4-methoxybenzalacetone, the cinnamic acid analog as a potential quorum sensing inhibitor against Chromobacterium violaceum and Pseudomonas aeruginosa.

The continuous increase in the incidence of infectious diseases and the rapid unchecked rise in multidrug-resistance to conventional antibiotics have led to the search for alternative strategies for treatment and clinical management of microbial infections. Since quorum sensing (QS) regulates numerous virulence determinants and pathogenicity in bacteria, inhibition of QS promises to be an attractive target for development of novel therapeutics. In this study, a series of cinnamic acid analogs and benzalacetone analogs were designed and synthesized, and their QS-inhibitory activities explored. We found that, among the test compounds, 4-methoxybenzalacetone (8) exhibited potent anti-quorum sensing property, as evidenced by inhibition of QS-controlled violacein production of Chromobacterium violaceum ATCC12472. The inhibitory activity of such a compound, which was the methyl keto analog of the corresponding cinnamic acid, was not only stronger than the parent cinnamic acid (1), but also superior to that of furanone, the reference drug. Based on our observations, its mechanism of quorum sensing inhibition is likely to be mediated by interference with N-acyl-homoserine lactones (AHL) synthesis. Moreover, 4-methoxybenzalacetone (8) also suppressed the production of pyocyanin, rhamnolipids and swarming motility of Pseudomonas aeruginosa, suggesting a broad spectrum of anti-QS activities of this compound. In terms of structure-activity relationship, the possible chemical substitutions on the scaffold of cinnamic acid required for QS inhibitory activity are also discussed. Since 4-methoxybenzalacetone (8) showed no toxicity to both bacteria and mammalian cells, our findings therefore indicate the anti-QS potential of this compound as a novel effective QS inhibitor.

Ethyl Rosmarinate Prevents the Impairment of Vascular Function and Morphological Changes in L-NAME-Induced Hypertensive Rats.

Background and Objectives: The potent, endothelium-independent, vasorelaxant effect of ethyl rosmarinate, an ester derivative of rosmarinic acid, makes it of interest as an alternative therapeutic agent for use in hypertension. This study was designed to investigate the effect of ethyl rosmarinate on Nω-nitro-L-arginine methyl ester (L-NAME)-induced hypertensive rats. Materials and Methods: L-NAME was given orally to male Wistar rats for 6 weeks to induce hypertension concurrently with treatment of ethyl rosmarinate at 5, 15, or 30 mg/kgor enalapril at 10 mg/kg Systolic blood pressure (SBP), heart rate, and body weight of all experimental groups were recorded weekly, while the vascular sensitivity and histological changes of the aorta were evaluated at the end of the experiment. Results: For all treatment groups, the data indicated that ethyl rosmarinate significantly attenuated the SBP in hypertensive rats induced by L-NAME, with no significant differences in heart rate and body weight. In addition, the response of vascular sensitivity to acetylcholine (ACh) was improved but there was no significant difference in the response to sodium nitroprusside (SNP). Furthermore, the sensitivity of the aorta to phenylephrine (PE) was significantly decreased. The thickness of the aortic wall did not differ between groups but the expression of endothelial nitric oxide synthase (eNOS) was increased in ethyl rosmarinate- and enalapril-treated groups compared with the hypertensive group. Conclusions: Ethyl rosmarinate is an interesting candidate as an alternative treatment for hypertension due to its ability to improve vascular function and to increase the expression of eNOS similar to enalapril which is a drug commonly used in hypertension.

Endothelium-dependent and endothelium-independent vasorelaxant effects of tiliacorinine 12'-O-acetate and mechanisms on isolated rat aorta.

