Background: HIV/AIDS remains a major health concern worldwide and the number of people infected in Sub-Saharan Africa continues to increase. This despite increased awareness and availability of HIV drugs in most countries. The success of current HIV-1 drugs is overshadowed by the emergence of drug resistant viral strains and the adverse side-effects they may cause. It is these limitations and many more that drives the continuous search for better HIV treatments. Research into drug discovery and development using natural products is becoming better established. With natural products, there are endless opportunities for discovering novel compounds which either ends up as final drugs or as backbones of drug leads. Methods: In this thesis, sixteen Lamiaceae (mint) plants were investigated for inhibitory properties against HIV-1 as well as for beneficial immune enhancing effects. This family of plants is commonly used in traditional medicine preparations for the treatment of various ailments including those that are virus induced. Cytotoxicity of the plant material was determined using tetrazolium dyes and the results subsequently confirmed with flow cytometry and real-time cell analysis. Direct enzyme assays were used to determine the inhibitory properties of the extracts and isolated compounds against HIV-1 protease (PR), reverse transcriptase (RT) and integrase (IN). The effect of the plant materials was also evaluated in an in vitro model of chronic and latent infection by measuring HIV-1 p24 protein secretion of an infected cell line (U1) following treatment. Most HIV-infected individuals only seek treatment during the chronic stages of disease and latent reservoirs of the virus perpetuate treatment. The immune modulating properties were determined by quantitating the effects of plant extracts/compounds on Th1/2/17 cytokine production in human mononuclear cells using the cytometric bead array technology. Finally, anti-oxidant and anti-inflammatory properties were also assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide colorimetric assays respectively. Results and discussion: The 50% cytotoxic concentration (CC50) of the extracts was between 4.2 and 100 μg/ml. Of the sixteen, extracts from six plants (Ocimum labiatum, Ocimum serratum, Plectranthus barbatus, Plectranthus neochilus, Salvia apiana and Stachys byzantina) were active against HIV-1. Four plants (P. neochilus, O. serratum, S. apiana and S. byzantina) demonstrated moderate inhibitory properties against HIV-1 PR, RT and IN (40-49%) and three of these plants (O. serratum, S. apiana and S. byzantina) significantly (p<0.05) suppressed HIV-1 replication in U1 cells. The most exciting data was obtained from extracts of P. barbatus and O. labiatum which demonstrated inhibition classified as good (>50%) against HIV-1 PR (IC50s 62 ±0.2 and 49.8 ±0.4 μg/ml), reduced the production of pro-inflammatory cytokines at non-cytotoxic concentrations and demonstrated strong antioxidant properties (IC50 values 13 ±0.8 and 15.8 ±0.3 μg/ml). O. labiatum extract also suppressed HIV-1 expression in U1 cells, significantly (p<0.05). In addition, one of the extracts (P. ciliatus) had anti-cancer potential with CC50 values <10 μg/ml. O. labiatum extract was purified to yield a chlorophyll derivative, pheophytin-a (phy-a); triterpene isomers (3-hydroxy-4,6a,6b,11,12,14b-hexamethyl- 1,2,3,4,6,6a,6b,7,8,8a,9,10,11,12,12a,14,14a,14b-octadecahydropicene-4,8adicarboxylic acid), amyrin and a labdane diterpenoid (labda-8(17),12E,14-triene-2R,18- diol). Phy-a inhibited HIV-1 PR with an IC50 value of 44.4 ±1.5 μg/ml. The triterpenes activated latent HIV-1 (a serious obstacle in the eradication of the virus) while the diterpenoid reduced the production of pro-inflammatory cytokines. These activities were observed at non-toxic concentrations of these compounds. There is an ongoing search for novel compounds that are able to activate latent HIV-1 to use in conjunction with HAART. If the triterpenes were to progress to clinical use, their use would be in activating latent virus for eradication by existing treatments. Conclusion: The findings presented in this thesis provide some scientific explanation for the anecdotal success of some Lamiaceae plants used traditionally to manage HIV/AIDS. The findings also conform to recommendations by the scientific community regarding the validation of the beneficial effects of plant products used traditionally. / Thesis (PhD)--University of Pretoria, 2013. / gm2014 / Biochemistry / unrestricted

Desde os primórdios da medicina, substâncias derivadas de animais, plantas e microrganismos têm sido utilizadas no tratamento e cura de diversas doenças. Especificamente o uso de plantas medicinais pela população como terapia alternativa no tratamento de doenças, tem sido uma prática comum desde milhares de anos antes da era presente (a.p.). Atualmente, sabe-se que muitos dos metabólitos secundários estão diretamente envolvidos nos mecanismos que permitem a adaptação das plantas ao seu habitat. Dessa forma, e pensando-se em espécies de uso medicinal, é esperado que o potencial biológico também sofra variações de acordo com os fatores bióticos e abióticos. Hyptis (Lamiaceae) constitui um gênero altamente promissor para estudos de prospecção de substâncias farmacologicamente ativas, favorecidos pela grande diversidade de espécies que ocorrem nos