mogu znaÄajno uticati na njihovu aktivnost i
pojavu neželjenih efekata. Jedna od in vitro metoda koja se najÄeÅ”Äe koristi za procenu
ovih osobina je PAMPA (Parallel Artificial Membrane Permeability Assay). Cilj ovog
rada je procena permeabilnosti i retencije u koži pet amida (MPMA, MPMAB, MPMAIB,
MPMGB i MPPA) i jednog tioestra (MPEMP) kortienske kiseline metilprednizolona
primenom PAMPA testa.
Procena permeabilnosti i retencije u koži izvrŔena je na hidrofobnoj Millipore
PVDF PAMPA mikrotitarskoj ploÄi sa 96 odeljaka. PraÄena je difuzija ispitivanih
jedinjenja kroz membranu koju Äine 70 % silikonsko ulje i 30 % izopropilmiristat.
Koncentracije u polaznim rastvorima, donorskim i akceptorskim odeljcima odreÄene su
primenom LCāMS/MS metode.
Primenom PAMPA testa odreÄeni su koeficijenti permeabilnosti (logPe) i
retencije (R). Vrednosti logPe su u opsegu od ā6,81 do ā5,09, dok su vrednosti R u
opsegu 1,52 ā 65,25. Jedinjenje sa najnižom vrednoÅ”Äu logPe je MPMGB, dok su za
MPEMP odreÄene najviÅ”e vrednosti parametara logPe i R, te se od ovog derivata mogu
oÄekivati najveÄa permeabilnost i retencija u koži.
Permeabilnost i retencija u koži pet amida (MPMA, MPMAB, MPMAIB, MPMGB i
MPPA) i jednog tioestra (MPEMP) kortienske kiseline metilprednizolona procenjeni su
primenom PAMPA testa. NajviŔe vrednosti parametara logPe i R su dobijene za MPEMP,
te se ovaj derivat izdvaja kao najpovoljniji za primenu na kožu.Amides or esters of methylprednisoloneāderived cortienic acid are potential soft glucocorticoids with fewer side effects in comparison to traditional glucocorticoids. Retention and permeability are important properties of soft glucocorticoids for local skin application which could significantly influence their activity and occurrence of side effects. PAMPA (Parallel Artificial Membrane Permeability Assay) is one of the mostly used in vitro methods for the estimation of these properties. The aim of this study was estimation of skin permeability and retention of five amides (MPMA, MPMAB, MPMAIB, MPMGB and MPPA) and one thioester (MPEMP) of metilprednisolone ā derived cortienic acid using PAMPA. Estimation of skin permeability and retention was performed on hydrophobic Millipore PVDF PAMPA microtiter 96āwell plate. Diffusion of tested compounds through membrane, consisting of 70% silicon oil and 30% isopropyl myristate, was tested. Concentrations in starting solutions, as well as in donor and acceptor compartments were determined using LCāMS/MS method. PAMPA permeability coefficients (logPe) and retention (R) were determined. logPe values ranged from ā6.81 to ā5.09, whereas R values ranged from 1.52 to 65.25. Derivative with the lowest value of logPe was MPMGB, whereas the highest values of logPe and R were determined for MPEMP. Therefore, highest skin permeability and retention could be expected from MPEMP. Skin permeability and retention of five amides (MPMA, MPMAB, MPMAIB, MPMGB and MPPA) and one thioester (MPEMP) of metilprednisolone ā derived cortienic acid was estimated by use of PAMPA. The highest values of logPe and R were calculated for MPEMP, which could be considered the best candidate for skin application.VII Kongres farmaceuta Srbije sa meÄunarodnim uÄeÅ”Äem
Zajedno stvaramo buduÄnost farmacije,
Beograd, 10-14. oktobar 2018
- ā¦