The visualization of the spatial distribution of Cocculus orbiculatus based on air flow-assisted desorption electrospray ionization mass spectrometry imaging.

Cocculus orbiculatus (C. orbiculatus), the root of plants belonging to the Menispermaceae family, has been extensively used to treat various diseases, including malaria and rheumatism. The main chemicals in these plants are alkaloids; however, the spatial distribution of these compounds within the plant roots remains undefined. This study aimed to visualize the spatial distribution of C. orbiculatus using air flow-assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI). In total, the spatial distribution of four aporphine alkaloids, five benzyltetrahydroisoquinoline alkaloids, six bisbenzylisoquinoline alkaloids, and one morphinane alkaloid in the cork layer, xylem, and ray of the root of C. orbiculatus was observed; the distribution characteristics of the different compounds in C. orbiculatus were significantly different. This study provides a visualized spatial distribution analysis method for the characterization of metabolites in the root tissue of C. orbiculatus and also provides valuable information for the specificity of the root of C. orbiculatus, which is beneficial for understanding its chemical separation, biosynthesis, and pharmacological activities.

Sinococuline, a bioactive compound of Cocculus hirsutus has potent anti-dengue activity.

Dengue virus (DENV) infection has increased worldwide, with over 400 million infections annually, and has become a serious public health concern. Several drug candidates, new and repurposed, have failed to meet the primary efficacy endpoints. We have recently shown that Aqueous Extract of the stem of Cocculus hirsutus (AQCH) was effective in vitro and in vivo against DENV and was safe in humans. We now report that an active ingredient of AQCH, Sinococuline, protects against the antibody-mediated secondary-DENV infection in the AG129 mouse model. DENV infection markers were assessed, viz. serum viremia and vital organs pathologies-viral load, proinflammatory cytokines and intestinal vascular leakage. The treatment with Sinococuline at 2.0 mg/kg/day; BID (twice a day), was the most effective in protecting the severely DENV-infected AG129 mice. Also, this dose effectively reduced serum viremia and tissue-viral load and inhibited the elevated expression levels of proinflammatory cytokines (TNF-α and IL-6) in several vital organs. Based on these findings, it could be explored further for pre-clinical and clinical developments for the treatment of dengue.

[Chemical constituents of Cocculus orbiculatus var. mollis root].

OBJECTIVE: To study the chemical constituents in the root of Cocculus orbiculatus var. mollis. METHODS: The compounds were isolated by silica gel chromatography, their structures were established by spectroscopic methods. RESULTS: Eleven compounds were isolated and identified as wattisine A (I), O-methylcocsoline (II), (+) cocsoline (III), (+) cocsuline (IV), magnoflorine (V), sino-coculine (VI), isosinococuline (VII), (-) coclaurine (VIII), daucosterol (IX), beta-sitosterol (X) and 1-oleioyl-3-(9Z, 12Z-arachoyl) glycerol (XI). CONCLUSION: Compound I is isolated from this genus for the first time,and compound II - XI are isolated from this plant for the first time.

Anticancer activity of Cocculus hirsutus against Dalton's lymphoma ascites (DLA) cells in mice.

CONTEXT: Cocculus hirsutus (L.) Diels (Menispermaceae) is used in Indian folk system of alternative medicine for rheumatism, eczema, diabetics, inflammation, and neuralgia. OBJECTIVE: To evaluate antitumor activities of C. hirsutus in vitro and in vivo. MATERIALS AND METHODS: C. hirsutus was successively extracted using hexane, petroleum ether, chloroform, ethyl acetate, methanol, and water. In vitro cytotoxicity was assessed by the MTT assay. Phytochemical analyses were conducted with methanol extract of C. hirsutus (MECH) and in vivo antitumor activity was carried out with MECH using Dalton's lymphoma ascites (DLA) mouse model. Antioxidant properties were assessed by estimating superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation. RESULTS AND DISCUSSION: Phytochemical studies indicated a high content of total alkaloid (165.6 mg/100 g), total phenolic (43.5 GAE mg/g), and total flavanoid (4.97 RE mg/g) in MECH. Anti-proliferative activity against the breast cancer cell line MCF-7 showed IC50 values of 221.5 ± 16.68, 255 ± 17.88, 213 ± 8.4, 147 ± 7.9, and 229 ± 8.02 µg/ml with hexane, petroleum ether, chloroform, ethyl acetate, methanol, and aqueous extracts, respectively. A significant (p < 0.01) decrease in packed cell volume, viable cell count, and increased lifespan (58 and 77%) was observed. Hematological and serum biochemical profiles were restored to normal levels in MECH-treated mice. MECH-treated group significantly (p < 0.001) decreased SOD, lipid peroxidation, and CAT towards normal. CONCLUSION: C. hirsutus exhibited significant in vitro and in vivo antitumor activities that are reasonably attributed to endogenous antioxidant mechanisms.

