SND1 is an oncoprotein found to be overexpressed in breast cancer, especially in triple-negative breast cancer(TNBC). In our previous study, a novel SND1-interacting peptide 4-2 was identified, exhibiting cytotoxicity to TNBC cells by inducing SND1 degradation. This study for the first time demonstrated the degradation of SND1 was proteasome-dependent. A series of peptide 4-2 derivatives were constructed using PROTAC technology. Among these, 4-2 VHL-recruiting PROTAC showed significantly increased SND1 degradation efficiency and higher anticancer activity to TNBC cells. The in vivo efficacy study suggested the D-isoform of 4-2VHL PROTAC suppressed the growth of TNBC cells in xenograft mouse model more effectively than peptide 4-2. Mechanistically, 4-2 VHL-recruiting PROTAC was demonstrated to induce pyroptosis of TNBC cells through Fas-mediated IL-17signaling. This study provides a new lead compound for the development of theSND1-targeted therapy via the proteolysis-targeting system.
Aberrant expression of mitochondrial quality regulation genes (MQRGs) is intricately linked to mitochondrial dysfunction and the progression of hepatocellular carcinoma (HCC), highlighting the urgent need for reliable prognostic biomarkers. In this study, we aimed to identify differentially expressed MQRGs from 20 candidate genes by analyzing transcriptomic profiles and clinical records from the TCGA (n = 371) and GEO (n = 167) datasets. The identified prognostic MQRGs, along with associated subtype differentially expressed genes (DEGs, n = 156), underwent LASSO and multivariate Cox regression analyses to construct a risk model, which was subsequently validated through time-dependent ROC analysis, Kaplan-Meier curves, and in vitro RT-qPCR. Our findings established a robust 4-MQRG signature comprising ANXA10, BAMBI, AKR1B15, and SPINK1, which revealed that patients classified as high risk had significantly shorter overall survival compared to their low-risk counterparts (p < 0.001). The predictive accuracy of this signature was noteworthy, yielding 1-, 3-, and 5-year AUCs of 0.725, 0.696, and 0.747 in the training cohort (n = 243) and 0.676, 0.627, and 0.592 in the testing cohort (n = 242), respectively. Furthermore, high-risk scores were associated with distinct immunosuppressive tumor microenvironments and varying sensitivity to systemic therapies. The dysregulated expression of the four genes was corroborated by RT-qPCR and analysis of the HPA database. In conclusion, this validated MQRG-based prognostic signature serves as an accurate tool for survival prediction and risk stratification, thus offering valuable biomarker support for personalized therapeutic approaches in HCC.
Dauriporphine is a monomer extracted from Menispermum dauricum DC, and it exhibits anti-cancer effect in non-small cell lung cancer (NSCLC). The regulatory mechanism of dauriporphine remains incompletely understood, and this study focused on its molecular targets in NSCLC progression. Cell viability, proliferation, apoptosis, invasion, migration, and stemness were evaluated using cell counting kit-8, ethynyl-2'-deoxyuridine assay, flow cytometry, transwell assay, scratch assay, and sphere formation assay, respectively. Bioinformatics analysis and weighted gene co-expression network analysis (WGCNA) were performed to identify targets of dauriporphine in NSCLC. The mRNA and protein expression was quantified using qPCR and Western blot. Co-immunoprecipitation was used to analyze protein interaction and ubiquitination regulation between ubiquitin-specific protease 7 (USP7) and calcium/calmodulin-dependent serine protein kinase (CASK). The role of dauriporphine in vivo was explored using xenograft tumor model. Dauriporphine restrained proliferation, invasion, migration, and stemness of NSCLC cells (p < 0.05). Bioinformatics analysis and WGCNA identified CASK as a core target of dauriporphine in NSCLC. CASK was highly up-regulated in NSCLC samples and cells (p < 0.05). Anti-cancer effects of dauriporphine on NSCLC cells were associated with reduced CASK expression (p < 0.05). USP7 stabilized CASK protein by inducing deubiquitination (p < 0.05). Silencing USP7 restrained NSCLC cell proliferation, metastasis, and stemness by inhibiting CASK (p < 0.05). Dauriporphine interacted with USP7, and then USP7 overexpression reversed the inhibition of dauriporphine in NSCLC cell malignant behaviors (p < 0.05). Dauriporphine reduced tumor growth in vivo and down-regulated USP7 and CASK expression (p < 0.05). This study suggested that dauriporphine blocked the key malignant phenotypes of NSCLC cells via inhibiting USP7-mediated deubiquitination of CASK, thereby promoting its proteasomal degradation. The study elucidates a molecular mechanism underlying anti-tumor role of dauriporphine and providing potential targets for dauriporphine treatment.
Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder for which single-target therapies often provide insufficient benefit, motivating the development of multi-target-directed ligands (MTDLs). In this study, a novel series of benzimidazolone-based hybrids incorporating piperazine, coumarin, and triazole moieties was designed, synthesized, and evaluated for inhibitory activity against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and monoamine oxidase-B (MAO-B), as well as antioxidant potential via on-line HPLC-based assays. Structures were confirmed by 1H-NMR, 13C-NMR (APT), and elemental analysis. Several compounds showed notable, micromolar-range inhibitory activity, though less potent than the reference drugs. Kinetic analysis showed that the leading compounds inhibited their respective enzymes via a mixed-type mechanism. Compound 1 showed the strongest AChE inhibition (IC50 = 0.777 ± 0.014 µM), compound 9b the highest BChE inhibition (IC50 = 0.659 ± 0.005 µM), and compound 9c the most potent MAO-B inhibition (IC50 = 2.431 ± 0.003 µM). Compound 8a showed the highest CUPRAC copper-reducing capacity, while 7c displayed the strongest DPPH radical-scavenging activity. Liposomal formulations of 4c, 7c, and 8a exhibited enhanced antioxidant responses relative to their free forms. In silico ADME predictions (SwissADME) indicated favorable drug-likeness and blood-brain barrier permeation for the compact scaffold (compound 1) and the piperazine-based hybrids, whereas the larger bis-conjugated derivatives were limited by high polarity and molecular weight. Overall, these benzimidazolone-based hybrids represent promising multi-target candidates for AD drug development.
Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder characterized by progressive cognitive decline, amyloid-β (Aβ) accumulation, oxidative stress, and excitotoxicity. Ferroptosis and N-methyl-D-aspartate (NMDA) receptor activity,may be interconnected in the pathogenesis of Aβ accumulation and associated neurodegeneration in AD. However, the interplay between these pathways remains poorly understood and underexplored for therapeutic intervention against the AD. The review aims to explore the shared molecular triggers of ferroptosis and NMDA receptor overactivation, including the roles of iron, glutamate overload, calcium dysregulation, and reactive oxygen species (ROS) accumulation. We further highlighted the convergent consequences of these processes on mitochondrial dysfunction, lipid peroxidation, and their impact on Aβ pathology. Particular attention is given to P-glycoprotein (P-gp), an efflux transporter involved in the Aβ clearance at the blood-brain barrier, whose expression and function may be modulated by oxidative stress, iron homeostasis, and NMDA receptor signaling. Emerging evidence indicated that ferroptosis and NMDA receptor activity may disrupt P-gp function, thereby impairing Aβ clearance and promoting its accumulation in the AD. Overall, the review elucidates the molecular mechanisms linking ferroptosis and NMDA receptor overactivation and their impact on P-gp-mediated Aβ transport in the AD, providing integrated mechanisms and harnessing their potential for AD therapeutics.
Aberrant activation of Aurora A kinase causes mitotic spindle assembly, chromosome segregation, and cell cycle progression, leading to genomic instability as well as disruption of several tumor suppressors. Furthermore, ERK has largely emerged as a survival signaling pathway controlling cell proliferation, differentiation, and metastasis. Unfortunately, this pathway is overexpressed in most of the human malignancies. In efforts to develop innovative inhibitors targeting Aurora A/ERK signaling pathway, a novel series of thiadiazolo-, triazolo-pyrimidine hybrids have been designed, synthesized, and assessed for their ability to block Aurora A/ERK and induce apoptosis. Cytotoxicity of the synthesized hybrids was examined against MCF-7, HCT-116 and A549 cell lines. Among the synthesized hybrids, 9a, 9c, and 14b demonstrated higher cytotoxic action than alisertib and GDC-0994 against the MCF-7 and A549 cancer cell lines. IC50 values for these hybrids were 2.59 ± 0.13, 4.63 ± 0.25, and 5.77 ± 0.38, respectively, against MCF-7 cell line and were 3.61 ± 0.19, 3.85 ± 0.21, and 4.23 ± 0.15, respectively, against A549. The selected hybrids significantly suppressed p-Aurora A kinase level as well as p-ERK1/2 level and its upstream regulators p-SRC, p-c-RAF, p-MEK1/2; meanwhile, ERK downstream effectors FOXO3a level was upregulated, and c-Myc was downregulated, in a dose-dependent manner. The selected hybrids significantly decreased the expression of Bcl-2 protein while increasing the levels of p53, caspase-7, caspase-9, and Bax. They effectively induced pre-G1 phase, G0/G1 phase apoptosis, and G2/M phase arrest. The synthesized hybrids possessed favorable binding interactions in the molecular docking investigations as well as appropriate drug-like characteristics.
