Cytotoxicity and antimicrobial activity of Teucrium scordium L. (Lamiaceae) extracts

Cytotoxicity and antimicrobial activity of cyclohexane, dichlormethane and methanol extracts of Teucrium scordium subspec. scordioides was studied. Cyclohexane and dichlormethane extracts of T. scordium possessed high citotoxicity against MDA-MB-361 cells (IC(50)=130.33+/-0.1 mu g/ml and IC(50)=189.89+/-3.99 mu g/ml, respectively). Dichlormethane extract was more effective against MDA-MB-453 cell line (IC(50)=130.33 +/- 0.1 mu g/ml). The methanol extract of T. scordium possessed no cytotoxicity against breast cancer cell lines, MDA-MB-361 and MDA-MB-453. Herb extracts of T. scordium have shown weak antibacterial activity on Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli and Bacillus subtilis with no activity against Staphylococcus aureus, S. epidermidis, Micrococcus luteus, Enterococcus faecalis, and Candida albicans

Антидијабетски потенцијал једноставних карбамата: компаративна експериментална и рачунарска студија

With the increasing global burden of diabetes mellitus type 2, the search for the new drugs, with better pharmacological profile is continued. As a part of this surge, the synthesis, pharmacological in vitro and computational evaluation of five, simple carbamate derivatives, against carbohydrate digestive enzyme α-glucosidase, is disclosed herein. Results of the experimental and computational assessment indicated that examined carbamates deterred the activity of α-glucosidase with acceptable IC50 values ranging from 65.34 to 79.89 μM compared to a standard drug acarbose (109.71 μM). Similarly, the studied compounds displayed in silico binding affinity for α-glucosidase enzyme with significant binding energies. Preliminary toxicity profiles of studied carbamates against three cancerous cell lines indicated their poor activity, suggesting that significant structural modifications have to be made to improve their anticancer efficiency. Results of the present study indicate that the examined carbamates were able to virtually or experimentally interact with an important target of diabetes mellitus type 2. Additionally, a new pharmacophore model is proposed featuring hydrogen bond donating carbamate –NH group, hydrogen bond accepting carbamate –OCH3 group and hydrophobic stabilization of aromatic moieties.Са порастом појаве дијабетеса типа 2 у свету, јавља се потрага за новим лековима са што ефикаснијим фармаколошким профилом. Као део овог истраживања, приказана је синтеза, фармаколошко in vitro и рачунарско испитивање пет карбамата једноставне структуре, као инхибитора – глукозидазе, ензима који учествује у дигестивном разлагању шећера. Резултати експерименталног испитивања показали су да испитивани карбамати инхибирају активност – глукозидазе са задовољавајућим IC50 вредностима у опсегу од 65,34 до 79,89 μM, а у поређењу са стандардним леком, акарбозом (109,71 μM). Такође, in silico методом добијене су значајне енергије везивања за активно место – глукозидазе. У прелиминарном испитивању цитотксичности према три типа канцерозних ћелија, карбамати су показали лошу активност, сугеришући да су потребне значајне структурне промене за побољшање њиховог антиканцерозног дејства. Уопштено говорећи, резултати ове студије показали су да су испитивани карбамати успешно виртуелно и експериментално интераговали са важном метом код дијабетеса типа 2. Такође је предложен и нови фармакофорни модел за -глукозидазу, који укључује карбаматну –NH групу као донора водоничне везе, затим карбаматну –OCH3 групу као акцептора водоничне везе, а такође и стабилизујуће хидрофобне интеракције ароматичних прстенова

Biological potential of marine macroalgae of the genus Cystoseira

In the present investigation the acetone extracts of three macroalgae, Cystoseira amentacea, Cystoseira barbata and Cystoseira compressa, were tested for antioxidant, antimicrobial and cytotoxic potential. As a result of the study, C. amentacea extract had more potent free radical scavenging activity (IC50 = 409.81 μg/mL) than C. barbata and C. compressa extracts. For reducing power, measured values of absorbance varied from 0.0352 to 0.8873. The IC50 values for superoxide anion scavenging activity for different extracts were 521.45–976.62 μg/mL. Total phenol and flavonoid contents in extracts were 39.96–81.28 μg PE/mg and 20.85–64.58 μg RE/mg respectively. Further, all three Cystoseira species exhibited a similar antimicrobial activity. The lowest MIC value (0.312 mg/mL) was shown in the extract obtained from C. compressa against Bacillus subtilis. Finally, extract of C. amentacea expressed the strongest cytotoxic activity toward tested cell lines with IC50 values ranging from 94.72 to 186.55 μg/mL. Based on these results, it can be stated that the tested macroalgae can be used as potential natural antioxidants and antimicrobial and cytotoxic agents

