Cancer incidence is increasing and novel anticancer drugs with new mechanisms of action are essential for future chemotherapeutic treatment. Natural products have historically played an important role in the development of anti-cancer drugs and have potential to do so also in the future. In this thesis two classes of natural products are identified as possible drug lead candidates, and the mechanisms of their action are elucidated. Initially, in a screening of a compound library for cytotoxic effects in colon cancer cells, natural products with potent activity were identified. Based on their potency, and on previously reported activities in cancer cells, two main groups of compounds, cardiac glycosides (CGs) and gambogic acid (GA) analogues, were selected for further in-depth studies. The concentration-dependent cytotoxicity was confirmed in cell lines of different origin. Cardiac glycosides were mainly evaluated for their activity in colon cancer cells and in leukemic cells, whereas the GA analogues were studied using a resistance-based panel of ten human cancer cell lines. Using activity profiles and the ChemGPS-NP model, the compounds were compared, structurally and mechanistically, to standard chemotherapeutic drugs. The results from these analyses suggested that the CGs and the GA analogues act by mechanisms different from those of antimetabolites, alkylating agents, topoisomerase I and II inhibitors, or tubulin-active agents. By analysis of drug-induced gene expression, one GA analogue, dihydro GA, was identified as a possible inhibitor of the ubiquitin-proteasome system (UPS), and the CGs showed similarities to protein synthesis inhibitors. Starting from these hypotheses, we further investigated the mechanisms of actions on a molecular level. The results showed that GA and dihydro GA act as inhibitors of the 20S proteasome chymotrypsin activity, leading to accumulation of ubiquitinated proteins. The CGs were confirmed to inhibit protein synthesis in colon cancer cell lines. However, interestingly, in leukemia cell lines, it seemed that the CGs act through a different, yet unexplored, mechanism of action. The leukemic cells (pre-B and T-ALL) were particularly susceptible to the cytotoxic effects of CGs, including at concentrations that may be achievable in the clinic.

Major depressive disorder affects people’s productivity and ability to function in everyday life. The disorder can be attributed to neurochemical imbalances of various neurotransmitters including but not limited to serotonin, dopamine, and norepinephrine. Conventional pharmacological treatments have focused primarily on these three neurotransmitters, and have been shown to be effective in alleviating most of the major symptoms of depression. Although these treatments are effective with most patients, they are known to have adverse side effects, causing patients to seek alternative treatments. Sceletium tortuosum, a succulent plant found in the Cape region of South Africa, has been shown to have anxiolytic effects when used recreationally. Studies have confirmed the presence of a family of alkaloids mesembrines that are present within the plant and believed to be responsible for the calming effects. Pharmacological analyses have revealed that individual members of the alkaloid family act as either serotonin reuptake inhibitors (SRI) or phosphodiesterase-4 (PDE4) inhibitors. The current study seeks to elucidate the antidepressant properties of the mesembrine alkaloids in a mouse model of depression. Isolated alkaloids were administered at a low dose (10 mg/kg) and a high dose (80 mg/kg) to BALB/c mice in the forced swim test a rodent model of behavioral despair. This was compared with paroxetine (Paxil) (1 mg/kg), a selective serotonin reuptake inhibitor with proven antidepressant efficacy, and 0.9% saline. Each trial of the forced swim test was administered for six minutes and the duration of swimming and immobility was measured. In order to assess any locomotor effects of the drug treatments, an open field exploration test was also employed one week following the forced swim task. Results from the forced swim test revealed a statistically significant reduction in the duration of immobility (behavioral despair) between the low dose of alkaloids and saline. No significant effects in immobility were found across the other drug treatment conditions (high dose mesembrine, paroxetine, and saline). Further, none of the treatment groups showed statistically significant locomotor interference effects in the open field exploration test. We conclude that the mesembrine alkaloids present in Sceletium tortuosum have antidepressant properties and may represent a suitable alternative for the treatment of major depressive disorder.

This study aimed to evaluate the antimicrobial efficacy of medicinal hemp plant extracts to determine the antibacterial effects of indigenous Sansevieria species and exotic Cannabis sativa phytotherapy varieties. This study also assessed whether aqueous o

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2007. / Härtill 5 uppsatser.

