Cannabichromene (CBC), a non-psychoactive cannabinoid found in Cannabis sativa, has recently been shown to possess several medicinal properties. However, how CBC produces anti-inflammatory effects and the mechanisms of this remain poorly studied. Therefore, we extracted and purified the CBC from the Cannabis sativa cv. pink pepper (hemp cultivar). The efficacy of CBC in reducing inflammation in RAW 264.7 macrophages and a λ-carrageenan-induced mouse model was then evaluated. CBC had no cytotoxicity up to a concentration of 20 μM and inhibited nitric oxide production by approximately 50% at a concentration of 20 μM. In addition, CBC treatment significantly inhibited causes of inflammation such as inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) at both the mRNA and protein levels. Moreover, CBC suppressed LPS-stimulated inflammation in RAW 264.7 cells by downregulating the nuclear factor kappa B (NF-kB) and mitogen-activated protein kinase pathways (MAPK). Furthermore, our in vivo experiments confirmed that the λ-carrageenan-induced increase in the levels of the cytokines iNOS, IL-1β, and IL-6 was abrogated following treatment with CBC. Therefore, CBC has potential anti-inflammatory effects and may be useful for preventing or treating inflammation.

Cannabis sativa L. contains more than 80 cannabinoids, among which cannabidiol (CBD) is the main neuroactive component. We aimed to investigate the anti-inflammatory efficacy of CBD in vitro and in vivo isolated from “Pink pepper”, a novel hemp cultivar, by repeating the method of selecting and cultivating individuals with the highest CBD content. We investigated the effects of CBD on inflammatory markers elevated by lipopolysaccharide (LPS) treatment in RAW 264.7 mouse macrophage cells through Western blot and RT-PCR. In addition, we confirmed these effects through the ELISA of inflamed paw tissue of a λ-carrageenan-induced mouse edema model that received an oral administration of CBD. CBD inhibited the LPS-induced phosphorylation of NF-κB and MAPK in RAW 264.7 and exhibited anti-inflammatory effects by participating in these pathways. In our in vivo study, we confirmed that CBD also inhibited the inflammatory mediators of proteins extracted from edematous mouse paw tissue. These results show that CBD isolated from “Pink pepper” exhibits potent anti-inflammatory effects. These anti-inflammatory effects of CBD have pharmacological and physiological significance, highlighting the industrial value of this novel cultivar.

Hemp, also known as Cannabis sativa L., contains over 80 cannabinoids, with cannabidiol (CBD) being the primary neuroactive component. CBD possesses various pharmacological properties and is considered a non-psychoactive compound, making it a promising component for various applications, such as pharmaceuticals, cosmetics, and nutraceuticals. The aim of this study was to identify the optimal conditions for extracting CBD from hemp using supercritical fluid extraction (SFE). Response surface methodology (RSM) was employed to optimize the SFE conditions. The Box–Behnken design and the central composite design were utilized to refine the extraction parameters, including extraction time, temperature, and pressure. The statistical significance and reliability of the optimized conditions were confirmed by the significant influence of these independent variables on CBD yield. The extracted CBD was purified to a high level of purity and converted from cannabidiolic acid (CBDA) through heat treatment and then analyzed using high-performance liquid chromatography (HPLC). The following extraction conditions were considered optimal and led to a CBD yield of approximately 70.46 g/kg: pressure of 48.3 MPa, temperature of 60 °C, and extraction time of 109.2 min. Validation experiments confirmed the accuracy of the model, with experimental values closely matching the predicted values (69.93 ± 0.88 g/kg). This study demonstrates that SFE is an efficient method for obtaining high-purity CBD from hemp, highlighting its potential for industrial applications. The findings suggest that optimizing SFE conditions through RSM can significantly enhance the efficiency and yield of CBD extraction, providing a robust framework for industrial-scale production.

Melanogenesis, the key biological process underlying skin hyperpigmentation, is tightly regulated by complex molecular signaling pathways. Consequently, targeting molecular regulators of this pathway is a crucial strategy for developing effective skin-whitening agents. Cannabinol (CBN), a minor cannabinoid, has been largely unexplored owing to its role in modulating skin pigmentation. This study aimed to elucidate the molecular mechanisms of CBN’s depigmenting effects using an α-MSH-induced B16F10 melanoma cell model. High-purity CBN was obtained via conversion of cannabidiol (CBD) and confirmed by HPLC. CBN significantly inhibited melanin synthesis and tyrosinase activity in a concentration-dependent manner, without any cytotoxicity. Furthermore, we investigated CBN’s impact on the melanogenesis signaling cascade. Our analysis revealed that CBN significantly downregulated the mRNA and protein levels of key melanogenic master regulators, including MITF, TYR, TYRP1, and TYRP2. Importantly, we also observed that CBN treatment selectively suppressed the protein phosphorylation of upstream signaling molecules such as p38 and JNK MAP kinases and NF-κB, while ERK phosphorylation remained unaffected. This finding indicates that its mechanism of action involves the selective modulation of pro-melanogenic signaling components. Collectively, these findings demonstrate that CBN effectively modulates the melanogenesis signaling pathway by targeting both upstream kinases and downstream melanogenic genes. These findings suggest that CBN holds great promise as a bioactive agent for skin-whitening applications and warrants further research to confirm its clinical efficacy and safety.

This study aimed to optimize fermentation conditions for maximizing ellagic acid accumulation/release from Juglans mandshurica (Manchurian walnut) fruit extract using Weissella confusa. A 20% ethanol extract was fermented across a central composite design spanning temperature (26.59–43.41 °C), pH (2.98–8.02), and time (3.91–24.09 h), and response surface methodology was used to determine the optimal conditions. The predicted optimum was 36.61 °C, initial pH 6.70, and 24.09 h, under which the ellagic acid yield reached 64.05 µg/mL, experimentally validated at this single optimum condition using three independent replicate fermentations (n = 3), giving 64.04 ± 0.95 µg/mL by HPLC. Antioxidant activity at the optimized condition was evaluated using total phenolic content, DPPH, and ABTS radical scavenging assays. The fermented extract showed markedly enhanced radical scavenging activity, with DPPH inhibition reaching 85.47% at 400 μg/mL and ABTS exceeding 99.99%, while the total phenolic content was 141.41 ± 2.32 mg gallic acid equivalents (GAE)/g. These results indicate that W. confusa-mediated fermentation enhances the ellagic acid content and antioxidant potential of J. mandshurica extract, presumably through enzymatic hydrolysis of ellagitannins involving tannase and β-glucosidase, although these enzyme activities were not directly measured. This approach offers a sustainable strategy for developing functional ingredients with natural antioxidant properties for food and nutraceutical applications.