In vitro screening of plant extracts traditionally used as cancer remedies in Ghana - 15-Hydroxyangustilobine A as the active principle in Alstonia boonei leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: Cancer represents a major health burden and drain on the global healthcare systems. Traditional African medicine widely use a variety of plant species for treatment of different kinds of cancer. A previous systematic survey by traditional healers in the Ashanti region of Ghana revealed a good overview on the plant species and herbal materials used for the different types of cancer. AIMS OF THE STUDY: The following study aimed to investigate 18 herbal materials from 10 plant species based on the cancer survey in Ghana regarding potential cytotoxicity against different cancer cell lines under in vitro conditions followed by subsequent bioassay-guided fractionation towards the active principle. MATERIALS AND METHODS: Ethanol-water (1:1) extracts were tested (1-100 µg/mL) against a panel of cancer cell lines according to their respective traditional use. Selected extracts with relevant cytotoxicity in this screening were also tested against common pediatric malignancies (leukemias (HL-60, REH) and Ewing sarcoma (RD-ES and CADO-ES1)). Bioassay-guided fractionation of the hydroalcoholic extract from Alstonia boonei was performed by liquid-liquid chromatography and preparative HPLC. Preliminary mechanistic studies on the mode of action were performed by flow cytometric cell cycle analysis as well as apoptosis and necrosis staining. RESULTS: Screening of plant extracts revealed relevant cytotoxicity against all tested cancer cell lines for Alstonia boonei leaves and stem of Paulinia pinnata. The A. boonei extract was additionally found to be active against common pediatric tumor types (leukemias and Ewing sarcoma). Bioassay-guided fractionation of the A. boonei extract revealed the presence of 15-hydroxyangustilobine A 1 as the active principle (IC50 26 µM against MCF-7 cells). This is the first report of this compound in A. boonei. 1 was shown to lead to cell cycle arrest in the G2/M-phase (MCF-7 cells), triggering cells at least partially into apoptosis. CONCLUSION: In summary, an appreciable in vitro activity was revealed for the leaf extract from A. boonei and the isolated vallesamine type indole alkaloid 1, which has to be investigated in future studies towards a potential clinical use.

Anthelmintic A-Type Procyanidins and Further Characterization of the Phenolic Composition of a Root Extract from Paullinia pinnata.

Extracts from the roots of Paullinia pinnata L. are used in West Africa as traditional remedies for a variety of diseases including infestations with soil-transmitted helminths. Based on the results of an ethnopharmacological survey in Ghana, an aqueous acetone (70%) extract was investigated for its anthelmintic and phytochemical properties. Partitioning of the crude extract followed by several fractionation steps of the ethyl acetate phase using Sephadex® LH-20, fast centrifugal partition chromatography, RP-18-MPLC and HPLC led to isolation of six oligomeric A-type procyanidins (1 to 6). To determine the anthelmintic activity, the crude extract, fractions and isolated compounds were tested in vitro against the model organism Caenorhabditis elegans. A significantly better activity was observed for the trimeric A-type procyanidin (1) compared to a B-type trimer. However, this effect could not be generalized for the tetrameric procyanidins, for which the type of the interflavan-linkage (4→6 vs. 4→8) had the greatest impact on the bioactivity. Besides the procyanidins, three novel compounds, isofraxidin-7-O-α-l-rhamnopyranosyl-(1″→6')-ß-d-glucopyranoside (17), 4-methoxycatechol-2-O-(5''-O-vanilloyl-ß-apiofuranosyl)-(1''→2')-ß-glucopyranoside (18) and a 6-(3-methoxy-4-hydroxyphenyl)-hexane-2,4-diol-2-O-hexoside (19) were isolated together with further ten known compounds (7 to 16), mainly coumarins and coumarinolignans. Except for 3-ß-d-glucopyranosyloxy-4-methyl-2(5H)-furanone (15), none of the isolated compounds has previously been described for P. pinnata. The anthelmintic activity was attributed to the presence of procyanidins, but not to any of the other compound classes. In summary, the findings rationalize the traditional use of P. pinnata root extracts as anthelmintic remedies.

