Effectiveness of Rhenium(I)-diselenoether Low Doses in a Triple-negative Breast Cancer Chicken Embryo Model.

BACKGROUND/AIM: Rhenium(I)-diselenoether (Re-diSe) is a promising anticancer agent composed of one rhenium and two selenium atoms. Its effectiveness was established in inhibiting cancer cells while maintaining low toxicity toward normal cells at a 5 µM dose for 120 hours in MDA-MB-231 cells. In MDA-MB-231 breast tumor-bearing mice, anti-tumor and anti-metastatic effects were observed at a 10 mg/kg dose. However, contradictory results were observed in the 4T1 breast cancer model, where a dose of 60 mg/kg had a pro-tumor effect. To address these discrepancies, the efficacy of Re-diSe at the effective 10 mg/kg dose was validated in a transplanted MDA-MB-231 breast tumor model using the chicken chorioallantoic membrane assay. MATERIALS AND METHODS: MDA-MB-231 cancer cells were xenografted onto the chicken chorioallantoic membrane (CAM), and daily drug administration was carried out for nine days at doses of 0.1, 1, and 10 mg/kg. At the study's conclusion, a standard histological analysis was conducted. RESULTS: The low dose of 0.1 mg/kg showed a significant reduction in tumor weights compared to controls. The 1 mg/kg dose resulted in an increased inflammation score but did not induce a significant difference in tumor weights compared to the 0.1 mg/kg dose. Notably, at the 10 mg/kg dose, six out of 11 treated embryos displayed no visible signs of tumors. These tumors exhibited extensive tumor necrosis and significant infiltration by inflammatory cells. CONCLUSION: In this particular model, the anticancer efficacy of Re-diSe was achieved at the low dose of 0.1 mg/kg. The higher dose of 10 mg/kg, while eliminating visible tumors, might have immune-mediated effects, as indicated by substantial tumor necrosis and infiltration by inflammatory cells. Overall, this study successfully demonstrated the effectiveness of Re-diSe as an anticancer agent.

Remarkable Effects of a Rhenium(I)-diselenoether Drug on the Production of Cathepsins B and S by Macrophages and their Polarizations.

BACKGROUND/OBJECTIVE: Tumor-associated macrophages (TAMs) produce an excessive amount of cysteine proteases, and we aimed to study the effects of anticancer rhenium(I)-diselenoether (Re-diSe) on the production of cathepsins B and S by macrophages. We investigated the effect of Re-diSe on lipopolysaccharides (LPS) induced M1 macrophages, or by interleukin 6 (IL-6) induced M2 macrophages. METHODS: Non-stimulated or prestimulated murine Raw 264 or human THP-1 macrophages were exposed to increasing concentrations of the drug (5, 10, 20, 50 and 100 µM) and viability was assayed by the MTT assay. The amount of cysteine proteases was evaluated by ELISA tests, the number of M1 and M2 macrophages by the expression of CD80 or CD206 biomarkers. The binding of Re-diSe with GSH as a model thiol-containing protein was studied by mass spectrometry. RESULTS: A dose-dependent decrease in cathepsins B and S was observed in M1 macrophages. There was no effect in non-stimulated cells. The drug induced a dramatic dose-dependent increase in M1 expression in both cells, significantly decreased the M2 expression in Raw 264 and had no effect in non-stimulated macrophages. The binding of the Re atom with the thiols was clearly demonstrated. CONCLUSION: The increase in the number of M1 and a decrease in M2 macrophages treated by Re-diSe could be related to the decrease in cysteine proteases upon binding of their thiol residues with the Re atom.

Enhancement of antifouling properties, metal ions and protein separation of poly(ether-ether-sulfone) ultrafiltration membranes by incorporation of poly ethylene glycol and n-ZnO.

