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Breast cancer remains a significant challenge in oncology, highlighting the need for alternative therapeutic strategies that target necroptosis to overcome resistance to conventional therapies. Recent investigations into natural compounds have identified 8,12-dimethoxysanguinarine (SG-A) from Eomecon chionantha as a potential necroptosis inducer. This study presents the first computational exploration of SG-A interactions with key necroptotic proteins-RIPK1, RIPK3, and MLKL-through molecular docking, molecular dynamics (MD), density functional theory (DFT), and molecular electrostatic potential (MEP) analyses. Molecular docking revealed that SG-A exhibited a stronger affinity for MLKL (-9.40 kcal/mol) compared to the co-crystallized ligand (-6.29 kcal/mol), while its affinity for RIPK1 (-6.37 kcal/mol) and RIPK3 (-7.01 kcal/mol) was lower. MD simulations further demonstrated the stability of SG-A within the MLKL site, with RMSD values stabilizing between 1.4 and 3.3 Å over 300 ns, indicating a consistent interaction pattern. RMSF analysis indicated the preservation of protein backbone flexibility, with average fluctuations under 1.7 Å. The radius of gyration (Rg) results indicated a consistent value of ~15.3 Å across systems, confirming the role of SG-A in maintaining protein integrity. Notably, SG-A maintains two critical H-bonds within the active site of MLKL, reinforcing the stability of the interaction. Principal component analysis (PCA) indicated a significant reduction in MLKL's conformational space upon SG-A binding, implying enhanced stabilization. Dynamic cross-correlation map (DCCM) analysis further revealed that SG-A induced highly correlated motions, reducing internal fluctuations within MLKL compared to the co-crystallized ligand. MM-PBSA revealed the enhanced binding efficacy of SG-A, with a significant binding free energy of -31.03 ± 0.16 kcal/mol against MLKL, surpassing that of the control (23.96 ± 0.11 kcal/mol). In addition, the individual residue contribution analysis highlighted key interactions, with ARG149 showing a significant contribution (-176.24 kcal/mol) in the MLKL-SG-A complex. DFT and MEP studies corroborated these findings, revealing that the electronic structure of SG-A is conducive to stable binding interactions, characterized by a narrow band gap (~0.16 units) and distinct electrostatic potential favourable for necroptosis induction. In conclusion, SG-A has emerged as a compelling inducer of necroptosis for breast cancer therapy, warranting further experimental validation to fully realize its therapeutic potential.

Euphorbia hirta is a plant belonging to the Euphorbiaceae family that is widely used in traditional medicine to treat various dermal diseases in tropical and subtropical countries. It has been found to have anti-inflammatory and wound-healing effects, which may make it a promising natural treatment for different medical conditions. The aim of the present study focuses on the investigation of the main active compounds associated with the wound-healing activity of plants. The plant powder of aerial part was sequentially extracted with n-hexane, chloroform, methanol, and water using the serial exhaustive extraction method.The extract with the best activities was further fractionated using various chromatographic techniques to isolate the active compounds responsible for wound healing. The isolated compounds were assessed for wound healing activity by measuring migration and proliferation against human dermal fibroblast cells. The structure elucidation of compounds was confirmed by IR, LC-MS, and NMR. These isolated compounds were subsequently identified as kaempferol and its glycoside, kaempferol-3-O-glucoside (astragalin). The findings also revealed that both isolated compounds were able to stimulate the cell population with no sign of toxicity and the highest cell migration ability shows after 48-hour treatment. This study provides new evidence to support the traditional use of E. hirta as a natural promising wound healer.

Introduction: Crocus sativus (C. sativus), known as saffron, has been recognized for its medicinal benefits since ancient times. The plant was first introduced to the Chongming Island region of Shanghai, China, in 1980. Chinese Saffron, derived from this plant, is one of China's most valuable traditional herbs, yielding about 1 ton in 2013. Traditional Chinese Medicine (TCM) has used this plant as an antianginal for a long time. The plant has been extensively studied for its pharmacological benefits in treating obesity and gastrointestinal problems. This study aimed to conduct a comprehensive phytochemical analysis of C. sativus stigma (saffron) ethanolic extract using LCMS/MS, quantify its major compound, i.e., trans-4-GG-crocin, and study its acute toxicity. In addition, this study aimed to investigate the effect of the extract on anti-obesity in a high-fat diet-induced zebrafish model. Methods: Before the anti-obesity study, we conducted a comprehensive profiling of saffron ethanolic extract using UHPLC-ESI-MS/MS in both positive and negative modes. Then, the principal constituent, i.e., trans-4-GG-crocin, was quantified using HPLC with a UV detector. The study was followed by an acute toxicity study for 72 h to evaluate the safety profile of this extract using five concentrations of extract, from 93.75–1500 mg/L. Six groups of wild-type zebrafish were then segregated according to food regimen, i.e., (1) control (5 mg artemia/fish/day), (2) overfeeding (60 mg of artemia/fish/day), (3–6) high-fat diet (5 mg artemia+30 mg egg yolk/fish/day), then starting week 9–16, groups 4–6 were received (4) 50 mg/L, (5) 100 mg/L and (6) 200 mg/L of saffron extract. Results: Negative mode analysis revealed twenty phytoconstituents, while twenty-two were found in positive mode LCMS/MS analysis. The extract contained 536.91 µg/g of trans-4-GG-crocin. The highest concentration tested in the acute toxicity killed 100% of the testing animals within 24 h, and probit analysis showed that the LC50 of the extract was 1021 mg/L. Eight weeks of treatment with 100 and 200 mg/L of saffron significantly reduced the body mass index of the high-fat diet-induced zebrafish model (p

