Chemical profiling of Thevetia peruviana leaves cytotoxic active extracts enhanced by microemulsion formulation

Abstract

Background: Natural products played an essential role as a complementary cytotoxic agent avoiding complications of other therapies. In phytoformulation research, herbal drugs of nano-size have attracted more attention for more bioavailability and less active dose. We aim in this work to introduce new non-mutagenic cytotoxic agent from Thevetia peruviana leaves extracts and potentiate the activity by loading upon microemulsion formulations as an advanced mode of drug delivery system. Results: Thevetia peruviana toxicity test revealed its safety (LD 50 = 3.083 g/kg). In vivo genotoxicity tests measuring chromosomal aberrations were done to prove non-mutagenicity of the plant for both somatic and germ cells. The plant showed antioxidant activity with 121.215, 128.925, and 120.098 mg/g extract, calculated as Trolox equivalent (TE/g) for DPPH, ABTS, and FRAP assays, respectively. Successive extracts of the plant were incorporated in microemulsion formulations (MEF) and characterized for their electrical conductivity, poly-dispersity index, and in vitro drug release action. Cytotoxic activity of successive extracts was investigated against breast MCF7 and liver HEPG2 carcinoma cell lines before and after loading upon MEs. Enhancement in activity was detected for both pet. ether and ethanol MEFs, as IC 50 of pet. ether extract decreased from 25 to 6.5 μg/ml against MCF7 cell line, while ethanol formulation recorded IC 50 = 3.58 μg/ml against HEPG2 cell line after showing no activity of the extract at the tested concentrations. Regarding metabolites of polar fraction, total ethanol extract was estimated for total phenolic and flavonoid contents to record 47.7941 mg GAE/g calculated as gallic acid equivalent and 32.7667 mg CE/g measured as catechin equivalent, respectively. HPLC analysis for polar fraction recorded twelve compounds identified according to the available authentics with rutin the major flavonoid (7.33 mg/g) and rosmarinic acid (13.48 mg/g) the most abundant phenolic acid. Phytoconstituents of non-polar fraction were investigated after saponification of pet. ether extract using GC/MS analysis revealed the identification of 88.02% of the total unsaponifiable matter and 89.17 % of the total saponifiable matter. Conclusion: Thevetia peruviana is a safe cytotoxic agent. Microemulsion formulations of active extracts potentiate the cytotoxic actions against HEPG2 and MCF7 cell lines with better bioavailability and less medicinal doses.

Thevetia peruviana (Pers.) K. Schum.: Poisonous and its potential

“…The bioactivity of T. peruviana as an anticancer has been reported by Cheng et al [29], Miyagawa et al [30], Haldar et al [31], Al-Rajhi et al [9], and El-Sawi et al [32]. T. peruviana seeds containing cardiac glycosides have cytotoxicity properties against cancer cell lines P15 (human lung cancer cells), MGC-803 (human gastric cancer cells), SW1990 (human pancreatic cancer cells), and normal hepatocyte cells LO2 indicates that compounds from T. peruviana can selectively inhibit cancer cell proliferation with IC50 from 0.05 to 0.15 M [29].…”

“…The anticancer potential of T. peruviana latex was recorded against PC-3 (97.11% toxicity) and MCF-7 (96.23% toxicity) at 1000 g/mL with IC50 of 48.26 g/mL and 40.31 g/mL, respectively [9]. Cytotoxic activity was increased in mice fed the T. peruviana emulsion fraction against MCF7 breast and liver carcinoma cell line HEPG2 [32]. The bioactivity of T. peruviana as an anti-cancer and antioxidant is thought to be related to its bioactive compounds.…”

“…The bioactivity of T. peruviana as an anti-cancer and antioxidant is thought to be related to its bioactive compounds. The polar fraction of T. peruviana contained rutin, major flavonoid (7.33 mg/g) and rosmarinic acid (13.48 mg/g) the most abundant phenolic acid [32]. Flowers of T. peruviana are palmitic acid, (E)β-ionone and 1,8-cineole [34], cardiac glycoside, thevetins a and b, peruvoside, theveside, theviridoside and terpenoids [18].…”

In vitro spermicidal effect of Thevetia Peruviana leaves on human spermatozoa

“…Phytoconstituents derived from bark extract of this plant were found to have antispermatogenic activity in the experimental animal model (Gupta et al, 2011). The Thevetin A and Thevetin B isolated from seed extract of T. peruviana trees act similar to cardiac glycoside (Kohls et al, 2012), but methanolic extract of T. peruviana leaves freed from cardiac glycosides (Samanta et al, 2021), and LD50 of this leaf extract is 3.083 g/Kg (El-Sawi et al, 2020). The standardisation of this plant extract has been performed by a preliminary study with other solvent extracts in our laboratory.…”

“…peruviana trees act similar to cardiac glycoside (Kohls et al, 2012), but methanolic extract of T . peruviana leaves freed from cardiac glycosides (Samanta et al, 2021), and LD50 of this leaf extract is 3.083 g/Kg (El‐Sawi et al, 2020). The standardisation of this plant extract has been performed by a preliminary study with other solvent extracts in our laboratory.…”