Toxicological Screening of Four Bioactive Citroflavonoids: In Vitro, In Vivo, and In Silico Approaches.

Rolffy Ortiz-Andrade,Victor Yáñez-Pérez,Jesús Alfredo Araujo-León,M A Lucio-Garcia,Avel Adolfo González-Sánchez,Gabriel Navarrete-Vazquez,Juan Carlos Sánchez-Salgado,Sergio Hidalgo-Figueroa,Ángel Josabad Alonso-Castro,Irma Aranda-González,Tania Isolina Coral-Martínez,Amanda Sánchez-Recillas,Emanuel Hernández-Núñez

doi:10.3390/molecules25245959

Abstract

Many studies describe different pharmacological effects of flavonoids on experimental animals and humans. Nevertheless, few ones are confirming the safety of these compounds for therapeutic purposes. This study aimed to investigate the preclinical safety of naringenin, naringin, hesperidin, and quercetin by in vivo, in vitro, and in silico approaches. For this, an MTT-based cytotoxicity assay in VERO and MDCK cell lines was performed. In addition, acute toxicity was evaluated on Wistar rats by OECD Guidelines for the Testing of Chemicals (Test No. 423: Acute Oral Toxicity-Class Method). Furthermore, we used the ACD/Tox Suite to predict toxicological parameters such as hERG channel blockade, CYP450 inhibition, and acute toxicity in animals. The results showed that quercetin was slightly more cytotoxic on cell lines (IC50 of 219.44 ± 7.22 mM and 465.41 ± 7.44 mM, respectively) than the other citroflavonoids. All flavonoids exhibited an LD50 value > 2000 mg/kg, which classifies them as low-risk substances as OECD guidelines established. Similarly, predicted LD50 was LD50 > 300 to 2000 mg/kg for all flavonoids as acute toxicity assay estimated. Data suggests that all these flavonoids did not show significant toxicological effects, and they were classified as low-risk, useful substances for drug development.

Highlights

Ethnomedicine is a millenary practice on bioactive compounds from plants and animals useful for therapeutics
The cytotoxicity assay was performed on VERO and Madin-Darby Canine Kidney Epithelial Cells (MDCK) cells using the MTT-based reduction method
HESP, NARGE, and NAR did not change the percentage of growth of both cells tested at concentrations ranging from 125 to 750 μg/mL (Figure 1)

Summary

Introduction

Ethnomedicine is a millenary practice on bioactive compounds from plants and animals useful for therapeutics. According to the World Health Organization (WHO), medicinal plants are underlying sources of many medicines, so they should be thoroughly investigated to demonstrate their safety and efficacy on humans [1]. In this context, one of the most studied medicinal plants is Citrus sp., which has prominent medicinal properties, such as digestive, antiseptic, diuretic, antibacterial, antiviral, and beneficial effects on cancer, cardiovascular disease (CVD), and diabetes [2]. Some studies have shown that daily consumption of orange preparations reduces the prevalence of CVD risk factors. The main bioactive compounds responsible for this effect are flavonoids [1,2,3,4]

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