Parkinson's disease (PD) is a neurodegenerative disease characterized by the degeneration of dopaminergic neurons in the substantia nigra and the presence of misfolded α-synuclein in the brain. Mitochondrial dysfunction and oxidative stress are factors contributing to the death of these neurons. Coenzyme Q10 (CoQ10) serve as an antioxidant and cofactor for mitochondrial enzymes, and its deficiency can exacerbate neurodegenerative processes in PD. However, the clinical efficacy of CoQ10 is limited by its low bioavailability and instability. Ubiquinol diacetate (CoQ10 Ac), an esterified form of CoQ10, shows improved pharmacokinetic properties and potential as a prodrug, converting into the reduced antioxidant form of CoQ10 by esterases in the body. This study aimed to investigate the antioxidant and neuroprotective effects of CoQ10 Ac compared to CoQ10 in SH-SY5Y cell line and Caenorhabditis elegans models of PD. CoQ10 Ac showed higher antioxidant activity than CoQ10 at both extracellular and intracellular levels, particularly in the membrane and cytosolic compartments. It exhibited superior neuroprotection against 6-hydroxydopamine toxicity, showing a greater ability to reduce the activation of caspase-3 and PARP1 compared to CoQ10. Both compounds decreased the increased ratio of mitochondrial fission protein, DRP1, to fusion protein, OPA1, induced by 6-hydroxydopamine in SH-SY5Y cells, enhancing OPA1 levels and promoting antiapoptotic death. However, CoQ10 Ac was more effective than CoQ10 in preserving mitochondrial structural integrity and mass. Additionally, both compounds significantly inhibited the aggregation of α-synuclein induced by 6-hydroxydopamine. Furthermore, CoQ10 Ac showed stronger neuroprotective effects than CoQ10 in C. elegans models of PD. It demonstrated greater anti-aggregant activity in C. elegans expressing human α-synuclein, suggesting higher bioavailability. These findings highlight CoQ10 Ac as a promising prodrug candidate and support further investigation in in vivo PD models.

Osteoarthritis (OA) is a chronic degenerative joint disease characterized by progressive cartilage breakdown driven by inflammatory and oxidative stress mechanisms. Blue turmeric (Curcuma caesia Roxb.), a rare and understudied spice, represents a source of bioactive compounds with anti-inflammatory properties. This study aimed to characterize the phenolic profile of a blue turmeric rhizome extract (BTE) and its bioaccessible fraction obtained after in vitro gastrointestinal digestion, and to assess their biological effects on human chondrocytes (C-28/I2) exposed to interleukin-1 β (IL-1β). HPLC-MS/MS analyses revealed a phytochemical profile rich in phenolic acids and flavan-3-ols, with partial stability maintained after in vitro digestion. Pre-treatment with both native BTE and digested BTE (dBTE) protected chondrocytes against IL-1β-induced cytotoxicity, mitochondrial dysfunction, and nuclear factor erythroid 2-related factor 2 (Nrf2) inactivation. Furthermore, BTE pre-treatment significantly reduced tumour necrosis factor-α (TNF-α) release and preserved cellular morphology. These findings demonstrate that BTE phenolic compounds are partially bioaccessible and exert chondroprotective effects, supporting further preclinical investigation of BTE as a potential functional ingredient for joint health maintenance and OA prevention strategies.

Vitamin E succinate and its derivatives have demonstrated encouraging cytotoxic potential in preclinical models of gastric cancer, paving the way for a novel class of therapeutics. Building on a previously described synthetic methodology based on the alkoxy-aryloxycarbonylation of alkenes, this study reports the investigation of the antitumor effects in gastric cancer models of mixed aryl-alkyl succinates esters featuring various substituents on the backbone. Nine compounds were screened for cytotoxicity against AGS and KATO III gastric cancer cell lines. The most active compounds - 2d (R = (CH2)4CH3), 2a (R = Ph), and 2c (R = CH2CH2Ph) - were further evaluated for efficacy, with 2d emerging as the most potent agent (IC50 30.9 μM in AGS; IC50 19 μM in KATO III). Mechanistically, 2d enhanced the expression of activated/cleaved Caspase-3, augmented PARP cleavage, and promoted LC3B lipidation - indicative of programmed cell death. Consistently, ultrastructural analysis of 2d-treated cancer cells revealed morphological hallmarks of both early and late apoptosis, including cytoplasmic vacuolization and autophagic vacuoles. Conversely, treatment with compound 2d did not affect the expression of apoptosis markers in healthy GES-1 gastric epithelial cells, suggesting a favorable safety profile. Overall, our findings provide insights into how specific structural features of succinate derivatives contribute to their antitumor activity, laying the groundwork for the design of more potent succinate-based agents.

