Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / This work describes the phytochemical investigation of two specimens of Hyptis: H. carvalhoi Harley and H. crassifolia Mart. ex Benth., whose purpose is to investigate northeastern Brazil plants of the Hyptis genus, in the search for bioactive compounds, especially with anticancer activity. The chemical analysis of both species resulted in the isolation and characterization of 10 substances for H. crassifolia and 12 substances for H. carvalhoi. Of the ethanol extract from roots of H. crassifolia were isolated nine diterpenes and the known triterpene betulinic acid. Of the nine diterpenes, four are abietanes: 12-hydroxy-8,11,13-abietatriene, 12-hydroxy-8,11,13-abietatrie-7-one, 11,12,15-tri-hydroxy-8,11,13-abietatrien-7-one and 6α,11,12,15-tetra-hydroxy-8,11,13-abietatrien-7-one, from which 11,12,15-tri-hydroxy-8,11,13-abietatrien-7-one is being reported for the first time in the literature as a new natural abietane diterpene and the 6α,11,12,15-tetra-hydroxy-8,11,13-abietatrien-7-one is unknow. Three have rearranged abietanes skeletons: 11,12,14,16-tetra-hydroxy-17 -abeo-abieta-8,11,13-trien-7-one, 11,12,16-tri-hydroxy-17 -abeo-abieta-8,11,13-trien-7-one and (16S)-12,16-epoxy-11,14-di-hydroxy-17 -abeo-abieta-8,11,13-trien-7-one, from which the 11,12,16-tri-hydroxy-17 -abeo-abieta-8,11,13-trien-7-one is unknown, and to the (16S)-12,16-epoxy-11,14-di-hydroxy-17 -abeo-abieta-8,11,13-trien-7-one is proposed a revision of the 1H and 13C NMR data previously reported in the literature. Two known labdane diterpenes: 11-oxomanoyl oxide and 11β-hydroxymanoyl oxide were also isolated. From the hexane extract of the roots of H. carvalhoi were isolated ten diterpenes, one compound from a mixed biosynthesis, 3β-[4â-acetoxyangeloiloxy]-tremetone, not yet reported for the Lamiaceae, and betulinic acid, a chemotaxonomic marker for the genus Hyptis. Of the ten diterpenes, five are abietane: 12-hydroxy-8,11,13-abietatrien-20-al, 8,11,13-abietatrien-12,20-diol, 11-hydroxy-12-metoxy-8,11,13-abietatrien-10-oic acid, 12-hydroxy-8,11,13-abietatrien-10-oic acid and 12-hydroxy-8,11,13-abietatriene, an unknown nor-diterpene, with a 20-nor-icetexane skeleton identified as 8(7),10(7)-di-epoxy-12-hydroxy-20-nor-8,11,13-abietatriene, a pimarane diterpene, 11-ketosandaracopimar-15-en-8β-ol, unknow in the Lamiaceae and three tanshinones: 7β-hydroxy-11,14-di-oxoabieta-8,13-diene, 7,12-di-hydroxy-8,12-abietadien-11,14-di-oxo-20-al and 7α-hydroxy-11,14-dioxoabieta-8,13-diene, from which the last is unknown. All compounds were isolated using successive chromatographic purification steps, including HPLC, and structural determination was performed by mean of spectroscopic techniques such as HRMS, IR, 1H and 13C NMR, including uni and bidimensional pulse sequences, and comparison with data from the literature. Among the isolated compounds, eighteen have been tested for cell-growth inhibition activity against several human cancer cell lines, and ten showed activity. From the compounds isolated from H. crassifolia, 12-hydroxy-8,11,13-abietatriene and 11,12,14,16-tetra-hydroxy-17 -abeo-abieta-8,11,13-trien-7-one showed a moderate cytotoxic activity, while 12-hydroxy-8,11,13-abietatrie-7-one exhibited a moderate, but selective activity against leukemia cell line. All diterpenes isolated from H. carvalhoi exhibited cytotoxic activity, but 7,12-di-hydroxy-8,12-abietadien-11,14-di-oxo-20-al, after 72 hours of incubation, showed IC50 values ranging from 3.91 to 32.01 μM in colon tumor (HCT-116) and leukemic (HL-60) cells, respectively. Its possible mechanism of action was then studied. / Este trabalho descreve a investigaÃÃo fitoquÃmica de dois espÃcimes de Hyptis: H. carvalhoi Harley e H. crassifolia Mart. ex Benth. O objetivo à investigar plantas do gÃnero Hyptis do Nordeste do Brasil, na busca por compostos bioativos, principalmente com atividade anticÃncer. A prospecÃÃo quÃmica relativa Ãs duas espÃcies