domínios fitogeográficos brasileiros, sendo, portanto, escolhido como modelo para responder às questões deste trabalho. O objetivo geral foi a expansão das análises fitoquímicas e do potencial biológico de quatro espécies de Hyptis pertencentes à seção Peltodon , além de verificar uma possível variação fitoquímica (quantitativa e/ou qualitativa) entre as espécies estudadas e coletadas não só em diferentes domínios, mas em diferentes fitofisionomias. Como resultado, Hyptis seção Peltodon apresentou composição química semelhante às espécies da subfamília Nepetoideae em relação à constituição fenólica, destacando a presença do ácido cafeico, ácido rosmarínico e nepetoidinas, corroborando estudos já existentes. Também reportamos que Hyptis seção Peltodon possui flavonoides derivados da flavona apigenina, sendo que identificação de flavonas C-glicosiladas nos extratos brutos sugerem o uso das mesmas como importantes marcadores taxonômicos no nível de seção. As espécies que apresentaram maior potencial antioxidante e anti-HIV-1 foram H. comaroides e H. meridionalis. Com relação às espécies coletadas nos dois domínios fitogeográficos propostos concluímos que para o grupo estudado e para as substâncias analisadas, populações que se encontrem no domínio Tropical Atlântico são as melhores candidatas como fonte de antioxidantes naturais. Entretanto, futuras pesquisas são necessárias a fim de investigar e compreender melhor os efeitos sinérgicos de múltiplos fatores ambientais no metabolismo secundário bem como no potencial biológico de espécies vegetais / Since the beginning of medicine, substances derived from animals, plants, and microorganisms have been used in the treatment and cure of various diseases. The use of medicinal plants by the population as an alternative therapy in the treatment of diseases has been a common practice for thousands of years before the present (b.p.). Nowadays we known that many of the plant secondary metabolites are directly involved in the mechanisms that allow adaptation to their habitat. Considering species of medicinal use, it is expected that the biological potential also suffers variations according to the biotic and abiotic factors. Hyptis (Lamiaceae) is a highly promising genus for prospective studies of bioactive substances, favored by the great diversity of species that occur in the Brazilian phytogeographical domains, being therefore chosen as a model to answer the questions of this work. The objective of this study was to contribute with chemical data and the biological potential of four Hyptis species belonging to the Peltodon section, as well as to verify a possible phytochemical variation (quantitative and/or qualitative) between species naturally occurring in two different phytogeographical domains. As results of this work, Hyptis section Peltodon presented phenolic composition similar to species of the subfamily Nepetoideae, with the presence of caffeic acid, rosmarinic acid, and nepetoidins, corroborating existing studies. We also reported that Hyptis section Peltodon possesses flavonoids derived from the flavone apigenin, and the identification of C-glycosylated flavones in the crude extracts suggest the use of these as important taxonomic markers at the section level. We observed that in general, the species that presented the highest antioxidant and anti-HIV-1 potential were H. comaroides and H. meridionalis. Regarding the influence of the phytogeographic domains we concluded that for the group studied and for the substances analyzed, populations that occurs in the Tropical Atlantic domain are the best candidates as source of natural antioxidants. Future research, however, is necessary to better understand the synergistic effects of multiple environmental factors on secondary metabolism, as well as the biological potential of plant species

Submitted by Verena Bastos (verena@uefs.br) on 2015-10-14T21:06:54Z No. of bitstreams: 1 Paloma_R._Meira-disserta??o_2013.pdf: 1508066 bytes, checksum: cc4d4d39667159152b8407fa887ad2fa (MD5) / Made available in DSpace on 2015-10-14T21:06:54Z (GMT). No. of bitstreams: 1 Paloma_R._Meira-disserta??o_2013.pdf: 1508066 bytes, checksum: cc4d4d39667159152b8407fa887ad2fa (MD5) Previous issue date: 2013-09-19 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / This paper describes the influence of abiotic and biotic factors in the concentration of secondary metabolites of pharmaceutical interest in medicinal plants, while it aims to answer how these factors influence the concentration of podophyllotoxin and yate?na Leptohyptis macrostachys in vitro, this goal. The specimen Leptohyptis macrostachys was established in vitro from seeds collected in the Chapada Diamantina-Ba; studying the rate of germination rate, the germination and mean germination of seeds grown on MS medium with different concentrations of salts constituents of this medium as well as different concentrations of sucrose and gibberellic acid. Was also studied, such as abiotic factors (culture medium, carbon source temperature, photoperiod and gas exchange) and biological (plant growth regulators) influenced the development (number of shoots and leaves and roots presence of friable callus, compact or oxidized) and production of podophyllotoxin and yate?na. To study the influence of these factors in getting these metabolites, plants were steeped at 40 ? C in