Chemical composition and bioactive properties of the lichen, Pleurosticta acetabulum

Purpose: To investigate the chemical composition and bioactivity of the acetone extract of Pleurosticta acetabulum lichen. Methods: Phytochemical analysis of the acetone extract of the lichen (Pleurosticta acetabulum) was carried out by high-performance liquid chromatography (HPLC). The antioxidant activity of the lichen extract was evaluated by determining the radical scavenging capacity on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and reducing power. To determine total phenolics and flavonoids, we used spectrophotometric methods. Antimicrobial activity was estimated by determination of the minimal inhibitory concentration using broth microdilution method. Anticancer activity of the lichen extract was tested using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT). Results: Salazinic, norstictic, protocetraric, evernic acid and atranorin were identified as compounds of lichen. P. acetabulum extract exhibited moderate free radical scavenging activity (half-maximum inhibitory concentration, IC50 of 151.7301 µg/mL). The spectrophotomeric absorbance of the extract for reducing power varied from 0.035 to 0.127, while the total phenolics and flavonoids in the extract were 35.39 µg PE/mg and 12.74 μg RE/mg, respectively. Minimum inhibitory concentration (MIC) was in the range of 1.25 to 20 mg/mL while cytotoxic activity (based on IC50 values) ranged from 24.09 to 45.94 µg/mL. Conclusion: The results confirm that lichen extract contains secondary metabolites that possess antioxidant, antimicrobial and anticancer activities, which opens up some possibilities for the extract to be developed as food supplements and pharmaceutical raw materials. Keywords: Pleurosticta acetabulum, Antioxidant activity, Antimicrobial activity, Cytotoxic activity, Liche

ACE and α‐glucosidase inhibitory activity of methanol extract of Alchemilla viridiflora Rothm. (Rosaceae)

Tanini, polifenolni biljni metaboliti, značajno smanjuju postprandijalnu hiperglikemiju inhibicijom α‐glukozidaze, i stoga mogu biti efikasna strategija u kontroli dijabetesa tipa 2. Takođe, dokazano je i da nespecifično inhibiraju aktivnost angiotenzin‐konvertujućeg enzima (ACE). Kako su tanini identifikovani samo u vrsti Alchemilla vulgaris L., cilj ovog istraživanja je da se odredi sadržaj tanina u do sada neistraženoj vrsti A. viridiflora Rothm. (Rosaceae), kao i inhibitorni uticaj na aktivnost ACE i α‐glukozidaze. Ukupni sadržaj tanina u metanolnom ekstraktu A. viridiflora određen je prema propisu Ph. Eur. 9.0. Suvi metanolni ekstrakt, enzimski rastvor (400 mU/ml α‐ glukozidaze u 0,1 M fosfatnom puferu) i supstrat, p‐nitrofenil α‐D‐glukopiranozid korišćeni su za kolorimetrijski test inhibitorne aktivnosti α‐glukozidaze. Kao pozitivna kontrola korišćena je akarboza. ACE inhibitorna aktivnost metanolnog ekstrakta ispitana je korišćenjem komercijalnog testa ACE Kit‐ WST (Dojindo Inc., Japan) prema uputstvu proizvođača. Procenat inhibicije enzima je izračunata IC50 vrednost, tj. procenjena koncentracija ekstrakta koja je izazvala 50% inhibicije aktivnosti enzima, koristeći linearnu regresionu analizu. IC50 vrednost metanolnog ekstrakta A. viridiflora, očitana sa dozno‐zavisne krive iznosi 2,6±0,5 μg/ml, i ekstrakt pokazuje bolju anti‐α‐glukozidaznu aktivnost od standarda akarboze (IC50=74,2±3,3 μg/ml). Takođe, ispitivani ekstrakt pokazuje doznozavisnu inhibiciju ACE pri IC50 2 μg/ml. Dobijeni rezultati su u korelaciji sa visokim sadržajem tanina u metanolnom ekstraktu A. viridiflora (3,74 %). Pokazane inhibicije angiotenzin‐konvertujućeg enzima i α‐glukozidaze čine metanolni ekstrakt vrste A. viridiflora pogodnim za dalje istraživanje u cilju pronalaženja novih prirodnih proizvoda značajnih za terapiju kardiovaskularnih bolesti i dijabetesa.Tannins, polyphenolic plant metabolites, significantly reduce postprandial hyperglycemia by inhibiting α‐glucosidase, and therefore can be an effective strategy for controlling type 2 diabetes. It has also been proven that they are non‐specific inhibitors of the activity of angiotensin‐converting enzyme (ACE). As tannins were identified only in the species Alchemilla vulgaris L., the aim of this study is to determine the content of tannins in the unexplored A. viridiflora Rothm. (Rosaceae), as well as the inhibitory effect on the activity of angiotensin‐converting enzyme and α‐glucosidase. The content of tannins in methanol extract of A. viridiflora was determined according to the Ph. Eur. 9.0. Dry methanol extract, enzyme solution (400 mU/ml of α‐ glucosidase in 0.1 M phosphate buffer) and substrate, p‐nitrophenyl α‐Dglucopyranoside were used for colorimetric α‐glucosidase inhibitory activity test. Acarbose was used as a positive control. The ACE inhibitory activity of the methanol extract was tested using the commercial ACE Kit‐WST (Dojindo Inc., Japan) according to the manufacturer's instructions. The percentage of enzyme inhibition is the calculated IC50 value, i.e. estimated concentration of the extract that caused 50% inhibition of enzyme activity using linear regression analysis. The IC50 of A. viridiflora methanol extract, read from the dose‐dependent curve, was 2.6±0.5 μg/mL, and this extract demonstrated better anti‐α‐glucosidase activity than standard acarbose (IC50=74.2±3.3 μg/mL). In addition, the examined extract shows a dose‐dependent inhibition of ACE with IC50 2 μg/mL. Obtained results were in correlation with high level of tannins in methanol extract of A. viridiflora (3.74%). The proven inhibitions of ACE and α-glucosidase make the methanol extract of A. viridiflora suitable for further scientific research in order to find a new natural product for the treatment of cardiovascular diseases and diabetes.VII Kongres farmaceuta Srbije sa međunarodnim učešćem: Zajedno stvaramo budućnost farmacije, Beograd, Srbija, 10-14. oktobar 2018.Usmeno izlaganje na skup

