Isolation and Characterisation of Compounds from ‘Naga Buana’ (Phyllanthus Pulcher) and ‘Similit Matinggi’ (Casearia Capitellata) and Their Cytotoxic Effects on Cancer Cell Lines

Phyllanthus pulcher from the family of Euphorbiaceae is used as a traditional medicine in Malaysia to treat stomachache and ulceration. Casearia species from the family of Flacourtiaceae are traditionally used as an antiseptic, cicatrizant and topical anesthetic agent. In a preliminary cytotoxic screening, P. pulcher and C. capitellata extracts were found to display remarkable activities against various in vitro tumour cell lines. On the basis of the screening results and literature review of the plants, P. pulcher and C. capitellata were selected for phytochemical investigations to identify compounds with cytotoxic activities. The investigations of the active dichloromethane extract of aerial parts from P. pulcher by a combination of different chromatographic techniques led to the isolation of three new natural products, bis(6-methylheptyl)phthalate (190), phyllanthal-A (192) and 3,4- dihydroxy-5-methoxy-3’,4’,5’-trihydroxyoxepino-chromene-2-one (196) and five other known compounds including stigmast-5-en-3-ol-oleate (189), diisobutyl adipate (195), β- sitosterol (193), 7-tridecanone (194) and β-sitosteryl-β-D-glucopyranoside (191). The isolation of stigmast-5en-3ol-oleate (189) is the first report from the genus Phyllanthus. All isolated compounds were tested for their cytotoxic activities against three human tumour cell lines: MCF-7 (breast), DU-145 (prostate) and H460 (lung). Compounds, 189, 190, 191 and 192 were found to exhibit significant antitumour activity against MCF-7 cells with the IC50 values of 17.6, 69.2, 54.1 and 47.7 μM, respectively. Among all the isolated compounds from P. pulcher aerial parts only the phyllanthal-A (192) exhibited cytotoxic activity against DU-145 cells with the IC50 value of 20.5 μM. Whereas all the isolated compounds from P. pulcher aerial parts failed to exhibit cytotoxic effect against H460 cells even at the highest concentration of 100 μM. Investigation on the active dichloromethane extract of P. pulcher roots resulted in the isolation of two new compounds, 12(13)-dehydro-3α-acetoxyolean-28-oic acid (199) and lupanol acetate (200) and three other known compounds including 3α-acetoxy-25- hydroxyolean-12-en-28-oic acid (197), glochidone (198) and glochidonol (201). 3α- acetoxy-25-hydroxyolean-12-en-28-oic acid (197) which was isolated for the first time in this genus showed cytotoxic activity against MCF-7 and DU-145 cell lines. Whereas 201 exhibited potent antitumour activity against all the tested cell lines with the IC50 values ranging 17.0 - 30.0 μM. Among all the isolated compounds from P. pulcher only the 201 exhibited antitumour activity against H460 cells with the IC50 value of 21.7 μM. Investigation on the bioactive phytochemicals of C. capitellata stem bark extracts yielded 7α-acetoxy-hop-12(13)-en-11-one (202), 5-methoxy-7-hydroxycoumarin (203), 5- methoxy-7-β-D-glucopyranosylcoumarin (204), 5,7-dimethoxycoumarin (205), casearine-A (206) and casearine-B (207). The isolation of 206 and 207 has never been reported from natural products before, whereas the isolation of 202, 203, 204, and 205 is the first report from this genus. Among all the isolated compounds, 206 and 207 exhibited significant antitumour activity against MCF-7 and DU-145 cell lines with the IC50 values ranging 5.0-14.0 μM. 206 and 207 also exhibited cytotoxic activity against H460 cells with the IC50 values of 77.0 and 56.3 μM, respectively. The structures of all isolated compounds were elucidated with the help of chemical and modern spectroscopic techniques (UV, IR, MS, 1H NMR, 13C NMR, DEPT, COSY, HSQC, and HMBC). In conclusion, some of these natural products, especially 189, 192, 201, 206, and 207, are potential lead molecules for future antitumour studies to discover potential clinical candidates

