isoliensinine and neferine

Topics: 5-alpha Reductase Inhibitors; Amino Acid Chloromethyl Ketones; Androgen Antagonists; Androgen Receptor Antagonists; Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; Autophagy; Benzylisoquinolines; Biological Products; Cell Line, Tumor; Cell Movement; Chromones; Heterocyclic Compounds, 3-Ring; Humans; Isoquinolines; Male; Morpholines; Nelumbo; Phenols; Phosphatidylinositol 3-Kinases; Prostatic Neoplasms; Proto-Oncogene Proteins c-akt; Signal Transduction

The coronavirus disease 2019 (COVID-19) pandemic has focused attention on the need to develop effective therapeutics against the causative pathogen, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and also against other pathogenic coronaviruses. In this study, we report on a kind of bisbenzylisoquinoline alkaloid, neferine, as a pan-coronavirus entry inhibitor. Neferine effectively protected HEK293/hACE2 and HuH7 cell lines from infection by different coronaviruses pseudovirus particles (SARS-CoV-2, SARS-CoV-2 [D614G, N501Y/D614G, 501Y.V1, 501Y.V2, 501Y.V3 variants], SARS-CoV, MERS-CoV) in vitro, with median effect concentration (EC

Topics: Antiviral Agents; Benzylisoquinolines; Calcium; Cell Line; Coronavirus; COVID-19; HEK293 Cells; Humans; Isoquinolines; Phenols; SARS-CoV-2; Virus Internalization

Cisplatin (CDDP) is a potent platinum-based chemotherapeutic agent used to treat solid tumors including colorectal cancer via inducing cytotoxicity. CDDP usage is limited due to the chemoresistance and associated adverse effects. A combinatorial regimen of phytochemicals with anticancer activity along with approved anticancer drugs seems to be a hopeful strategy against cancer treatment. Lotus-derived compounds such as neferine and isoliensinine have proven significant chemosensitizing activity in different cancer cells. Present study aims to compare chemosensitizing activity/anticancer potential of neferine/isoliensinine in combinatorial regimen with CDDP. Results documented that neferine/isoliensinine with CDDP augmented 'intracellular uptake of cisplatin' consequently apoptosis in HCT-15 cells exemplified by 'apoptotic morphological changes', 'S phase cell cycle arrest', 'ROS mediated oxidative stress' with the concomitant escalation in intracellular calcium & dissipation of MMP and activation of MAPK/PI3K/AKT pathway'. Furthermore, isoliensinine combination with CDDP exclusively enhanced CDDP uptake and induced more ROS-mediated apoptosis compared to other treatment regimens. Combination regimens induced downregulation of Bcl2 and upregulation of cytochrome c, caspase 3, 9, PARP cleavage indicating apoptosis induction through the intrinsic pathway. Thus, the results of the present study suggest that CDDP combination with neferine/isoliensinine augments the anticancer potential of CDDP in an additive manner and decrease CDDP dose requirement.

Topics: Antineoplastic Agents; Apoptosis; Benzylisoquinolines; Cell Line, Tumor; Cisplatin; Drug Combinations; Drug Synergism; Humans; Isoquinolines; MAP Kinase Signaling System; Membrane Potential, Mitochondrial; Reactive Oxygen Species

A new skeleton benzylisoquinoline (BI) named neoliensinine (1) was isolated from embryos of lotus seed (Nelumbo nucifera Gaertn.), a traditional Chinese herb. The tribenzylisoquinoline (TBI) structure of 1 was confirmed by interpreting spectroscopic data of UV, IR, MS, 1D and 2D NMR. The stereo-configurations of the new compound, together with two known bisbenzylisoquinolines (BBI), neferine and isoliensinine were established by analyzing

Topics: Alkaloids; Animals; Benzylisoquinolines; Female; In Vitro Techniques; Isoquinolines; Male; Mice, Inbred C57BL; Molecular Structure; Muscle, Smooth; Nelumbo; Neuromuscular Agents; Phenols; Seeds