Acetylcholinesterase (AChE) inhibitors remain the primary therapeutic drug that can alleviate Alzheimer’s disease’s (AD) symptoms. Several Cassia species have been shown to exert significant anti-AChE activity, which can be an alternative remedy for AD. Cassia timoriensis and Cassia grandis are potential plants with anti-AChE activity, but their phytochemical investigation is yet to be further conducted. The aims of this study were to identify the phytoconstituents of C. timoriensis and C. grandis and evaluate their inhibitory activity against AChE and butyrylcholinesterase (BChE). Two compounds were isolated for the first time from C. timoriensis: arachidyl arachidate (1) and luteolin (2). Five compounds were identified from C. grandis: β-sitosterol (3), stigmasterol (4), cinnamic acid (5), 4-hydroxycinnamic acid (6), and hydroxymethylfurfural (7). Compound 2 showed significant inhibition towards AChE (IC50: 20.47 ± 1.10 µM) and BChE (IC50: 46.15 ± 2.20 µM), followed by 5 (IC50: 40.5 ± 1.28 and 373.1 ± 16.4 µM) and 6 (IC50: 43.4 ± 0.61 and 409.17 ± 14.80 µM) against AChE and BChE, respectively. The other compounds exhibited poor to slightly moderate AChE inhibitory activity. Molecular docking revealed that 2 showed good binding affinity towards TcAChE (PDB ID: 1W6R) and HsBChE (PDB ID: 4BDS). It formed a hydrogen bond with TYR121 at the peripheral anionic site (PAS, 2.04 Å), along with hydrophobic interactions with the anionic site and PAS (TRP84 and TYR121, respectively). Additionally, 2 formed three H-bonds with the binding site residues: one bond with catalytic triad, HIS438 at distance 2.05 Å, and the other two H-bonds with GLY115 and GLU197 at distances of 2.74 Å and 2.19 Å, respectively. The evidence of molecular interactions of 2 may justify the relevance of C. timoriensis as a cholinesterase inhibitor, having more promising activity than C. grandis.

Despite being widely used traditionally as a general tonic, especially in South East Asia, scientific research on Cassia timoriensis, remains scarce. In this study, the aim was to evaluate the in vitro activities for acetylcholinesterase (AChE) inhibitory potential, radical scavenging ability, and the anti-inflammatory properties of different extracts of C. timoriensis flowers using Ellman’s assay, a DPPH assay, and an albumin denaturation assay, respectively. With the exception of the acetylcholinesterase activity, to the best of our knowledge, these activities were reported for the first time for C. timoriensis flowers. The phytochemical analysis confirmed the existence of tannins, flavonoids, saponins, terpenoids, and steroids in the C. timoriensis flower extracts. The ethyl acetate extract possessed the highest phenolic and flavonoid contents (527.43 ± 5.83 mg GAE/g DW and 851.83 ± 10.08 mg QE/g DW, respectively) as compared to the other extracts. In addition, the ethyl acetate and methanol extracts exhibited the highest antioxidant (IC50 20.12 ± 0.12 and 34.48 ± 0.07 µg/mL, respectively), anti-inflammatory (92.50 ± 1.38 and 92.22 ± 1.09, respectively), and anti-AChE (IC50 6.91 ± 0.38 and 6.40 ± 0.27 µg/mL, respectively) activities. These results suggest that ethyl acetate and methanol extracts may contain bioactive compounds that can control neurodegenerative disorders, including Alzheimer’s disease, through high antioxidant, anti-inflammatory, and anti-AChE activities.

Seventeen methanol extracts from different plant parts of five different Cassia species, including C. timorensis, C. grandis, C. fistula, C. spectabilis, and C. alata were screened against acetylcholinesterase (AChE). C. timorensis extracts were found to exhibit the highest inhibition towards AChE whereby the leaf, stem, and flower methanol extracts showed 94–97% inhibition. As far as we are aware, C. timorensis is one of the least explored Cassia spp. for bioactivity. Further fractionation led to the identification of six compounds, isolated for the first time from C. timorensis: 3-methoxyquercetin (1), benzenepropanoic acid (2), 9,12,15-octadecatrienoic acid (3), β-sitosterol (4), stigmasterol (5), and 1-octadecanol (6). Compound 1 showed moderate inhibition towards AChE (IC50: 83.71 μM), while the other compounds exhibited poor to slightly moderate AChE inhibitory activity. Molecular docking revealed that the methoxy substitution of 1 formed a hydrogen bond with TYR121 at the peripheral anionic site (PAS) and the hydroxyl group at C5 formed a covalent hydrogen bond with ASP72. Additionally, the OH group at the C3′ position formed an interaction with the protein at the acyl pocket (PHE288). This possibly explains the activity of 1 in blocking the entry of acetylcholine (ACh, the neurotransmitter), thus impeding the hydrolysis of ACh.