Current Pharmaceutical Design - Online First

Citrus reticulata (CR) has a long-standing role in traditional medicine, primarily due to its bioactive constituents such as hesperidin and narirutin, which are known for their antioxidant, anti-inflammatory, antibacterial, and anticancer properties.

This study investigates the anti-proliferative activity of CR water extracts against DU-145 prostate cancer cells and explores the therapeutic potential and underlying molecular mechanisms of hesperidin and narirutin in prostate cancer (PCa) and benign prostatic hyperplasia (BPH) through network pharmacology and molecular docking approaches.

Cytotoxicity assays were employed to determine the anti-cancer efficacy (IC50) of processed CR water extracts in DU-145 cells. Targets related to hesperidin, narirutin, PCa, and BPH were identified using bioinformatics platforms. Network pharmacology was applied to construct compound-target interaction networks and perform enrichment analyses (GO, KEGG, and DisGeNET) to elucidate key signalling pathways. Molecular docking was conducted to validate compound-target interactions.

Soil-processed CR extracts exhibited the strongest anti-cancer activity (IC50 = 1.789 mg/mL). Enrichment analyses identified significant pathways, including AGE-RAGE signalling, p53 signalling, inflammation, angiogenesis, and apoptosis. Molecular docking confirmed strong binding affinities of hesperidin and narirutin to the predicted targets.

Based on in vitro assays and in silico analyses, processed Citrus reticulata peel extracts may exert beneficial effects in both prostate cancer and benign prostatic hyperplasia. The soil-processed water extract demonstrated the most significant potential. The results suggest that the major compounds may act on several key proteins and pathways related to apoptosis and inflammation. However, further experimental and in vivo studies are needed to confirm their efficacy and safety.

Anti-proliferative assays, network pharmacology, and molecular docking collectively demonstrate that hesperidin and narirutin from CR show strong therapeutic potential against PCa and BPH. The findings highlight the involvement of AGE-RAGE and p53 signalling pathways and support the potential of these compounds in future drug development.

The current study aimed to synthesize and identify the biological activities of pyrazine-piperidine amide pharmacophore derivatives against non-small lung carcinoma (Calu-6) cells.

The combinatorial formulation was prepared by an active mixture of different chemical substituents, and five (6A-E) different molecules were synthesized. The chemical structures were confirmed by Fourier transform infrared (FT-IR) spectroscopy and proton nuclear magnetic resonance (H1) spectroscopy.

These compounds were also screened for cytotoxicity against the Calu-6 cell line. Compounds 6B and 6D displayed potent cytotoxicity, with IC50 Values of 45.21 µM and 89.64 µM, respectively. Cellular uptake and apoptotic studies using compound microscopy and flow cytometry revealed that cell damage gradually increased, leading to cell death. Compound 6B at 25 µM and 50 µM had 75.3% and 65.3% viability, 8.61% and 9.85% apoptotic effects, 12.05% and 21.4% late apoptosis, and 4.02% and 3.4% necrosis, respectively.

Compound 6B was found to significantly enhance cell cycle arrest at the G2/M phase. Additionally, real-time RT-PCR and western blot analyses further confirmed the enhanced expression of apoptotic markers, such as caspase-3 and 8, as well as the antiproliferative gene p53.

These findings indicate that compound 6B has a promising anticancer effect on lung cancer.

Jiuzhuan Huangjing Pills (JHP) have been shown to exert therapeutic effects on metabolic dysfunction-associated fatty liver disease (MAFLD), with a stronger intervention effect than single herbs. The purpose of this study was to elucidate the chemical constituents and mechanisms of JHP and its raw materials, Polygonati Rhizoma (PR) and Angelicae Sinensis Radix (ASR), in the treatment of MAFLD.

Serum pharmacochemistry and metabolomics were performed to examine drug-derived and endogenous components in MAFLD rats. In addition, network pharmacology was used to predict the key active components and targets of JHP, PR, and ASR in MAFLD mitigation, followed by molecular docking. ELISA kits were used to detect the levels of LCAT, GPCPD1, NNMT, NMRK1, ADO, and CSAD in liver tissues, while Western blotting was applied to determine the expression of CYP7A1 and CYP27A1.

A total of 22, 8, and 10 compounds from JHP, PR, and ASR, respectively, were identified in serum. Meanwhile, 15, 5, and 7 compounds from JHP, PR, and ASR, respectively, were detected in rat tissues. Moreover, 157, 131, and 114 differential metabolites involved in 27, 6, and 9 pathways were found to be altered by JHP, PR, and ASR, respectively. JHP, PR, and ASR regulated LCAT and GPCPD1 in glycerophospholipid metabolism. JHP and ASR regulated NNMT and NMRK1 in nicotinic and nicotinamide metabolism. JHP further regulated ADO and CSAD in taurine and hypotaurine metabolism, as well as CYP7A1 and CYP27A1 in primary bile acid biosynthesis. Ten components of JHP acted on 12 targets to regulate 12 pathways in MAFLD treatment. Three components of PR acted on seven targets to regulate four pathways, while five components of ASR acted on five targets to regulate three pathways. The binding energies between these drug-derived compounds and their targets were all less than −5 kcal·mol⁻¹.

These findings provide a theoretical foundation for the clinical application of JHP in MAFLD and underscore the value of traditional Chinese medicine formulas in addressing complex metabolic diseases through synergistic regulation. However, the intervention effects of JHP-derived components on MAFLD and their potential mechanisms of action on specific targets and metabolites require further investigation.

Our study found that JHP was associated with more components, targets, and pathways, which may be the mechanisms of JHP synergism.