Black Trumpet, Craterellus cornucopioides (L.) Pers.: Culinary Mushroom with Angiotensin Converting Enzyme Inhibitory and Cytotoxic Activity

Nutritional value and chemical composition, including the content of vitamins, fatty acids, 5'-nucleotides and nucleosides and amino acids, as well as biological activities, including antioxidant, angiotensin converting enzyme (ACE) inhibitory and cytotoxic activity of black trumpet (Craterellus cornucopioides (L.) Pers.) were tested in vitro. C. cornucopioides was low in energy, fat and carbohydrate contents, but rich in dietary fibre, especially β-glucan as well as niacin and α-tocopherol. The content of essential and non-essential free amino acids was 1.49 and 5.48 mg/g dry weight (dw). The nucleosides and 5'-nucleotides were determined at 1.84 and 3.99 mg/g dw, respectively. The share of unsaturated fatty acids (UFAs) was 75.92% with oleic acid as the major UFA. Cyclohexane and dichloromethane extracts expressed significant cytotoxic activity against selected cell lines, human epithelial cervical cancer cells (HeLa), adenocarcinomic human alveolar basal epithelial cells (A549), colorectal cancer cells (LS174) and normal MRC-5 human embryonic lung fibroblast cells (IC50 of 78.3-155.6 μg/mL). ACE inhibitory activity of the aqueous extract was strong with an IC50 of 0.74 μg/mL. It can be concluded that black trumpet is a good source of nutrients, such as vitamins, dietary fibres, amino acids, nucleotides and fatty acids, which contribute to the overall nutritional value of this fungus with potential for ACE inhibitory activity and use in anti-hypertensive diet

Chemical composition and antiproliferative potential of dried wild apple and pear tea before and after in vitro simulated digestion

Decoctions obtained from dried apple and pear fruits were subjected to in vitro digestion in the presence of a food matrix in order to determine changes in the polyphenol content and antiproliferative activity. The total phenolic and total proanthocyanidins contents were determined using spectrophotometrical methods, and the phenolic compounds were analyzed by RP-HPLC/DAD before and after digestion. Chlorogenic acid and phlorizin dihydrate were the major identified compounds. The addition of a food matrix immediately decreased the contents of individual and total phenolics. After digestion, they were slightly elevated but still lower than in the initial samples. Antiproliferative activity was investigated on human epithelial carcinoma cell line (HeLa), human colon carcinoma (LS174) and human foetal lung fibroblast (MRC-5) cells. The exhibited growth inhibition was not correlated with the content of phenolics in the tested samples, indicating that it could not be explained solely by their content. With the exception of the apple decoction effect on HeLa cells, the digestion process resulted in significant increases in the antiproliferative activity

