Cannabis sativa L. and Moringa oleifera Lam. Leaf Extracts and Cannabidiol as Multi-Target Inhibitors: Lipoxygenase, Free Radical Scavenging Activity, and in Silico Studies

Abstract Effective treatments are needed for inflammatory diseases but conventional medications have side effects. The exploration of natural compounds for therapeutic purposes has gained significant attention in recent years, particularly concerning their potential as multi-target inhibitors in various biological processes and as a safer alternative. Among these compounds, Cannabis sativa L. ( C. sativa ) and Moringa oleifera Lam. ( M. oleifera ) have emerged as promising due to their rich phytochemical profiles. The aim of this study was to determine if solvent extracts of C. sativa and M. oleifera and cannabidiol (CBD) alone or in combination had anti-inflammatory and antioxidant properties. This study employed anti-inflammatory (LOX inhibition test) and antioxidant (FRAP and ABTS assay) assays to evaluate the biological potential of two plant species. The primary reason for using combined plant extracts and cannabidiol was to assess their potential synergistic effect, where the combined effect of two or more compounds is greater than the sum of their individual effects owing to interactions among diverse bioactive compounds. Outcomes revealed, the highest total flavonoid content was found in the hexane extracts of C. sativa and M. oleifera (88.68 ± 2.40 µg/mL and 159.91 ± 1.53 µg/mL) while the highest phenolic content was recorded in the propanol extract of C. sativa (57.21 ± 0.79 µg/mL), and methanolic extract of M. oleifera (71.50 ± 1.07 µg/mL) leaves. C. sativa propanol and M. oleifera hexane leaf extracts showed maximum ABTS, Fe³⁺-TPTZ scavenging activity, and 15-lipoxygenase inhibition at 100 µg/mL. The minimum IC 50 values were: C. sativa propanol extract (IC 50 ; 457.65 ± 0.33 µg/mL for ABTS: IC 50 ; 69.00 ± 0.35 µg/ mL for Fe³⁺-TPTZ: IC 50 ; 57.95 ± 0.58 µg/mL for 15-LOX initiation), M. oleifera hexane extract (IC 50 ; 340.76 ± 0.23 µg/mL for ABTS: IC 50 ; 22.94 ± 0.61 µg/mL for 15-LOX inhibition). This suggests that the extraction solvent used significantly influenced the bioactive compounds available for antioxidant activity. The combination (1:1) of extracts from C. sativa and M. oleifera or with CBD enhanced the overall Fe 3 +-TPTZ and ABTS + radical scavenging effect, suggesting potential synergistic effects. The maximum synergistic effect was observed for C. sativa methanol + M. oleifera aqueous extracts (IC 50 ; 291.45 ± 0.33 µg/mL for ABTS) and C. sativa propanol extract + CBD (IC 50 ; 21.65 ± 0.28 µg/mL for Fe³⁺-TPTZ), and C. sativa + M.oleifera aqueous extracts + CBD (IC 50 ; 11.58 ± 0.33 µg/mL). Molecular docking studies revealed that catechin and quercetin exhibited the highest binding interactions with 15-LOX. Experimental results show CBD as a strong inhibitor, but in silico findings suggest a higher affinity for quercetin and catechin due to experimental conditions, metabolism, and concentration. Overall, the study depicted propanol extract of C. sativa and the hexane extract of M. oleifera exerted maximum anti-inflammatory and anti-oxidant effects owing to a considerable amount of phenolic and flavonoid contents. In conclusion, the combination of C. sativa and M. oleifera along with CBD is a suitable option to enhance the efficacy of extracts and might help to overcome multidrug resistance. To validate their use in combination with treating inflammatory diseases, further preclinical trials are required.