Aging is a complex biological process influenced by genetic, environmental, and lifestyle factors, including diet and exercise. This process not only leads to visible signs of skin aging, such as wrinkles and elasticity loss, but also predisposes individuals to a spectrum of age-related diseases. Oxidative stress and inflammation are key events driving this phenomenon. This study aims to delve into the antioxidative and protective potential of a rhizome extract from Curcuma caesia Roxb., a less explored species within the turmeric family, focusing on its effects on HaCaT cells subjected to UV radiation. Curcuma caesia Roxb. rhizome was extracted for a phytocomplex (CCRE). Through HPLC-ESI-MS/MS, the chemical analysis identified key phenolic compounds, including (-)-epicatechin, procyanidin B2, and p-coumaric acid. Human immortalized keratinocyte cell lines (HaCaT) were treated with CCRE in healthy and UVB-induced conditions to assess their impact on oxidative stress and aging. Results demonstrated a significant reduction in mitochondrial superoxide anion levels without affecting mitochondrial membrane potential, indicating enhanced cellular resilience to oxidative stress. Additionally, CCRE decreased UVB-induced IL-6 expression and IKK phosphorylation, which play a crucial role in inflammaging. Notably, CCRE treatment also improved cell viability upon UVB exposure and mitigated UVBinduced cell apoptosis, further underscoring its potential to preserve cellular integrity and function in relation to environmental stressors. CCRE chemical composition and biological effects position it as a potential ingredient for formulations and functional foods to combat the signs of skin aging and enhance overall skin health.

Endoplasmic reticulum oxidoreductin 1 alpha (ERO1A) is a disulfide oxidase that facilitates oxidative protein folding by reoxidizing protein disulfide isomerase (PDI), a process essential for maintaining endoplasmic reticulum (ER) homeostasis. Under ER stress, ERO1A expression is upregulated via the unfolded protein response (UPR), promoting cell survival. However, sustained ERO1A activity can impair proteostasis and contribute to disease. Notably, ERO1A is overexpressed in triple-negative breast cancer (TNBC), where it supports tumor growth and adaptation to hypoxia, and in SEPN1-related myopathy, a rare congenital muscle disorder linked to ER and oxidative stress. To investigate ERO1A as a therapeutic target, we conducted a structure-activity relationship (SAR) study of EN460-based pyrazolone inhibitors. Forty derivatives and three EN460 salts were synthesized to optimize potency and solubility. In vitro and cell-based assays revealed that effective inhibition required covalent binding to Cys397, interactions with Arg287 and Trp200, and distortion of the phenyl ring. While sulfonic acid substitution improved solubility, it abolished activity by disrupting key interactions. The most potent compound, I29, featuring a mono ortho-fluorine substitution, demonstrated improved inhibitory activity (IC₅₀ = 2.6 µM) and efficacy in preclinical models of TNBC and SEPN1-related myopathy. These findings highlight ERO1A's pathological role in cancer and congenital muscle disease and support its inhibition as a promising therapeutic strategy for conditions characterized by chronic ER and oxidative stress.

Blackthorn (Prunus spinosa L.) juice, a polyphenol-rich blend derived from the Marche region of Italy, was chemically characterized and evaluated for its selective effects against gastric cancer cells. The juice contained a mix of polyphenols, including neochlorogenic acid, cyanidin-3-glucoside, rutin, and smaller molecules - such as gallic, caffeic, and vanillic acids - characterized by high absorption rates. In vitro, the juice significantly reduced the viability of AGS and KATO III gastric cancer cells, while sparing non-tumorigenic GES-1 cells. In malignant cells, the juice activated Caspase-3, upregulated Bax, and downregulated Bcl-xL, while also modulating autophagy by increasing LC3B-II levels and decreasing Beclin-1. Mitochondrial impairment was confirmed by cardiolipin loss and cytochrome c release into the cytoplasm. These findings support the potential of Prunus spinosa L. juice as a selective, innovative functional beverage for gastric cancer prevention, acting beyond nutrition but before the need for medical intervention.

