Genotoxic effects of (-)-cubebin in somatic cells of mice

(?)‐Cubebin belongs to the dibenzylbutyrolactone lignan group, which is widely distributed in the plant kingdom. Because this compound shows interesting biological activities, it is extremely important to evaluate its possible genotoxic activity to allow its safe use in humans. Thus, the present study was performed to investigate the genotoxicity potential activity of (?)‐cubebin assessed by two assays: micronucleus in bone marrow cells and comet test in peripheral blood leukocytes of Swiss mice. In the (?)‐cubebin dose range‐finding assays, the maximum tolerated dose was greater than 2000 mg kg^?1. The compound was administered by an oral route at single doses of 250, 500 and 2000 mg kg^?1 body weight. Cytotoxicity was assessed by scoring 200 consecutive total polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). Under our experimental conditions, micronucleus and comet assays, respectively, showed that (?)‐cubebin caused dose‐related clastogenic and genotoxic effects in the somatic cells investigated. PCE/NCE ratio showed no cytotoxicity for the three doses of the compound. The data suggest caution in the ingestion of (?)‐cubebin by humans, especially at high doses. Copyright © 2010 John Wiley & Sons, Ltd.     

The Lignan (‐)‐Cubebin Inhibits Vascular Contraction and Induces Relaxation Via Nitric Oxide Activation in Isolated Rat Aorta

Cubebin, the most abundant lignan in Piper cubeba, has been described as having several effects as trypanocidal, antimycobacterial, antispasmodic, antimicrobial, anti‐inflammatory, and analgesic. This study investigated the vasorelaxant effect produced by (‐)‐cubebin in isolated rat aortic rings pre‐contracted with phenylephrine (Phe), and the possible mechanism involved in this event was evaluated. Endothelium‐dependent relaxation was evoked by acetylcholine and (‐)‐cubebin in intact aortic rings, while endothelium‐independent vasorelaxation was elicited by sodium nitroprusside and (‐)‐cubebin in denuded rings. Cumulative concentration–response curves for Phe (10^?10–10^?5 M) were determined for endothelium‐intact and endothelium‐denuded aortic rings in either the presence or absence of (‐)‐cubebin. Dose–response curves were also constructed for pre‐incubation of vascular rings with Nω‐nitro‐L‐arginine methyl ester (L‐NAME) (a non‐specific nitric oxide synthase inhibitor), indomethacin (an unspecific cyclooxygenase inhibitor), and 1H‐[1,2,4] oxadiazolo [4,3‐a]quinoxalin‐1‐one (ODQ) (a guanylyl cyclase inhibitor). (‐)‐Cubebin was found to exert a vasorelaxant effect irrespective of the presence of endothelium, which was abolished by pretreatment with L‐NAME and ODQ, but not with indomethacin. In addition, (‐)‐cubebin was able to reduce Phe contraction in the case of intact rings. These results suggest that (‐)‐cubebin promotes vasorelaxation via NO/cGMP pathway in rat aorta, without prostacyclin involvement. Copyright © 2013 John Wiley & Sons, Ltd.     

Analgesic and anti-inflammatory activities evaluation of (-)-O-acetyl, (-)-O-methyl, (-)-O-dimethylethylamine cubebin and their preparation from (-)-cubebin

The anti-inflammatory and antinociceptive effects of the acetylated (2), methylated (3) and aminated (4) derivatives of cubebin (1), obtained by its reaction with acetic anhydride, methyl iodide and dimethylethylamine chloride, respectively, were investigated, using different animal models. The compound (2) was the most effective anti-inflammatory one in the carrageenin-induced paw edema in rats and was the only one which showed dose-response correlation for this assay with r = 0.993 and Y = 64.58x + 0.22. Besides, compounds (2) and (4) were more effective than cubebin in inhibiting acetic acid-induced writhing in mice, producing dose-response correlation with doses of 10, 20 and 30 mg/kg, respectively. Regarding the hot plate and the cell migration tests in rats, none of the four tested compounds showed activity. Overall, the results showed that the acetylation and amination of cubebin were efficient in enhancing its analgesic activity, as well as its anti-inflammatory activity.     

基于网络药理学和分子对接技术探讨柴银颗粒抗冠状病毒感染潜在分子机制

目的基于网络药理学和分子对接分析柴银颗粒抗冠状病毒感染的分子机制。方法从TCMSP数据库中获取柴银颗粒成分,其潜在靶点以及冠状病毒作用靶点分别利用SwissTargetPrediction数据库和GeneCards数据库获取。通过绘制韦恩图发现柴银颗粒抗冠状病毒感染的潜在药效成分和作用靶点。潜在药效成分-作用靶点网络和潜在作用靶点的互作网络可视化由Cytoscape 3.7.0软件完成。GO功能和KEGG通路分析在String数据库中进行。潜在药效成分和关键靶点的分子对接由autodock vina 1.1.2实现。结果发现了柴银颗粒抗冠状病毒感染的51个潜在药效成分和14个潜在作用靶点。生物信息学分析发现PI3K-Akt、IL-17等信号通路与柴银颗粒抗冠状病毒感染的分子机制相关。分子对接结果显示柴银颗粒中的黄芩苷等成分与NTRK2等靶点的亲和力强。结论柴银颗粒中的黄芩苷、荜澄茄素、黄连碱等作用于NTRK2、PRKCα、TNF等,调节PI3K-Akt/m TOR、Erb B/Ras和IL-17等信号通路抑制冠状病毒的侵袭和复制,增强宿主免疫能力,实现抗冠状病毒感染。     

Evaluation of piper cubeba extract, (-)-cubebin and its semi-synthetic derivatives against oral pathogens

The activities of the crude ethanol extract from Piper cubeba seeds, (-)-cubebin and its semi-synthetic derivatives were evaluated against oral pathogens. The crude ethanol extract was more active against Streptococcus salivarius (MIC value of 80 microg/mL). (-)-Cubebin displayed MIC values ranging from 0.20 mm for Streptococcus mitis to 0.35 mm for Enterococcus faecalis. The natural product (-)-cubebin and its semi-synthetic derivative (-)-hinokinin displayed bacteriostatic activity at all evaluated concentrations, as well as fungicidal activity against Candida albicans at 0.28 mm. The O-benzyl cubebin derivative showed fungistatic and fungicidal effects against C. albicans at 0.28 mm and 0.35 mm, respectively. Also, the other dibenzylbutyrolactone derivatives [(-)-6,6'-dinitrohinokinin and (-)-O-(N,N-dimethylaminoethyl)-cubebin] displayed bacteriostatic and fungistatic effects at the evaluated concentrations. Moreover, the semi-synthetic derivative (-)-6,6'-dinitrohinokinin was the most active compound against all the evaluated microorganisms. Therefore, it may be suggested that the presence of the carbonyl group at C-9 plus the introduction of polar groups in the aromatic rings improve the antimicrobial activity of dibenzylbutyrolactone compounds.