Composite polymeric membranes with enhanced anti-fouling properties, antimicrobial activities and flux were produced via the phase inversion technique using poly (ether-ether-sulfone) (PEES)/polyethylene glycol (PEG) and n-ZnO. SEM and ATR-FTIR spectroscopy were used to study the morphological and chemical properties of the resulting ultrafiltration membranes. PEG and n-ZnO concentration has an effect on membrane morphologies, ultrafiltration performance, thermal characteristics, metal ion separation studies, surface hydrophilicity and anti-fouling capabilities. The permeate flux increased when the PEG concentration was raised. This results revealed that adding PEG and n-ZnO to membranes increased their surface hydrophilicity and anti-fouling properties. The inclusion of 1.5 wt % n-ZnO and 5 wt % PEG to the pristine PEES membrane resulted in a higher flux of 233.76 L m-2 h-1, 70.09 % of water content, 47.46° of contact angle, the porosity of 30.20 %, and hydraulic resistance of 0.22 kPa/Lm-2h-1. Anti-fouling properties of the fabricated membrane were assessed using a model foulant BSA, which revealed a high flux recovery ratio value. As a result, the PEG and n-ZnO incorporated membrane is more hydrophilic than the virgin membrane. In addition, the prepared PEES/PEG/n-ZnO membrane showed a significant increase in metal ions and protein rejection. Furthermore, an antibacterial test of the membrane revealed that the PEG and n-ZnO composite membrane outperformed the bare PEES membrane in terms of antibacterial capabilities. Overall, the findings reveal that combining n-ZnO and PEG resulted in a membrane with improved anti-fouling capabilities and hydrophilicity, making it suitable for water purification.

In vitro and in silico anti-leukemic activity of 2-amino-6-nitro-4-(4-oxo-2-thioxothiazolidin-5-yl)-4H-chromene-3-carbonitrile (ANC) through inhibition of anti-apoptotic Bcl-2 proteins.

An array of 4H-chromene derivatives have been reported for anticancer properties but their selectivity and mode of anticancer activity are unexplored. In this context, we have investigated a biologically active synthetically designed 4H-Chromene carbonitrile derivative, 2-amino-6-nitro-4-(4-oxo-2-thioxothiazolidin-5-yl)-4H-chromene-3-carbonitrile (ANC) that is strongly and selectively inhibited Bcl-2 over expressing human leukemic (HL-60 and K562) cells for its interaction and elucidated the mode of action. The interaction of ANC was investigated against the antiapoptotic proteins such as Bcl-2, Bax, Bcl-xL and Bcl-w that were overexpressed in leukemic cells using in silico and fluorescent spectroscopic studies. Fluorescent spectroscopic based interaction studies showed that the derivative had strong interaction with Bcl-xL followed by Bcl-2/Bax and least interaction with Bcl-w. Based on the results, the ANC had strong interactions with antiapoptotic Bcl-2 and Bax proteins than the Bcl-xL and Bcl-w proteins. The in vitro biological validation of ANC treated leukemic cells showed downregulation of Bcl-xL than Bcl-2 but least effect on Bcl-w proteins. Furthermore, the ANC had possible four isomers as RR, RS, SR and SS isomers. Among them, RS isomer of ANC had shown more active that correlated with biological interactions and gene expression studies of ACN with oncoproteins. These results confirmed the induction of apoptosis by RS-ACN isomer through inhibition of antiapoptotic machineries of leukemic cells confirming the antiapoptotic Bcl-2 inhibitory activities.Communicated by Ramaswamy H. Sarma.

Targeting Staphylococcus aureus and its biofilms with novel antibacterial compounds produced by Lactiplantibacillus plantarum SJ33.

The emergence of microbial resistance to conventional antibiotics, partially attributed to biofilm formation, has urged for new antimicrobial compounds. Here, we reported two novel low molecular weight (LMW) compounds from Lactiplantibacillus plantarum SJ33 and evaluated their biofilm inhibitory effects on Staphylococcus aureus. The compounds C1 and C2 were purified by RP-HPLC and structurally identified as 3-amino-5-hydroxy-6-(hydroxymethyl)-4-(1-hydroxyprop-2-yn-1-yl)-3,3a,4,5,6,7a-hexahydro-7H-indazol-7-one and 1-(dimethylamino)-3-hydroxy-3-((2-hydroxypropan-2-yl)oxy)-1-(methylamino)-butan-2-one by spectroscopic techniques. High ESI-MS data confirmed the molecular weight of C1 and C2 as 254.1141 and 234.1658 Da, respectively. Time-kill assay demonstrated bactericidal action of compounds, whereas scanning electron microscopy revealed morphological changes in treated S. aureus MTCC96 and methicillin-resistant S. aureus (MRSA) cells. The antibacterial compounds reduced biofilm formation in S. aureus MTCC96 and MRSA by crystal violet assay. Further, fluorescence and scanning electron microscopic images exhibited biofilm formation by pathogens and biofilm inhibition by compounds treatment. The Quantitative RT PCR revealed the down-regulation of icaC and icaD genes involved in intercellular adhesion of biofilms. The results confirmed the anti-biofilm activity of novel LMW compounds by eliminating preformed biofilms formed by S. aureus MTCC96 and MRSA.