Plant-derived nanoparticle treatment with cocc 30c ameliorates attention and motor abilities in sleep-deprived rats.

Sleep is an essential physiological process that underlies crucial cognitive functions as well as emotional reactivity. Thus, sleep deprivation (SD) may exert various deleterious effects. In this study, we aimed to examine the adverse behavioral and hormonal effects of SD and a potential treatment with Plant-derived nanoparticle treatment - cocc 30c. The study was a 4-arm trial with randomization and double-blinding of verum and placebo treatments. SD was induced by using the Multiple Platform Method for 48 h. The effects of SD were evaluated behaviorally (pre-pulse inhibition (PPI), startle response and rotor-rod) at baseline as well as at 6, 12, 24h, and 14 days post deprivation. cocc 30c treatment was administrated Per Os every three hours starting immediately after baseline tests and for a period of 24h. On day 14, blood samples were taken and serum levels of corticosterone, testosterone, serotonin and leptin were tested. We found that cocc 30c improved PPI 12 and 24h post deprivation, likewise, cocc 30c improved motor learning. On day 14 SD led to increased startle response that was ameliorated by cocc 30c. Likewise, SD led to increased levels of corticosterone and serotonin while decreasing testosterone and leptin. Interestingly, cocc 30c treatment has moderated these hormonal alterations. We conclude that the treatment with cocc 30c recovers both short-term behavioral and the long-term hormonal modulations following SD.

Nephroprotective activity of Cocculus hirsutus leaf extract in 5/6 nephrectomized rat model.

BACKGROUND: Cocculus hirsutus, commonly known as broom creeper, belongs to the family Menispermaceae. It is widely used in folk medicine to treat leprosy, skin diseases, dyspepsia, etc. Hence, an effort has been made to investigate the nephroprotective potential of C. hirsutus. METHODS: The nephroprotective activity of ethanolic C. hirsutus leaf extract (ECHE) in the 5/6 nephrectomized rat model was investigated. Different parameters like postoperative survival rate, change in body weight and levels of red blood corpuscles (RBCs), hemoglobin (Hb), triglyceride, cholesterol, creatinine, urea and uric acid were estimated in experimental rats. RESULTS: The findings revealed the postoperative survival rate of rats in the investigated novel method to be 100%. Change in the body weight of ECHE-treated groups I and II was found to be 32 and 30 g, respectively. These values suggested that ECHE treatment normalized the elevated body weight levels in experimental rats. Furthermore, ECHE treatment normalized the decreased RBC levels and the elevated Hb, triglyceride, cholesterol, creatinine, urea and uric acid levels in experimental rats. CONCLUSIONS: The results indicate that C. hirsutus has strong nephroprotective activity. However, further scrutiny is essential for isolation and characterization of the active components that can be employed to allay various human maladies.

Simultaneous determination of structurally diverse compounds in different Fangchi species by UHPLC-DAD and UHPLC-ESI-MS/MS.