Xihuangcao is a folk medicinal herb used in southern China with three botanical origins: Isodon lophanthoides (IL), I. lophanthoides var. graciliflorus (ILG) and I. serra (IS). They are often used indiscriminately, numerous commercially available herbal products list Xihuangcao as an ingredient without listing the source. This situation has led to a growing concern about the differentiation and quality evaluation of Xihuangcao. To address this concern, a systematic study was conducted to identify the origin. The study is divided into five parts, which aimed to establish and apply the authentication methods of the origins. Four Isodon species were recorded in research papers as the plant sources. However, a new classification suggested in 2004 and two of the IL varieties were merged. In the ancient herbal documents, ILG was first recorded as the origin plant. IL was the major species in the ancient texts, IS was only listed as an additional sources in recent herbal references. The“yellow juices which proven to be the exudates of glandular scales was the key identification features recorded. Macroscopic and microscopic studies provided identification features of the three Isodon species. IL and ILG share very similar features, but IS can be easily distinguished. By morphological features, IL and ILG can be distinguished by the shape of leaves, which IL has a broader leaves than ILG; IS can be identified by its very bitter taste and broadly winged petioles. By microscopic features, IL and ILG have a tiny difference in the shape of epidermal cells of leaf, and IS can be recognized by small raphides of calcium oxalate. In the UPLC-MS fingerprinting and tissue-specific profiling, the chemical profiles the three species were revealed. The chemical profiles of IL and ILG were similar, while IS has its specific chemical profiles. Twenty-seven characteristic peaks were chosen and showed a good distinction of the three species. The tissue-specific profiling of leaves showed the diterpenoids of all the species were accumulated only in the glandular scales. Lipidomics study on IL, ILG and IS was also conducted. A total of 92 lipids were identified. The variation of lipid profiles of the three Isodon species was further quantified, the results showed that the contents of the lipids in the three Isodon species varied. Statistical analyses showed IS has distinctly different lipid profile, while that of IL and ILG are very similar. Finally, the methods of macroscopic microscopic authentication and UPLC-MS fingerprinting were applied in identifying the source species of commercial Xihuangcao products. Twenty-seven batches of Xihuangcao decoction pieces were identified, results showed ILG is the major source of the collected samples. The ingredients in eight Xihuangcao herbal tea bags were also identified. IS is the major species, and none of the samples match their labels. The study provided valuable information on the authentication and quality control of folk medicinal herb Xihuangcao. The work also provided fundamental information on further studies on the chemical constituents of IL and ILG, also and role of lipids in the production of bioactive diterpenoids in Isodon species

La pharmacognosie, science multidisciplinaire étudiant les produits naturels, n'a eu de cesse de s'adapter et d'évoluer parallèlement aux autres disciplines, en particulier les techniques d'analyse chimique et les technologies de l'information. De nouvelles approches complémentaires, comme la pharmacognosie inverse allant des molécules vers les organismes sources, un cheminement en sens inverse de la pharmacognosie, ont été développées. Les données nécessaires à ces études n'ont jamais été aussi nombreuses et hétérogènes. Molécules, activités, cibles, organismes sources et leurs usages traditionnels, sont autant d'informations essentielles à la pharmacognosie. Le but de cette thèse a été de développer une plateforme informatique, regroupant toutes ces informations diverses au sein d'une vaste base de données, accompagnée d'une interface web dédiée et d'outils de chémoinformatique et de fouille de données. Parallèlement, un enrichissement constant de cette base a été réalisé. Cette plateforme permet de rechercher et de consulter les informations, mais également de les croiser afin d'en faire émerger de nouvelles connaissances. Réellement optimisée pour la pharmacognosie, elle simplifie et améliore l'étude des produits naturels en permettant une approche sur de multiples niveaux (ethnopharmacologie, botanique, chimie), offrant de nouvelles possibilités d'interaction entre les données de ces différents domaines comme l'organism hopping (sauts entre organismes), une approche originale de comparaison des organismes selon leurs profils chimiques. / Pharmacognosy, a multidisciplinary science which studies natural products, has greatly evolved in parallel with other disciplines particularly analytical chemistry and information technologies. New complementary approaches like reverse pharmacognosy, from molecules to source organisms (in contrary of pharmacognosy), have been developed. Data necessary for these studies are overwhelming and heterogeneous. Molecules, activities, targets, organisms, traditional uses, are some of essential information for pharmacognosy. The main goal of this thesis was to develop an information platform, including a large database, to collect the diverse data, along with a specific web interface and also mining and cheminformatics tools. A constant enrichment of this database has also been realized. This platform allows to query and browse data, but also to cross them in order to discover new knowledge. Truely optimised for pharmacognosy, it makes easier and enhances the study of natural products, due to a multi-level approach (ethnopharmacology, botany, chemistry). It offers new possibilities of interaction between data from different domains, such as organism hopping, a new concept of organism comparisons according to their chemical profiles.