Cyclooxygenase (COX) plays a crucial role in the inflammatory response, making selective COX-2 inhibition a significant strategy for developing safer anti-inflammatory medications. Accordingly, developing new pharmacotherapies is a critical objective in anti-inflammatory drug discovery. In this study, a new pyrazole-linked pyridine derivatives 2-7 were synthesized through the reaction of the 4-(pyridin-1-yl)benzohydrazide derivative 1 with substituted-ketenes, activated olefinic dimers, and α,β-unsaturated carbonyl compounds. The synthesized derivatives were characterized and subsequently assessed in vitro for their inhibitory effects on COX-2. The results of the COX-2 inhibition assay revealed a broad range of activity, with IC50 values ranging from 0.70 ± 0.02 to 63.49 ± 2.32 µM. Among the derivatives, 3 and 7b demonstrated the most significant inhibitory effects, with IC50 values of 1.11 ± 0.04 and 0.70 ± 0.02 µM, respectively, compared to celecoxib (IC50 = 0.87 ± 0.03 µM). Furthermore, these promising derivatives exhibited COX-1 inhibitory activity, with IC50 values of 16.08 ± 0.54 µM (SIcox-2 = 14.48) and 2.77 ± 0.09 µM (SIcox-2 = 3.95), respectively, compared to indomethacin (0.199 ± 0.01 µM) and celecoxib (21.62 ± 0.73 µM). Given the extensive COX activity of 7b, further investigations were conducted, as it demonstrated immunomodulatory potential through the suppression of IL-6 (IC50 = 167.65 ± 6.65 pg/mL) and TNF-α (IC50 = 816.59 ± 28.01 pg/mL) compared to celecoxib's IC50 (129.50 ± 4.44 pg/mL) and (399.7 ± 13.7 pg/mL), respectively. Additionally, pyrazole-linked pyridine derivative 7b significantly reduced PGE2 levels to 539.49 ± 17.87 pg/mL and NO production to 4.230 ± 0.159 nM, compared to celecoxib 256.79 ± 8.50 pg/mL for PGE2 and 3.997 ± 0 nM for NO, respectively, suggesting that compound 7b can attenuate both cytokine and eicosanoid signaling, exhibit extensive anti-inflammatory activity, and may serve as a potential multi-target therapeutic agent. The molecular docking simulation revealed that compound 7b exhibited a significant binding affinity through various interactions. Finally, the DFT calculation indicated that 7b has a low energy band gap, which suggests lower hardness and higher softness that implies a greater ability to redistribute electron density during binding, favoring the formation of stable polar interactions with protein residues. Overall, these findings demonstrate that compound 7b is a promising multi-target anti-inflammatory lead drug with strong COX-2 inhibition and immunomodulatory properties. Moreover, further studies will concentrate on in vivo anti-inflammatory evaluation and pharmacokinetic profiling.
This research employs a molecular hybridization strategy to repurpose the pyrazoline scaffold 6a, 6c, 7a-c, 11a, and 12b, transforming it into a high-efficiency conjugate designed to tackle the multifaceted pathology of neuroinflammation and epilepsy. By integrating a selective phenoxyacetic acid moiety. Our findings identified compound 7c as a potential lead candidate for the development of novel anticonvulsant agents. In vivo trials demonstrated that 7c offers a superior therapeutic window compared to valproic acid, yielding 90% seizure protection in PTZ models and a remarkable 212.27% delay in seizure onset within the pilocarpine model, alongside 100% survival. Beyond mere symptomatic suppression, 7c re-engineers the hippocampal environment by slashing glutamate-driven excitotoxicity by 64.23% and silencing the cytokine-glial activation axis (TNF-alpha, IL-6, GFAP, and Iba-1). Supported by ADME profile confirming optimal BBB permeability and molecular docking indicating a robust binding affinity of -10.3 kcal/mol, this study positions 7c as a versatile, non-toxic, and repurposed-ready hybrid candidate for advanced neuroprotective intervention.
