Biotransformation of the terpenes ?-caryophyllene and 3?-acetoxy-copalic acid found in Copaifera sp oleoresins using filamentous fungi and bacteria from gastrointestinal tract and cytotoxic activity evaluation of the obtained derivatives

Author(s)

Tatiane Cruz de Carvalho

• Document type: Doctoral Thesis
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC)
• Defense date: 2016-05-09

Examining board members

Niege Araçari Jacometti Cardoso Furtado; Hosana Maria Debonsi; Carlos Henrique Gomes Martins; Jonas Augusto Rizzato Paschoal; Rodrigo Cassio Sola Veneziani

• Advisor: Niege Araçari Jacometti Cardoso Furtado

Abstract

Oleoresins from Copaifera sp species are widely used in Brazilian folk medicine and chemical and biological studies showed that diterpenes and sesquiterpenes are among the main bioactive compounds of oleoresins. An important factor for assessing the safety and efficacy of drugs is the knowledge about their metabolism, which can be acquired through biotransformation studies using filamentous fungi and human gut bacteria. Filamentous fungi are eukaryotic organisms and the enzymatic apparatus is similar to those of mammals and may contribute to the elucidation of metabolic pathways providing information on the production of pharmacologically active, inactive or toxic compounds to the organism. These studies may also contribute to provide new compounds with potential biological activity. The use of human gut bacteria in metabolism studies is also important because drugs and foods, when ingested, can be metabolized by bacteria that colonize the gastrointestinal tract. The aim of this study was to evaluate the potential of filamentous fungi and human gut bacteria to biotransform 3?-acetoxy-copalic acid and ?-caryophyllene found in oleoresins from Copaifera as well as to isolate the biotransformation products for evaluation of their cytotoxicity. Biotransformation experiments were carried out with 10 filamentous fungi strains and 8 human gut bacteria strains. The ethyl acetate extracts obtained from the biotransformation processes of diterpene were analyzed by HPLC/DAD/CAD and HPLC-MS and n-hexane and ethyl acetate extracts obtained from the biotransformation processes of sesquiterpene, as well as the volatile fraction collected by solid phase microextraction headspace, were analyzed by GC-MS. All tested microorganisms were able to biotransform the 3?-acetoxy-copalic acid and 9 biotransformation products were isolated, their chemical structures were identified, from which six are new and all the products were evaluated for cytotoxic activity against normal mammary gland (MCF-10A) and breast adenocarcinoma (MCF-7) cell lines. All metabolites of 3?-acetoxy-copalic acid showed lower cytotoxicity against the evaluated cell lines than the 3?-acetoxy-copalic acid. The fungi A. niger, A. brasiliensis and the three Cunninghamella strains were able to biotransform ?-caryophyllene. However, as good yields were not achieved in these processes, it was not possible to isolate the biotransformation products. The obtained data became possible to just make a proposal for the chemical structure of a product from the ?-caryophyllene biotransformation process performed by C. echinulata ATCC 9245. (AU)

• FAPESP's process: 12/01766-4 - Biotransformations of ²-caryophyllene and acid 3-acetoxy-copalic: use of filamentous fungi and bacteria from gastrointestinal tract for in vitro metabolism studies and evaluation of cytotoxic activity of the obtained derivatives.
• Grantee: Tatiane Cruz de Carvalho
• Support Opportunities: Scholarships in Brazil - Doctorate