The in vivo effects of traditional herbal medicines are generally mediated by multiple bioactive components. The main constituents of Lotus Plumule are alkaloids such as liensinine, isoliensinine and neferine. In this study, a simple, sensitive, and robust analytical method based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) has been developed for the determination of the three alkaloids in rat plasma using carbamazepine as internal standard (IS).. After precipitation of the proteins with acetonitrile, chromatography was performed on an Acquity UPLC BEH C18 column (2.1mm×50mm, 1.7μm particle size) using a gradient elution with 0.1% formic acid in water and acetonitrile. Mass spectrometry involved positive electrospray ionization and multiple reaction monitoring (MRM) of the transitions at m/z 611.7→206.2 for liensinine, 611.3→192.2 for isoliensinine, 625.2→206.1 for neferine and m/z 237.1→194.2 for IS.. The method was linear over the concentration range 5-1000ng/mL with a lower limit of quantifof 5ng/mL for each alkaloid. Inter- and intra-day precision (RSD%) were all within 11.4% and the accuracy (RE%) were equal or lower than 10.4%. Recoveries were more than 75.3% and matrix effects were not significant. Stability studies showed that the three alkaloids were stable under a variety of storage conditions.. The method was successfully applied to a pharmacokinetic study involving intravenous administration of liensinine, isoliensinine and neferine to rats.

Topics: Administration, Intravenous; Animals; Benzylisoquinolines; Chromatography, Liquid; Isoquinolines; Male; Phenols; Rats, Sprague-Dawley; Tandem Mass Spectrometry

Isoliensinine, liensinine and neferine are major bisbenzylisoquinoline alkaloids in the seed embryo of lotus (Nelumbo nucifera), and exhibit potential anti-cancer activity. Here, we explored the effects of these alkaloids on triple-negative breast cancer cells and found that among the three alkaloids isoliensinine possesses the most potent cytotoxic effect, primarily by inducing apoptosis. Interestingly, isoliensinine showed a much lower cytotoxicity against MCF-10A, a normal human breast epithelial cell line. Further studies showed that isoliensinine could significantly increase the production of reactive oxygen species (ROS) in triple-negative breast cancer cells, but not in MCF-10A cells. The isoliensinine-induced apoptosis could be attenuated by radical oxygen scavenger N-acetyl cysteine, suggesting that the cytotoxic effect of isoliensinine on cancer cells is at least partially achieved by inducing oxidative stress. We found that both p38 MAPK and JNK signaling pathways were activated by isoliensinine treatment and contributed to the induction of apoptosis. Furthermore, inhibitors or specific siRNAs of p38 MAPK and JNK could attenuate apoptosis induced by isoliensinine. However, only the p38 inhibitor or p38-specific siRNA blocked the elevation of ROS in isoliensinine-treated cells. Our findings thus revealed a novel antitumor effect of isoliensinine on breast cancer cells and may have therapeutic implications.

Topics: Apoptosis; Benzylisoquinolines; Cell Line, Tumor; Enzyme Activation; Female; Humans; Isoquinolines; MAP Kinase Signaling System; MCF-7 Cells; p38 Mitogen-Activated Protein Kinases; Phenols; Reactive Oxygen Species; Triple Negative Breast Neoplasms

We attempted to ascertain if bisbenzylisoquinoline alkaloids, liensinine and isoliensinine from Nelumbo nucifera Gaertner have antidepressant-like effects and compare the effects with those previously obtained by their analogue neferine.. Using mice, the forced swimming test (FST) was carried out after treatment with liensinine, isoliensinine and neferine.. Liensinine and isoliensinine elicited antidepressant-like effects in mice after the FST. Anti-immobility effects of liensinine and isoliensinine were antagonized by the 5-hydroxytryptamine1 A (5-HT1 A ) receptor antagonist WAY 100635, but not by the α1 -adrenoceptor antagonist prazosin. The anti-immobility effects of liensinine, isoliensinine and neferine were blocked by pretreatment with p-chlorophenylalanine (PCPA), which depletes serotonin (5-HT).. These data suggest that liensinine and isoliensinine from Nelumbo nucifera Gaertner have antidepressant-like effects and that antidepressant-like effects of liensinine and its analogues are closely related to serotonergic mechanisms.