Diabetes mellitus (DM) related erectile dysfunction (ED) is a common complication in males. We aim to explore the specific regulatory mechanisms of TEA domain family member 1 (TEAD1) in DMED rats. Diabetic ED in rats was induced by streptozotocin and evaluated by the intracavernosal pressure response to electrical stimulation and the apomorphine test. Corpus cavernosum smooth muscle cells (CCSMCs) were isolated to explore the role of TEAD1 in phenotypic transformation. TEAD1 knockdown was performed in DMED rats and CCSMCs using the CRISPR/Cas9 technology. Hematoxylin-Eosin and Masson staining were applied to observe the penile tissue pathology. The CCSMC function was evaluated by detecting viability and apoptosis. Protein and gene expression levels were examined by Western blot and real-time quantitative polymerase chain reaction. The TEAD1 gene knockdown ameliorated erectile function in DMED rats. Knockdown of TEAD1 in CCSMCs of DMED rats enhanced their viability and inhibited their apoptosis, promoting the transformation of CCSMCs from a synthetic to a contractile phenotype. Furthermore, bioinformatics analysis identified the calcium signaling pathway as a candidate pathway warranting further experimental investigation. TEAD1 knockdown was associated with altered expression of calcium-related proteins and elevated intracellular Ca2+ levels. The calcium channel blocker Nimodipine could reverse the improvement of erectile function in DMED rats and the transformation of CCSMC phenotype caused by TEAD1 knockdown. TEAD1 knockdown promotes the transformation of CCSMCs from a synthetic to a contractile phenotype and is associated with modulation of calcium signaling-mediated contractile machinery, contributing to the relief of DMED.
Among adolescents worldwide, osteosarcoma (OS) is one of the most frequently occurring cancers. DSCAM-AS1, a recognized lncRNA, has been noted for its abnormal expression in the development of certain cancers, but its role in OS is still unclear. Quantification of DSCAM-AS1, miR-211-5p, and PDCD6 expression was carried out via qRT-PCR or western blotting in human osteosarcoma cell lines (HOS, MG63, U2OS, SaOS-2) and a normal osteoblastic cell line (hFOB 1.19) using quantitative real-time polymerase chain reaction (qRT-PCR) for RNA detection and western blotting for protein detection, respectively. Cellular proliferation, apoptosis, migration, and invasion in OS cell models were comprehensively evaluated through a combination of experimental approaches, including EdU incorporation assay, CFSE labeling coupled with flow cytometric analysis, as well as transwell invasion and migration assays. To shed light on the molecular mechanisms of interactions among pertinent RNA molecules, researchers conducted luciferase reporter assays, RNA pull-down experiments, and RIP assays. Expression of DSCAM-AS1, miR-211-5p, and PDCD6 was analyzed based on the GEO database, and their correlations were evaluated. The levels of DSCAM-AS1and PDCD6 were predominantly overexpressed, while miR-211-5p was apparently lowly expressed in OS cells. Functional loss-of-function experiments demonstrated that silencing (knockdown) of DSCAM-AS1 expression substantially diminished OS cell proliferation, migration, and invasion, while simultaneously promoting cellular apoptosis in vitro. Mechanistically. DSCAM-AS1 acted as an upstream factor for miR-211-5p and could increase the expression of miR-211-5p-targeted PDCD6. DSCAM-AS1 facilitated the advancement of OS by increasing PDCD6 levels through miR-211-5p sponging. This observation could shed light on a novel therapeutic target option for OS.