Toxicity and Activity of Ethanolic Leaf Extract of Paullinia pinnata Linn (Sapindaceae) in Shigella flexneri-Induced Diarrhea in Wistar Rats.

Herbal products from Paullinia pinnata Linn are widely used in African folk medicine to treat several infectious diseases. Although the extracts from this plant has been shown to possess antimicrobial potential, their activity in infectious diarrhea is less reported. Diarrhea was induced by oral administration of 1.2 × 109 CFU/mL of Shigella flexneri to the rats. The infected rats were treated for 5 days with the doses of 111.42, 222.84, and 445.68 mg/kg of P pinnata. The level of biochemical parameters was assessed and histology of organs examined by 14 days subacute toxicity. S flexneri stool load was considerably reduced after 4 days of treatment with the dose of 445.68 mg/kg, 5 days at the dose of 222.84 mg/kg for the extract, and 2 days with ciprofloxacin. The dose of 111.42 mg/kg appeared efficient after 5 days of treatment. The creatinine level increased at the dose of 445.68 mg/kg in both male and female rats and decrease at the dose of 222.84 mg/mL in female rats while an increase was noted in the male rats. Liver and kidney histology were modified at the dose of 445.68 mg/kg while no change was observed at the doses of 111.42 and 222.84 mg/kg. P pinnata leaf extract is efficient against infectious diarrhea at 111.42 mg/kg without side effect.

Aqueous and methanol extracts of Paullinia pinnata L. (Sapindaceae) improve inflammation, pain and histological features in CFA-induced mono-arthritis: Evidence from in vivo and in vitro studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Paullinia pinnata L. (Sapindaceae) is an African woody vine, traditionally used for the treatment of itch and pain-related conditions such as rheumatoid arthritis. AIM: This work evaluates, in vitro and in vivo, the anti-inflammatory and analgesic effects of aqueous (AEPP) and methanol (MEPP) extracts from Paullinia pinnata leaves. METHODS: AEPP and MEPP (100, 200 and 300 mg/kg/day) were administered orally in monoarthritic rats induced by a unilateral injection of 50 µl of Complete Freund's Adjuvant (CFA) in the ankle joint. During the 14 days of treatment, pain and inflammation were evaluated alternatively in both ankle and paw of the CFA-injected leg. Malondialdehyde (MDA) and glutathione (GSH) levels were assessed in serum and spinal cord. Histology of soft tissue of the ankle was also analyzed. For in vitro studies, AEPP and MEPP (10, 30 and 100 µg/ml) were evaluated against nitric oxide (NO) production by macrophages that were either non-stimulated or stimulated with LPS, 8-Br-AMPc and the mixture of both substances after 8 h exposure. These extracts were also evaluated on TNF-α and IL-1ß production in cells stimulated with LPS for 8 h. Finally, the ability of the extracts to bind to neuroactive receptors was evaluated in vitro using competitive binding assays with >45 molecular targets. RESULTS: AEPP and MEPP significantly reduced by 20-98% (p < 0.001) the inflammation and pain sensation in both the ankle and paw. AEPP significantly increased glutathione levels (p < 0.05) in serum. Both extracts reduced MDA production in serum and spinal cord (p < 0.001), and significantly improved tissue reorganization in treated arthritic rats. P. pinnata extracts did not affect NO production in non-stimulated macrophages but significantly reduced it by 47-88% in stimulated macrophages. AEPP and MEPP also significantly inhibited TNF-α (35-68%) and IL-1ß (31-36%) production in LPS stimulated macrophages. No cytotoxic effect of plant extracts was observed. MEPP showed concentration-dependent affinity for Sigma 2 receptors with an IC50 of 50 µg/ml. CONCLUSION: These results demonstrate the analgesic and anti-inflammatory effects of P. pinnata extracts on monoarthritis and further support its traditional use for pain and inflammation. These activities are at least partly due to the ability of these extracts to inhibit the production of NO, TNF-α, IL-1ß and to likely modulate Sigma 2 receptors.