Two bisbenzylisoquinoline alkaloids, two morphine alkaloids, one aporphine alkaloid, syringaresinol and aristolochic acid І were selected as marker compounds and simultaneously analyzed using an ultra-high pressure liquid chromatography-diode array detection (UHPLC-DAD) method. These marker compounds were used for the quality control of Fangchi species of different origins, including Sinomenium acutum, Stephania tetrandra, Cocculus trilobus and Aristolochia fangchi. A reversed-phase UHPLC-DAD method was developed and validated for the simultaneous quantification of structurally diverse markers in different Fangchi species. In addition, an UHPLC-electrospray ionization tandem mass spectrometry (ESI-MS/MS) method was used for marker identification in Fangchi species, which provided diagnostic MS/MS spectral patterns that were dependent upon the marker structures. The UHPLC-MS/MS data were used to confirm and complement the UHPLC-DAD quality evaluation results. Additionally, magnoflorine and syringaresinol were observed for the first time in S. tetrandra and C. trilobus, respectively. Twenty different Fangchi species samples were analyzed for aristolochic acid I, syringaresinol and the alkaloids using the UHPLC-DAD and MS/MS method. Based on the levels of markers and principal component analysis (PCA), this method allowed for the clear classification of the samples into four different groups representing samples originating from the four species.

Evaluation of repellent properties of botanical extracts against Culex tritaeniorhynchus Giles (Diptera: Culicidae).

In recent times, there were considerable efforts made to promote the use of environmentally friendly and biodegradable natural insecticides and repellents, particularly from botanical sources. The present study explored the effects of crude leaf ethyl acetate, acetone, and methanol extracts of Aegle marmelos (L.) Correa ex Roxb, Andrographis lineata Wallich ex Nees., Andrographis paniculata (Burm.f.) Wallich ex Nees., Cocculus hirsutus (L.) Diels, Eclipta prostrata L., and Tagetes erecta L. on repellent activity against Culex tritaeniorhynchus Giles. The maximum repellent activity was observed at 500 ppm in methanol extracts of A. marmelos, ethyl acetate extracts of A. lineata, C. hirsutus, and E. prostrata and the mean complete protection time ranged from 120 to 150 min with the different extracts tested. The ethyl acetate extract of A. lineata showed 100% repellency in 120 min; acetone extracts of A. marmelos and C. hirsutus and methanol extract of T. erecta showed complete protection in 90 min at 250 ppm, respectively. These results suggest that the leaf extracts of A. marmelos, A. lineata, and C. hirsutus have the potential to be used as an ideal eco-friendly approach for the control of the C. tritaeniorhynchus. Therefore, this study provides first report on the repellent activity against Japanese encephalitis, C. tritaeniorhynchus of plant extracts from Southern India.

Efficacy of botanical extracts against Japanese encephalitis vector, Culex tritaeniorhynchus.

The purpose of the present study was to assess the effect of leaf hexane and chloroform extracts of Aegle marmelos, Andrographis lineata, Andrographis paniculata, Cocculus hirsutus, Eclipta prostrata, and Tagetes erecta on repellent, ovicidal, and oviposition-deterrent activities against Culex tritaeniorhynchus Giles (Diptera: Culicidae). The repellent action of the plant extracts tested varied depending on the plant species, part, solvent used in extraction, and the extract dose. The hexane extract of A. paniculata was more effective in exhibiting the repellent action against the mosquito as compared with A. lineata extract. Complete protections for 150 min were found in hexane extract of A. paniculata at 500 ppm against mosquito bites. Mean percent hatchability of the ovicidal activity was observed 24 h after treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. No hatchability was observed with hexane, and chloroform extracts of A. lineata, A. paniculata, and hexane extract of T. erecta were exerted at 1,000 ppm. The percentage of effective oviposition repellency were 95.90, 94.75, 95.04, 90.58, 87.93, 87.14, 94.82, 95.71, 92.26, 90.58, 83.35, and 78.16 at 500 ppm, and the lowest repellency was 69.93, 53.06, 64.81, 70.06, 51.82, 54.54, 48.31, 66.71, 68.82, 61.85, 34.84, and 39.53 at 31.25 ppm in hexane and chloroform extracts of A. marmelos, A. lineata, A. paniculata, C. hirsutus, E. prostrata, and T. erecta, respectively. The oviposition activity index values revealed that the solvent plant extracts have deterrent effect, and they caused a remarkable negative response resulting in oviposition of very few eggs. These results clearly reveal that the hexane extracts of A. marmelos and A. paniculata served as a potential repellent, ovicidal, and oviposition deterrent against Japanese encephalitis vector, C. tritaeniorhynchus.

