Avalia????o do potencial farmacol??gico de derivados semissint??ticos de terpenos obtidos do ??leo de copa??ba (Copaifera spp.- Fabaceae)

Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2019-07-10T19:59:15Z No. of bitstreams: 2 Reprodu????o N??o Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2019-07-19T13:55:09Z (GMT) No. of bitstreams: 2 Reprodu????o N??o Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Made available in DSpace on 2019-07-19T13:55:09Z (GMT). No. of bitstreams: 2 Reprodu????o N??o Autorizada.pdf: 47716 bytes, checksum: 0353d988c60b584cfc9978721c498a11 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-12-17 92 981241860 Copaiba oil from several species of the genus copaifera has been used for many years, commercially, because of its possible modulatory action on inflammatory processes. Phytochemical studies of the species, which elucidate their chemical composition, fractionated in sesquiterpenes and diterpenes, have been performed. Although crude oil is well studied, it is scarce to know the effect of its isolated substances, and there are few studies on the structural modifications of these diterpene acids, seeking to improve their biological activities. In this sense, the present work had as objectives the isolation of the majority diterpenic acids present in the Copaifera multijuga oil: copalic and 3-hydroxy-copalic acids and Copaifera reticulata: hardwickiic acid, with structural modifications in the 2 (two) lowcost and optimized chemistries, as well as in vitro and in vivo assays of the substance with the best anti-inflammatory screening and subsequent in silico studies. After isolation, the substances were semi-synthesized and their structures elucidated by 1H and 13C NMR analysis, and therefore, tested in cell viability assay and TNF-?? inhibition activity check test in vitro. The in vivo model of pleurisy/pleuritis in balb-c mice was performed with the best anti-inflammatory substance (ACD13B-esterification of ibuprofen with 3-hydroxy-copalic acid), as well as expression assay of IKB-?? protein. In addition, the gastrointestinal test was also performed with the same substance. The semi-synthesized derivatives were: ACD4- epoxidation of copalic acid, ACD13B-esterification of ibuprofen with 3-hydroxy-copalic acid and ACD51-acylation of phthalic anhydride with 3-hydroxy-copalic acid. ACD13B showed no cytotoxic potential in the THP-1 line when tested at the concentration of 2 ??M. The other substances did not present significant cytotoxicity at concentrations tested. In addition, ACD13B significantly inhibited TNF-?? production in LPS-stimulated THP-1 cells. In animals with induced pleurisy, ACD13B reduced the total number of leukocytes by 40% and the number of mononuclear cells at a dose of 100 mg/kg intraperitoneally (IP) by up to 50%. There was also inhibition of pIKB-??. ACD13B presented a lower gastrointestinal effect when compared to ibuprofen in mice. The docking study revealed the affinity and mode of interaction of the substance ACD13B in COX-2 and COX-1, identifying hydrophobic and hydrophilic interactions. The results indicate that the ibuprofen derivative with 3-hydroxy copalic acid is a prototype of an anti-inflammatory drug with a lower gastrointestinal effect than ibuprofen, which may be explored in the future for other functional and therapeutic properties. O ??leo de copa??ba oriundo de diversas esp??cies do g??nero copaifera tem sido utilizado h?? muitos anos, comercialmente, em virtude de sua poss??vel a????o modulat??ria sobre processos inflamat??rios. Estudos fitoqu??micos das esp??cies, que elucidam sua composi????o qu??mica, fracionada em sesquiterpenos e diterpenos, tem sido realizados. Apesar do ??leo bruto ser bastante estudado, o conhecimento do efeito de suas subst??ncias isoladas ?? escasso, e poucos s??o os estudos sobre modifica????es estruturais desses ??cidos diterp??nicos, buscando melhorar suas atividades biol??gicas. Neste sentido o presente trabalho teve como objetivos o isolamento dos ??cidos diterp??nicos majorit??rios presentes no ??leo de Copaifera multijuga: ??cidos cop??lico e 3-hidr??xi-cop??lico e Copaifera reticulata: ??cido hardw??ckiico, realizando-se modifica????es estruturais nos 2 (dois) primeiros, por rea????es qu??micas de baixo custo e otimizadas, bem como ensaios in vitro e in vivo da subst??ncia com melhor screening antiinflamat??rio e posteriormente estudo in silico. Ap??s o isolamento, as subst??ncias foram semissintetizadas e suas estruturas elucidadas por an??lise de RMN 1H e 13C, e por conseguinte, testadas em ensaio de viabilidade celular e teste de verifica????o da atividade de inibi????o do TNF-?? in vitro. O modelo in vivo de pleurisia/pleurite em camundongos balb-c foi realizado com a subst??ncia de melhor screening anti-inflamat??rio (ACD13B-esterifica????o do ibuprofeno com o ??cido 3-hidr??xi-cop??lico), assim como ensaio de express??o da prote??na IKB-??. Alem disso, o ensaio de gastrotoxicidade tamb??m foi procedido com a mesma subst??ncia. Os derivados semissintetizados foram: ACD4-epoxida????o do ??cido cop??lico, ACD13B-esterifica????o do ibuprofeno com o ??cido 3-hidr??xi-cop??lico e ACD51-acila????o do anidrido ft??lico com o ??cido 3-hidr??xi-cop??lico. O ACD13B n??o apresentou potencial citot??xico na linhagem THP-1 quando testado na concentra????o de 2 ??M. As demais subst??ncias n??o apresentaram citotoxicidade nas concentra????es testadas. Al??m disso, o ACD13B inibiu significativamente a produ????o de TNF-?? em c??lulas THP-1 estimuladas por LPS. Em animais com pleurisia induzida o ACD13B reduziu em torno de 40% o total de leuc??citos e em at?? 50% o n??mero de mononucleares na dose de 100 mg/Kg por via intraperitonial (IP). Houve tamb??m inibi????o da pIKB-??. O ACD13B apresentou menor efeito gastrot??xico quando comparado ao ibuprofeno em camundongos. O estudo de docking revelou a afinidade e o modo de intera????o da subst??ncia ACD13B em COX-2 e COX-1, identificando intera????es hidrof??bicas e hidrof??licas. Os resultados obtidos indicam que o derivado do ibuprofeno com o acido 3-hidr??xi cop??lico apresenta-se como um prot??tipo de f??rmaco anti-inflamat??rio com menor efeito gastrot??xico que o ibuprofeno, o qual pode ser futuramente explorado quanto a outras propriedades funcionais e terap??uticas.