Britannin suppresses MCF-7 breast cancer cell growth by inducing apoptosis and inhibiting autophagy.

Objective: Breast cancer is the main reason for cancer-related death in women. Britannin is a sesquiterpene lactone compound derived from Inula aucheriana with anti-tumor properties. We aimed to explore the impacts of britannin on apoptosis and autophagy in MCF-7 breast cancer cell line. Materials and Methods: The cytotoxic influences of britannin on MCF-7 cells were estimated by the MTT method. The expression levels of apoptosis-associated genes such as CASP3, BCL2, BCL2L1, STAT3, and JAK2 and transcripts of autophagy markers including ATG1, ATG4, ATG5, ATG7, ATG12, BECN1, and MAP1LC3A were quantified using quantitative real time-PCR (qRT-PCR). Western blotting method was used to evaluate the amount of caspase 3, phosphorylated JAK2, phosphorylated STAT3, ATG1, ATG4, ATG5, Beclin1, and LC-III. Results: Treatment of MCF-7 cells with various concentrations of britannin remarkably hindered the viability of these cells compared to the controls. This compound significantly elevated the expression of pro-apoptotic caspase-3 but did not influence the levels of anti-apoptotic BCL2 and BCL2L1. Britannin decreased the levels of phosphorylated forms of JAK2 and STAT3 proteins causing the blockage of the JAK/STAT pathway. Four autophagy factors expressions, including ATG4, ATG5, Beclin1, and LCIII, were reduced due to the effect of britannin on MCF-7 cells. Conclusion: Britannin triggered apoptosis in MCF-7 cells by a mechanism that led to the blockade of the JAK/STAT pathway. Moreover, britannin prohibited autophagy in these cancer cells. This may suggest britannin as an agent for the suppression of breast tumors or as an adjutant for the enhancement of anti-breast cancer drugs effect.

Targeting of human fibroblast growth factor receptor 2 by a novel specific nanobody.

Inhibition of FGFR2 signaling is promising in targeted therapy of FGFR2-related tumors. In this study, anti-FGFR2 nanobodies (Nbs) were isolated through screening of an immune camelid phage display library. Four rounds of biopanning were carried out with commercial human FGFR2 antigen and enrichment was assessed by ELISA and phage titration. The gene of Nb was sub-cloned into the expression vector, and the recombinant vector was transformed into Escherichia coli WK6 cells. The recombinant protein was purified using Ni-NTA affinity chromatography. The anti-FGFR2 Nb (C13) was characterized by SDS-PAGE, western blotting, competitive inhibition ELISA, flow cytometry, MTT, and migration assay. C13 Nb recognized FGFR2 with high specificity and no cross-reactivity was observed with other tested antigens. The affinity of C13 Nb was calculated to be 1.5 × 10-9 M. Results of cytotoxicity showed that C13 Nb (10 µg/ml) inhibited 85% of the proliferation of T-47D cells (p < 0.001). In addition, C13 inhibited the migration of 68% of T-47D toward the source of the growth factor (p < 0.01). The flow cytometry showed that C13 Nb bound to the surface of FGFR2+ cells, T-47D cell line (96%). Results indicate the potential of anti-FGFR2 Nb for targeted therapy of FGFR2-overexpressing tumors after complementary investigations.

Biological Activities and the Essential Oil Analysis of Cousinia harazensis and C. calocephala.

This research aimed to evaluate the cytotoxicity, anti-bacterial, anti-fungal and heme polymerization inhibition activities, as well as the detection of the chemical composition of essential oils and measurement of the amount of total phenol and flavonoids of Cousinia harazensis and C. calocephala. In-vitro growth inhibitory effects of methanol extracts on A2780, T-47D, A549 and Hep-G2 cells were evaluated by MTT assay. MIC and MBC/MFC were determined by the agar dilution method. The anti-malarial activity of herbs was assessed with an inhibition test of heme detoxification (ITHD). Total phenol and flavonoids content measured by Folin-Ciocalteu method. The essential oils from two herbs were extracted by hydro-distillation, and GC/MS analyzed their compositions. Cell studies against selected cell lines growth in MTT assay were related to C. harazensis on Hep-G2 with IC50 of 4.521 µg/mL. The MIC of anti-bacterial and anti-fungal effects is related to C. harazensis extract on Staphylococcus epidermidis and Aspergillus fumigatus with 15.62 and 62.5 mg/mL, respectively. Both extracts do not have anti-malarial activity. C. harzensis content was richer in total phenol and flavonoids rather than the other herb. m-benzyl benzyl alcohol (46.7%) and butyl phthalate (14.7%) are the major compounds of C. harazensis; main components of C. calocephala are 3-methyl-tetrahydrofuran (24.6%) and oleic acid (15.4%). In conclusion, C. harazensis with more phenol and flavonoids content showed better results in terms of biological activities.

Antimalarial Activities of (Z)-2-(Nitroheteroarylmethylene)-3(2H)-Benzofuranone Derivatives: In Vitro and In Vivo Assessment and ß-Hematin Formation Inhibition Activity.

