title: In vitro basal cytotoxicity assay applied to estimate acute oral systemic toxicity of grandisin and its major metabolite.
abstract: Preclinical investigations can start with preliminary in vitro studies before using animal models. Following this approach, the number of animals used in preclinical acute toxicity testing can be reduced. In this study, we employed an in-house validated in vitro cytotoxicity test based on the Spielmann approach for toxicity evaluation of the lignan grandisin, a candidate anticancer agent, and its major metabolite, the 4-O-demethylgrandisin, by neutral red uptake (NRU) assay, on mouse fibroblasts Balb/c 3T3 cell line. Using different concentrations of grandisin and its major metabolite (2.31; 1.16; 0.58; 0.29; 0.14; 0.07; 0.04; 0.002 μM) in Balb/c 3T3-A31 NRU cytotoxicity assay, after incubation for 48 h, we obtained IC(50) values for grandisin and its metabolite of 0.078 and 0.043 μM, respectively. The computed LD(50) of grandisin and 4-O-demethylgrandisin were 617.72 and 429.95 mg/kg, respectively. Both were classified under the Globally Harmonized System as category 4. Since pharmacological and toxicological data are crucial in the developmental stages of drug discovery, using an in vitro assay we demonstrated that grandisin and its metabolite exhibit distinct toxicity profiles. Furthermore, results presented in this work can contribute to reduce the number of animals required in subsequent pharmacological/toxicological studies.
authors.affiliation: Laboratório de Farmacologia e Toxicologia Celular, Faculdade de Farmácia, Universidade Federal de Goiás, Praça Universitária, No 1166, Setor Universitário, CEP 74605-220 Goiânia, GO, Brazil.
authors.forename: Marcelo de Sousa
authors.initials: Mde S
authors.lastname: Vieira
authors.forename: Valéria
authors.initials: V
authors.lastname: de Oliveira
authors.forename: Eliana Martins
authors.initials: EM
authors.lastname: Lima
authors.forename: Massuo Jorge
authors.initials: MJ
authors.lastname: Kato
authors.forename: Marize C
authors.initials: MC
authors.lastname: Valadares
mesh_terms: D000826:Animal Testing Alternatives / Q000379:methods*; D000818:Animals; D041702:BALB 3T3 Cells; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005663:Furans / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000633:toxicity*; D017705:Lignans / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000633:toxicity*; D051379:Mice; D015394:Molecular Structure; D031701:Piper / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry; D023382:Toxicity Tests, Acute / Q000379:methods*
chemical_list: C561033:4-O-demethylgrandisin; D005663:Furans; D017705:Lignans; D010936:Plant Extracts; C074346:grandisin