ethanol to produce the extract. This was partitioned with water and after a specific treatment, injected into the HPLC. We established a protocol for Leptohyptis macrostachys growing from seeds in ? MS medium supplemented with 1.5% sucrose and 11.55 mM of GA3, there was no significant formation of callus in treatments applied and the concentration of podophyllotoxin was favored when L. macrostachys was grown in ? MS medium supplemented with 1.5% sucrose, closed with PVC film and kept in a growth room at 30 ? C. / Este trabalho descreve a influ?ncia de fatores abi?ticos e bi?tico na concentra??o de metab?litos secund?rios de interesse farmacol?gico em plantas medicinais, ao mesmo tempo que se prop?e a responder como esses fatores influenciam na concentra??o de podofilotoxina e yate?na em Leptohyptis macrostachys in vitro, objetivo deste. A esp?cime Leptohyptis macrostachys foi estabelecida in vitro a partir de sementes coletadas na regi?o da Chapada Diamantina-Ba; estudando o ?ndice de velocidade de germina??o, a germinabilidade e o tempo m?dio de germina??o de sementes cultivadas em meio MS com diferentes concentra??es de sais constituintes deste meio, bem como diferentes concentra??es de sacarose e ?cido giber?lico. Foi estudado tamb?m, como fatores abi?ticos (meio de cultivo, fontes de carbono temperatura, fotoper?odo e trocas gasosas)e bi?tico (reguladores vegetais) influenciaram no desenvolvimento (quantidade de brotos folhas e ra?zes e presen?a de calos fri?veis, compactos ou oxidados) e na produ??o de podofilotoxina e yate?na. Para estudar a influ?ncia estes fatores na obten??o destes metab?litos, as plantas foram maceradas ? 40?C em etanol para a produ??o do extrato. Este foi particionado com ?gua e, ap?s tratamento espec?fico, injetado no CLAE. Foi estabelecido um protocolo de cultivo para Leptohyptis macrostachys a partir de sementes em meio MS ? suplementado com 1,5% de sacarose e 11,55?M de GA3; n?o houve forma??o significativa de calos nos tratamentos aplicados e a concentra??o da podofilotoxina foi favorecida quando L. macrostachys foi cultivada em meio MS ? suplementado com 1,5% de sacarose, fechada com filme de PVC e mantida em sala de crescimento a 30?C.

Pollination data is provided for a third of the Plectranthus species in southern Africa. In the largest genus of Lamiaceae in the region (53 species), 18 were studied, plus two species of allied genera (Pycnostachys urticifolia and Aeollanthus parvifolius). Study of these 20 species aimed to describe the groups of pollinators that have driven pollinator specialisation. Case histories are provided upon which future studies of Lamiaceae pollination, breeding systems and speciation may be based. Bees (Apidae) and flies (Nemestrinidae, Acroceridae and Tabanidae) are the main pollinating insect groups. Seven straight-tubed Plectranthus species show a match between corolla tube- and proboscis length of nectar-feeding pollinators. Long-proboscid nemestrinid flies are specialised on long-tubed Plectranthus species (P. ambiguus, P. hilliardiae, P. reflexus and P. saccatus), while shorter-proboscid flies of all three families are important pollinators of straight-tubed species with medium- and short corolla tubes. Seven species with sigmoid corolla tubes are bee-pollinated, with fly-pollination prevalent in some. Bent corolla tubes, coupled with length, act as barriers to illegitimate nectarfeeders and ensures alignment of pollinators for effective pollen placement and carryover. It is suggested that straight-tubed species may have evolved from sigmoid-tubed species. Long-tubed species with straight corollas in other Lamiaceae may show convergent pollination by long-proboscid flies, with the guild being dependent on habitat and distribution of plants and flies. Formal establishment of the Stenobasipteron wiedemanni pollination guild extends the study from Plectranthus to selected Acanthaceae, Orchidaceae, Balsaminaceae, Gesneriaceae and Iridaceae, occurring in forested habitat along the Eastern seaboard. Micro-distillation of essential oils confirmed parentage of a putative natural hybrid; once established, hybrid data allows studies of the importance of natural hybridisation events in explaining pollinator fidelity. Nectar sugar studies in Plectranthus mostly showed sucrose dominance; cases of hexose dominance are noted and discussed. Nectar volume and concentration proved variable and do not fit any trends. Pollination by medium-proboscid acrocerid flies has importance for ‘medium-tubed’ plants, since six of the Plectranthus species are solely or partially reliant on Acroceridae for pollination. An appendix with consolidated data describes the 20 study species i.t.o. morphology, habitat, study sites, field work, pollinator observations and insect vouchers. