ACS Pharmacology and Translational Science最新文献

Kratom (Mitragyna speciosa Korth.) has gained increasing scientific interest for its potential in pain management and addiction treatment. This study employs a green chemistry approach to optimize the extraction of kratom leaves by using Accelerated Solvent Extraction (ASE) with an ethanol–water binary solvent system. The goal was to improve the yield and potency of key bioactive compounds, especially mitragynine. Optimization was performed using One-Factor-at-a-Time (OFAT) analysis and Response Surface Methodology (RSM) employing a Box-Behnken Design (BBD). The optimal extraction conditions were determined to be an 8 min extraction time, 60 °C temperature, and 40% ethanol concentration, which resulted in mitragynine content of 4.66%, total phenolic content of 212.69 GAE mg/g, and total flavonoid content of 126.13 QE mg/g. The safety profile of the optimized ASE kratom leaf extract was evaluated using MTT cytotoxicity assay, which revealed selective cytotoxicity against HepG2 liver cancer cells (IC₅₀ = 7.69 μg/mL), while showing no cytotoxicity toward HL-7702 normal liver cells (IC₅₀ > 200 μg/mL). Antinociceptive activity was tested in BALB/c albino mice using the hot-plate test, where the optimized ASE kratom leaf extract demonstrated analgesic effects at dosages of 100, 200, and 500 mg/kg. Phytochemical profiling combining NMR and UPLC-ESI-QTOF-MS/MS identified several known kratom constituents, including mitragynine and its congeners as well as bioactive flavonoids such as isoquercitrin and rutin. The optimized ASE method using a green ethanol–water system produces kratom extracts with promising safety and therapeutic potential, though further work is needed to refine and scale the approach for broader phytopharmaceutical use.

Fatty acid esters of hydroxy fatty acids (FAHFAs) and alkyl-diacylglycerols (TG(O)s) are endogenous lipids that exert protective effects against diabetes, inflammation, and the development of childhood obesity. They have been identified as minor lipids in human breast milk, and an accurate characterization of FAHFAs and TG(O)s could further our understanding of changes in human milk lipid profiles across different phenotypes. However, due to differences in their polarities and chemical properties, current studies on FAHFAs and TG(O)s tend to be conducted independently. The study of these compounds is further complicated by a shortage of validated methods for the isolation of FAHFAs and TG(O)s from complex, lipid-rich biological matrices, such as human breast milk. Additionally, cohort studies on human tissues, including breast milk, are often confronted with limited sample availability. In this study, we describe the systematic validation of a workflow to reliably extract, isolate, and characterize both FAHFAs and TG(O)s from 1.6 mL of human breast milk. We have applied this method to the measurement of 16 and 19 endogenous FAHFA and TG(O) species originating from two cohort studies, respectively. Our validated workflow also includes the application of liquid chromatography-multistage mass spectrometry approaches to identify FAHFA and TG(O) molecular species as well as FAHFA regioisomers. This method has the potential to be adapted for the isolation and characterization of other minor lipids with different polarities from limited sample volumes of complex biological matrices.

Hepatocellular carcinoma (HCC) is a major contributor to cancer-related deaths worldwide. Among the emerging therapies, ultrasound-mediated sonosensitizers have shown promise in the treatment of HCC. Sonosensitizers exposed to low-intensity ultrasound energy can generate reactive oxygen species (ROS), which can lead to cancer cell death. Indocyanine green (ICG) is a commonly used sonosensitizer that can be used for treating HCC. However, ICG has a short half-life and needs a carrier to improve its therapeutic efficacy. Herein, we report the development of ICG-loaded liposomes with controlled size distribution using a simple one-step microfluidic device-based strategy. We evaluated the stability of ICG-loaded liposomes (lipo-ICG) by subjecting them to various storage conditions. The designed lipo-ICGs are stable and capable of inducing liver cancer cell death (HepG2 cells) upon ultrasound exposure. Last but not least, the designed lipo-ICGs are cytocompatible to both cancerous (HepG2 cells) and noncancerous cells (HHSteC) without ultrasound exposures. Taken together, our findings highlight the potential of this microfluidic platform for the efficient production of lipo-ICG nanoparticles and demonstrate the promise of ultrasound-mediated therapy as a targeted, minimally invasive strategy for treating HCC.