Cordia ecalyculata Vell. (Boraginaceae), espécie medicinal vulgarmente conhecida como porangaba e chá-de-bugre, é utilizada popularmente como emagrecedor, diurético, para curar tosses catarrais e reumatismos além de ser usada no tratamento de úlceras externas em forma de banhos. No Brasil existe grande confusão com relação aos nomes populares das espécies medicinais, fato que motivou a realização do presente trabalho. A espécie estudada é confundida com outras espécies vegetais, particularmente com Casearia sylvestris Swartz (Flacourtiaceae), também conhecida pelos nomes de guaçatonga e erva-de-bugre. O estudo farmacobotânico de C. ecalyculata revelou estruturas papilosas na epiderme e células esclereificadas no parênquima cortical, que não haviam sido descritas anteriormente. Foram listadas as principais diferenças morfoanatômicas entre as folhas de C. sylvestris e C. ecalyculata. auxiliares na diagnose da droga vegetal comercializada. As amostras adquiridas no comércio como porangaba não apresentaram características de C. ecalyculata, mas sim, de C. sylvestris, como: duas a três camadas de parênquima paliçádico; cavidades secretoras; drusas e cristais prismáticos em abundância; estômatos paracíticos; células epidérmicas poligonais; ausência de litocistos e areia cristalina. Fotomicrografias ilustram o trabalho. A triagem fitoquímica do extrato hidroetanólico liofilizado (EHEL) de C. ecalyculata apresentou resultado positivo para taninos, flavonóides e saponinas. A análise cromatográfica permitiu verificar a presença de substância coincidente com a alantoína no extrato de C. ecalyculata, além de possibilitar o desenvolvimento do perfil cromatográfico, auxiliar na diferenciação de C. sylvestris. Os ensaios de quantificação de alantoína no EHEL foram realizados por meio de dois métodos espectrofotométricos, cujos valores encontrados foram 0,68% e 0,70% de alantoína. O EHEL não apresentou róxicidade no ensaio de toxicidade aguda, na dose de 5 g/kg de massa corpórea do animal, por via oral. O experimento de atividade antiúlcera do EHEL de C. ecalyculata não apresentou diferenças significativas entre os animais controle e tratados. / Cordia ecalyculata Vell. (Boraginaceae) is a medicinal species most known as porangaba and chá-de-bugre. It is commonly used as diuretic, weight controller, for phleem cough healing, rheumatism healing and also in baths for externai ulcers treatment. The present work aims at clarifying the popular names of medicinal species often misnamed in Brazil. For example, the species cited above has always been mistaken as Casearia sylvestris Swartz. (Flacourtiaceae), also known as guaçatonga and erva-de-bugre. The pharmacological-botanical study of C. ecalyculata revealed papillary structures on the epidermis and sclerenchymatous cells on the cortical parenchyma which had not yet been described in the literature. The main morphological-anatomical differences between the C. sylvestris and the C. ecalyculata leaves were listed. These differences have been of great help concerning the diagnosis of commercialized vegetal drug. The commercialized samples do not show the features of the C. ecalyculata, but the features of the C. sylvestris, such as: two or three palisade parenchyma layers; secretive cavities; plenty of druses and prismatic crystals; paracytic stomata; polygonal epidermical cells; absence of lythocysts and crystal-sand, as it can be seen in the illustrative photomicrographies. The phytochemical screening of the lyophilized hydroethanolical extract (EHEL) of the C. ecalyculata showed a positive outcome for tannins, flavonoids and saponins. The chromatographic analysis not only allowed the evaluation of the coincident substance with the allantoin in the C. ecalyculata extract, but also enable the development of its chromatographic aspect, what eventually helped to differenciate it from C. sylvestris. The quantification assays of allantoin in the EHEL extract of the C. ecalyculata were performed through two spectrophotometric methods and the values found were 0,68% and 0,70% of allantoin. The EHEL extract displayed no toxicity in the acute toxicity trial in the 5 g/kg oral dosage per animal body weight. The anti-ulcer assay of this extract showed no meaningful differences among the subject and the treated animais.

