Evaluation of the Safety of Three Phenolic Compounds from <i>Dipteryx alata</i> Vogel with Antiophidian Potential

Abstract

Phenolic compounds from Dipteryx alata Vogel were assayed against the in vitro neurotoxic effect induced by Bothrops jararacussu (Bjssu) venom. Mutagenicity was assessed by the Ames test using Salmonella typhimurium strains TA98, TA97a, TA100, and TA102, in experiments with and without metabolic activation. Anti-bothropic activity was obtained by using mouse phrenic nerve-diaphragm (PND) preparation and myographic technique. Control experiments with physiological Tyrode solution were used for keeping the PND preparations alive (n = 4). Concentrations of phenolic compounds were as follow: protocatechuic and vanillic acids (200 µg/mL, n = 4), vanillin (50 µg/mL, n = 4). These compounds were used alone or pre-incubated with the venom (40 µg/mL), 30 min prior the addition to the organ bath (n = 4). Phenolic compounds significantly inhibited the neuromuscular blockade of Bjssu in the following order of potency: vanillic acid > protocatechuic = vanillin. Vanillic acid added 10 min after the Bjssu venom was also able to avoid the venomblockade evolution. The mutagenicity assay indicated that all phytochemicals were unable to increase the number of revertants, demonstrating the absence of mutagenic activity. This study * Corresponding author. E. H. Yoshida et al.2 demonstrated both the safety and therapeutical potential of the three phenolic compounds as novel complementary anti-bothropic agents.

Anti-Inflammatory and Antibothropic Properties of Jatropha Elliptica, a Plant from Brazilian Cerrado Biome

“…Besides, it showed the ability of the TE to avoid paralysis evolution, suggesting antibothropic properties, such as Casearia gossypiosperma, 12 Vellozia flavicans, 19 and phenolics from Dipteryx alata. 20 Some constituents of our J. elliptica extract, such as flavonoids (apigenin, quercetin, and rutin), probably have a role in the protection against the paralysis induced by Bjssu, although they were found in a very small content (0.08%). Previously, quercetin showed some protection against the neuromuscular blockade of Bjssu in the same experimental conditions.…”

“…Polyphenol content: Polyphenols were determined by a modified method by Hagerman. 13 The percentage of polyphenols was determined by a calibration curve (5, 10, 15,20,25,30,35, and 40 μg/mL) of gallic acid (Sigma). …”

The antimutagenic potential of Vanillin and Betulin based on the antimutagenicity of Resveratrol

“…Previously, the Ames test showed that vanillin and betulin are devoid of mutagenicity in the presence (+S9) as in the absence (-S9) of metabolic activation (Yoshida et al, 2015;, becoming strong candidates for the antimutagenicity test. On the other hand, resveratrol has already been tested in the same mutagenicity assay in Salmonella/microsome bacteria (Langová et al, 2005;Uenobe et al, 1997), comet assay and chromosomal aberration showing no genotoxicity in any of these assays (Jeong et al, 2014;Karthika et al, 2022), corroborated by other assays as Allium cepa assay (Botti and Irazusta, 2018).…”

“…In addition to the three phytochemicals identified by Coelho Kaplan et al (1966), a further 15 phytochemicals were isolated and identified by Puebla et al (2010), therefore, a total of 18 substances are known, namely: four triterpenes, lupeol, lupenone; 28-hydroxy-lup-20(29)-en-3one and betulin; nine flavonoids, 8-O-methylretusin, 7-hydroxy-5,6,4'-trimethoxysoflavone, afrormosin, 7-hydroxy-8,3',4'-trimethoxyisoflavone, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, odoratin, 7,8,3'-trihydroxy-4'-methoxy isoflavone, 7,8,3'-trihydroxy-6,4'-dimethoxyisoflavone, dipterixin; a chalcone, isoliquiritigenin; an aurone, sulphidethine; and three phenolic compounds, vanillic acid, vanillin and protocatechuic acid. Among these substances from D. alata, protocatechuic acid, vanillic acid, vanillin (Yoshida et al, 2015) and betulin (Yoshida et al, 2016) were assayed to assess mutagenicity using the Ames test. All these phytochemicals have no mutagenicity.…”

Evaluation of Betulin Mutagenicity by Salmonella/Microsome Test

“…Ames (Berkeley, CA, USA), with and without metabolization by the preincubation method. 15 This assay was made as described by Yoshida et al, 16 as following: the strains from frozen cultures were grown overnight for 12-14 h in Oxoid Nutrient Broth No. 2.…”