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Researches involving plant derived products aiming to discover extractives useful for different purposes have been growing, and among them, essential oils (EO) are noteworthy. One of the main sources of EO is the Lamiaceae family. Belonging to this family, Hesperozygis ringens (Benth.) Epling is a native species from the Pampa biome, popularly known as espanta-pulga . This study aimed to evaluate the biological potential of EO from its leaves as allelopathic and larvicidal, as well as to check the seasonal and individual variability of yield and chemical composition of this oil. EO of four specimens were obtained from leaves collected in different seasons of a year and had their yield (% m/m) and chemical compositions determined. EO were extracted by hidrodestillation in Clevenger apparatus and analyzed by gas chromatography coupled to mass spectrometry (GC-MS) and gas chromatography with flame ionization detector (GC-FID). The allelopathic potential of the EO was tested against seeds of Dicots and Monocots classified as bioindicators [Avena strigosa Schreb. and Lactuca sativa L.], weeds [Lolium multiflorum Lam and Bidens pilosa L.] and crops [Oryza sativa L. and Glycine max (L.) Merr.] by observing the following variables: germination, emergence velocity index, root length, shoot lenght and total length of seedlings. The tested concentrations were 0.25, 0.50, 1.00, 2.50 and 5.00%.The EO of H. ringens had also its larvicidal activity tested against larvae of Odonata in order to determine its 50% lethal concentration (LC50). Regarding the allelopathic tests on Dicotyledons, EO with 82.02% of pulegone inhibited completely the germination of B. pilosa at 5.00%, without affecting significantly L. sativa, but slowing the emergence and damaging root lengths and seedlings of G. max. Regarding the monocots, the same concentration strongly impaired L. multiflorum, without affecting the germination of O. sativa, but it influenced negatively the other evaluated parameters. In addition, the development of the seeds of A. strigosa was completely inhibited at the concentration of 2.50%. The yield of EO proved to be influenced by seasonality, being higher in spring, summer, and autumn. The yield also showed individual variability (1.15 a 4.38%), proving to be higher in specimen 4. The chemical composition of EO obtained from leaves collected in autumn, spring, and summer showed similar results. The production of the major constituent (pulegone) and the predominant class (oxygenated monoterpenoids) were favored in spring, summer, and autumn. These seasons proved to be more indicated for the EO extraction, since they enabled concomitantly higher yields and higher contents of oxygenated monoterpenoids and pulegone. Regarding the larvicidal activity, EO containing 95.18% of pulegone showed a LC50 of 69.05 µL L-1 against the Odonata larvae. / Pesquisas sobre produtos derivados de plantas visando à descoberta de extrativos úteis para diferentes fins são crescentes e, dentre estes, destacam-se os óleos essenciais (OE), tendo como uma de suas principais fontes a família Lamiaceae. Pertencente a ela, Hesperozygis ringens (Benth.) Epling é uma espécie nativa do bioma Pampa, conhecida popularmente por espanta-pulga. O presente trabalho teve como objetivos avaliar o potencial biológico do OE de suas folhas como alelopático e larvicida, bem como verificar as variabilidades sazonal e individual do rendimento e composição química desse óleo. O OE de quatro indivíduos foi obtido de folhas coletadas nas diferentes estações de um ano, determinando-se o rendimento do OE (% m/m) e as composições químicas das amostras. O OE foi extraído por hidrodestilação, em aparelho Clevenger e teve sua composição química analisada por cromatografia gasosa acoplada à espectrometria de massas (CG-EM) e cromatografia gasosa com detector de ionização em chama (CG-DIC). O potencial alelopático do OE foi testado em sementes de Dicotiledôneas e Monocotiledôneas, classificadas como bioindicadoras [Avena strigosa Schreb. e Lactuca sativa L.], plantas daninhas [Lolium multiflorum Lam. e Bidens pilosa L.] e culturas [Oryza sativa L. e Glycine max (L.) Merr.], através da avaliação das seguintes variáveis: germinação, índice de velocidade de emergência e comprimentos das raízes, partes aéreas e total das plântulas. As concentrações testadas foram: 0,25; 0,50; 1,00; 2,50; 5,00%. O OE de H. ringens também teve sua atividade larvicida testada em larvas de Odonata, a fim de determinar sua concentração letal 50% (CL50). Em relação aos testes alelopáticos sobre Dicotiledôneas, o OE com 82,02% de pulegona inibiu completamente a germinação de B. pilosa na concentração de 5,00%, sem afetar significativamente L. sativa, porém retardando a emergência e prejudicando os comprimentos de raiz e plântula de G. max. Em relação às monocotiledôneas, a mesma concentração prejudicou fortemente L. multiflorum, sem afetar a germinação de O. sativa, porém influenciando negativamente os demais parâmetros. Já o desenvolvimento das sementes de A. strigosa foi completamente inibido na concentração de 2,50%. O rendimento do OE demonstrou ser influenciado pela sazonalidade, sendo maior na primavera, verão e outono. O rendimento também apresentou variabilidade individual (1,15 a 4,38%), demonstrando ser mais elevado no indivíduo 4. Em relação à composição química, os OE obtidos de folhas coletadas no outono, primavera e verão apresentaram resultados semelhantes. A produção do constituinte majoritário (pulegona) e da classe predominante (monoterpenoides oxigenados) foram favorecidos naprimavera, verão e outono. Estas estações demonstraram ser as mais favoráveis para extração de OE por apresentar concomitantemente maiores rendimentos e teores mais elevados de monoterpenoides oxigenados e de pulegona. Quanto à atividade larvicida, o OE contendo 95,18% de pulegona apresentou uma CL50 de 69,05 µL L-1 frente a larvas de Odonata.