Effects of Rhenium(I)-diselenoether and of its Diselenide Ligand on the Production of Cathepsins B and S by MDA-MB231 Breast Malignant Cells.

BACKGROUND/AIM: Rhenium(I)-diselenoether (Re-diSe) is a drug under development for the treatment of metastatic cancers, with selective inhibitory effects on MDA-MB231 cancer cells compared to normal HEK-293 cells, and with greater effects than its diselenide (di-Se) ligand. Rhenium (Re) compounds inhibit cathepsins, which are important proteolytic enzymes in cancer. This study investigated the effects of Re-diSe and di-Se on the production of cathepsins B and S in MDA-MB231 malignant and HEK-293 normal cells and their inhibitory effects following treatment with different doses for 72 h. MATERIALS AND METHODS: Elisa tests were used to assay the amount of cathepsins B and S in the medium of cultures. RESULTS: Re-diSe, but not diSe affected the viability of malignant cells and the expression of cathepsins B and S. CONCLUSION: To the best of our knowledge, this is the first demonstration that Re-diSe may decrease the production of cathepsins B and S in cancer cells at doses as low as 10 µM.

Atranorin, an antimicrobial metabolite from lichen Parmotrema rampoddense exhibited in vitro anti-breast cancer activity through interaction with Akt activity.

Atranorin (ATR), lichenized secondary metabolite and depside molecule with several biological potentials such as antimicrobial, anticancer, anti-inflammatory, antinociceptive, wound healing and photoprotective activities. Cytotoxic reports of ATR are documented in several cancer cells and in vivo models but its molecular interaction studies are poorly understood. Therefore, in this present investigation, we have used the in silico studies with biological validation of the molecular targets for the anti-breast cancer mechanism of ATR. The molecular docking studies with the breast cancer oncoproteins such as Bcl-2, Bax, Akt, Bcl-w and Bcl-xL revealed the highest interaction was observed with the Akt followed by Bax, Bcl-xL and Bcl-2 & least with the Bcl-w proteins. The cytotoxicity studies showed ATR selectively inhibited MDA MB-231 and MCF-7 breast cancer cells in differential and dose-dependent manner with the IC50 concentration of 5.36 ± 0.85 µM and 7.55 ± 1.2 µM respectively. Further mechanistic investigations revealed that ATR significantly inhibited ROS production and significantly down-regulated the anti apoptotic Akt than Bcl-2, Bcl-xL and Bcl-w proteins with a significant increase in the Bax level and caspases-3 activity in the breast cancer cells when comparison with Akt inhibitor, ipatasertib. In vitro biological activities well correlated with the molecular interaction data suggesting that atranorin had higher interaction with Akt than Bax and Bcl-2 but weak interaction with Bcl-w and Bcl-xL. In this present study, the first time we report the interactions of atranorin with molecular targets for anti-breast cancer potential. Hence, ATR represents the nature-inspired molecule for pharmacophore moiety for design in targeted therapy.Communicated by Ramaswamy H. Sarma.

A New Model Applied for Evaluating a Rhenium-diselenium Drug: Breast Cancer Cells Stimulated by Cytokines Induced from Polynuclear Cells by LPS.

BACKGROUND/AIM: New anticancer drugs are usually tested on cancer cells in culture in a standard medium. We stimulated immune polynuclear cells by lipopolysaccharides to obtain an enriched medium (EM) containing inflammatory cytokines more closely reflecting the tumor microenvironment and tested a rhenium-diselenium (Re-diSe) drug in this new model. Concentrations of cytokines were compared with a control medium (CM). MATERIALS AND METHODS: Human-derived breast cancer cells were grown in culture either in CM or EM with or without Re-diSe. Assays of tumor necrosis factor alpha (TNFα), interleukin 6 (IL6), intereukin 1 beta (IL1ß), transforming growth factor-beta (TGFß), insulin growth factor 1 (IGF1) and vascular epidermal growth factor A (VEGFA) were performed by enzyme-linked immunosorbent assays. The production of reactive oxygen species (ROS) was determined by 2,7-dichlorofluorescein test. The cell growth was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tests. RESULTS: Concentrations of TNFα, IL6 and Il1ß were observed to be significantly higher in EM than in CM. There was no difference for TGFß, IGF1 and VEGFA. The cells were sensitive to Re-diSe, with reduced concentrations of TGFß, IGF1, VEGFA and ROS, but the half-maximal inhibitory concentration was significantly higher in EM than in CM. CONCLUSION: The efficacy of the Re-diSe drug was confirmed in this model of aggressive cancer.

