Background Pancreatic ductal adenocarcinoma (PDAC) harbors TP53 mutations at high frequency, yet therapeutic strategies that specifically target mutant p53 remain limited. Purpose This study aimed to identify S100A6, a calcium-binding protein frequently upregulated in TP53-mutant PDAC, as a critical regulator of mutant p53 stability and tumor progression, and to explore potential S100A6-targeting agents for therapeutic intervention. Methods We integrated computer-assisted drug screening with transcriptomics, acetylation omics, and molecular biology techniques to identify Agrimol B (AgrB), a bioactive compound derived from the traditional Chinese herb Agrimonia pilosa Ledeb., as a potential S100A6-targeting agent. Results High S100A6 expression was closely associated with poor prognosis in patients with TP53-mutant PDAC, whereas S100A6 depletion markedly suppressed PDAC cell growth and metastatic potential. Mechanistically, AgrB enhanced the interaction between S100A6 and the deacetylase HDAC2, leading to reduced acetylation of mutant p53 at lysine 382. This disruption activated autophagy-dependent cell death and thereby inhibited PDAC progression. Conclusion Our findings reveal an S100A6-HDAC2-mutant p53 acetylation axis that regulates TP53-mutant pancreatic tumorigenesis, providing mechanistic evidence supporting S100A6 as a therapeutic vulnerability and highlighting AgrB as a promising natural-product-derived candidate for further development against this aggressive malignancy.

The development of dual inhibitors of histone deacetylases (HDACs) and enhancer of zeste homologue 2 (EZH2) is an efficient strategy that not only synergistically suppresses critical pathways in tumorigenesis but also circumvents the potential risks of drug cocktails. In this study, a series of pyridone derivatives were rationally designed via pharmacophore merging, and N1-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-N8-hydroxyoctanediamide (15c) was identified as the most potent compound against hematological tumor cells MV4-11 and SU-DHL-10, with IC50 values in the submicromolar range. 15c also effectively inhibited HDAC1 and EZH2 with IC50 values of 9.2 nM and 311.1 nM, respectively. Molecular simulations revealed key interactions between 15c and both targets. These findings indicated that compound 15c warrants further investigation as a novel dual HDAC/EZH2 agent.

Prenatal cannabis use is on the rise, and observational studies suggest that such use results in neurodevelopmental deficits in the offspring. Because observational studies can be confounded by unaccounted factors, we studied the neurodevelopmental consequences, at the molecular level, of prenatal cannabis use. We applied an integrated multi-omics approach, combining transcriptomics and global proteomics, to first trimester (T1) and second trimester (T2) human fetal brains from pregnancies with and without documented maternal cannabis exposure and no use of drugs of abuse. Prenatal cannabis exposure produced minimal molecular effects in female T1 fetal brains but induced pronounced system-level disruption in male T2 fetal brains. These disrupted pathways have molecular signatures linked to neurodevelopmental and neuropsychiatric disorders, including autism spectrum disorder, schizophrenia-related pathology, and disorders of cortical connectivity, raising significant concerns of prenatal cannabis use.

The interaction between the gut microbiome and drug metabolism is bidirectional and can influence the pharmacokinetics of certain drugs. In mice, the gut microbiome has been shown to influence Cyp3a11. However, evidence for microbial regulation of human CYP3A4 is lacking. We aimed to bridge this gap by manipulating the microbiome of a humanized mouse model expressing CYP3A4, CYP3A7, PXR and CAR. Three groups of male and female humanized mice were studied: conventional (CV), germ-free (GF), and germ-free mice conventionalized using sex-matched pooled human fecal samples (GFCV). The presence of microbiome upregulated CYP3A4 expression by 7.6-fold in male CV mice (p < 0.001) but downregulated CYP3A4 expression by 1.69-fold in female CV mice (p = 0.012) compared to GF mice. The human fecal microbiome transplant to sex-matched GF mice resulted in decreased microbial diversity (p < 0.05 in males and p < 0.01 in females) and was not effective in restoring CYP3A4 expression, suggesting complex underlying microbe-CYP3A4 interactions. We show that the hepatic CYP3A4 mRNA and protein expression were strongly correlated (R = 0.91; p = 2.6 x 10-6). A total of 57 bacterial species from the mouse gut microbiome were identified to be significantly correlated with CYP3A4 protein expression (p < 0.05). Five bile acids and no short chain fatty acids were correlated with CYP3A4 protein expression. In summary, alterations in the gut microbiome influenced hepatic CYP3A4 in humanized mice in a sex-dependent manner, with distinct microbes strongly correlating with this regulatory pattern. Significance Statement This study is the first to evaluate the expression of CYP3A4 under different microbial conditions in a humanized mouse model, including conventionalization of germ-free mice using pooled sex-matched human feces. Alterations in the gut microbiome influenced hepatic CYP3A4 in a sex-dependent manner and were strongly correlated with microbial species.