Salvia officinalis (Lamiaceae), conhecida popularmente como sálvia, tem sido muito utilizada no sul do Brasil como condimento nos alimentos, tintura hidroalcoólica e chá para o tratamento de vários distúrbios de saúde. As propriedades dessa planta são estudadas por apontar possíveis mecanismos de ação antioxidante e antitumoral. Neste trabalho, o extrato hidroalcoólico e aquoso de sálvia foram analisados quimicamente por meio de cromatografia gasosa acoplada a espectrômetro de massas (GC-MS) e electrospray de alta resolução acoplada a espectrômetro de massas (ESI-QTOF MS/MS) em modo negativo. A identificação química mostrou a presença de ácidos como caféico, rosmarínico, málico, succínico, tartárico, cítrico, ursólico e compostos como luteolina-7-O-glucoronide, eucaliptol, β-tujona, β-cariofileno, α-cariofileno, α- tujona, cânfora viridiflorol, mannol, rosmanol e seus isômeros metilcarnosato e ácido 12-metoxicarnosinico. Os extratos apresentaram compostos polifenólicos com capacidade de varrer os radicais 2,2-difenil-1-picrilhidrazil (DPPH•) e 2,2’- azino-bis(3-etilbenzotiazolina-6-ácido sulfônico) (ABTS•+), além de atividade catalase (CATlike) e superóxido dismutase (SOD-like). Ambos os extratos apresentaram menor citotoxicidade em linhagens não tumorais (HeK-293 e MRC-5) e seletividade para linhagens tumorais (Hep-2, HeLa, A-549, HT-29, A-375 e HepG2). Além disso, verificou-se que o aumento do tempo de exposição ao extrato hidroalcoólico diminuiu a viabilidade em linhagens tumorais. Foram observadas alterações morfológicas por coloração de Giemsa e a avaliação da apoptose com anexina V e iodeto de propídeo mostraram a maioria das células tumorais em estágios finais do processo de apoptose e necrose após 24h de tratamento em ambos os extratos. Sugere-se que o extrato de Salvia officinalis (L.) possui atividade biológica em células tumorais, podendo ser objetivo de mais estudos com a finalidade de comprovar sua eficácia como possível agente para o tratamento do câncer. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2015-02-12T12:00:17Z No. of bitstreams: 1 Dissertacao Charlene Silvestrin Celi Garcia.pdf: 2151872 bytes, checksum: 7bfd73bdf5fd152982466e3f75044d21 (MD5) / Made available in DSpace on 2015-02-12T12:00:17Z (GMT). No. of bitstreams: 1 Dissertacao Charlene Silvestrin Celi Garcia.pdf: 2151872 bytes, checksum: 7bfd73bdf5fd152982466e3f75044d21 (MD5) / Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul, FAPERGS / Salvia officinalis (Lamiaceae), popularly known as sage, has been used in south of Brazil as a condiment in foods, as tincture and tea for the treatment various health disorders. The properties of this plant have been studied and suggest possible mechanisms of antioxidant and antitumor action. Here, the sage hydroalcoholic and aqueous extract were chemically analyzed by gas chromatography-mass spectrometry (GC-MS) and by high-resolution electrospray ionization mass spectrometry (ESI-HRMS) in negative mode. The chemical identification showed the presence of acids as caffeic, rosmarinic, malic, succinic, tartaric, citric, ursolic and compouns like luteolin-7-O-glucoronide, eucalyptol, β-thujone, β-caryophyllene, α-caryophyllenen and α-thujone, camphor, viridiflorol, mannol, rosmanol and its isomers methylcarnosate and 12-methoxycarnosinic acid. The extracts showed the content of polyphenolic compounds, ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), moreover catalase (CAT-like) and superoxide dismutase (SOD-like) activity. Both extracts showed lower cytotoxicity in non-tumor lines (HEK-293 and MRC-5) and selectivity for tumor cell lines (Hep-2, HeLa, A-549, HT-29, A-375 e HepG2). Furthermore, it was found that increasing the treatment exposure time of the hydroalcoholic extract decreases the tumor cell viability. Morphological changes by giemsa were observed and staining for annexin V and propidium iodide showed majority of tumor cells at late stages of the apoptotic process and necrosis after 24h treatment with both extracts. The results of this study suggest that the extract of Salvia officinalis (L.) has biological activity against tumor cell and may be the objective of further studies in order to prove its effectiveness as a possible agent for the treatment of cancer.