Tiliacorinine 12'-O-acetate is a modified analog of Tiliacorinine, a major compound in Tiliacora triandra. The present study explored the vasorelaxation property of tiliacorinine 12'-O-acetate and its mechanism in isolated rat aorta using the organ bath technique. Tiliacorinine 12'-O-acetate exhibited concentration-dependent (10-15-10-3.5 M) vasorelaxation in endothelium-intact rings (Emax = 93.53 ± 2.79%) and endothelium-denuded rings (Emax = 74.31 ± 5.09%). The effects of tiliacorinine 12'-O-acetate were attenuated by pre-incubation with N(ω)-nitro-l-arginine methyl ester (L-NAME, endothelium nitric oxide synthase inhibitor) (100 µM), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, soluble quanylylcyclase inhibitor) (1 µM), and 4-aminopyridine (1 mM, Kv channel blocker). However, this effect was not impacted by indomethacin (10 µM, cyclooxygenase inhibitor), tetraethylammonium (5 mM, Kca channel blocker), barium chloride (1 mM, KIR channel blocker), or glibenclamide (10 µM, KATP channel blocker). Moreover, pretreatment with tiliacorinine 12'-O-acetate reduced the effect of L-NAME (100 µM) on acetylcholine-induced vasorelaxation. Tiliacorinine 12'-O-acetate showed inhibitory effects on CaCl2-induced contracted rings and reduced the contraction induced by phenylephrine (10 µM) and caffeine (20 mM) in a Ca2+-free solution. The results of this study suggest that tiliacorinine 12'-O-acetate induced endothelium-dependent vasorelaxation through the eNOS/NO/sGC pathway, and also induced endothelium independent vasorelaxation involving the modulation of sGC activity, Kv channels, Ca2+ influx through Ca2+ channels and intracellular Ca2+ release. The data concerning the benefits of tiliacorinine 12'-O-acetate might be further investigated for the application of tiliacorinine 12'-O-acetate as an antihypertensive compound.

Ethyl rosmarinate relaxes rat aorta by an endothelium-independent pathway.

Ethyl rosmarinate is an ester derivative of rosmarinic acid, a major constituent of Hyptis suaveolens. The present study investigated the vasorelaxant mechanism of ethyl rosmarinate in isolated rat aortic rings using an organ bath system. Ethyl rosmarinate (0.1 µM-3mM) produced concentration-dependent relaxation in aortic rings pre-contracted with phenylephrine (10 µM), exhibiting a pD2 value of 4.56 ± 0.08 and an Emax value of 93.82 ± 5.00% (in endothelium-intact rings), as well as a pD2 value of 4.42 ± 0.05 and an Emax value of 92.10 ± 3.78% (in endothelium-denuded rings). In the endothelium-denuded rings, the vasorelaxant effect of ethyl rosmarinate was reduced by only 4-aminopyridine (1mM); however, this was not the case with tetraethylammonium (5mM), glibenclamide (10 µM), barium chloride (1mM), and 1H-[1,2,4] oxadiazolo [4,3-a]quinoxalin-1-one (ODQ, 1 µM). Ethyl rosmarinate also reduced the contraction induced by phenylephrine (10 µM) and caffeine (20mM) in a Ca(2+)-free solution, and inhibited the contraction induced by increasing extracellular Ca(2+) influx, which was induced by KCl (80 mM). Ethyl rosmarinate (10 µM) inhibits concentration-response curves for phenylephrine, while in the same concentration of ethyl rosmarinate has no effect on contractions induced by increasing concentrations of calcium in the presence of high extracellular potassium. Our results suggests that ethyl rosmarinate induces relaxation in aortic rings via an endothelium-independent pathway, which involves the opening of voltage-gated potassium (Kv) channels and the blockade of both Ca(2+)release from intracellular stores and extracellular Ca(2+) influx. Moreover, ethyl-rosmarinate acts on the extracellular Ca(2+) influx inhibition by interacting with voltage-operated calcium channels (VOCCs) and receptor-operated calcium channels (ROCCs).

In vitro and in vivo antitumor activity of tiliacorinine in human cholangiocarcinoma.

Cholangiocarcinoma (CCA) is a fatal cancer with poor prognosis and less than 10% of CCA patients can be offered surgical cure. Conventional chemotherapy results in unfavorable outcomes. At present, plant-derived compounds are gaining interest as potential cancer therapeutics, particularly for treatment-refractory cancers. In this study, antitumor activity of tiliacorinine, the major alkaloid isolated from a tropical plant, on CCA was first demonstrated. Antiproliferative effects of tiliacorinine on human CCA cell lines were investigated using SRB assays. Acridine orange/ethidium bromide staining, flow cytometric analysis and DNA laddering assays were used for apoptotic determination. Apoptosis-related proteins were verified by Western blotting and antitumor activity of tiliacorinine in vivo was demonstrated in CCA xenografted mice. Tiliacorinine significantly inhibited proliferation of human CCA cell lines with IC50 4.5-7 µM by inducing apoptosis through caspase activation, up- regulation of BAX, and down-regulation of BclxL and XIAP. Tiliacorinine considerably reduced tumor growth in CCA xenografted mice. These results demonstrated antitumor effects of tiliacorinine on human CCA in vitro and in vivo. Tiliacorinine may be an effective agent for CCA treatment.