This study investigates the chemopreventive potential of a grape pomace extract (GPE) derived from Verdicchio grapes, aligning with circular economy principles to repurpose winery waste into a nutraceutical targeting gastric cancer prevention. Soxhlet extraction yielded a bioactive-rich extract. Comprehensive chemical characterization via HPLC/ESI/Q-TOF identified 39 metabolites spanning key chemical classes. Anthocyanins were predominant, with malvidin glucoside (12,546 mg/kg DM; 36.3%), malvidin coumaroyl glucoside (9941 mg/kg DM; 28.8%), and malvidin acetylglucoside (7189 mg/kg DM; 20.8%) as the most abundant compounds. Carboxylic acids included tartaric, malic, isocitric, aconitic, and succinic acids, while lipid molecules, such as phytosphingosine and stearic acid, and amino acids like proline, valine, leucine, and phenylalanine further enriched the extract's chemical heterogeneity. Biological evaluations revealed GPE's selective cytotoxicity against AGS (IC50: 13.64 μg/mL) and KATO III (IC50: 7.11 μg/mL) gastric cancer cells, sparing GES-1 cells. Mechanistically, GPE-induced apoptosis through caspase-3 activation and mitochondrial dysfunction, as evidenced by inner membrane disruption and cardiolipin peroxidation. TEM and CLSM morphological analyses revealed hallmark apoptotic features, including chromatin condensation, micronuclei formation, and apoptotic bodies. Additionally, GPE impaired autophagy, marked by Beclin-1 downregulation and LC3B-II upregulation. The accumulation of degradative vacuoles indicated disrupted autophagosome clearance, shifting autophagy from a survival mechanism to a cell death-promoting pathway. These findings highlight GPE's dual impact on apoptosis and autophagy in gastric cancer cells, underscoring its potential as a dietary intervention for gastric cancer prevention.

Cancer cells adapt to harsh environmental conditions by inducing the Unfolded Protein Response (UPR), of which ERO1A is a mediator. ERO1A aids protein folding by acting as a protein disulfide oxidase, and under cancer-related hypoxia conditions, it favors the folding of angiogenic VEGFA, leading tumor cells to thrive and spread. The upregulation of ERO1A in cancer cells, oppositely to the dispensability of ERO1A activity in healthy cells, renders ERO1A a perfect target for cancer therapy. Here, we report the upregulation of ERO1A in a cohort of aggressive triple-negative breast cancer (TNBC) patients in which ERO1A levels correlate with a higher risk of breast tumor recurrence and metastatic spread. For ERO1A target validation and therapy in TNBC, we designed new ERO1A inhibitors in a structure-activity campaign of the prototype EN460. Cell-based screenings showed that the presence of the Micheal acceptor in the compound is necessary to engage the cysteine 397 of ERO1A but not sufficient to set out the inhibitory effect on ERO1A. Indeed, the ERO1 inhibitor must adopt a non-coplanar rearrangement within the ERO1A binding site. I2 and I3, two new EN460 analogs with different phenyl-substituted moieties, efficiently inhibited ERO1A, blunting VEGFA secretion. Accordingly, in vitro assays to measure ERO1A engagement and inhibition confirmed that I2 and I3 bind ERO1A and restrain its activity with a IC50 in a low micromolar range. EN460, I2 and I3 triggered breast cancer cytotoxicity while specifically inhibiting ERO1A in a dose-dependent manner. I2 more efficiently impaired cancer-relevant features such as VEGFA secretion and related cell migration. I2 also acted on the tumor microenvironment and viability of xenografts and syngeneic TNBC. Thus, small molecule-mediated ERO1A pharmacological inhibition is feasible and promises to lead to effective therapy for the still incurable TNBC.