resultou no isolamento de 10 substÃncias para H. crassifolia e 12 substÃncias para H. carvalhoi. Do extrato etanÃlico das raÃzes de H. crassifolia foram isolados nove diterpenos e o triterpeno conhecido como Ãcido betulÃnico. Dos nove diterpenos, quatro sÃo abietanos: 12-hidroxi-8,11,13-abietatrieno, 12-hidroxi-8,11,13-abietatrien-7-ona, 11,12,15-tri-hidroxi-8,11,13-abietatrien-7-ona e 6α,11,12,15-tetra-hidroxi-8,11,13-abietatrien-7-ona, dos quais a 11,12,15-tri-hidroxi-8,11,13-abietatrien-7-ona està sendo relatada pela primeira vez como um novo diterpeno abietano natural e a 6α,11,12,15-tetra-hidroxi-8,11,13-abietatrien-7-ona à inÃdita na literatura. TrÃs apresentam esqueletos abietanos rearranjados: 11,12,14,16-tetra-hidroxi-17 -abeo-abieta-8,11,13-trien-7-ona, 11,12,16-tri-hidroxi-17 -abeo-abieta-8,11,13-trien-7-ona e (16S)-12,16-epoxi-11,14-di-hidroxi-17 -abeo-abieta-8,11,13-trien-7-ona, sendo a 11,12,16-tri-hidroxi-17 -abeo-abieta-8,11,13-trien-7-ona inÃdita, e para a (16S)-12,16-epoxi-11,14-di-hidroxi-17 -abeo-abieta-8,11,13-trien-7-ona està se propondo uma revisÃo dos dados de RMN de 1H e 13C relatados na literatura. Foram isolados tambÃm dois diterpenos labdanos conhecidos: Ãxido de 11β-hidroximanoila e Ãxido de 11-oxomanoila. Do extrato hexÃnico das raÃzes de H. carvalhoi foram isolados dez diterpenos, uma substÃncia de biossÃntese mista, denominada de 3β-[4â-acetoxiangeloiloxi]-tremetona, ainda nÃo relatada na famÃlia Lamiaceae e o Ãcido betulÃnico, marcador quimiotaxonÃmico no gÃnero Hyptis. Dos dez diterpenos, cinco sÃo abietanos: 12-hidroxi-8,11,13-abietatrien-20-al, 8,11,13-abietatrien-12,20-diol, Ãcido 11-hidroxi-12-metoxi-8,11,13-abietatrien-10-Ãico, Ãcido 12-hidroxi-8,11,13-abietatrien-10-Ãico e 12-hidroxi-8,11,13-abietatrieno, um nor-diterpeno, com esqueleto 20-nor-icetexano inÃdito, denominado 8(7),10(7)-diepoxi-12-hidroxi-20-nor-8,11,13-abietatrieno, um diterpeno pimarano, o 11-cetosandaracopimar-15-en-8β-ol, inÃdito na famÃlia Lamiaceae e trÃs tanshinonas: 7β-hidroxi-11,14-dioxoabieta-8,13-dieno, 7,12-di-hidroxi-8,12-abietadien-11,14-di-oxo-20-al e 7α-hidroxi-11,14-dioxoabieta-8,13-dieno, sendo a Ãltima inÃdita na literatura. Todos os compostos foram isolados utilizando sucessivos fracionamentos cromatogrÃficos, incluindo CLAE e a determinaÃÃo estrutural foi realizada atravÃs de tÃcnicas espectroscÃpicas como EMAR, IV, RMN de 1H e 13C, incluindo sequÃncias de pulsos uni e bidimensionais, e comparaÃÃo com dados descritos na literatura. Dentre os compostos isolados, dezoito foram testados com relaÃÃo a inibiÃÃo do crescimento celular de quatro linhagens de cÃlulas humanas cancerÃgenas e dez mostraram atividade. Dos compostos isolados de H. crassifolia o 12-hidroxi-8,11,13-abietatrieno e a 11,12,14,16-tetra-hidroxi-17 -abeo-abieta-8,11,13-trien-7-ona apresentaram atividade citotÃxica moderada, enquanto a 12-hidroxi-8,11,13-abietatrien-7-ona apresentou uma atividade citotÃxica moderada, porÃm seletiva contra cÃlulas tumorais leucÃmicas. Dos compostos isolados de H. carvalhoi todos os diterpenos apresentaram atividade citotÃxica, mas a 7,12-di-hidroxi-8,12-abietadien-11,14-di-oxo-20-al, apÃs 72 horas de incubaÃÃo, apresentou valores de CI50 que variaram de 3,91 a 32,01 μM em cÃlulas tumorais de cÃlon (HCT-116) e leucÃmicas (HL-60), respectivamente. O seu possÃvel mecanismo de aÃÃo, foi entÃo estudado.
| Identifer | oai:union.ndltd.org:IBICT/oai:www.teses.ufc.br:8649 |
| Date | 28 August 2014 |
| Creators | Karisia Sousa Barros de Lima |
| Contributors | Edilberto Rocha Silveira, Mary Anne Sousa Lima, Telma Leda Gomes de Lemos, Nilce Viana Gramosa Pompeu de Sousa Brasil, Angelo da Cunha Pinto |
| Publisher | Universidade Federal do CearÃ, Programa de PÃs-GraduaÃÃo em QuÃmica, UFC, BR |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFC, instname:Universidade Federal do Ceará, instacron:UFC |
| Rights | info:eu-repo/semantics/openAccess |
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