Antidiabetic and in vitro antioxidant effects of hydromethanol extract of Paullinia pinnata root bark in alloxan-induced diabetic rat.

Background The study evaluated phytochemical composition, antidiabetic, oral glucose tolerance test and in vitro antioxidant activities of hydromethanol extract of Paullinia pinnata root bark. Methods Cold maceration method was used in extract preparation and scavenging of 2,2-diphenyl-1-picrylhydrazyl radicals was used to evaluate antioxidant properties of the extract. Diabetes was induced with alloxan at the dose of 160 mg/kg. The antidiabetic activity of the extract was tested at doses of 50, 100 and 200 mg/kg, and glibenclamide was used as reference drug. Results Phytochemical analysis of the extract showed the presence of alkaloids, flavonoids, glycosides, tannins, saponins and terpenes/sterols. The extract produced a significant (p<0.05) time-dependent decrease in the fasting blood glucose (FBG) levels in the treated rats when compared with the distilled water treated rats, but did not produce dose-dependent effects. The extract 50, 100 and 200 mg/kg and glibenclamide (2 mg/kg) caused 83.62 %, 60.66 %, 47.77 % and 68.52 % reduction respectively in FBG at 6 h post-treatment while the distilled water (5 mL/kg) produced 8.12 % reduction in FBG at 6 h post treatment. The extract (50 mg/kg) and glibenclamide (2 mg/kg) produced a significant (p<0.05) oral glucose tolerance effect in both normoglycemic and diabetic rats. The extract produced concentration-dependent increase in antioxidant activity and had its optimum effect at 400 µg/mL concentration. Conclusions This study suggests that P. pinnata root bark has potent antidiabetic and antioxidant activities and also validates its use in folkloric medicine in the management of diabetes-related conditions.

Chemical constituents and insecticidal activity of essential oil of Paullinia pinnata L (Sapindaceae) / Compuestos químicos y actividad insecticida del aceite esencial de Paullinia pinnata L (Sapindaceae)

The chemical constituents and insecticidal activity of essential oil obtained by hydrodistillation of the leaves of Paullinia pinnata Linn (Sapindaceae) are being reported. The essential oil were analysed by using gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS). Different concentrations (50, 100, 150, 200 and 250 mg/mL) of P. pinnata essential oil prepared separately and diluted in dimethyl sulfoxide (DMSO) were tested on the maize weevil, Sitophilus zeamais. The main constituents of the oil were pentadecanoic acid (17.9%), isoaromadendrene epoxide (11.5%) and wine lactone (11.2%). Other significant compounds of the essential oil were eremophilene (6.9%) and phytol (6.2%). The essential oil displayed 100% mortality (fumigant toxicity) against S. zeamais adults at tested concentration of 150 mg/mL with lethal concentrations (LC50) of 51.87 mg/mL air. This is the first report on the chemical constituents and insecticidal activity of essential oil of P. pinnata and may be explore as a potential natural herbal plant for the control of insect pest.

Se informan los componentes químicos y la actividad insecticida del aceite esencial obtenido por hidrodestilación de las hojas de Paullinia pinnata Linn (Sapindaceae). El aceite esencial se analizó mediante cromatografía de gases (GC) y cromatografía de gases acoplada con espectrometría de masas (GC-MS). Se ensayaron diferentes concentraciones (50, 100, 150, 200 y 250 mg/ml) de aceite esencial de P. pinnata preparado separadamente y diluido en dimetilsulfóxido (DMSO) en el gorgojo de maíz, Sitophilus zeamais. Los componentes principales del aceite fueron ácido pentadecanoico (17,9%), isoaromadendreno epóxido (11,5%) y vino lactona (11,2%). Otros compuestos significativos del aceite esencial fueron eremophilene (6,9%) y phytol (6,2%). El aceite esencial mostró una mortalidad del 100% (toxicidad fumigante) contra los adultos de S. zeamais a una concentración de 150 mg/ml con concentraciones letales (CL50) de 51,87 mg/ml de aire. Este es el primer informe sobre la composición del aceite esencial de P. pinnata y su actividad insecticida. Puede ser explorado como una potencial planta herbácea natural para el control de la plaga de insectos.