Swarming Inhibitory Potential of Cinnamtannin B1 from Cinnamomum tamala T. Nees and Eberm on Pseudomonas aeruginosa.

In a preliminary screening, the methanol extract of Cinnamomum tamala leaves was found to inhibit the swarming motility of Pseudomonas aeruginosa. Bioassay-guided fractionation by silica gel column chromatography led to the identification of cinnamtannin B1 (1) as one of the active components of the extract. It inhibited the swarming motility (at 12.5 µg/mL) and biofilm formation (at 25 µg/mL) ofP. aeruginosa. Comparative gene expression analysis revealed downregulation of rhlA and fliC genes upon treatment with the tannin. The tannin may be affecting rhamnolipid and flagellin production. Thus, cinnamtannin B1 is an active component of C. tamala responsible for inhibiting the swarming motility of P. aeruginosa.

The rhenium(I)-diselenoether anticancer drug targets ROS, TGF-ß1, VEGF-A, and IGF-1 in an in vitro experimental model of triple-negative breast cancers.

The rhenium(I)-diselenoether complex (Re-diSe) is a rhenium tricarbonyl-based drug chelated by a diselenoether ligand. In this work, we compared its inhibitory effects on the hormone-independent MDA-MB231cancer line and other different cancer cell lines after an exposure time of 72 h by MTT assays. The sensitivity of MDA-MB231 was in the same range than the hormone-dependent MCF-7 breast cancer, the PC-3 prostate and HT-29 colon cancer cells, while the A549 lung and the HeLa uterine cancer cells were less sensitive. We compared the inhibitory effects of Re-diSe and of its diselenide ligand (di-Se) on MDA-MB231 and a normal HEK-293 human embryonic cell line, after 72 h and 120 h of exposure. The cytotoxicity was also studied by flow cytometry using ethidium bromide assays, as well as the effects on the ROS production by DFCA-test, while the levels of TGF-ß1, VEGF-A, IGF-1 were addressed by ELISA tests. The dose required to inhibit 50% of the proliferation (IC50) of MDA-MB231 breast cancer cells decreased with the time of exposure to 120 h, while the free ligand (di-Se) was found poorly active, demonstrating the important role of Re in this Re-diSe combination. The cytotoxic effects of Re-diSe were highly selective for cancer cells, with a significant increase of the number of dead cancer cells at 5 µM for an exposure time of 120 h, while normal cells were not affected. A remarkable and significant decrease of the production of ROS together with a decrease of VEGF-A, TGF-ß1, and IGF-1 by the cancer cells were also observed when cancer cells were exposed to Re-diSe.

Purification and identification of 4-allylbenzene-1,2-diol: an antilisterial and biofilm preventing compound from the leaves of Piper betle L. var Pachaikodi.

Antibiotic-resistant food-borne Listeriosis has been rising with up to 30% mortality threat in humans since several decades. Hence, discovering antilisterial from the extracts of ethnomedicinal plants may be of value as a novel antidote. In our preceding study, we reported that ethanolic extract of Piper betle L. var Pachaikodi leaves exhibited antibacterial activity towards Listeria monocytogenes MTCC 657. Consequently in the present study, the bioactive molecule responsible for anti-Listeria activity was purified and identified as 4-allylbenzene-1,2-diol. This identified bioactive compound may have significance while used as antimicrobials and/or food additives in food processing sector as evidenced by dual action: biofilm inhibition and pore formation on cell membrane.

The cooperative effect of Lewis pairs in the Friedel-Crafts hydroxyalkylation reaction: a simple and effective route for the synthesis of (±)-carbinoxamine.

An efficient C-C bond formation strategy between aromatic/heteroaromatic π-nucleophiles and Lewis acid activated aldehydes is described. This aromatic electrophilic substitution reaction of arenes or heteroarenes is facilitated by Lewis acid AlBr3. Aromatic rings with electron donating substituents are excellent nucleophilic counterparts in this reaction, generating carbinols in excellent yields (61-94%). The formation of triarylmethanes is also witnessed in the case of certain reactive aldehydes and aromatic π-nucleophiles through reactive carbocation formation. The formation of triarylmethane is reduced to a greater extent via retardation of the second π-nucleophile addition through a Lewis base, for example, pyridine, coordination with an aluminium alkoxide intermediate. Various aliphatic aldehydes also underwent Friedel-Crafts type hydroxyalkylation and generated the expected carbinols in moderate yields (41-53%) in the presence of AlBr3. This protocol has been successfully applied to the synthesize of the (±)-carbinoxamine, a therapeutically important histamine H1 antagonist, in a one-pot manner.