Oviposition-deterrent, ovicidal, and repellent activities of indigenous plant extracts against Anopheles subpictus Grassi (Diptera: Culicidae).

Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. The leaf acetone, ethyl acetate, and methanol extracts of Aegle marmelos (Linn.) Correa ex Roxb, Andrographis lineata Wallich ex Nees, and Cocculus hirsutus (L.) Diels were tested for oviposition-deterrent, ovicidal, and repellent activities against Anopheles subpictus Grassi (Diptera: Culicidae). The percentage of effective oviposition repellency of 92.60 , 93.04, 95.20, 88.26, 92.80, 94.01, 95.77, 96.93, and 92.54 at 500 ppm and the lowest repellency of 47.14, 58.00, 56.52, 64.93, 71.09, 66.42, 50.62, 57.62, and 65.73 at 31.25 ppm in acetone, ethyl acetate, and methanol extracts of Aegle marmelos, Andrographis lineata, and Cocculus hirsutus, respectively. The oviposition activity index (OAI) value of acetone, ethyl acetate, and methanol extracts of Aegle marmelos, Andrographis lineata, and Cocculus hirsutus at 500 ppm were -0.86, -0.87, -0.90, -0.78, -0.87, -0.86, -0.91, -0.94, and -0.86 respectively. The OAI values revealed that the solvent plant extracts have deterrent effect, and they caused a remarkable negative response resulting in oviposition of very few eggs. Mean percent hatchability of the ovicidal activity was observed 24 h after treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Mortality of 100% with ethyl acetate extract of Aegle marmelos, methanol extracts Aegle marmelos, Andrographis lineata, and Cocculus hirsutus were exerted at 1,000 ppm. The maximum repellent activity was observed at 500 ppm in methanol extracts of Aegle marmelos, Andrographis lineata, and ethyl acetate extract of Cocculus hirsutus, and the mean complete protection time ranged from 90 to 120 min with the different extracts tested. These results suggest that the leaf extracts of Aegle marmelos, Andrographis lineata, and Cocculus hirsutus have the potential to be used as an ideal ecofriendly approach for the control of the Anopheles subpictus. Therefore, this study provides first report on the oviposition, ovicidal, and repellent activities against malaria vector, Anopheles subpictus of plant extracts from Southern India.

Bisbenzylisoquinoline alkaloids from Cocculus pendulus.

A new bisbenzylisoquinoline alkaloid, 1,2-dihydrokurramine (1), and four known bisbenzylisoquinoline alkaloids 2-5, along with a morphinan alkaloid 6 were isolated from Cocculus pendulus. The structures for these compounds were deduced by spectroscopic methods. All bisbenzylisoquinoline alkaloids showed inhibitory activities against acetyl- and butyrylcholinesterases.

Acute and chronic diuretic effect of ethanolic extract of leaves of Cocculus hirsutus (L.) Diles in normal rats.

OBJECTIVES: The objective of this study was to evaluate the acute and chronic diuretic effect of the ethanolic extract of the leaves of Cocculus hirsutus (L.) Diles. METHODS: The ethanolic extract was administered (100, 200 and 400 mg/kg, p.o.) in Wistar rats. In the acute study, rats received drugs orally and urine was collected after 1, 2, 3, 4, 5 and 6 h. The chronic study involved repeated administration of ethanolic extract for 28 days and urine was collected on day 1, 7, 14, 21 and 28. The parameters were total urine volume, concentration of Na+, K+ and Cl- ions, creatinine in urine and serum. Urine output, electrolytes (Na+, K+ and Cl- ions) and creatinine were determined on day 7, 14, 21 and 28. KEY FINDINGS: The highest dose (400 mg/kg) of the ethanolic extract significantly (P < 0.01) enhanced urine output. Excretion of cations (Na+ and K+ ions) and anions (Cl- ions) increased significantly with respect to the control (gum acacia 2% dissolved in saline, 10 ml/kg) group. The increase of cations in the urine after treatment with ethanolic extract was dose dependent. The ethanolic extract of the leaves of C. hirsutus (100, 200 and 400 mg/kg) and furosemide (10 mg/kg) did not significantly change the concentration of Na+, K+ and Cl- ions in serum. The ethanolic extract of the leaves of C. hirsutus (100, 200 and 400 mg/kg) and furosemide (10 mg/kg) increased the excretion of creatinine in urine but with a corresponding decrease in serum. CONCLUSIONS: It was concluded that the ethanolic extract of the leaves of C. hirsutus (400 mg/kg) had significant diuretic effect in rats.

