Antimicrobial and cytotoxic effects of the Copaifera reticulata oleoresin and its main diterpene acids

J Ethnopharmacol. 2019 Apr 6;233:94-100. doi: 10.1016/j.jep.2018.11.029. Epub 2018 Nov 23.


Ethnopharmacological relevance: The oleoresin of Brazilian Copaifera reticulata is a traditional remedy used for the treatment of skin and urinary tract infections, respiratory diseases, rheumatism, ulcer and tumours; thus, playing an important role in the primary health care of the indigenous population.

Aim: As most previous pharmacological tests used the crude oleoresin and only a few studies so far dealt with enriched fractions or pure chemically defined compounds, the aim of this study was to systematically evaluate the antimicrobial and cytotoxic properties of the Copaifera reticulata oleoresin and to assign traditional uses to specific secondary metabolites.

Materials and methods: The oleoresin, as well as its neutral and acidic fractions were tested for their activity against six cancer cell lines, two clinically relevant bacterial strains, and two dermatophytes. Both fractions were analysed by GC-MS and UHPLC-ELSD, respectively. The antibacterial acidic phase was further fractionated by preparative chromatography to purify and characterize the compounds responsible for the observed pharmacological effect.

Results: Whereas no cytotoxic activity was detected, the crude oleoresin and its acidic fraction showed antibacterial activity against gram-positive bacteria Enterococcus faecium (IC50 values 4.2 and 4.8 µg/mL, respectively) and methicillin-resistant Staphylococcus aureus (MRSA, IC50 values 5.3 and 7.2 µg/mL, respectively). Purification of the acidic fraction of the C. reticulata oleoresin yielded two dicarboxylic diterpene acids and the four main diterpene acids, comprising three different diterpene scaffolds. Interestingly, the activity was not restricted to a particular diterpene-type but rather depended on the compounds' lipophilicity, with the most active constituent showing IC50 values of 1.6 (E. faecium) and 2.5 µg/mL (MRSA), respectively. Furthermore, ent-polyalthic acid, the major diterpenoid, was significantly active against dermatophytes with IC50 values of 6.8 µg/mL (Trichophyton rubrum) and 4.3 µg/mL against (T. mentagrophytes).

Conclusion: The present study proved the antimicrobial effects of the C. reticulata oleoresin and its diterpenoid constituents, confirming its wide use in folk medicine for the treatment of skin and urinary tract infections. The inhibitory activity of copaiba diterpenoids against dermatophytic fungi as well as the gram-positive bacteria E. faecium and MRSA is being reported for the first time, providing potential lead structures for the treatment of these clinically relevant bacterial strains.

Keywords: 1:ent-agathic acid; 2: (13E)-ent-labd-8(17)-en-15,18-dioic acid; 3:ent-polyalthic acid; 4:ent-kaurenoic acid; 5: kolavenic acid; 6: (13E)-ent-labda-7,13-dien-15-oic acid; Copaiba oil; Copaifera reticulata; Dermatophytes; Diterpenoid; Enterococcus faecium; Kaurenoic acid; Kolavenic acid; MRSA; Natural product; Polyalthic acid.

MeSH terms

  • Anti-Infective Agents / chemistry
  • Anti-Infective Agents / pharmacology*
  • Bacteria / drug effects
  • Bacteria / growth & development
  • Cell Line
  • Cell Survival / drug effects
  • Diterpenes / analysis
  • Diterpenes / pharmacology*
  • Fabaceae
  • Humans
  • Plant Extracts / chemistry
  • Plant Extracts / pharmacology*
  • Trichophyton / drug effects
  • Trichophyton / growth & development


  • Anti-Infective Agents
  • Diterpenes
  • Plant Extracts
  • oleoresins