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-04-11T18:07:24Z No. of bitstreams: 1 Tietbohl, Luis Armando Candido [Dissertação, 2014].pdf: 8958883 bytes, checksum: 41b0a4341bcfc24a0012440a901a42f7 (MD5) / Made available in DSpace on 2017-04-11T18:07:24Z (GMT). No. of bitstreams: 1 Tietbohl, Luis Armando Candido [Dissertação, 2014].pdf: 8958883 bytes, checksum: 41b0a4341bcfc24a0012440a901a42f7 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A espécie Myrciaria floribunda (H.West ex Willd.) O. Berg., Myrtaceae, popularmente conhecida como camboim-amarelo, foi coletada no Parque Nacional da Restinga de Jurubatiba (RJ, Brasil). Esta espécie é rica em estruturas secretoras de óleo essencial. Neste trabalho, é descrita a análise da composição química dos óleos essenciais de folhas, caules e flores. O monoterpeno 1,8 cineol apresenta-se como componente majoritário nos óleos de folhas (38,4%) e flores (22,8%). No óleo de caules a substância majoritária foi o acetato de farnesila (19,9%). Além das substâncias já descritas para o óleo essencial de folhas desta espécie, foram identificadas pela primeira vez: α-pineno (0,4%), mirceno (0,7%), (Z)-β-ocimeno (0,8%), γ-terpineno (0,8%), acetato de geranilo (1,5%), γ-himachaleno (7,0%), valenceno (1,8%), δ-amorpheno (4,0%), zonareno (4,6%), selina-3.7(11)-dieno (3,8%), neo-intermedeol (0,8 %) e (2E, 6Z)-farnesol (1,4%). As análises dos óleos essenciais de caules e flores ainda não haviam sido descritas para esta espécie. Também foram isolados e identificados pela primeira vez na espécie o triterpeno ácido betulínico e o flavonoide miricetina 3-O-β-galactosídeo da fração acetato de etila de folhas. O óleo essencial de folhas foi avaliado quanto as atividades: anticolinesterásica, antimicrobiana e inseticida. Os óleos de folhas, flores e caules foram avaliados quanto a atividade anticolinesterásica, mostrando um IC50 de 1583 e 681 μg/mL para flores e folhas, respectivamente. O óleo de caules não apresentou atividade. O teste de susceptibilidade antimicrobiana do óleo essencial das folhas utilizando cepas de Staphylococcus aureus ATCC 29213, apresentou um CMI de 250 μg/mL. A atividade inseticida do óleo essencial das folhas foi avaliada frente aos fitófagos Oncopeltus fasciatus e Dysdercus peruvianus e os óleos essenciais das folhas e flores foram avaliados frente ao percevejo hematófago da espécie Rhodnius prolixus. Os tratamentos dos insetos com os óleos essenciais de M. floribunda induziram mortalidade, atraso no desenvolvimento e inibição de muda. Para avaliar a atividade hipoglicemiante e integridade do fêmur em ratos machos, foi utilizado o extrato aquoso das folhas de M. floribunda a uma concentração de 5 % (p/v), onde foi possível verificar ao final dos 42 dias de tratamento uma diferença significativa na glicemia e resultados similares para o fêmur. O grupo tratado apresentou resultados significativamente aumentados de colesterol, proteína total, fósforo, magnésio e aspartato aminotransferase, massa de fígado menor e massa dos rins maiores quando comparados com o grupo controle. / The species Myrciaria floribunda (H.West ex Willd.) O. Berg., Myrtaceae, popularly known as “camboim-amarelo”, was collected in the “Parque Nacional da Restinga de Jurubatiba” (RJ, Brazil). This species is rich in secretory structures essential oil. This work describes the analysis of the chemical composition of the essential oil of leaves, stems and flowers in that the monoterpene 1.8-cineole present as the major component in the oil leaves and flowers corresponding 38.4% 22.8%, respectively. Already at oil from stems the substance majority was farnesyl acetate (19.9%). In addition to the substances already described for the essential oil from leaves of the species were first identified the following substances : α-pinene (0.4%), myrcene (0.7%), (Z)-β-ocimene (0.8%), γ-terpinene (0.8%), geranyl acetate (1.5%), γ-himachalene (7.0%), valencene (1.8%), δ-amorphene (4.0%), zonarene (4.6%), 3.7-selina (11) -diene (3.8%), neo-intermedeol (0.8%) and (2E, 6Z)-farnesol (1.4%). The analysis of essential oils from stems and flowers had not yet been described. Were also isolated and identified for the first time in the kind triterpene betulinic acid and flavonoid myricetin 3-O-β-galactoside of ethyl acetate fraction of leaves. The essential oil from leaves was evaluated for activities: acetylcholinesterase, antimicrobial and insecticidal. The oils from leaves, flowers and stems were evaluated for anticholinesterase activity, showing an IC50 of 1583 and 681 mg/mL for flowers and leaves, respectively, and stems showed no activity. The antimicrobial susceptibility testing essential oil from leaves using Staphylococcus aureus ATCC 29213, filed an IMC 250 μg/mL. Insecticidal activity of the essential oil from the leaves was evaluated against phytophagous Oncopeltus fasciatus and Dysdercus peruvianus, and the essential oil from the leaves and flowers were evaluated against the Rhodnius prolixus. All treatments of insects with essential oils of M. floribunda induced mortality, developmental delay and inhibition of molting. To evaluate the hypoglycemic activity and femur health in male rats, were evaluated with the aqueous extract of leaves of M. floribunda at a concentration of 5%, which was verified at the end of the 42 days of treatment the infusion decreased the glycemia e maintenance results similar to the femur. Treated group apresented results significantly increased to cholesterol, total protein, phosphorus, magnesium and aspartate aminotransferase, mass minor liver and kidney mass was increased compared with the control group