Hematological effects of repeated graded doses of the methanol extract of Paullinia pinnata (Linn.) leaves in Wistar albino rats.

BACKGROUND: Paullinia pinnata is a medicinal plant used for the treatment of various diseases, including anemia in West Africa. AIM: This study was carried out to investigate the effect of increasing doses of the methanolic leaves extract of P. pinnata on hematological parameters in rats. MATERIALS AND METHODS: Thirty-six male Wistar albino rats were grouped into six groups of six animals each. Five doses; 50,100, 200, 400 and 800 mg/kg body weight of the P. pinnata extract were administered separately to five groups. The sixth group served as a control and received only the vehicle (70% physiological saline: 30% Tween 80 [v/v]). Administration was done orally daily for 28 days at 24 h interval. On day 29, the animals were made inactive, blood was then collected from the heart and various hematological parameters were evaluated. STATISTICAL ANALYSIS: Analysis of variance was employed. RESULTS: The packed cell volume and red blood cell count increased significantly (P < 0.05) in the treatment groups except at 200 mg/kg dose. The hemoglobin concentration increased in all the treatment groups. The values for the neutrophils at 50, 100, 200 and 800 mg/kg doses were higher than that of the control. The white blood cell count increased significantly (P < 0.05) at 50 and 400 mg/kg doses compared to the control and exceeded the normal physiological range. CONCLUSION: The maximum tolerable dose is 200 mg/kg body weight of the methanolic leaves extract of P. pinnata and the extract has anti-anemic property with the ability to increase neutrophils count.

A new antimicrobial and radical-scavenging glycoside from Paullinia pinnata var. cameroonensis.

A new glycoside, pinnatoside A (1), together with two known compounds (2 and 3), were isolated from the stems of Paullinia pinnata. Their structures were elucidated on the basis of extensive spectroscopic analysis and chemical methods. Compound 1 showed significant antibacterial activity with a minimum inhibitory concentration (MIC) value of 1.56 µg/mL against Escherichia coli, and 2 displayed significant antibacterial activity with a MIC value of 1.56 µg/mL against Enterobacter aerogenes and E. coli. Equally, compound 1 exhibited the best radical-scavenging activity (RSa50 = 25.07 ± 0.49 µg/mL).

An ethnopharmacological survey and in vitro confirmation of the ethnopharmacological use of medicinal plants as anthelmintic remedies in the Ashanti region, in the central part of Ghana.

ETHNOPHARMACOLOGICAL RELEVANCE: Infections with helminths are still a big problem in many parts of the world. The majority of the people in West Africa treat such infections with medicinal plants related to the local traditional medicine. The present study aims at identifying medicinal plants traditionally used for worm infections in the Ashanti region, Ghana. In vitro screening of selected extracts from plants on which scientific knowledge is limited was to be performed. MATERIALS AND METHODS: Validated questionnaires were administered to 50 traditional healers in the Ashanti region, Ghana. Interviews and structured conversations were used to obtain relevant information. Quantitative and qualitative evaluation was performed additionally to structured cross-referencing of the data using SciFinder(®) data base. Selected plant species were used for in vitro testing on anthelmintic activity against the free-living model nematode Caenorhabditis elegans. RESULTS: 35 plant species were recorded for the use in humans and 6 for the use in animals. Plant material most frequently used were the seeds from Carica papaya, mentioned by nearly all healers. The plausibility of most plants used for treatment of infections with helminths was given in most cases by documentation of potential anthelmintic activity in recent scientific literature. 9 species from plants not or scarcely described in literature for this indication were investigated on in vitro activity. A hydroethanolic (1:1) extract of Combretum mucronatum was most active with a survival rate of nematodes of 89% at 0.1mg/mL and 58% at 1mg/mL respectively (levamisole 16%). Extracts of Paullinia pinnata and Phyllanthus urinaria were also assessed to exhibit a minor (85% and 89% respectively at 1mg/mL), but still significant activity. CONCLUSION: Traditional use of anthelmintic plants from Ghana can be well rationalized by cross-referencing with published literature and phytochemical/pharmacological plausibility.The in vitro investigations of extracts from Combretum mucronatum, Paullinia pinnata and Phyllanthus urinaria exhibited significant effects against nematodes. The anthelmintic activity of these plants should be investigated in detail for pinpointing the respective lead structures responsible for the activity.