Phytochemicals from Phyllanthus niruri Linn. and their pharmacological properties: a review

This review discusses the medicinal plant Phyllanthus niruri Linn. (Euphorbiaceae), its wide variety of phytochemicals and their pharmacological properties. The active phytochemicals, flavonoids, alkaloids, terpenoids, lignans, polyphenols, tannins, coumarins and saponins, have been identified from various parts of P. niruri. Extracts of this herb have been proven to have therapeutic effects in many clinical studies. Some of the most intriguing therapeutic properties include anti-hepatotoxic, anti-lithic, anti-hypertensive, anti-HIV and anti-hepatitis B. Therefore, studies relating to chemical characteristics and structural properties of the bioactive phytochemicals found in P. niruri are very useful for further research on this plant as many of the phytochemicals have shown preclinical therapeutic efficacies for a wide range of human diseases, including HIV/AIDS and hepatitis B

Fatty acid composition and antioxidant activity of oils from two cultivars of Cantaloupe extracted by supercritical fluid extraction

The effect of supercritical fluid extraction (SFE) fractionation of three oil fractions (1st, 2nd, 3rd fraction) on the fatty acid composition and antioxidant activity of oils from two cultivars of cantaloupe were investigated. Rock melon oil (RMO) and Golden Langkawi oil (GLO) were extracted using SFE and the major fatty acids for both cultivars were linoleic, oleic, palmitic, and stearic acid. The SFA decreased from 15.78 to 14.14% in RMO 1st fraction, and MUFA decreased from 18.30 to 16.56% in RMO 2nd fraction, while PUFA increased from 65.9 to 69.30% in RMO 3rd fraction. On the other hand SFA decreased from 16.35 to 13.91% in GLO 1st fraction, and MUFA decreased from 17.50 to 15.57% in GLO 2nd fraction, while PUFA increased from 66.15 to 70.52% in GLO 3rd fraction. The different fractions of the two oils showed high antioxidant activity in reducing the oxidation of pi-carotene in beta-carotene bleaching assay (BCB) and the quenching of 1,1-diphenyl-2-picrylhydrazyl (DPPH)

Vanillin rich fraction regulates LDLR and HMGCR gene expression in HepG2 cells.

Vanillin and its analogs have been exploited for their various health benefits. This work aimed to investigate the antioxidant properties and regulatory effects of vanillin rich fraction (VRF) extracted from vanilla pods using the supercritical fluid extraction (SFE) and commercial vanillin on low density lipoprotein receptor (LDLR) and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) gene expression in HepG2 cells. The vanillin content in the VRF was 2.6% (w/w) obtained at a temperature of 80 °C and a pressure of 600 bar. The VRF exhibited better antioxidant activity compared to the vanillin in DPPH and BCB tests. LDLR mRNA level was increased significantly by 2, 3 and 1.3 fold in the VRF treated cells at 100, 200 and vanillin treated cells at 100, respectively, compared with untreated cells. On the other hand, the HMGCR mRNA level was decreased significantly by 14, 58 and 13% respectively, in the VRF treated cells at 100, 200 and V treated cells at 100, respectively, compared with untreated cells. The VRF showed potential antioxidant activity and regulated genes involved in cholesterol metabolism including LDLR and HMGCR in dose-dependent manner

Actividad antioxidante y composición de ácidos grasos de aceites de semilla de melón de dos variedades extraídos mediante extracción con fluido supercrítico

