Coffee presents beneficial health properties, including antiobesity effects. However, its effects on inflammation are controversial. Hydroxycinnamic acids are the main coffee phenolic bioactive compounds. In human bioavailability studies carried out with coffee, among the most abundant compounds found in urine and plasma were the colonic metabolites, dihydrocaffeic (DHCA), dihydroferulic (DHFA), and hydroxyhippuric (HHA) acids. To understand the hepato-protective potential of these three compounds, we tested whether treatment with realistic concentrations (0.5–10 µM) were effective to counteract inflammatory process and oxidative status induced by tumor necrosis factor α (TNF-α). First, we established a novel model of inflammation/oxidation using TNF-α and HepG2 cells. Afterwards, we evaluated the activity of DHCA, DHFA, and HHA against the inflammatory/oxidative challenge through the determination of the inflammatory mediators, interleukins (IL)-6, and IL-8 and chemokines, monocyte chemoattractant protein-1, and macrophage inflammatory protein-1, as well as the levels of biomarkers of oxidative stress, such as reactive oxygen species, reduced glutathione, and the antioxidant enzymes glutathione peroxidase and reductase. Results showed that all three compounds have a potential hepato-protective effect against the induced inflammatory/oxidative insult.

Obesity is coupled with an altered redox state and low-level inflammation. Oxidative stress may increase pre-adipocyte proliferation, adipocyte differentiation and mature adipocyte size. Regarding inflammation, the dysregulation of cytokine production by adipose tissue takes place in obesity, which is promoted by oxidative stress. Polyphenols may exert a positive effect on obesity, not only by modulating the redox state, but also due to their anti-inflammatory activity. Coffee, which is one of the most consumed beverages, is very rich in phenolic compounds. Bioavailability studies on coffee phenols have shown that the most abundant group of metabolites in plasma and urine are dihydrocaffeic (DHCA), dihydroferulic (DHFA), and hydroxyhippuric (HHA) acids, the three acids of colonic origin. To better understand the antioxidant and anti-inflammatory properties of DHCA, DHFA, and HHA, an inflammation/oxidation model was set up in the pre-adipocyte 3T3-L1 cell line using tumor necrosis factor-α (TNF-α). After the exposure of 3T3-L1 cells to 0.5, 1, 5, and 10 µM of TNF-α at different times, the cell viability, interleukin (IL)-6 secretion, and the production of reactive oxygen species (ROS) and glutathione (GSH) were determined. Using the TNF-α prooxidant and proinflammatory conditions established (10 µM, 24 h), it was observed that the physiological concentrations (0.5, 1, 5, and 10 µM) of DHCA, DHFA, and HHA induced dose-dependent antioxidant effects according to the ROS, GSH, and antioxidant enzyme (glutathione peroxidase) results. In addition, reductions in the IL-1β, IL-6, and monocyte chemoattractant protein-1 (MCP-1) concentrations were observed to different extents depending on the metabolite (DHFA, HHA, or DHCA) and the concentration used. In conclusion, the main colonic metabolites from coffee chlorogenic acids may counteract TNF-α-induced inflammation and oxidative stress in the 3T3-L1 cell line, and thus, they present antiobesity potential.

Green coffee has weight management properties, yet its effects on appetite and satiety remain unclear as few, mainly acute, studies perform objective measurements. Therefore, the influence on appetite/satiety of acute and regular consumption of two nutraceuticals, a decaffeinated green coffee phenolic extract (GC) alone or combined with oat β-glucans (GC/BG), with known satiating properties, has been analysed subjectively using visual analog scales (VAS) and objectively measuring actual food intake and postprandial appetite and satiety hormones. A randomised, cross-over, blind trial was carried out in 29 overweight volunteers who consumed GC or GC/BG twice a day for 8 weeks. After acute (day = 0) and regular consumption (day = 56) of the nutraceuticals, satiety was measured at 30, 60, 90, 150, and 210 min, as well as food intake at breakfast (30 min) and lunch (300 min). Additionally, in a subgroup of participants (n = 9), cholecystokinin, peptide-YY, glucagon-like-peptide-1, ghrelin and leptin concentrations were analysed in blood samples taken at the same time-points. According to VAS results, GC/BG reduced hunger more efficiently than GC. However, there were no statistically significant differences in food intake. Comparing the effects of the acute consumption of GC/BG and GC, leptin concentration at 150 min was higher after GC/BG intake vs. GC. Moreover, when comparing the effects of regularly consuming the two nutraceuticals, maximum ghrelin level decreased with GC/BG vs. GC. In conclusion, acute and regular effects of the nutraceuticals on appetite/satiety differed, and subjective and objective results partially agreed; GC/BG may reduce hunger more efficiently than GC.