Topics: Animals; Antidepressive Agents; Behavior, Animal; Benzylisoquinolines; Disease Models, Animal; Dose-Response Relationship, Drug; Isoquinolines; Male; Mice, Inbred ICR; Motor Activity; Nelumbo; Phenols; Phytotherapy; Plant Extracts; Plants, Medicinal; Seeds; Serotonergic Neurons; Serotonin Agents; Stress, Psychological; Swimming

To identify the metabolites of norcoclaurine,liensinine, isoliensinine and neferine in Caco-2 cells by LC/ MS/MS.. After Caco-2 cells were treated with norcoclaurine, liensinine, isoliensinine or neferine for 3, 6 and 12 h, samples were collected, purified and then analyzed by LC/MS/MS. The structures of the metabolites were elucidated by molecular masses, retention times, MS and MS/MS spectra comparing with those of the parent drug.. The procedure identified that the major metabolites of norcoclaurine were methylnorcoclaurine and norcoclaurine-glucuronide, the major metabolite of liensinine was demethyl-liensinine, the major metabolite of isoliensinine was demethyl-isoliensinine, the major metabolites of neferine were liensinine, isoliensinine and their further demethylation products.. LC/MS/MS is simple, rapid and sensitive for the metabolites identification. Methylation, demethylation and glucuronidation are main metabolic pathways of alkaloids from Nelumbinis Plumula in Caco-2 cells.

Topics: Alkaloids; Benzylisoquinolines; Caco-2 Cells; Chromatography, Liquid; Drugs, Chinese Herbal; Humans; Isoquinolines; Magnoliopsida; Metabolic Networks and Pathways; Methylation; Molecular Weight; Phenols; Tandem Mass Spectrometry; Tetrahydroisoquinolines

Lotus plumule, the dried young cotyledon and radicle of the Nelumbo nucifera Gaertn. (Fam. Nymphaeaceae) ripe seed, is a famous Traditional Chinese Medicine to remove heat from the heart, anchor the mind, improve seminal emission, and arrest bleeding for centuries in China. Liensinine and its analogs neferine and isoliensinine are the major active components in lotus plumule. Aim of the study is to investigate the association of liensinine, neferine, and isoliensinine with efflux transporters.. Caco-2, MDCK, MDCK-MDR1, and MDCK-MRP2 were used as cell models for the transcellular transport and accumulation studies.. The results obtained in Caco-2 cells suggested that P-glycoprotein (P-gp) might be involved in transcellular transport. Cellular accumulation and transport experiments were further performed in MDCK-MDR1 cells. GF120918 and cyclosporine A were found to completely inhibit the efflux, and the net efflux ratios of these alkaloids exhibited saturation over the concentration range. No significant differences in liensinine accumulation and transport were observed between MDCK and MDCK-MRP2 cells.. These results demonstrated that liensinine, neferine, and isoliensinine are substrates of P-gp, whereas MRP2 is not involved in the transport process, suggesting that P-gp might be responsible for the absorption and distribution of the 3 alkaloids.

Topics: Absorption; Animals; ATP Binding Cassette Transporter, Subfamily B, Member 1; Benzylisoquinolines; Biological Transport; Caco-2 Cells; Cell Survival; Dogs; Humans; Isoquinolines; Madin Darby Canine Kidney Cells; Multidrug Resistance-Associated Protein 2; Multidrug Resistance-Associated Proteins; Phenols

This study was conducted to investigate the effect of Nelumbinis Plumula total alkaloid (NPA) and its main alkaloid components on oxidative stress induced by tert-butyl hydroperoxide (t-BHP) in the human hepatocellular HepG2 cell line. According to HPLC analysis, several major alkaloid compounds such as liensinine, isoliensinine and neferine were present in NPA. The cytotoxic effects in 0.55 mM t-BHP-induced HepG2 cells were significantly inhibited by NPA and the major compound in NPA, neferine, showed the strongest activities. The protective effect of neferine against oxidative stress induced by t-BHP may be associated with decreased ROS formation, TBARS generation, LDH release and increased GSH levels, suggesting their involvement of the cytoprotective on oxidative stress. The effects were comparable with quercetin, which was used as positive control. Overall, total alkaloid and alkaloid compounds from Nelumbinis Plumula displayed a significant cytoprotective effect against oxidative stress. Further study is needed to elucidate the relationship between the chemical structures of the components in NPA and their protective effect on oxidative stress.