Cognitive impairment is a common symptom of many psychiatric disorders and can significantly reduce the patients' quality of life. Vindeburnol, a synthetic derivative of eburnamine–vincamine alkaloids, has shown potential in models of depression and neurodegeneration; however, its comprehensive properties and molecular mechanisms of action have remained unexplored. We conducted a comprehensive preclinical assessment of vindeburnol, including a subchronic toxicity study (20 and 80 mg/kg/day, orally, 14 days), pharmacokinetic analysis (40 mg/kg), behavioral tests for memory and learning, and transcriptomic profiling of the locus coeruleus (LC) in mice. Vindeburnol demonstrated a good safety profile at 20 mg/kg, while the 80 mg/kg dose caused 20% mortality and hepatotoxicity. The compound exhibited high oral bioavailability (75%) and a long half-life (7.58 h). Behavioral analysis revealed a specific improvement in episodic memory in the novel object recognition test without affecting motor activity. Transcriptomic analysis of the LC identified 10 differentially expressed genes, including the upregulation of Npas3 and Cfap69, key regulators of neurogenesis and synaptic plasticity, and the downregulation of genes associated with neuroinflammation (Ctss, Hspa1b). Our study is the first to comprehensively characterize vindeburnol as a promising compound with proven procognitive effects. The data reveal its unique mechanism of action, which promotes neuroplasticity and counteracts pathological processes by modulating key transcriptional programs in the locus coeruleus, supporting its further investigation as a potential neurotropic agent.

Background and Purpose: SKF-96365 is widely used as a broad-spectrum inhibitor of calcium entry. It was initially identified as a Receptor-Operated Ca²⁺ Entry (ROCE) blocker and was later shown to inhibit ORAI1–STIM1-mediated Store-Operated Ca²⁺ Entry (SOCE). However, its selectivity for TRPC versus the ORAI channels remains unclear. Experimental Approach: To examine the selectivity of SKF-96365, we evaluated its effects on SOCE and ROCE in wild-type and TRPC hepta-KO mouse embryonic fibroblasts (MEFs), as well as on TRPC-mediated OAG-induced calcium entry in HEK293 cells overexpressing TRPC3, TRPC6, or TRPC7. Additional assays were conducted on HEK293 cells expressing the muscarinic M5 receptor (M5R). Half-maximal inhibitory concentration (IC₅₀) values were determined under all conditions. Key Results: SKF-96365 suppressed thapsigargin (Tg)-induced SOCE similarly in wild-type and TRPC hepta-KO MEFs, with IC₅₀ values around 4–5 μM. Comparable inhibition was observed for carbachol (CCh)-activated ROCE in TRPC-deficient cells. In contrast, OAG-activated Ca²⁺ entry by TRPC3/6/7 was only weakly inhibited, with IC₅₀ values exceeding 100 μM. Notably, TRPC-mediated Ca²⁺ entry was unaffected by CRAC channel blockers or ORAI coexpression, confirming its independence of SOCE mechanisms Conclusions and Implications: Our findings demonstrate that ORAI-mediated SOCE is approximately 25-fold more sensitive to SKF-96365 than TRPC-mediated calcium entry. GSK-7975A further confirmed the ORAI selectivity by blocking SOCE without affecting TRPC channels. These results clarify the pharmacological profile of SKF-96365, confirming its primary action on ORAI channels and highlight the need for concentration-aware interpretation in calcium signaling studies, particularly in the context of CRAC channel-related disorders.

Osteoarthritis (OA) is a leading cause of physical disability, psychological distress, and a significant economic burden worldwide. Current treatments alleviate symptoms; however, disease progression remains largely uncontrolled, highlighting the urgent need for investigation of disease-modifying therapies. Symptomatic slow-acting drugs for osteoarthritis (SYSADOAs), such as glucosamine (GlcN), chondroitin sulfate (CS), and hyaluronic acid (HA), have gained increasing attention for their potential benefits in alleviating pain and mitigating the inflammatory and degenerative processes that characterize OA. These compounds modulate several homeostatic mechanisms, promoting anti-inflammatory, antioxidant, antiapoptotic, and anabolic countermensuring effects. Nevertheless, debates regarding their long-term efficacy and safety remain controversial, which explains why major osteoarthritis societies do not provide the same recommendations for the pharmacological treatment of OA. In this context, this review critically evaluates the current evidence surrounding HA, GlcN, and CS, highlighting their safety, mechanisms of action, and promising therapeutic perspectives for modifying the natural course of knee, hand, and hip OA.

