Extraction, purification and in vitro assessment of the antioxidant and anti-inflammatory activity of policosanols from non-psychoactive Cannabis sativa L

Clarissa Caroli; Giovanna Baron; Giorgio Cappellucci; Virginia Brighenti; Larissa Della Vedova; Francesca Fraulini; Simonetta Oliaro-Bosso; Andrea Alessandrini; Alfonso Zambon; Gigliola Lusvardi; Giancarlo Aldini; Marco Biagi; Lorenzo Corsi; Federica Pellati

doi:10.1016/j.heliyon.2024.e30291

Extraction, purification and in vitro assessment of the antioxidant and anti-inflammatory activity of policosanols from non-psychoactive Cannabis sativa L

Heliyon. 2024 Apr 26;10(9):e30291. doi: 10.1016/j.heliyon.2024.e30291. eCollection 2024 May 15.

Authors

Clarissa Caroli^{1

2}, Giovanna Baron³, Giorgio Cappellucci⁴, Virginia Brighenti¹, Larissa Della Vedova³, Francesca Fraulini⁵, Simonetta Oliaro-Bosso⁶, Andrea Alessandrini^{7

8}, Alfonso Zambon⁵, Gigliola Lusvardi⁵, Giancarlo Aldini³, Marco Biagi⁹, Lorenzo Corsi^{1

8}, Federica Pellati¹

Affiliations

¹ Department of Life Sciences, University of Modena and Reggio Emilia, Via Giuseppe Campi 103-287, 41125, Modena, Italy.
² Clinical and Experimental Medicine PhD Program, University of Modena and Reggio, Via Giuseppe Campi 287, 41125, Modena, Italy.
³ Department of Pharmaceutical Sciences, University of Milan, Via Mangiagalli 25, 20133, Milan, Italy.
⁴ Department of Physical Sciences, Earth and Environment, University of Siena, Via Laterina, 8, 53100, Siena, Italy.
⁵ Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Via Giuseppe Campi 103, 41125, Modena, Italy.
⁶ Department of Drug Science and Technology, University of Turin, Via Pietro Giuria 9, 10125, Turin, Italy.
⁷ Department of Physics, Informatics and Mathematics, University of Modena and Reggio Emilia, Via Giuseppe Campi 213/A, 41125, Modena, Italy.
⁸ National Institute of Biostructures e Biosystems (INBB), 00136, Roma, Italy.
⁹ Department of Food and Drug, University of Parma, Parco Area delle Scienze 27/A, 43124, Parma, Italy.

Abstract

Policosanols (PCs) are bioactive compounds extracted from different natural waxes. In this work, the purification, characterization and assessment of the antioxidant and anti-inflammatory activity was carried out on PCs from an innovative source, i.e. a waxy material from supercritical-fluid extraction (SFE) of non-psychoactive Cannabis sativa L. (hemp) inflorescences. Starting from this material, PCs were obtained by microwave-assisted trans-esterification and hydrolysis, followed by preparative liquid chromatography under normal phase conditions. The purified product was characterized using high-performance liquid chromatography (HPLC) with an evaporative light scattering detector (ELSD). In vitro cell-free and cell-based antioxidant and anti-inflammatory assays were then performed to assess their bioactivity. HPLC-ELSED analysis of the purified mixture from hemp wax revealed C₂₆OH and C₂₈OH as the main compounds. In vitro assays indicated an inhibition of intracellular reactive oxygen species (ROS) production, a reduction of nuclear factor kappa B (NF-κB) activation and of the activity of the neutrophil elastase. Immunoblotting assays allowed us to hypothesize the mechanism of action of the compounds of interest, given the higher levels of MAPK-activated protein kinase 2 (MK2) and heme oxygenase-1 (HO-1) protein expression in the PC pretreated HaCaT cells. In conclusion, even if more research is needed to unveil other molecular mechanisms involved in hemp PC activity, the results of this work suggest that these compounds may have potential for use in oxinﬂammation processes.

Keywords: Anti-inflammatory activity; Antioxidant activity; Cannabis sativa L.; Extraction; HPLC; Policosanols.