Topics: Alkaloids; Benzylisoquinolines; Biphenyl Compounds; Drug Evaluation, Preclinical; Free Radical Scavengers; Free Radicals; Glutathione; Hep G2 Cells; Humans; Isoquinolines; L-Lactate Dehydrogenase; Malondialdehyde; Nelumbo; Oxidants; Oxidative Stress; Phenols; Picrates; Plant Extracts; Reactive Oxygen Species; tert-Butylhydroperoxide; Thiobarbituric Acid Reactive Substances

A rapid and specific HPLC method has been developed and validated for the simultaneous determination of liensinine (CAS 2586-96-1), isoliensinine (CAS 6817-41-0) and neferine (CAS 2292-16-2) in rat plasma. The sample was prepared by a liquid-liquid extraction with diethyl ether and the recovery was above 80% from the plasma for the three compounds. Chromatographic separation was achieved with a Hypersil BDS C18 column (4.0 mm x 250 mm, particle size 5 microm). A mobile phase consisting of methanol: 0.2 M KH2PO4:0.2 M NaOH:triethylamine (71:17:12:0.002, v/v/v/v, pH 9.2-9.3) was slowly delivered at 0.8 ml/min in isocratic mode with a detection wavelength of 282 nm. The linearity of calibration curves were good (r > 0.999) in the concentration range of 0.031-2.00 microg/ ml. The lower limit of quantification can reach 0.03 microg/ml for the three compounds. The intra-day and inter-day variations estimated with QC samples were less than 8% for the three tested concentration levels. This developed method was applied in the plasma pharmacokinetic study of total bisbenzylisoquinoline alkaloids (TAL) of the lotus flower (Lian Zi Xin) following a single oral and intravenous administration of TAL in rats.

Topics: Administration, Oral; Animals; Area Under Curve; Benzylisoquinolines; Calibration; Chromatography, High Pressure Liquid; Female; Half-Life; Injections, Intravenous; Isoquinolines; Male; Nelumbo; Phenols; Rats; Rats, Wistar; Reference Standards; Reproducibility of Results; Seeds; Solutions; Spectrophotometry, Ultraviolet

A preparative high-speed counter-current chromatography method for separation and purification of liensinine, isoliensinine and neferine from seed embryo of Nelumbo nucifera GAERTN was successfully established by using n-hexane-ethyl acetate-methanol-water (5:8:4:5, v/v, containing 0.5% NH(4)OH) as the two-phase solvent system. From 200 mg of crude extract, 18.4 mg of liensinine, 19.6 mg of isoliensinine and 58.4 mg of neferine were obtained with the purity of 96.8, 95.9, and 98.6%, respectively. The identification of the three alkaloids was performed with (1)H NMR and (13)C NMR.

Topics: Benzylisoquinolines; Chromatography, High Pressure Liquid; Countercurrent Distribution; Isoquinolines; Magnetic Resonance Spectroscopy; Nelumbo; Phenols; Plant Extracts; Seeds

An application of mass spectrometric methods has been developed to characterize, prepare and quantitatively analyze three bisbenzylisoquinoline alkaloids (liensinine, isoliensinine and neferine) from embryo of the seed of Nelumbo nucifera Gaertn. Initially, an analytical method based on liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) with positive ionization mode using a MonoChrom C18 column (4.6 mm x 250 mm i.d. 10 microm) has been developed to characterize liensinine, isoliensinine and neferine, and then scaled up to purify them on a 21.4 mm x 250 mm preparative column. The structures of liensinine, isoliensinine and neferine were elucidated by NMR. Finally, a LC-MS/MS determination method, successfully applied to separation within 3 min, was developed for high throughput simultaneous measurement of liensinine, isoliensinine, and neferine in the extract samples. Multiple reaction monitoring (MRM) was used to monitor the transition of the protonated molecules m/z 611, 611, 625 [M+H]+ to the product ions m/z 206, 192, 206 for analysis of liensinine, isoliensinine and neferine. The LC-MS/MS system was linear in the concentration range of 0.0247-6.02 microg/ml with correlation coefficients of r2>0.992. The quantitative method was validated, with an S/N=3 detection limit of 0.15 ng for liensinine, 0.19 ng for isoliensinine and 0.1 2ng for neferine. The mass fractions of liensinine, isoliensinine, and neferine in the crude extract and the phenolic alkaloid sample of embryo of the seed of N. nucifera Gaertn. were 16.5+/-1.1 and 228.6+/-11.9 for liensinine (mg/g), 45.7+/-1.8 and 640.7+/-15.2 for isoliensinine (mg/g), 59.7+/-6.4 and 58.8+/-9.8 for neferine (mg/g).