For centuries, Traditional Chinese Medicine has been a subject of extensive research for its healing properties, including its effects against viruses. The pollen of Typha angustifolia emerges as a notable natural source of antiviral agents, with earlier investigations focusing on its antioxidant and anti-inflammatory properties, which are associated with flavonoids and phenolics that facilitate electron transfer. These bioactive compounds could potentially disrupt viral entry and replication, thereby necessitating further studies.

Molecular docking analysis was conducted on 11 compounds from T. angustifolia targeting the entry protein of dengue virus, the NS5B polymerase of hepatitis C virus, and the RdRp of Japanese encephalitis virus. The binding affinity was evaluated through LibDock score assessments, and simulations of molecular dynamics (RMSD and RMSF) were performed to analyze the stability of the complexes.

Naringenin was consistently identified as one of the highest binders for all three viral proteins, achieving the top score for the RdRp of Japanese encephalitis (129.288). Isorhamnetin showed the greatest binding affinity for the hepatitis C NS5B polymerase (120.827), exceeding that of sofosbuvir (120.629), while isorhamnetin-3-O-rutinoside displayed strong binding to the dengue viral entry protein (97.0838). Molecular dynamics confirmed the stability of ligand-protein interactions, underlined by sustained van der Waals and electrostatic forces.

These findings underscore naringenin as a versatile antiviral candidate, with other flavonoids exhibiting specific effectiveness that could facilitate multitarget inhibition approaches. This polypharmacological potential of flavonoids aligns with their established antiviral properties, although confirmatory experimental studies are critical.

Naringenin emerged as the most potent and reliable antiviral agent among the compounds of T. angustifolia, particularly against the RdRp of Japanese encephalitis. These computational insights validate T. angustifolia pollen as a promising natural antiviral resource, warranting further validation through in vitro and in vivo studies.

Myocarditis (MC) is an inflammatory cardiomyopathy with high morbidity and mortality. Current treatment options for MC have limitations and side effects, necessitating the exploration of new therapies. Traditional Chinese Medicine (TCM), particularly Huaiqihuang Granules (HQH), has shown promise due to its anti-inflammatory, antioxidative, and anti-apoptotic properties. However, the application in cardiovascular diseases remains underexplored.

We employed network pharmacology, molecular docking, and Molecular Dynamics (MD) simulations to evaluate HQH’s effects on MC. This involved identifying bioactive components and therapeutic targets, conducting enrichment analyses, and performing molecular docking and MD simulations to validate the interactions between HQH components and MC-related targets.

A total of 57 bioactive components in HQH and 143 potential therapeutic targets for MC were identified. Enrichment analyses revealed that HQH’s potential treatment effects on MC involve various processes and pathways, including response to lipopolysaccharide, peptidase activity, the extracellular region, and pathways in cancer. Molecular docking indicates that Physalin A, sibiricoside A_qt, zhonghualiaoine 1, and methylprotodioscin_qt, along with ALB, PTGS2, AKT1, ESR1, and MMP9, may serve as key therapeutic components and targets. MD simulations confirmed strong interactions between HQH’s core components and MC-related targets, supporting their potential therapeutic effects.

This study suggests that HQH exerts therapeutic effects against MC through multi-target mechanisms and stable targets. These findings provide valuable insights into alternative treatment strategies for MC, offering a foundation for further research and clinical exploration.

This study confirms that HQH can influence MC through various active components and multiple therapeutic targets.

Diabetic Nephropathy (DN) is a Chronic Kidney Disease (CKD), and its main pathological changes are renal tubular injury and glomerulosclerosis. Semen Ziziphi Spinosae (SZS) is the seed of Ziziphus jujuba var. spinosa (Bunge) Hu ex H.F. Chow. As a triterpene saponin, Jujuboside A (Ju A) is the main active substance isolated from SZS. This study sought to investigate the potential effect and mechanism of Jujuboside A against DN.

The anti-apoptotic effects of Ju A on renal parenchymal cells of DN were examined by in vivo and in vitro studies. Molecular docking and Molecular Dynamics (MD) simulation revealed that Ju A could bind to TNF-α and Caspase-3 via forming stable receptor-ligand complexes, respectively. Immunofluorescence (IF) staining and ELISA detection were carried out to investigate the potential mechanisms by which Ju A exerted its amelioration effect on DN.

Our study showed that, accompanied by the restored renal function, Ju A inhibited apoptosis of renal tubules and glomeruli in vivo and in vitro. Network pharmacology revealed that 42 overlapping targets were related to Ju A and DN. Among them, IL6, IL1B, TNF, VEGFA, EGFR, ALB, IGF1, FGF2, CASP3, and ESR1 were the top 10 targets. Ju A could bind to TNF-α and Caspase-3 via forming stable receptor-ligand complexes, respectively, as demonstrated by molecular docking and MD simulation. Ju A decreased the protein levels of TNF-α and IL-1β in renal tubules and glomeruli of diabetic mice, and in HG-cultured HK-2 cells and podocytes, leading to the alleviation of inflammation. Besides, the up-regulated relative phosphorylation levels of NF-κB p65 and cleaved caspase-3 were also down-regulated by Ju A in vivo and in vitro.

The research showed that Ju A had a high affinity for Caspase-3 and TNF-α, and the underlying mechanism of Ju A against DN was the inhibition of apoptosis in renal tubular epithelial cells and podocytes. These findings strengthened the evidence that Ju A could be a potential treatment strategy for DN and offered opportunities for therapeutic advances in the field.

Ju A could inhibit apoptosis and alleviate inflammation of renal parenchymal cells by inactivating the TNF-α/NF-κB p65/Caspase-3 signaling pathway, exerting renal protective effect against DN.