Inhibitors of the Bruton’s tyrosine kinase (BTK) are of broad utility in the treatment of multiple diseases including several B-cell malignancies via effective blockade of oncogenic B-cell receptor (BCR) signaling. BTK is a cytoplasmic tyrosine kinase which harbors a targetable cysteine residue (Cys481) and the majority of BTK inhibitors are covalent modifiers directed at this position. Despite possessing a common mechanism of action, BTK inhibitors differ in key attributes including off-target kinome profiles, tolerability, pharmacokinetics and the underlying BTK inhibition kinetics. These characteristics play a significant role in the ultimate utility of these drugs. Herein, we compare several clinically active BTK inhibitors in biochemical and in vitro assays to gain a broader appreciation of the similarities and differences that govern the success of this important drug class. The combined datasets highlight that each agent has excellent on-target potency and good BTK selectivity. The data further suggests an association between optimized BTK inhibition kinetics and in vitro cytotoxicity profiles.

Gastric ulcers (GUs), a frequent gastrointestinal condition, cause mucosal injury and inflammation. We aimed to investigate the protective effects and underlying mechanisms of ezetimibe, statins, and their combination in indomethacin-induced GUs. Rats were assigned into six groups (n = 8, each): (I) normal (control), (II) gastric ulcer induced with a single oral dose of indomethacin (30 mg/kg body weight), (III) rats received oral simvastatin (40 mg/kg/day) for 14 days, (IV) rats received oral ezetimibe (10 mg/kg/day) for 14 days, (V) the combination group received both oral simvastatin and ezetimibe, and (VI) standard group received oral famotidine (20 mg/kg). On day 14, gastric ulcers were induced by a single oral dose of indomethacin (30 mg/kg), and the animals were sacrificed 6 h later for sample collection and tissue analysis. Tissue levels of malondialdehyde (MDA), tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), Kelch-like ECH-associated protein 1 (Keap1), superoxide dismutase (SOD), nuclear erythroid factor 2 (Nrf-2) and hem-oxygenase 1 (HO-1) were measured. Ezetimibe, simvastatin, and their combination prevented GU. The combination therapy significantly reduced MDA, TNF-α, Keap1, IL-1, and serum C-reactive protein (CRP). However, Nrf-2, HO-1, and SOD were significantly increased when compared to the GU group and monotherapy. Histological investigations demonstrated that the combination therapy reduced GU severity and preserved stomach tissue. Simvastatin plus ezetimibe exerted synergistic gastroprotective effects in rats, associated with Nrf2/HO-1 activation and suppression of Keap1, oxidative stress, and pro-inflammatory cytokines. This combination may represent a novel therapeutic approach for preventing NSAID-induced GUs, meriting further mechanistic and translational studies.

Histone deacetylases (HDACs) regulate gene expression and are promising targets in oncology. Especially the class I isoforms HDAC1 and HDAC2 are overexpressed in cancer. However, while ortho-aminoanilides with a suitable (het)aryl substitution are well-characterized HDAC1/HDAC2 inhibitors, the corresponding phenol analogs have not been sufficiently investigated so far. To this end, we compared the ortho-hydroxyanilide derivative ST13 with the pan-HDAC inhibitor vorinostat and Cpd-60, an ortho-aminoanilide with high HDAC1/HDAC2 selectivity. ST13 was further developed into a light-activatable prodrug (ST17) by masking its zinc-binding group with a photoremovable 4,5-dimethoxy-2-nitrobenzyl protecting group. Overall, we verified that ST13 is a selective, slow- and tight-binding HDAC1/HDAC2 inhibitor with antiproliferative activity. Furthermore, we demonstrated that the light-activatable prodrug ST17 readily releases ST13 upon irradiation, thereby allowing to precisely control its antiproliferative properties. These findings validate ortho-hydroxyanilides as effective HDAC1/HDAC2-selective inhibitors and highlight photocaging as a promising strategy to achieve spatiotemporal control of epigenetic therapies in cancer.

Sulforaphane (SFN) is a phytoderived compound abundant in cruciferous plants that possesses a broad spectrum of anticancer properties. We showed that SFN-induced caspase-mediated apoptosis in grade IV bone metastasis-derived androgen-insensitive PC-3 (IC₅₀ = 4.2 μM), and lymph node metastasis-derived androgen-sensitive LNCaP (IC₅₀ = 2.8 μM) prostate adenocarcinoma cells. SFN-mediated cardiotoxic side effects were tested in a preclinical in vitro model that enables the study simultaneously of the impact of drugs on cancer cell death and contractile properties of engineered heart tissues generated from human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM EHT). Thereby, SFN exposure induced PC-3 cell death without affecting the contractile force of hiPSC-CM EHT. Interestingly, the irregular beating pattern of hiPSC-CM EHT observed in the presence of PC-3 coculture was normalized compared to vehicle treatment. Overall, this in vitro coculture model of hiPSC-CM EHT and cancer cells could facilitate the study of cardiotoxic cancer drug side-effects.