Topics: Alkaloids; Benzylisoquinolines; Calibration; Chromatography, High Pressure Liquid; Isoquinolines; Mass Spectrometry; Nelumbo; Phenols; Plant Extracts; Reproducibility of Results; Seeds; Spectrometry, Mass, Electrospray Ionization; Spectrophotometry, Ultraviolet

To determine the optimum technical parameters through controlling the different factors and the content of Dripping Pill.. Time of dissolution, spherical degree and pill weight as parameters, the influential factors were investigated by orthogonal test and then to determine the content of liensinine, isoliensinine, neferine in plumula nelumbinis Alkaloid Dripping Pill.. The optimum condition: 75% PEG4000 as matrix, methyl-silicon oil as refrigerant, 4 cm dripping distance, 10 degrees C refrigerant, 30 drippings per minute.. The good quality pills can be produced through this process and the HPLC determination method is simple, reliable and can be used in the quality control of plumula nelumbinis Alkaloid Dripping Pill.

Topics: Alkaloids; Benzylisoquinolines; Chromatography, High Pressure Liquid; Isoquinolines; Microspheres; Nelumbo; Particle Size; Phenols; Plants, Medicinal; Polyethylene Glycols; Quality Control; Seeds; Solubility; Technology, Pharmaceutical

To develop a RP-HPLC method for determination of three alkaloids in total alkaloids exfracted from Lotus plumule.. A Hypersil C18 column was used with the mobile phase of methanol-phosphate (73:27, pH 9.0) at the detection wavelength of 230 and 286 nm. The flow rate was 1 mL min(-1).. The linear range was 20-700 microg mL(-1), the recovery of three alkaloids were 97.9%, 98.4% and 98.1%. The RSD of intra-day and inter-day was 0. 24% -4. 83%. Different groups of samples were determined by this method. The result showed that the extraction method with methanol was prior to the extraction method with alcohol.. The method is simple, rapid and suitable for the determination of three alkaloids in total alkaloids exfracted from L. plumule.

Topics: Alkaloids; Benzylisoquinolines; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Isoquinolines; Lotus; Phenols; Plants, Medicinal; Reproducibility of Results; Seeds

The present study utilized LC-MS and HPLC approaches to characterize the metabolites of neferine in rat liver after an oral administration of 20 mg x kg(-1), and investigated the involvement of CYP450 isoforms in the metabolism of neferine by their selective inhibitors in vitro, separately. In positive ionization mode, besides neferine, four metabolites (M1-M4) were detected. M2 (the major metabolite) and M4 were identified as liensinine and isoliensinine by comparison with reference substances. Moreover, according to the analysis of metabolic rule of parent drug (neferine), M1 and M3 may be desmethylliensinine and desmethyl-isoliensinine, respectively. Furthermore, the metabolism of neferine in rat liver microsomes showed that the percentage inhibition of the major metabolism (liensinine) formation was 80.5% by quinidine (10 micromol x L(-1), selective CYP2D1 inhibitor) and 25.7% by ketoconazole (1 micromol x L(-1), selective CYP3A1 inhibitor). Neferine was mainly metabolized by CYP2D1 or CYP3A1 to liensinine, isoliensinine, desmethyl-liensinine and desmethyl-isoliensinine.

Topics: Administration, Oral; Alcohol Oxidoreductases; Animals; Aryl Hydrocarbon Hydroxylases; Benzylisoquinolines; Chromatography, High Pressure Liquid; Cytochrome P-450 CYP3A; Cytochrome P450 Family 2; Isoquinolines; Ketoconazole; Male; Microsomes, Liver; Nelumbo; Phenols; Plants, Medicinal; Quinidine; Rats; Seeds; Spectrometry, Mass, Electrospray Ionization

Neferine and isoliensinine are the two major compositions of the Chinese medicine lotus plumule. They have extensive cardiovascular activity, such as antiarrhythmic, antithrombic and antihypertensive. In order to confirm the structures reported in the literature by explicit 1H and 13C assignments, we used a series of NMR experiments including 1H, 1H-COSY, HSQC and HMBC. The absolute configuration of neferine (C-1 and C-1') was determined as R and S and that of isoliensinine (C-1 and C-1') was determined as R and R.

Topics: Benzylisoquinolines; Drugs, Chinese Herbal; Isoquinolines; Magnetic Resonance Spectroscopy; Molecular Structure; Nelumbo