The interaction between the gut microbiome and drug metabolism is bidirectional and can influence the pharmacokinetics of certain drugs. In mice, the gut microbiome has been shown to influence Cyp3a11. However, evidence for microbial regulation of human cytochrome P450 3A4 (CYP3A4) is lacking. We aimed to bridge this gap by manipulating the microbiome of a humanized mouse model expressing CYP3A4, CYP3A7, pregnane X receptor and constitutive androstane receptor. Three groups of male and female humanized mice were studied: conventional (CV), germ-free (GF), and germ-free mice conventionalized (GFCV) using sex-matched pooled human fecal samples. The presence of microbiome upregulated CYP3A4 expression by 7.6-fold in male CV mice (P < .001) but downregulated CYP3A4 expression by 1.69-fold in female CV mice (P = .012) compared with GF mice. The human fecal microbiome transplant to sex-matched GF mice resulted in decreased microbial diversity (P < .05 in males and P < .01 in females) and was not effective in restoring CYP3A4 expression, suggesting complex underlying microbe-CYP3A4 interactions. We show that the hepatic CYP3A4 mRNA and protein expression were strongly correlated (R = 0.91; P = 2.6 × 10-6). A total of 57 bacterial species from the mouse gut microbiome were identified to be significantly correlated with CYP3A4 protein expression (P < .05). Five bile acids and no short-chain fatty acids were correlated with CYP3A4 protein expression. In summary, alterations in the gut microbiome influenced hepatic CYP3A4 in humanized mice in a sex-dependent manner, with distinct microbes strongly correlating with this regulatory pattern. SIGNIFICANCE STATEMENT: To the best of our knowledge, this study is the first to evaluate the expression of cytochrome P450 3A4 under different microbial conditions in a humanized mouse model, including conventionalization of germ-free mice using pooled sex-matched human feces. Alterations in the gut microbiome influenced hepatic cytochrome P450 3A4 in a sex-dependent manner and were strongly correlated with microbial species.

The Ganoderma fungus has been revered for centuries in traditional Chinese medicine and foods with high nutritional value for its purported health benefits.The extracts of Ganoderma applanatum(Pers.)Pat.showed promising protective activity against neurological damage caused by scopolamine in zebrafish.Eleven highly oxygenated lanostane triterpenoids with one or more α,β-unsaturated conjugated moiety,including three previously undescribed compounds appterpenactones A-C(1-3),and a nor-abietane diterpenoid,ganorphenol(4),together with related congeners were obtained from the fruiting bodies of G.applanatum.The structures and absolute configurations of the previously undescribed compounds were elucidated by analysis of nuclear magnetic resonance(NMR)spectra,quantum chemical calculations of NMR and electronic circular dichroism spectra,and X-ray crystallographic data.Compound 1 shares an unusual 6/6/5/6/5 ring system scaffold,while compound 4 represents a rearranged aromatic diterpenoid with rare ortho-dimethyl groups.All compounds were evaluated for their neuroprotective activity in glutamate-induced mouse hippocampal neuronal cell line(HT-22 cells).Ganorphenol(4)and applanoic acid C(6)demonstrated significant neuroprotective activity in inhibiting glutamate-induced HT22 cell death.Biochemical assays suggest that applanoic acid C may exert protective effects by activating the Nrf2/HO-1 pathway.Moreover,the further mechanistic study revealed that ganorphenol(4)can exert neuroprotective effects through the PI3K-AKT-mTOR pathway.Collectively,comprehensive experiments and molecular docking studies demonstrated that compounds 4 and 6 are potential neuroprotective agents.These findings provide valuable insights into the actions of trace bioactive components from edible materials and contribute to the broader understanding of neuroprotective compounds.