Magister Scientiae - MSc / This study aimed to evaluate the antimicrobial efficacy of medicinal hemp plant extracts to determine the antibacterial effects of indigenous Sansevieria species and exotic Cannabis sativa phytotherapy varieties. This study also assessed whether aqueous o / South Africa

Leaf and bark extracts of a Mexican medicinal plant, Byrsonima crassifolia (Malpighiaceae), exhibited spasmogenic effects on isolated rat fundus and biphasic effects on jejunum and ileum. Preliminary evaluations using rat fundus in Krebs solution indicated that the activity of a 2% acetic acid extract of eaves (HOAcE) could be split into two types: (i) high-efficacy, low-potency, n-butanol-extracted, pargyline- and 1-(1-naphtylpiperazine) (1-NP)-sensitive, atropine-insensitive activity, and (ii) low-efficacy, high-potency, ethyl acetate-extracted, pargyline-insensitive, atropine- and 1-NP-sensitive activity. HOAcE lacked muscarinic and nicotinic effects on rat jejunum and frog rectus abdominis. Serotonin (5-HT) and HOAcE curves in fundus were parallel and 5-HT potency was 6,037 times that of HOAcE (95% confidence limits: 4,624-7,852). The pD2 (affinity constant) for 5-HT was 7.96 (7.92-8.00) with pargyline added to the medium. 5-HT receptor-interaction kinetics using cholinergics and 5-HT agonists and antagonists was carried out. 1-NP competitively antagonized 5-HT. The 5-HT, antagonist s-(-)propranolol did not significantly antagonize 5-HT. The 5-HT2 blocker ketanserin noncompetitively antagonized 5-HT and α-Me-5-HT (pD'2 = 5.6 and 6.7, respectively). The 5-HT3 antagonist MDL-72222 inactivated only a small proportion of receptors (pD'2 = 6.46). Atropine did not significantly modify the curve of 5-HT while fluoxetine noncompetitively antagonized 5-HT (pD'2 = 5.8). 5-HT and α-Me-5-HT curves were biphasic indicating two receptor interactions (high and low affinity). High-affinity pD2 values for six different 5-HT agonists and 1-NP on rat fundus correlate well with reported rat brain (radioligand binding) pKd values at the 5-HT1C receptor (r = 0.94). Large scale extraction and fractionation of a methanol extract of leaves yielded two peaks of activity (Peak 1, lipophilic; Peak 2, polar). Peak 1 contained Compounds 1 to 7 (C1-C7); Peak 2 included C8-C15. Compound 1, C2, C3, C4, C10 and C11 were inactive while C8, C12 and C13 showed equivocal effects. Compound 5, C6, C7, C9, C14, C15, quercetin and gallic acid were active. Potencies were: C5 > C6 > C7 = quercetin > C9 > gallic acid. Efficacy (IA) was: C15 ≥ C14 > gallic acid > C9 > C5 > C7 > quercetin > C6. Compound 9 and its aglycone quercetin were partial agonists (C9 IA = 70%, pD2 = 6.35; quercetin IA = 60%, pD2 = 6.58). Compound 9 noncompetitively antagonized 5-HT (pD'2 = 6.10), while quercetin did not. Compound 14 and C15, the most active compounds, had similar response curves but these curves were not parallel to 5-HT. Spasmogenic ED50 values for C14 = 0.76 (0.38-1.54) μg/mL and C15 = 0.76 (0.41-1.42) μg/mL. Gram for gram 5-HT was 181 x C14 and 182 x C15.