Association between TGFB1 C-509T polymorphism and acute toxicity after radiotherapy for prostate cancer

Background: Radiotherapy (RT) for prostate cancer (PCa) is associated with a spectrum of side effects (toxicity) in the surrounding normal tissues. TGFB1 is a key cytokine associated with inflammation and fibrosis, but its role in acute toxicity is unclear. The presence of T allele at −509 bp of the promoter region of TGFB1 gene is related to higher concentrations of TGFB1 than C allele. We aimed to investigate association between C-509T polymorphism (rs1800469) and RT-induced acute toxicity. Patients and methods: Eighty six patients who had a localized or locally advanced PCa were treated with radical (72 Gy)(n=49) or postoperative/salvage (66 Gy)(n=37) RT without previous hormonal therapy. TGFB1 C-509T was determined by PCR-RFLP analysis on DNA from PBMC. The differences in the distribution of genotypes between patients with or without acute genitourinary (GU) or gastrointestinal (GI) toxicity were tested by χ2 and Fisher’s exact test. P values ≤0.05 were considered statistically significant. The genotype-specific associations with toxicity were estimated as odds ratios (OR) with 95% confidence intervals (CIs) for dominant, recessive, codominant and overdominant genetic model. Results: Heterozygote carriers of TGFB1 C-509T had statistically significant lower rate of acute GU and GI toxicity then homozygotes (CC plus TT) (p=0.048; p=0.047). Additionally, the OR indicated lower risk for acute toxicity development in heterozygote than homozygote patients (OR (95%CI) were: 0.12 (0.01- 1.11) for GU and 0.19 (0.03- 1.02) for GI). Conclusion: The obtained data indicate that CT genotype of TGFB1 C-509T could be potential biomarkers of lower risk for acute RT-induced toxicity.5th Congress of the Serbian Association for Cancer Research with International Participation SDIR-5, Virtual event, December 3, Belgrade, 2021

Supplementary data for article: Pantelic, N.; Stanojkovic, T. P.; Zmejkovski, B. B.; Sabo, T. J.; Kaluerovic, G. N. In Vitro Anticancer Activity of Gold(III) Complexes with Some Esters of (S, S)-Ethylenediamine-N, N G2-Di-2-Propanoic Acid. European Journal of Medicinal Chemistry 2015, 90, 766–774. https://doi.org/10.1016/j.ejmech.2014.12.019

Supplementary material for: [https://doi.org/10.1016/j.ejmech.2014.12.019]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1651

Hypericum perforatum L. extracts exert cytotoxic effects and show different miRNA signatures in PC-3 and DU 145 prostate cancer cells

Phytochemicals and bioactive substances derived from a wide range of plant extracts have been reported to exert various anticancer effects. Prostate cancer is one of the leading causes of cancer-related deaths within the male population. Prostate cancer-specific miRNA signatures were associated with cancer formation and progression, with various subtypes, and response to therapy. MicroRNA levels of expression were shown to change after the treatment of various compounds and substances extracted from natural products. Natural herbal compounds were shown to induce variations in miRNA expression levels in cancer cells. The aims of this study were to investigate the cytotoxic effects of methanol, ethyl-acetate, and hexane extracts obtained from branch-body part and flowers of Hypericum perforatum L. against humane PC-3 and DU 145 and to test potential miRNA-128/133b/155/193a/206/21/335 signature changes and differences between the two prostate cancer cell lines. Cytotoxic activity of H. perforatum extracts, their effects on cell cycle distribution, and miRNA expression levels were examined in humane PC-3 and DU 145 prostate cancer cells by MTT cell survival assay, flow cytometry, and quantitative real-time PCR. Hexane extract of flowers showed the strongest intensity of cytotoxic activity against PC-3 and DU 145 cells. The highest increase in the percentage of PC-3 cells in the subG1 phase was observed in cell samples treated with hexane extract of flowers and branch-body part. Significant differences in miRNA-128/133b/155/193a/206/21/335 levels were observed between PC-3 and DU 145 cell lines, especially in samples treated with flower extracts compared with the branch-body part. Conclusions: Investigated extracts have significant anticancer potential not only from the aspects of cytotoxicity and cell cycle effects but also from the aspect of lowering oncogenic or increasing tumor-suppressive miRNAs. The best effect might be the increase of tumor-suppressive miR-128 (accompanied by miR-193a) induced by the hexane extract of the flowers, which also exerted the highest cytotoxic activity. Hexane extract of flowers may be the candidate for further investigation for improving the efficiency of standard therapies for PCa. A miRNA signature might be cell-type specific after the treatment with H. perforatum extracts