A phytochemical study of Ganoderma applanatum identified four predominant triterpenoids, with ganoapplanilactone C (GATC) exhibiting the most significant lipid-reducing effects in high-fat diet-fed zebrafish, surpassing atorvastatin at 5 μM. Histopathological analysis confirmed GATC’s protective effects on the liver against high-fat diet-induced damage. The Enzyme-Linked Immunosorbent Assay (ELISA) results showed a positive correlation between GATC treatment and liver health markers, as well as antioxidant enzymes, while they revealed a negative correlation with triglycerides and inflammatory cytokines. Metabolomic profiling demonstrates GATC’s impact on metabolites such as amino acids, fatty acids, and the mechanistic Target of Rapamycin (mTOR) signaling pathway, suggesting its role in regulating multiple metabolic processes. The increase in Adenosine Monophosphate-activated protein kinase (AMPK) phosphorylation in the GATC-treated groups indicates the activation of the AMPK/mTOR pathway, a key mechanism in lipid metabolism and liver protection. Molecular docking studies highlighted the importance of GATC’s spirocyclic ketone system and hydroxyl group in binding to target proteins. These findings underscore GATC’s potential as a therapeutic agent for metabolic dysfunction-associated steatotic liver disease (MASLD), emphasizing its superior efficacy compared to other triterpenoids due to its unique C-23 spiro 5/7 system. This study provides valuable insights into the prevention and treatment of MASLD using G. applanatum-derived compounds.

Background P. ginseng is prized for its nutritional and medicinal properties, primarily due to the presence of ginsenosides. During thermal processing, ginsenosides undergo complex chemical reactions, including hydrolysis and elimination, generating rare ginsenosides with enhanced bioactivity. Deciphering these reaction patterns and mechanisms is crucial for optimizing processing techniques. Methods P. ginseng samples were processed at various temperatures and for different durations, and ginsenoside content was analyzed using UPLC. Thermodynamic parameters were calculated using Gaussian software, and molecular mechanics and frontier orbital theories were applied to predict reaction sites and explore mechanisms. Simulated reactions of reference solutions were used to verify the quantitative rules. Results Significant variations in ginsenoside content were observed across different temperatures and durations. Rb1 transformation peaks under moderate heat, selectively forming Rg5. Rg1 was readily transformed, producing Rh4 under harsh conditions and Rk3 under mild ones, allowing for product-specific tuning. Thermodynamic calculations revealed that most reactions had negative Gibbs free energy changes (ΔG), indicating spontaneity, with ΔG decreasing at higher temperatures. The HOMO-LUMO energy gaps of ginsenosides were less than 0.4, indicating high ginsenoside reactivity, and the energy difference between the LUMO of ginsenosides and the HOMO of H2O influenced hydrolysis reaction rates. Conclusions This study provides valuable insights into the chemical reaction patterns of ginsenosides during thermal processing. Temperature significantly impacts reaction direction, extent, and product selectivity. The findings establish a mechanistic foundation for optimizing P. ginseng processing conditions to achieve the desired ginsenoside profiles.

We previously showed that THC is effluxed in the perfused human placenta cotyledons, but surprisingly this efflux was not inhibited by valspodar, a P-gp and BCRP inhibitor. P-gp and BCRP have multiple binding sites, and therefore THC may be binding to a transport site not blocked by valspodar. To test this hypothesis, we perfused human placenta cotyledons with THC in the absence and presence of a cocktail of P-gp and BCRP inhibitors. The inhibitor cocktail significantly increased the unbound maternal-to-fetal THC clearance index, indicating that P-gp and/or BCRP are likely involved in determining fetal THC exposure.