The effect of supercritical fluid extraction (SFE) fractionation of three oil fractions (1st, 2nd, 3rd fraction) on the fatty acid composition and antioxidant activity of oils from two cultivars of cantaloupe were investigated. Rock melon oil (RMO) and Golden Langkawi oil (GLO) were extracted using SFE and the major fatty acids for both cultivars were linoleic, oleic, palmitic, and stearic acid. The SFA decreased from 15.78 to 14.14% in RMO 1st fraction, and MUFA decreased from 18.30 to 16.56% in RMO 2nd fraction, while PUFA increased from 65.9 to 69.30% in RMO 3rd fraction. On the other hand SFA decreased from 16.35 to 13.91% in GLO 1st fraction, and MUFA decreased from 17.50 to 15.57% in GLO 2nd fraction, while PUFA increased from 66.15 to 70.52% in GLO 3rd fraction. The different fractions of the two oils showed high antioxidant activity in reducing the oxidation of β-carotene in beta-carotene bleaching assay (BCB) and the quenching of 1,1-diphenyl-2-picrylhydrazyl (DPPH).El efecto del fraccionamiento mediante extracción con fluido supercrítico de tres fracciones (fracción 1ª, 2ª y 3ª) sobre la composición de ácidos grasos y actividad antioxidante de aceites de dos variedades de melón fué investigado. Aceites de melón de los cultivares Rock (RMO) y Golden Langkawi GLO) fueron extraídos usando SFE y los principales ácidos grasos en cada cultivar fueron ácido linoleico, oleico, palmítico y esteárico. Los ácidos grasos saturados (SFA) disminuyeron desde 15.78 a 14.14% en la 1ª fracción de RMO y los ácidos grasos monoinsaturados (MUFA) disminuyeron desde 18.30 a 16.56% en la 2ª fracción de RMO, mientras que los ácidos grasos poliinsaturados (PUFA) aumentaron de 65.9 a 69.30% en la 3ª fracción de RMO. Por otra parte, SFA disminuyo de 16.35 a 13.91% en la primera fracción de GLO y MUFA disminuyo de 17.50 a 15.57% en la 2ª fracción de GLO, mientras que PUFA aumento de 66.15 a 70.52% en la 3ª fracción de GLO. Las diferentes fracciones de los dos aceites mostraron una alta actividad antioxidante al reducir la oxidación del β-caroteno en el ensayo de decoloración de beta-carotene (BCB) y en la eliminación del radical 1,1-difenil-2-picrilhidrazilo (DPPH)

Isolating the Effect of Off-Road Glance Duration on Driving Performance: An Exemplar Study Comparing HDD and HUD in Different Driving Scenarios

Objective: We controlled participants’ glance behavior while using head-down displays (HDDs) and head-up displays (HUDs) to isolate driving behavioral changes due to use of different display types across different driving environments. Background: Recently, HUD technology has been incorporated into vehicles, allowing drivers to, in theory, gather display information without moving their eyes away from the road. Previous studies comparing the impact of HUDs with traditional displays on human performance show differences in both drivers’ visual attention and driving performance. Yet no studies have isolated glance from driving behaviors, which limits our ability to understand the cause of these differences and resulting impact on display design. Method: We developed a novel method to control visual attention in a driving simulator. Twenty experienced drivers sustained visual attention to in-vehicle HDDs and HUDs while driving in both a simple straight and empty roadway environment and a more realistic driving environment that included traffic and turns. Results: In the realistic environment, but not the simpler environment, we found evidence of differing driving behaviors between display conditions, even though participants’ glance behavior was similar. Conclusion: Thus, the assumption that visual attention can be evaluated in the same way for different types of vehicle displays may be inaccurate. Differences between driving environments bring the validity of testing HUDs using simplistic driving environments into question. Application: As we move toward the integration of HUD user interfaces into vehicles, it is important that we develop new, sensitive assessment methods to ensure HUD interfaces are indeed safe for driving

Isolation and cytotoxicity of triterpenes from the roots of Phyllanthus pulcher Wall. ex Müll. Arg. (Euphorbiaceae).