Made available in DSpace on 2015-05-14T12:59:55Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 8172962 bytes, checksum: ccd3348e9b6251ca6982caf521b2f2ce (MD5) Previous issue date: 2013-11-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Lamiaceae family has 295 genera and about 7,775 species with cosmopolitan distribution. It is estimated that 22 genera and 402 species occur in Brazil. This study describes the phytochemical study of two species of the genus Hyptis found in semiarid Paraiba, Hyptis macrostachys Benth. and Hyptis umbrosa Salzm. ex Benth. Leaves and stems of the species studied were subjected to extraction and subsequently processes of filtration and chromatography of their extracts. Posteriorly, chemical structures of the isolated compounds were identified and/or determined by spectroscopic methods. The crude ethanol extract (CEE) of Hyptis macrostachys obtained phases hexane, and dichlorometane and ethyl acetate. In study of dichlorometanic phase from H. macrostachys were isolated five substances by conventional chromatographic methods: two diterpenes, beier-15-en-17-ol (erytroxilol B) encoded as Hm-1, isolated as before and ent-labda (8)-17.14-dien,9,13-epoxy-12α-hydroxy-19-oic acid (Hm-3) unpublished literature (hyptenol); two triterpenes, 3-β-acetoxy-olean-12-en-28-oic (acetate oleanolic acid) and 3β-hydroxy-lup-20 (29)-on-28-oic acid (betulinic acid), have previously been isolated from species of Hyptis (Hm-2 and Hm-4, respectivety), plus a pyrone 6R-[(5'S, 6'S-diacetoxy)-1'Z, 3'E-heptenyl]-5,6-dihydro-2H-pyran-2-one (Hm-5), unpublished literature (hyptenolide). From ethyl acetate phase two triterpens have been isolated, a mixture of ursolic and oleanolic acids (Hm-6) and 2α,3β,19α-trihydroxy-urs-12-en-28-oic acid (tormentic acid), encoded as Hm-7. The CCE from Hyptis umbrosa was subjected to medium pressure chromatography. The fractions were monitored by TLC and assembled according to their chromatographic profiles. In CEE of Hyptis umbrosa, two diterpens were isolated, pomifrim D (Hu-1) first reported in this species, but its NMR data were completely reassignment and salviol (Hu-2), previously isolated from other species of Lamiaceae family. How many species of Lamiaceae are known to have significant amount of essential oils, it was decided to analyze the fresh leaves of Hyptis macrostachys and Hyptis umbrosa was extracted by hydrodistillation in clevenger apparatus and characterized by GC-MS. The chemical constitution of the essential oils of Hyptis macrostachys and Hyptis umbrosa was 98.37 and 92.9%, respectively, of monoterpenes and sesquiterpenes. Being 1,8-cineole (36.3%) the major constituent for Hyptis macrostachys and fenchona (24.8%) for Hyptis umbrosa, consistent with the chemical composition of essential oils of other species of Lamiaceae. These results contributed to the phytochemical study of the species of the semiarid of Paraíba, H. macrostachys and H. umbrosa with the structural identification of nine substances, two of the unpublished (hyptenol and hyptenolide). / A família Lamiaceae possui 295 gêneros e cerca de 7.775 espécies com distribuição cosmopolita. Estima-se que 22 gêneros e 402 espécies ocorrem no Brasil. Neste trabalho, realizou-se um estudo fitoquímico de duas espécies do gênero Hyptis encontradas no semiárido paraibano, Hyptis macrostachys Benth. e Hyptis umbrosa Salzm. ex Benth. As folhas e os caules das espécies em estudo foram submetidas a processos de extração, de filtração e cromatografia de seus respectivos extratos. Posteriormente, as estruturas químicas das substâncias isoladas foram identificadas e/ou determinadas por métodos espectroscópicos. Do extrato etanólico bruto (EEB) de Hyptis macrostachys obteve-se as fases hexânica, diclorometano e acetato de etila. No estudo da fase diclorometânica de H. macrostachys foram isoladas cinco substâncias através de métodos cromatográficos convencionais, sendo dois diterpenos: o beier-15-en-17-ol (eritroxilol B), codificado como Hm-1, isolado anteriormente e o ácido ent-labda(8)-17,14-dien,9,13-epóxi,12α-hidroxi-19-oico (Hm-3), inédito na literatura (hiptenol); dois triterpenos: 3-β-acetoxi-olean-12-en-28-oico (acetato de ácido oleanólico) e o ácido 3β-hidroxi-lup-20(29)-en-28-oico (ácido betulínico), isolados anteriormente de espécies de Hyptis (Hm-2 e Hm-4, respectivamente); além de uma pirona 6R-[(5 S,6 S-diacetoxi)-1 Z,3 E-heptenil]-5,6-diidro-2H-pirano-2-ona (Hm-5), inédita na literatura (hiptenolídeo). Da fase acetato de etila foram isolados dois triterpenos, sendo uma mistura dos ácidos oleanólico e ursólico (Hm-6) e o ácido 2α,3β,19α-triidroxi-urs-12-en-28-oico (ácido tormêntico), codificado como Hm-7. O EEB de Hyptis umbrosa foi submetido à cromatografia de média pressao. As frações obtidas foram monitoradas por CCDA e reunidas de acordo com os seus respectivos perfis cromatográficos. Do EEB de Hyptis umbrosa, foram isolados dois diterpenos: o pomiferin D (Hu-1), relatado pela primeira vez nesta espécie, porém seus dados de RMN foram completamente reassinalados, e o salviol (Hu-2), isolado anteriormente em outras espécies da família Lamiaceae. Como várias espécies de Lamiaceae são conhecidas por apresentarem quantidade significativa de óleos essenciais, decidiu-se analisar as folhas frescas de Hyptis macrostachys e Hyptis umbrosa, onde os óleos essenciais foram extraídos por hidrodestilação em aparelho de clevenger e caracterizados por cromatografia gasosa acoplada à espectroscopia de massas (CG-EM). A constituição química dos óleos essenciais de Hyptis macrostachys e Hyptis umbrosa foi de 98,37 e 92,9%, respectivamente, de monoterpenos e sesquiterpenos. Sendo o 1,8-cineol (36,3%) o constituinte majoritário para Hyptis macrostachys e a fenchona (24,8%) para Hyptis umbrosa, compatíveis com a composição química de óleos essenciais de outras espécies de Lamiaceae. Esses resultados contribuiram com o estudo fitoquímico das espécies do semiárido paraibano, H. macrostachys e H. umbrosa, com a identificação estrutural de nove substâncias, sendo duas inéditas na literatura (hiptenol e hiptenolídeo).