Antityphoid and radical scavenging properties of the methanol extracts and compounds from the aerial part of Paullinia pinnata.

Paullinia pinnata Linn (Sapindaceae) is a medicinal plant, locally used in the West Region of Cameroon for the treatment of typhoid fever. This work was designed to evaluate the antityphoid and antioxidant activities of the extracts and compounds of P. pinnata. The methanol extracts of the leaves and stems were tested for antityphoid and antioxidant activities. Compounds were isolated, and their structures elucidated by analysis of spectroscopic data in conjuction with literature data and tested for the same activities. The leaf extract was also tested in vivo for its antityphoid potential in a Salmonella typhimurium-induced typhoid fever model in Wistar rats. Seven known compounds: methylinositol (1), ß-sitosterol (2), friedelin (3), 3ß-(ß-D-Glucopyranosyloxy)stigmast-5-ene (4), (3ß)-3-O-(2'-Acetamido-2'-deoxy-ß-D-glucopyranosyl) oleanolic acid (5), (3ß,16α-hydroxy)-3-O-(2'-Acetamido-2'-deoxy-ß-D-glucopyranosyl) echinocystic acid (6) and (3ß,)-3-O-[ß-D-glucopyranosyl-(1″-3')-2'-acetamido-2'-deoxy-ß-D-galactopyranosyl]oleanolic acid (7) were isolated. Compounds 5 and 1 showed the highest antibacterial (MIC = 0.781-1.562 µg/ml) and DPPH radical scavenging (RSa50 = 19.27 ± 4.43 µg/ml) activities respectively. The maximum extract dose (446.00 mg/kg bw) had comparable activity with ciprofloxacin (7.14 mg/kg bw) and oxytetracycline (5 mg/kg bw). The extract induced significant dose-dependent increase of WBCs and lymphocytes. These results support the ethnomedicinal use of P. pinnata and its isolated Compounds could be useful in the standardization of antityphoid phytomedicine from it.

AN INVESTIGATION INTO THE ANTIMALARIAL EFFECT OF METHANOLIC EXTRACT OF PAULLINIA PINNATA LEAVES IN PLASMODIUM BERGHEI INFECTED MICE AND COURSE OF INFECTION.