A series of (Z)-2-(nitroheteroarylmethylene)-3(2H)-benzofuranones possessing nitroheteroaryl groups of nitroimidazole, nitrofuran, and nitrothiophene moieties was screened for antiplasmodium activity against a drug-sensitive strain (3D7 strain) and a multidrug-resistant (chloroquine [CQ] and pyrimethamine) strain (K1 strain) of Plasmodium falciparum. 5-Nitroimidazole and 4-nitroimidazole analogs were highly selective and active against resistant parasites, while 5-nitrofuran and 5-nitrothiophene derivatives were more potent against the 3D7 strain than against the K1 strain. Among the synthetic analogues, (Z)-6-chloro-2-(1-methyl-5-nitroimidazol-2-ylmethylene)-3(2H)-benzofuranone (compound 5h) exhibited the highest activity (50% inhibitory concentration [IC50], 0.654 nM) against the K1 strain and (Z)-7-methoxy-2-(5-nitrothiophen-2-ylmethylene)-3(2H)-benzofuranone (10g) showed the highest activity (IC50, 0.28 µM) against the 3D7 strain in comparison with the activities of CQ (IC50s of 3.13 and 206.3 nM against 3D7 and K1 strains, respectively). The more active compounds, with IC50s lower than 5 µg/ml (∼20 µM), were further studied for their cytotoxicity responses using KB cells. From these studies, 5-nitroimidazole, 4-nitroimidazole, and 5-nitrofuran analogues were shown to be cytotoxic against KB cells, while 5-nitrothiophene analogues were shown to have the least cytotoxic effects. To gain some insight into their potential contributing mechanisms of action, three derivatives, 10e, 10g, and 10h (from the nitrothiophene subgroup, possessing 6-methoxy, 7-methoxy, and 6,7-dimethoxy substituents, respectively, on their benzofuranone moieties), showing the least toxicity and highest selectivity indices were assessed for their ß-hematin formation inhibition activity. Compound 10g demonstrated the highest inhibition activity (IC50, 10.78 µM) in comparison with that of CQ (IC50, 2.63 µM) as the reference drug. Finally, these three analogues (10e, 10g, and 10h) were further evaluated for their in vivo activities against the Plasmodium berghei/albino mouse model (Peter's test). The tested analogues were shown to be active, reducing the percentages of erythrocytes that contained parasites by 53.4, 48.8, and 32.4%, respectively.

Synthesis of novel amodiaquine analogs and evaluation of their in vitro and in vivo antimalarial activities.

BACKGROUND & OBJECTIVES: Due to the rapid increase of drug resistance in Plasmodium parasites, there is a pressing need of developing new antiplasmodial drugs. In this study, new amodiaquine (AQ) analogs were synthesized, followed by an evaluation of their antiplasmodial activity. METHODS: A new series of quinoline derivatives containing N-alkyl (piperazin-1-yl)methyl benzamidine moiety was synthesized by reacting 4-[(4-(7-chloroquinolin-4-yl)piperazin-1-yl)methyl]benzonitrile with appropriate primary amines. The synthesized compounds were investigated for inhibitory activity by inhibition test of heme detoxification (ITHD). Their antiplasmodial activity was then evaluated using the classical 4-day suppressive test (Peter's test) against Plasmodium berghei-infected mice (ANKA strain). RESULTS: The results showed that the percentage of heme detoxification inhibition in the active compounds was 90%. The most promising analogs, N-butyl-4-[(4-(7-chloroquinolin-4-yl)piperazin-1-yl)methyl]benzamidine (compound 1e), and 4-[(4-(7-chloroquinolin-4-yl)piperazin-1-yl)methyl)]-N-(4-methylpentan-2-yl)benzamidine (compound 1f) displayed 97.65 and 99.18% suppressions at the doses of 75 and 50 mg/kg/day, respectively. Further, the mean survival time of the mice treated with these compounds was higher than that of the negative control group. INTERPRETATION & CONCLUSION: The newly synthesized amodiaquine analogs presented sufficient antiplasmodial activity with excellent suppressions and high in vitro heme detoxification inhibition. Higher mean survival time of the mice treated with synthetic compounds further confirmed the in vivo antimalarial activity of these new AQ analogs. Therefore, these compounds have the potential to replace common drugs from 4-aminoquinoline class. However, further investigations such as pharmacokinetic evaluations, cytotoxicity, toxicity, and formulation seem to be necessary.

Leaves Antimicrobial Activity of Glycyrrhiza glabra L.

Licorice (Glycyrrhiza glabra L.) is an important medicinal plant. In this study, the antimicrobial activities of ethanolic and aqueous extracts from licorice leaves were studied compared to root extracts activities. Bacillus subtilis, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli, and Candida albicans were used as test organisms. Antimicrobial activity was tested by paper disc agar diffusion and serial dilution methods in order to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The root and leave extracts showed activity against Candida albicans, and tested gram-positive bacteria in a dose dependent manner. The ethanolic extract of the leaves was the most active extract against gram-positive bacteria. Its effectiveness against strains provides hope that it can serve as an alternative therapeutic agent.