BackgroundMitochondrial dysfunction and oxidative stress are central mechanisms in the progression of neurodegenerative diseases. This study first evaluated the toxicity of Q-Der (Q10-diacetate), a derivative of Coenzyme Q10, in HT22 hippocampal neurons under normal and oxidative stress conditions.MethodsHT22 cells were treated with Q-Der at 2.5, 5 and 10 µM with and without rotenone. Mitochondrial superoxide production (Mitosox), gene expression (via qRT-PCR), and protein levels (via Western blot) were measured. Morphological analyses were performed using transmission (TEM) and scanning (SEM) electron microscopes.ResultsQ-Der significantly reduced mitochondrial superoxide levels, particularly at 5 μM, and upregulated key mitochondrial biogenesis genes, including PGC-1α and TFAM. Additionally, it restored the expression of MT-ND1 and MT-COI, which were downregulated by rotenone. Western blot results showed a significant recovery in CV-ATP5A (complex V) expression (p < 0.05), preserving mitochondrial ATP production. Morphological analyses further confirmed Q-Der’s ability to maintain cellular and mitochondrial structure under stress conditions.ConclusionThese findings suggest that Q-Der is non-toxic under normal conditions and protects against oxidative stress, supporting its potential as a therapeutic agent for neurodegenerative diseases.

The design of inhaled selective phosphatidylinositol 3-kinase delta (PI3K delta) inhibitors for the treatment of inflammatory lung diseases was pursued. Knowledge-based design of a novel isocoumarin scaffold that was able to adopt a propeller-shape topology ensured the desired PI3K delta selectivity. Achievement of low nanomolar cellular potencies through hinge binder group optimization, reduction of intrinsic permeability through head group optimization to extend lung retention, and screening of crystalline forms suitable for administration as dry powders culminated in the identification of compound 18. This novel inhaled selective PI3K delta inhibitor displayed durable anti-inflammatory activity in a disease-relevant rat model of Th-2-driven acute lung inflammation and safe in vitro and in vivo preclinical profiles. Therefore, compound 18 showed the appropriate discovery profile and was progressed to clinical trials in healthy volunteers and chronic obstructive pulmonary disease (COPD) patients as CHF-6523.

Gastric cancer (GC), the third leading cause of cancer-related death globally, is complex and heterogeneous. This review explores multidisciplinary investigations of traditional Chinese medicine (TCM) combined with Western medical practices, emphasizing the development of nutraceuticals for cancer prevention. Using advanced analytical chemistry and food chemistry techniques, this study investigated how TCM components may be optimized for nutraceutical development. Focusing on molecular interactions with GC pathways, particularly the NF-κB, PI3K/Akt, and Wnt/β-catenin pathways, we examined the effects of TCM polyherbal formulas, extracts, and isolated compounds. These agents modulate apoptosis and cellular proliferation, underscoring their potential in preventive strategies. The convergence of nutraceutical and medicine food homology studies highlights a significant shift towards integrating TCM-derived compounds in a preventive health framework. This approach aims not only to enhance efficacy and reduce side effects but also to champion a preventive paradigm using personalized medicine to advance proactive health maintenance and disease prevention. The combination of TCM and western medical practices offers promising avenues for future research and practical applications in GC prevention.

A selective, mild, convenient, and green protocol for the preparation of S-acyl and N-acyl glutathiones is described involving the chemical modification of glutathione (GSH) with N-acyl imidazoles at room temperature in water. The syntheses of S-acyl glutathiones were achieved in very high yields using 1 equiv. of an N-acyl imidazole in water at room temperature, without the need of a base. Double acylation of GSH with various N-acyl imidazoles in weakly basic aqueous media in the presence of N-hydroxysuccinimide (HOSu) as the activating reagent followed by selective deprotection of the S-acyl group with aqueous ammonia at room temperature gave high yields of N-acyl glutathiones. Moreover, the reaction could accommodate a diverse range of carboxylic acids such as (hetero)benzoic acids, phenylacetic acids, aliphatic acids from short-chain fatty acids (including acetic acid), long-chain polyunsaturated fatty acids, secondary or tertiary amino acids, and carboxylic acids containing clickable functional groups, fluorescent probes, or drugs.