The epidermal growth factor receptor (EGFR) is frequently expressed in ovarian cancer, yet its potential as a therapeutic target remains underexplored. We investigated EGFR expression and its therapeutic exploitation using antibody-drug conjugates (ADCs) based on the clinical anti-EGFR antibody cetuximab. EGFR protein and mRNA levels were evaluated in patient-derived tumors and ovarian cancer cell lines by western blotting and qPCR, and cell surface localization was analyzed by flow cytometry. Several cetuximab-based ADCs were generated using clinically validated cytotoxics: DM1, deruxtecan (DXd), and monomethyl auristatin F (MMAF), with cleavable or non-cleavable linkers. Their structural integrity, antiproliferative activity, internalization dynamics, and mechanisms of action were examined in vitro, and in vivo efficacy and pharmacokinetics were assessed in SKOV3 and OVCAR8 xenograft models. All ADCs showed successful conjugation and preserved antibody integrity. MMAF-conjugated ADCs displayed superior antiproliferative effects, particularly the cleavable cetuximab-vc-MMAF, while A2780 and PEO4, which showed low EGFR expression, were less responsive. Cetuximab alone or free MMAF were markedly less active. Mechanistic studies revealed rapid internalization and lysosomal trafficking of cetuximab-vc-MMAF, leading to G2/M arrest, increased mitotic markers, spindle defects, DNA damage, and apoptosis. In vivo, cetuximab-vc-MMAF significantly inhibited tumor growth and prolonged survival without systemic toxicity. Pharmacokinetic analyses confirmed selective intratumoral accumulation of the ADC and its payload with minimal off-target distribution. Treated tumors exhibited reduced proliferation and increased markers of mitotic arrest, DNA damage, and apoptosis. These results demonstrate that cetuximab-based EGFR-targeting ADCs exert potent and selective antitumor activity in ovarian cancer, supporting their further preclinical and clinical development.

Background: Resistance to HER2-targeted therapies remains a major limitation in the treatment of HER2-positive breast cancer, where disease progression inevitably occurs in advanced stages. Development of next-generation strategies that retain activity in resistant disease is therefore a critical priority. Disitamab vedotin (RC48) is a novel antibody–drug conjugate (ADC) targeting HER2 that couples a humanized anti-HER2 antibody to the potent microtubule-disrupting agent monomethyl auristatin E. Methods: We compared the activity and mechanism of action of RC48 with that of trastuzumab emtansine (T-DM1) across HER2-positive and HER2-low cellular models, including multiple sublines resistant to current HER2-targeted agents. Results: In HER2-overexpressing breast cancer cell lines, RC48 consistently demonstrated superior antiproliferative effect with respect to T-DM1. Treatment with RC48 induced G2/M arrest and apoptotic cell death, associated with increased pHistone-H3 and cyclin B1 and downregulation of Wee1, consistent with blockade of cell cycle progression in mitosis. Although RC48 and T-DM1 internalized similarly, RC48 displayed more efficient intracellular payload release, providing a mechanistic explanation for its enhanced efficacy. Notably, RC48 retained strong activity in BT474-derived sublines resistant to T-DM1, lapatinib, or neratinib, inducing cell cycle arrest, apoptosis, and caspase activation in all resistant models. In contrast, T-DM1 exhibited only partial effects in resistant cells and was completely ineffective in a T-DM1-refractory clone. Conclusions: Together, these findings identify disitamab vedotin as a potent next-generation HER2-targeting ADC with the unique capacity to overcome acquired resistance to HER2-directed therapies. RC48 represents a promising therapeutic strategy for patients with refractory HER2-positive breast cancer and warrants further clinical investigation.