Kaurenoic Acid from Sphagneticola trilobata Inhibits Inflammatory Pain: Effect on Cytokine Production and Activation of the NO–Cyclic GMP–Protein Kinase G–ATP-Sensitive Potassium Channel Signaling Pathway

Abstract

Kaurenoic acid [ent-kaur-16-en-19-oic acid (1)] is a diterpene present in several plants including Sphagneticola trilobata. The only documented evidence for its antinociceptive effect is that it inhibits the writhing response induced by acetic acid in mice. Therefore, the analgesic effect of 1 in different models of pain and its mechanisms in mice were investigated further. Intraperitoneal and oral treatment with 1 dose-dependently inhibited inflammatory nociception induced by acetic acid. Oral treatment with 1 also inhibited overt nociception-like behavior induced by phenyl-p-benzoquinone, complete Freund's adjuvant (CFA), and both phases of the formalin test. Compound 1 also inhibited acute carrageenin- and PGE(2)-induced and chronic CFA-induced inflammatory mechanical hyperalgesia. Mechanistically, 1 inhibited the production of the hyperalgesic cytokines TNF-α and IL-1β. Furthermore, the analgesic effect of 1 was inhibited by l-NAME, ODQ, KT5823, and glybenclamide treatment, demonstrating that such activity also depends on activation of the NO-cyclic GMP-protein kinase G-ATP-sensitive potassium channel signaling pathway, respectively. These results demonstrate that 1 exhibits an analgesic effect in a consistent manner and that its mechanisms involve the inhibition of cytokine production and activation of the NO-cyclic GMP-protein kinase G-ATP-sensitive potassium channel signaling pathway.

Development and Validation of a Rapid and Reliable RP-HPLC-PDA Method for the Quantification of Six Diterpenes in Copaifera duckei, Copaifera reticulata and Copaifera multijuga Oleoresins

“…2,9,10,35 Despite the importance of sesquiterpenes on the chemical and biological characterization of Copaifera oleoresins, [1][2][3][17][18][19] recent reports have established that several non-volatile diterpenes, such as (-)-copalic acid, ent-kaurenoic acid and (-)-polyalthic acid, are also responsible for important biological properties displayed by these balsams. 3,11,12,14,15,22 Even though and as stated before, there are only two analytical methods to characterize these diterpene acids in Copaifera oleoresins. 9,21 Concerning to diterpenes, the literature 11,17,36 reported that (-)-copalic acid (4) can be found in all oleoresins of Copaifera species and that it has been proposed as the chemical marker of the genus.…”

“…Acid diterpenes stands out for a wide range of the biological properties reported for these oleoresins, such as: antimicrobial, antitumoral, antinociceptive and anti-inflammatory, among others. 3,4,6,[11][12][13][14][15] Despite this fact, most of the studies reporting the chemical identification and quantification of the diterpenes of Copaifera oleoresins have been performed by using gas chromatographic hyphenated 9 only comprises the analysis of one standard ((-)-copalic acid) in commercial samples of oleoresins and the study published by Mangabeira et al…”

“…Acid diterpenes stands out for a wide range of the biological properties reported for these oleoresins, such as: antimicrobial, antitumoral, antinociceptive and anti-inflammatory, among others. 3,4,6,[11][12][13][14][15] Despite this fact, most of the studies reporting the chemical identification and quantification of the diterpenes of Copaifera oleoresins have been performed by using gas chromatographic hyphenated techniques and mass spectra comparison with spectral libraries, 2,[16][17][18][19][20] which are more adequate for analyses of volatile compounds like sesquiterpenes. 9 In the literature, there are only two studies describing the development and validation of analytical methods for the analysis acid diterpenes 9,21 in Copaifera species.…”

“…Acid diterpenes stands out for a wide range of the biological properties reported for these oleoresins, such as: antimicrobial, antitumoral, antinociceptive and anti-inflammatory, among others. 3,4,6,[11][12][13][14][15] Despite this fact, most of the studies reporting the chemical identification and quantification of the diterpenes of Copaifera oleoresins have been performed by using gas chromatographic hyphenated 9 only comprises the analysis of one standard ((-)-copalic acid) in commercial samples of oleoresins and the study published by Mangabeira et al21 is based on ultra-performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS).Based on the need of new analytical validated methods, and as part of our ongoing efforts to explore the potential of Copaifera oleoresins, we are describing, in this paper, the development and validation of a simple and novel analytical method using reversed-phase high-performance liquid chromatography (RP-HPLC). The developed method is capable of quantifying six diterpenes in the non-volatile fractions of the oleoresins obtained from the most commercially explored species: Copaifera duckei, Copaifera reticulata and Copaifera multijuga.…”

Immunomodulatory and Antioxidant Properties of Kaurenoic Acid on Macrophages of BALB/c <i>in Vitro</i>

“…Among the biological effects of this diterpene, studies have shown analgesic activity (Mizokami et al, 2012), antiinflammatory effects in asthma models (Cho et al, 2010), antitumoral, antimicrobial and antiprotozoal actions (Costa-Lotufo et al, 2002;Wilkens et al, 2002;Izumi et al, 2012;Santos et al, 2013).…”