Salvia officinalis (Lamiaceae), conhecida popularmente como sálvia, tem sido muito utilizada no sul do Brasil como condimento nos alimentos, tintura hidroalcoólica e chá para o tratamento de vários distúrbios de saúde. As propriedades dessa planta são estudadas por apontar possíveis mecanismos de ação antioxidante e antitumoral. Neste trabalho, o extrato hidroalcoólico e aquoso de sálvia foram analisados quimicamente por meio de cromatografia gasosa acoplada a espectrômetro de massas (GC-MS) e electrospray de alta resolução acoplada a espectrômetro de massas (ESI-QTOF MS/MS) em modo negativo. A identificação química mostrou a presença de ácidos como caféico, rosmarínico, málico, succínico, tartárico, cítrico, ursólico e compostos como luteolina-7-O-glucoronide, eucaliptol, β-tujona, β-cariofileno, α-cariofileno, α- tujona, cânfora viridiflorol, mannol, rosmanol e seus isômeros metilcarnosato e ácido 12-metoxicarnosinico. Os extratos apresentaram compostos polifenólicos com capacidade de varrer os radicais 2,2-difenil-1-picrilhidrazil (DPPH•) e 2,2’- azino-bis(3-etilbenzotiazolina-6-ácido sulfônico) (ABTS•+), além de atividade catalase (CATlike) e superóxido dismutase (SOD-like). Ambos os extratos apresentaram menor citotoxicidade em linhagens não tumorais (HeK-293 e MRC-5) e seletividade para linhagens tumorais (Hep-2, HeLa, A-549, HT-29, A-375 e HepG2). Além disso, verificou-se que o aumento do tempo de exposição ao extrato hidroalcoólico diminuiu a viabilidade em linhagens tumorais. Foram observadas alterações morfológicas por coloração de Giemsa e a avaliação da apoptose com anexina V e iodeto de propídeo mostraram a maioria das células tumorais em estágios finais do processo de apoptose e necrose após 24h de tratamento em ambos os extratos. Sugere-se que o extrato de Salvia officinalis (L.) possui atividade biológica em células tumorais, podendo ser objetivo de mais estudos com a finalidade de comprovar sua eficácia como possível agente para o tratamento do câncer. / Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul, FAPERGS / Salvia officinalis (Lamiaceae), popularly known as sage, has been used in south of Brazil as a condiment in foods, as tincture and tea for the treatment various health disorders. The properties of this plant have been studied and suggest possible mechanisms of antioxidant and antitumor action. Here, the sage hydroalcoholic and aqueous extract were chemically analyzed by gas chromatography-mass spectrometry (GC-MS) and by high-resolution electrospray ionization mass spectrometry (ESI-HRMS) in negative mode. The chemical identification showed the presence of acids as caffeic, rosmarinic, malic, succinic, tartaric, citric, ursolic and compouns like luteolin-7-O-glucoronide, eucalyptol, β-thujone, β-caryophyllene, α-caryophyllenen and α-thujone, camphor, viridiflorol, mannol, rosmanol and its isomers methylcarnosate and 12-methoxycarnosinic acid. The extracts showed the content of polyphenolic compounds, ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), moreover catalase (CAT-like) and superoxide dismutase (SOD-like) activity. Both extracts showed lower cytotoxicity in non-tumor lines (HEK-293 and MRC-5) and selectivity for tumor cell lines (Hep-2, HeLa, A-549, HT-29, A-375 e HepG2). Furthermore, it was found that increasing the treatment exposure time of the hydroalcoholic extract decreases the tumor cell viability. Morphological changes by giemsa were observed and staining for annexin V and propidium iodide showed majority of tumor cells at late stages of the apoptotic process and necrosis after 24h treatment with both extracts. The results of this study suggest that the extract of Salvia officinalis (L.) has biological activity against tumor cell and may be the objective of further studies in order to prove its effectiveness as a possible agent for the treatment of cancer.