The synthesis and characterization of a novel bis-urea-based cage receptor for anions (3S,15S)-3,15,20,25-tetramethyl-1,4,6,12,14,17,20,25-octaazatricyclo[15.5.5.17.11]octacosa-7(28),10-diene-2,5,13,16-tetraone (L) is reported. L is a macro-bicyclic ligand built on the 1,7-dimethyl-1,4,7,10-tetraazacyclododecane scaffold to obtain a cage topology in which two ureido moieties have been inserted as binding sites for anions. L can interact with anion guests (G) via H-bonding; in particular, it binds both spherical (Cl-) and V-shaped anions (AcO-) as well as more complex carboxylate anions, such as the norfloxacin (Nor-). NMR experiments highlight that the interaction between L and G mainly occurs at the ureido moieties. L forms L-G adducts of 1 : 1 ([LG]-) and 1 : 2 ([LG2]2-) stoichiometry with Cl- and AcO-. Otherwise, in the case of Nor- only the formation of the [LG]- complex is observed. L shows higher formation constants values for [LAcO]- (2.9) and [LNor]- (3.6) than [LCl]- suggesting a stronger interaction with the carboxylate anions. In the solid state, three crystal structures of the HL⋅G species were obtained (G=Cl-, AcO-, ClO4 -) highlighting the H-bonding interaction between the chloride, acetate or perchlorate anions and the -NH functions of the ureido fragment. The comparison between the two parent open chain receptors (Lb-c) and L has been reported and discussed.

This research investigates boronated tryptophans as potential boron delivery agents for boron neutron capture therapy (BNCT) of cancer. We synthesized both enantiomers of 5- and 6-boronotryptophans (1a and 1b) using simple and inexpensive methods. Their uptake was assessed in two human cancer cell lines, CAL27 (head and neck cancer) and U87-MG (brain cancer), and compared to l-p-boronophenylalanine (l-BPA) as a reference. To determine whether these tryptophan derivatives are substrates for large amino acid transporter 1, we performed molecular dynamics simulations to explore their transport mechanism. Our findings reveal differences in boron compound accumulation between the cancer cell lines, indicating that tryptophan derivatives could serve as effective boron carriers when the clinically used boron carrier, BPA, is ineffective.

The labeled ligand commonly employed in competition binding studies for melatonin receptor ligands, 2-[125I]iodomelatonin, showed slow dissociation with different half-lives at the two receptor subtypes. This may affect the operational measures of affinity constants, which at short incubation times could not be obtained in equilibrium conditions, and structure-activity relationships, as the Ki values of tested ligands could depend on either interaction at the binding site or the dissociation path. To address these issues, the kinetic and saturation binding parameters of 2-[125I]iodomelatonin as well as the competition constants for a series of representative ligands were measured at a short (2 h) and a long (20 h) incubation time. Concurrently, we simulated by molecular modeling the dissociation path of 2-iodomelatonin from MT1 and MT2 receptors and investigated the role of interactions at the binding site on the stereoselectivity observed for the enantiomers of the subtype-selective ligand UCM1014. We found that equilibrium conditions for 2-[125I]iodomelatonin binding can be reached only with long incubation times, particularly for the MT2 receptor subtype, for which a time of 20 h approximates this condition. On the other hand, measured Ki values for a set of ligands including agonists, antagonists, nonselective, and subtype-selective compounds were not significantly affected by the length of incubation, suggesting that structure-activity relationships based on data collected at shorter time reflect different interactions at the binding site. Molecular modeling simulations evidenced that the slower dissociation of 2-iodomelatonin from the MT2 receptor can be related to the restricted mobility of a gatekeeper tyrosine along a lipophilic path from the binding site to the membrane bilayer. The enantiomers of the potent, MT2-selective agonist UCM1014 were separately synthesized and tested. Molecular dynamics simulations of the receptor-ligand complexes provided an explanation for their stereoselectivity as due to the preference shown by the eutomer at the binding site for the most abundant axial conformation adopted by the ligand in solution. These results suggest that, despite the slow-binding kinetics occurring for the labeled ligand, affinity measures at shorter incubation times give robust results consistent with known structure-activity relationships and with interactions taken at the receptor binding site.