Atopic dermatitis (AD) is a systemic immune disease that primarily affects infants and children, characterized by recurring severe pruritus and chronic eczema. Studies have demonstrated that histone deacetylase 6 inhibitors (HDAC6is) can exhibit anti-inflammatory activities by regulating the acetylation level of target proteins. Building on these findings, our research focused on a synthetic diphenylpyrimidine derivative, specifically 15b, which we identified as a potent HDAC6i and an effective anti-inflammatory agent. This designation was determined by its safety profile, HDAC6 inhibitory activity, selectivity, and its anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In 2,4-dinitrochlorobenzene (DNCB)-induced AD mice, daily intraperitoneal injections of 15b significantly alleviated symptoms such as skin edema, dryness, crusting, and peeling, and reduced the frequency of scratching. Moreover, 15b mitigated ear swelling, addressed the increase in epidermal thickness, and reduced mast cell infiltration. Further mechanistic studies revealed that 15b selectively inhibited HDAC6, enhanced the acetylation of α-tubulin and heat shock protein 90 (HSP90) in RAW264.7 cells and BALB/c mice back skin tissue, and attenuated the activation of TLR4/MAPK, STAT3, NF-κB pathways. Consequently, both inflammatory cytokines (IL-4 and IFN-γ) and proteins (iNOS and COX-2) were dose-dependently decreased. These findings suggest that the HDAC6 inhibitor 15b can serve as a potential anti-inflammatory agent for the treatment of AD.

Targeting the WDR5-MLL1 protein-protein interaction (PPI) is considered to be an effective approach for the treatment of MLL-rearranged leukemia. However, interfering with WDR5-MLL1 PPI reduces methylated H3K4 levels and induces a decline in acetylated H3 levels, which may contribute to the suboptimal cellular efficacy of WDR5 inhibitors. We observed that cotreatment with WDR5-MLL1 PPI and HDAC inhibitors augmented the antiproliferative effect in MV-4-11 cells. Thus, a series of dual-target inhibitors was developed by merging the pharmacophores of the WDR5 and HDAC inhibitors. Among the developed inhibitors, compound 32d displayed an 89-fold increase in antiproliferative efficacy and induced potent cell apoptosis by impeding the DNA damage repair signaling pathway. Furthermore, the administration of 30 mg/kg of compound 32d was well tolerated, inhibiting MV-4-11 xenograft growth by 87.1%. Our investigation established the therapeutic effectiveness of the developed WDR5-MLL1/HDAC dual-target inhibitor against acute myeloid leukemia, providing a valuable tool for further exploration of crosstalk between the two targets.

Three new compounds (1-2, 4) along with ten known ones (3, 5-13), were isolated from the whole plant of Erigeron breviscapus. Compounds 1 and 2, two novel C-10 acetylenic acids and compound 4, a jasmone glucoside were elucidated by the detailed analysis of 1D and 2D NMR, HRESIMS spectra, and experimental and calculated electronic circular dichroism (ECD). Compounds 1-3 represent the first example of acetylenic acids incorporating C-10 skeleton from E. breviscapus. In addition, the antioxidant effects of all compounds were evaluated by ferric reducing power, 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate acid) (ABTS) and 2.2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. Our results indicated the significant antioxidant activity of caffeoylquinic acids. Additionally, compounds 10-11 and 13 played protective role on alcoholic liver injury cells in a dose-dependent manner.

Δ9-Tetrahydrocannabinol (THC) is the primary psychoactive component of cannabis which is being increasingly consumed by pregnant people. In humans, THC is sequentially metabolized in the liver to its circulating metabolites 11-hydroxy-THC (11-OH-THC, psychoactive) and 11-nor-9-carboxy-THC (THC-COOH, non-psychoactive). Human and macaque data show that fetal exposure to THC is considerably lower than the corresponding maternal exposure. Through perfused human placenta studies, we showed that this is due to the active efflux of THC (fetal-to-maternal) by a placental transporter(s) other than P-glycoprotein or breast cancer resistance protein. The identity of this placental transporter(s) as well as whether THC or its metabolites are substrates or inhibitors of hepatic solute carrier transporters is unknown. Therefore, we investigated whether 5 μM THC, 0.3 μM 11-OH-THC, and 2.5 μM THC-COOH are substrates and/or inhibitors of placental or hepatic solute carrier transporters at their pharmacologically relevant concentrations. Using HEK cells overexpressing human OATP1B1, OATP1B3, OATP2B1, OCT1, OCT3, OAT2, OAT4, or NTCP, and prototypic substrates/inhibitors of these transporters, we found that THC and THC-COOH were substrates but not inhibitors of OCT1. THC-COOH was a weak substrate of OCT3 and a weak inhibitor of OAT4. THC, 11-OH-THC, and THC-COOH were found not to be substrates/inhibitors of the remaining transporters investigated.