La croissance démographique, la consommation incontrôlée des ressources naturelles ainsi que l’activité industrielle jouent un rôle déterminant dans la dégradation de l'environnement mondial. A ce sujet et en se basant sur le concept que les consommateurs partagent la responsabilité de la pollution et de l'épuisement des ressources et leur coût, la mise en oeuvre des ressources renouvelables dans les processus industriels apparait comme une clé décisive. La renouvelabilité, la recyclabilité, la durabilité, la biodégradabilité sont les concepts de base sur lesquels nous pouvons compter pour protéger la nature et l’environnement. Dans ce contexte, une nouvelle approche de la bioraffinage a été développée dans cette thèse, elle a été appliquée à cinq espèces appartenant à 3 familles différentes (Foeniculum vulagre, Cuminum cyminum and Carum carvi (Apiaceae), Salvia hispania (Lamiaceae) et Amaranthus cruentus (Amaranthaceae)). Ces espèces partagent une caractéristique commune, elles peuvent être définies comme Aroma Tincto Oleo Crops (ATOC) puisqu’elles sont une source d'huile végétale et d'huile essentielle (ou de colorant). Notre objectif est donc de déterminer la faisabilité biologique et technologique de l'application du concept ATOC-bioraffinage basé sur le développement, l'optimisation et la production de nouveaux produits biologiques, des nouvelles molécules biosourcées et d'autres technologies innovantes. Notre étude met l'accent sur l'évaluation de divers paramètres influençant le rendement et la qualité de l’huile dans les graines d'ATOC, y compris leurs origines géographiques et le procédé d'extraction, et leur impact sur l'activité biologique des extraits de résidus. La valorisation des huiles pour des applications dans l’industrie cosmétique a été realisée en les utilisant comme (i) additifs dans des formulations de crème hydratante. De plus, l’utilité du tourteau d'extraction comme (ii) substrat pour la fabrication d'agromatériaux par thermopressage et comme (iii) un ingrédient dans des formulations de pain de blé entier a été examinée. / In an age where the risk of environmental degradation have become more apparent due to a combination of several factors including the sheer rise of world population, consumption, industrial activity, etc. With regard to this and since the concept that consumers share responsibility for pollution and resources depletion and their cost has been increasingly believed, the implementation of renewable resources in industrial processes could be a decisive key. Renewability, recyclability, sustainability, biodegradability are the basic concepts on which we can rely to make a difference in the environment today and tomorrow. In this context, a new biorefinery approach was developed in this thesis and applied to five plant species belonging to 3 different plant families (Foeniculum vulagre, Cuminum cyminum and Carum carvi (Apiaceae family), Salvia hispania (Lamiaceae family) and Amaranthus cruentus (Amaranthaceae family). These species share a common characteristic, they can be defined as Aroma Tincto Oleo Crops (ATOC) as they are a source of both vegetable oil and essential oil or color. Our aim is thus to determine the biological and technological feasibility of application of ATOC-biorefinery concept based on the development, optimization and production of new bio-based products and other innovative technologies. Our study focus on the evaluation of major various parameters influencing oil’s yield and quality in ATOC seeds including their geographic origins and extraction process, and their impact on biological activity of residues extracts. The feasability of valorization of oils as (i) additive in cosmetic formulas, and of extraction cake as (ii) substrate for agromaterial manufacturing by thermopressing or as (iii) ingredient in whole wheat bread formulations were also stated.

O número de pessoas infectadas por protozoários dos gêneros Leishmania e Trypanosoma cruzi, no mundo, permanece preocupante, havendo cerca de 12 milhões de casos de leishmaniose e 7 a 8 milhões referentes à doença de Chagas. Face ao grave quadro destas doenças e às limitações da terapêutica atual, a busca de novos fármacos é urgente. Considerada uma das principais estratégias disponíveis, a pesquisa a partir de espécies vegetais contendo constituintes bioativos é promissora fonte de moléculas potencialmente ativas. Neste contexto, espécies do gênero Lavandula mostraram atividade anti-Leishmania in vitro, tendo motivado a seleção de L. angustifolia (Lamiaceae). Embora tenham sido comprovadas diversas atividades farmacológicas desta espécie, a atividade antiprotozoária frente aos agentes avaliados ainda não havia sido estudada. Além disto, a produção em larga escala e o seu uso bastante difundido no mundo, sobretudo nas indústrias cosméticas, de perfumes e de domissanitários, despertou interesse. O presente trabalho visou à análise da composição do óleo volátil de uma variedade de L. angustifolia aclimatada ao Brasil e à avaliação de sua atividade antiprotozoária in vitro, frente a cepas de Leishmania infantum, L. amazonensis e de Trypanosoma cruzi. A análise do óleo volátil das inflorescências e das folhas, por CG-EM, indicou a predominância de monoterpenos, tendo-se estabelecido análise comparativa entre estes órgãos. De forma geral, o óleo foi inativo frente às espécies de Leishmania, tendo apresentado promissora atividade anti-Trypanosoma. / The number of people infected by Leishmania and Trypanosoma cruzi protozoa around the world has increased and raised concerns that approximately 12 million cases of leishmaniasis and 7 to 8 million cases of Chagas disease have been reported. The alarming statistics involving these diseases and limitations of therapeutics currently in use point to an urgent need for new drugs. Considered one of the main strategies available, research on plants containing bioactive constituents is a promising source of potentially active molecules. Species of Lavandula genus have shown in vitro anti-Leishmania activity and this encouraged the present study on L. angustifolia (Lamiaceae) species. Although several pharmacological activities have been identified, the antiprotozoal activity against these specific agents has not yet been approached. Moreover, the possibility of a large-scale production and its quite comprehensive use worldwide, especially for the manufacturing of cosmetics, perfumes and household sanitizing products have attracted our interest. This study aimed at the analysis of the volatile oil compounds of the L. angustifolia species acclimatized in Brazil as well as the evaluation of the in vitro antiprotozoal activity against the strains of Leishmania (L.) infantum, L. (L.) amazonensis and Trypanosoma cruzi. The CG-EM analysis of the volatile oil from the inflorescences and leaves has shown monoterpenes as majority constituents also allowing a comparative analysis between these organs. Overall, the oil was not active against the Leishmania species but presented promising anti-Trypanosoma activity.