Total synthesis and antimicrobial activity of +/--laurelliptinhexadecan-1-one and +/--laurelliptinoctadecan-1-one.

The structures previously assigned to (+)-laurelliptinhexadecan-1-one (1a) and (+)-laurelliptinoctadecan-1-one (1b) from Cocculus orbiculatus (L.) DC. (Menispermaceae) have been confirmed by total synthesis of the racemic alkaloids. The key step of the synthesis involved formation of ring C of the aporphines by a radical-intiated cyclisation. Both (+/-)-laurelliptinhexadecan-1-one (1a) and (+/-)-laurelliptinoctadecan-1-one (1b) were inactive against Staphylococcus aureus ATCC25932, Escherichia coli ATCC10536 and Candida albicans ATCC90028.

The effect of cinnarizine and cocculus indicus on simulator sickness.

Pensacola Simulator Sickness Questionnaire (SSQ) is a valuable method to analyse symptoms evoked by exposure to a flight simulator environment that can also be adopted to evaluate the effectiveness of preventive tools, aiming at reducing simulator sickness (SS). In this study we analysed SSQ data in subjects undergoing a standard ground based spatial disorientation training inside a flight simulator, in order to evaluate the SS prevention obtained with two different pharmacological tools. Twelve males volunteers participated to an experimental design based on a double-blind, balanced administration of either 30 mg cinnarizine (CIN), or Cocculus Indicus 6CH (COC), or placebo (PLC) before one trial of about one hour spent inside a spatial disorientation trainer. All subjects underwent the three different conditions (CIN, COC, PLC) during 3 non-consecutive days separated by at least 2 weeks. During each experimental day, all subjects filled in SSQ. In addition, both postural instability (with the use of a static stabilometric platform), and sleepiness symptoms were evaluated. All the tests were performed before and after the simulated flight, at different times, in one-and-half-hour intervals. Results indicated a strong increase of sickness after flight simulation that linearly decreased, showing pre-simulator scores after 1.30 hours. In contrast to both PLC and COC, CIN showed significant side effects immediately following flight simulation, with no benefit at the simultaneous SSQ scores. Globally, no highly significant differences between COC and PLC were observed, although a minor degree of postural instability could be detected after COC administration. As far as the present exposure to a simulator environment is concerned, none of the pharmacological tools administered in this study resulted effective in reducing SS symptoms as detected by the SSQ. Moreover, CIN significantly increased sleepiness and postural instability in most subjects.

New bisbenzylisoquinoline alkaloids from Cocculus laurifolius.

Two new bisbenzylisoquinoline alkaloids, alpha, alpha'-dioxo-7'-O-demethylstebisimine (1) and 7'-O-demethylstebisimine (2), along with 14 known alkaloids, were isolated from the roots of Cocculus laurifolius. These alkaloids were characterized mainly by spectroscopic methods.

Alkaloid and phenolic compounds of Galanthus caucasicus, Magnolia obovata, Cocculus laurifolius, and Veratrum lobelianum grown in Georgia.

A collection of plants from the country of Georgia (Galanthus caucasicus, Magnolia obovata, Cocculus laurifolius, and Veratrum lobelianum) has been studied for their alkaloid content. Our studies led to the isolation and identification of 24 alkaloids, three of which are being identified and reported for the first time.