The dried powdered roots of Phyllanthus pulcher Wall. ex Müll. Arg. (Euphorbiaceae), were sequentially extracted with dichloromethane (DCM), ethyl acetate (EtOAc) and methanol (MeOH). The extracts were tested for cytotoxic activity against three human cancer cell lines: MCF-7 (breast), NCI-H460 (lung) and DU-145 (prostate). The DCM extract exhibited the strongest cytotoxic activity compared with EtOAc and MeOH extracts. Hence from the DCM extract, five pentacyclic triterpenes, 3α-acetoxyl-25-hydroxyolean-12-en-28-oic acid (1), glochidone (2), 12(13)-dehydro-3α-acetoxyolean-28-oic acid (3), lupanyl acetate (4) and glochidonol (5) were isolated and identified by spectroscopic analyses (1H NMR, 13C NMR, FT-IR, UV, DEPT, HMQC, HMBC and HREIMS). This is the first report on the isolation of 4 from a natural source, whereas 1 and 3 have already been isolated from the families Hamamelidaceae and Compositae (Asteraceae), respectively. However this is the first study reporting the presence of 1 and 3 in the Euphorbiaceae family. The isolated tritepenes 1-5 were tested against the three human tumour cell lines as stated above. Only compounds 1 and 5 exhibited cytotoxic activity, 5 being most potent with IC50 values ranging 7.5–13.4 µg/mL (17.1–30.5 µM)

Development of orodispersible films with selected Indonesian medicinal plant extracts

This study focused on the incorporation into orodispersible films (ODFs) of the dried extracts of five selected Indonesian medicinal plants: Lagerstroemia speciosa (L.) Pers. (LS), Phyllanthus niruri L. (PN), Cinnamomum burmanii Blume (CB), Zingiber officinale Roscoe (ZO) and Phaleria macrocarpa (Scheff.) Boerl (PM). Suitable formulae for solvent casting were developed to produce extract containing films with either a combination of hypromellose (HPMC) with carbomer 974P or only hydroxypropyl cellulose (HPC) as film forming agent. Each extract and dose in a formulation rendered different ODF characteristics. Extracts of ZO and CB and a low dose of PM extract (5 mg) could be formulated into an ODF containing HPMC with carbomer 974P. For extracts of LS, PN and high doses of PM extract HPC were the most suitable film forming agents. For each extract a different maximum load in a film was found, up to maximum 30 mg for extracts of LS and PN. Good products were obtained with 5 mg and 10 mg of each extract. The quality of the produced ODFs was tested organoleptically, and characterized by determination of uniformity of weight, thickness, disintegration time, surface pH, crystallinity, mechanical properties, water content, residual ethanol, dynamic vapour sorption, physical stability and control of the qualitative profiling of extract composition in the film. Thin layer chromatography indicated that all five extracts remained chemically unaffected during ODF production. In conclusion, ODFs are a suitable novel dosage form for herbal extracts, provided that tailor-made formulations are developed for each extract and each dose

Etnobotânica e medicina popular no tratamento de malária e males associados na comunidade ribeirinha Julião – baixo Rio Negro (Amazônia Central)

RESUMO A utilização de plantas medicinais para o tratamento de doenças tropicais como a malária na Amazônia Central é de suma importância, principalmente em locais onde o sistema único de saúde não se encontra presente como na maioria das comunidades ribeirinhas desta região. Sendo assim, investigar e resgatar o conhecimento popular a respeito de plantas medicinais utilizadas no tratamento de malária e males associados pelos moradores da comunidade Julião situada na Reserva de Desenvolvimento Sustentável do Tupé, Manaus-AM, torna-se importante no registro de como as populações locais se previnem e tratam essa doença tão prevalente e perigosa na região. O trabalho foi conduzido na forma de oficinas participativas, segregadas por gênero e complementadas com entrevistas semiestruturadas aliadas à técnica da turnê-guiada nos quintais e floresta adjacente à comunidade. Foram calculados os índices de diversidade de Shannon-Wiener, equitabilidade e concordância quanto ao uso principal (CUP). A partir da colaboração efetiva de 13 comunitários foram registradas 62 espécies vegetais pertencentes a 53 gêneros e 34 famílias botânicas que resultaram em índice de diversidade (H’) de 1,62 decits e equitabilidade de 0,9. As famílias mais representativas foram: Fabaceae (7 espécies), Asteraceae e Lamiaceae (4 espécies cada) e Solanaceae e Rutaceae (3 espécies cada). Vale destacar que 16 espécies (25,8%) foram citadas para tratamento de malária e males associados pela primeira vez em estudos etnobotânicos realizados na América Latina