AIM: The aim of this study was to investigate the antimalarial activity of methanolic leaves extract of Paullinia pinnata on chloroquine-sensitive Plasmodium berghei NK 65 infected mice. METHODOLOGY: The curative study was conducted in thirty-six Wistar albino mice of both sexes which were divided into six groups of six animals each. The animals were infected with P. berghei NK 65. Group I was the negative control and received the vehicle (10% DMSO). Group II received no treatment. Groups III and IV were the positive controls and received chloroquine (CQ) (10mg/kg) and artesunate (4 mg/kg)-amodiaquine (10mg/kg) combination (ACT) respectively. Groups V and VI received 100mg/kg and 200mg/kg doses of the extract respectively. Administration was done orally once for three or four days for the standard drugs or the extract/vehicle respectively. The percentage parasitaemia, packed cell volume (PCV), body weight and death was monitored on days 0, 1, 2, 3, 4 and 11 (7 day post administration). The study of the course of infection of P. berghei was monitored in eighteen Wistar albino mice of both sexes which were similarly grouped, infected and treated for 3 days. Group A received the vehicle (distilled water) only. Group B was treated with CQ (10 mg/kg) and Group C with ACT. The percentage parasitaemia and death was monitored from day 0 to day 30 (27 day post administration). RESULTS: In the curative study, the extract suppressed parasitaemia at both doses on day 4. The group treated with 200mg/kg dose showed a higher percentage chemosuppression though not significant. The course of infection study revealed that recrudescence occurred on day 8 in the CQ treated group which lasted until day 23 after which the recrudescence was lost without re-treatment. A similar result was observed in the ACT group. CONCLUSION: The methanolic leaves extract of Paullinia pinnata has weak anti-malarial property. Chloroquine-sensitive P. berghei NK65 loses credibility and needs to be revalidated biannually.

Paullinia cupana para o controle de fogachos em pacientes com câncer de mama: um estudo piloto / Paullinia cupana for control of hot flashes in breast cancer patients: a pilot study

OBJETIVO: Avaliar se a Paullinia cupana diminui o número e a gravidade dos fogachos em mulheres após diagnóstico de câncer de mama. MÉTODOS: Estudo piloto prospectivo fase II realizado com mulheres que sobreviveram ao câncer de mama, que completaram o tratamento pelo menos 3 meses antes e que apresentavam ao menos 14 episódios de fogachos por semana. Utilizando o desenho de Simon para que a primeira etapa fosse considerada positiva, ao menos 9 de 15 mulheres deveriam ter a gravidade dos fogachos diminuída em pelo menos 50%. As pacientes receberam 50mg do extrato seco de Guaraná oralmente 2 vezes por dia por 6 semanas. Foram avaliadas, a gravidade e a frequência dos fogachos. RESULTADOS: Dezoito pacientes iniciaram o tratamento com Paullinia cupana e 15 completaram o estudo. Três pacientes deixaram o estudo imediatamente após iniciarem o tratamento em razão de dificuldade na participação e não adesão. Das 15 pacientes que completaram o estudo, 10 obtiveram diminuição de mais de 50% dos índices de gravidade de fogachos. Durante as 6 semanas de tratamento, diminuições estatisticamente significativas foram observadas tanto no número de fogachos (p=0,0009), quanto nos índices de gravidade (p<0,0001). Paullinia cupana foi bem tolerada, e não houve relato de toxicidade como causa de saída do estudo. CONCLUSÕES: Paullinia cupana pareceu promissora para o controle de fogachos. Estudos mais extensivos são necessários.

OBJECTIVE: To evaluated whether Paullinia cupana decrease number and severity of hot flashes in breast cancer survivors. METHODS: This was a prospective phase II pilot study. We studied female breast cancer survivors who had completed the cancer treatment 3 months previously and who were experiencing at least 14 hot flashes per week. At least 9 of the 15 patients were required to have a decrease of at least 50% in hot flash severity score in keeping with the Simon Design. Patients received 50mg of dry extract of Paullinia cupana orally twice a day for 6 weeks. We assessed both frequency and severity of hot flashes. RESULTS: A total of 18 patients started the Paullinia cupana treatment, and 15 completed the study. Three patients left the study immediately after starting the treatment because of personal difficulties in participation or noncompliance. Of the 15 patients who completed the study 10 had a decrease of more than 50% in hot flash severity scores. During the 6 weeks of treatment, statistically significant decreases were seen in both numbers of hot flashes (p=0.0009) and severity scores (p<0.0001). Paullinia cupana was well tolerated, and there were no instances of discontinuation because of toxicity. CONCLUSIONS: Paullinia cupana appears promising for controlling hot flashes. More extensive studies seem warranted.