Tumor drug resistance and metastasis are leading causes of cancer‑related mortality, both of which are tightly governed by multiple signaling pathways. The AKT‑GSK‑3β axis is a critical regulator of tumor progression, mediating drug resistance and epithelial‑mesenchymal transition (EMT) through its downstream targets. Aberrantly activated AKT‑GSK‑3β signaling modulates the expression and degradation of the drug efflux pump P-gp, which expels chemotherapeutic agents, including paclitaxel (PTX), from cancer cells, resulting in chemotherapy failure and drug resistance. Moreover, hyperactivated AKT‑GSK‑3β signaling drives EMT, a key process closely linked to tumor metastasis and malignant progression. Dioscin (Dio), a natural steroidal saponin, exhibits significant anti‑tumor activity in multiple cancers. However, whether Dio reverses chemoresistance and inhibits tumor growth by targeting the AKT‑GSK‑3β pathway to promote P-gp degradation and suppress EMT remains elusive, which is the central focus of this study. To explore whether Dio enhances the sensitivity of drug-resistant cancer cells to PTX and inhibits cancer metastasis and the EMT process, as well as its potential mechanism(s). Paclitaxel-resistant TE-1/PTX and HeLa/PTX cells were subjected to SRB, colony formation, Rh123 accumulation, wound-healing, and Transwell assays to evaluate Dio's chemosensitizing, anti-EMT, and anti-metastatic effects. Network pharmacology, molecular docking, CETSA, and proteolysis assays verified a direct Dio-AKT1 interaction. Western blotting, Co-IP, and MG132 and MK2206 rescue experiments clarified AKT/GSK3β-dependent P-gp ubiquitin-proteasomal degradation and EMT suppression. HeLa/PTX xenograft models were generated; H&E staining and immunoblotting were used to assess tumor growth, biosafety, and intratumoral protein profiles for in vivo validation. Dio sensitized PTX-resistant cells to paclitaxel, increased intracellular Rh123 accumulation, and inhibited cell migration and invasion. Mechanistically, Dio directly bound AKT1 to suppress AKT/GSK3β signaling, promoted ubiquitin-proteasomal degradation of P-gp, and reversed EMT by upregulating epithelial markers and repressing mesenchymal markers and EMT transcription factors. In vivo, Dio restrained xenograft tumor growth with negligible systemic toxicity, and intratumoral expression patterns of AKT/GSK3β, P-gp, and EMT-related proteins mirrored in vitro findings. Dio has the potential to be a safe and effective agent for drug-resistant cancer therapy.
MDR ESKAPE pathogens are the leading cause of hospital-acquired infections (HAIs) that resist most antibiotics and form biofilms. Biofilm formation is dependent on the two-component system (TCS), which regulates virulence traits including adhesion to host tissues, evasion of innate immunity, the synthesis of exopolysaccharides, and antibiotic resistance. TCS sense environmental stimuli such as pH, osmotic pressure and antimicrobial peptides. They regulate gene expression to promote bacterial survival and persistence during infection. TCS represents a contemporary and novel pathway for the advancement of targeted, adjuvant-based options to fend off infections caused by ESKAPE pathogens. Targeting TCS may improve drug penetration and reduce resistance pressure, although challenges remain due to their structural conservation, regulatory complexity, and mutation-driven resistance. However, there are several limitations in targeting TCS for drug development like TCS not always being essential for bacterial viability, structural homologies between TCS are not sufficient for broad-spectrum inhibitors, sometimes TCS can be involved in complex and essential regulatory networks and can evolve resistance mechanisms by mutations. To further investigate this paradox, we implemented comprehensive computational and phylogenetic analyses of selected ESKAPE TCS. The findings from this investigation form the basis for our in-depth review of the significance and challenges of TCS as emerging drug targets.
A novel series of 5-chloro-N-alkyl-1',1″-dimethyl-4'-aryldispiro[indene-2,3'-pyrrolidine-2',3″-indoline]-1,2″(3H)-diones (4a-r) was rationally designed and synthesized via a one-pot multicomponent reaction of N-alkylated 5-chloroisatin derivatives, 2-(arylmethylidene)-2,3-dihydro-1H-inden-1-ones (2a-i), and sarcosine (3). To assess their potential therapeutic efficacy, the entire library of synthesized compounds was screened for its inhibitory profiles against both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), where most derivatives exhibited moderate to potent cholinesterase inhibitory activity. Notably, derivatives 4b, 4f, and 4o emerged as the most effective dual inhibitors, displaying strong potent activity against AChE (IC50 = 0.91 ± 0.02, 0.89 ± 0.01, and 0.74 ± 0.01 µM, respectively; donepezil IC50 = 0.68 ± 0.005 µM) alongside significant BChE inhibition (IC50 = 10.19 ± 0.16, 9.59 ± 0.08, and 9.11 ± 0.01 µM, respectively; donepezil IC50 = 2.97 ± 0.01 µM). The structure of the most active derivatives (4b, 4f, and 4o) was further confirmed by X-ray crystallographic analysis. In addition, antioxidant evaluation of compounds 4b, 4f, and 4o demonstrated that derivative 4f possessed the most superior radical scavenging profile (IC50 = 23.18 ± 0.33 µM), representing approximately a 5.5-fold enhancement in potency relative to ascorbic acid (IC50 = 128.20 ± 0.82 µM). Molecular docking studies revealed favorable binding interactions of the lead derivatives within the catalytic binding pockets of both AChE and BChE, with docking scores comparable to those of the standard inhibitor. Furthermore, in silico ADME profiling demonstrated promising pharmacokinetic behavior, characterized by robust gastrointestinal absorption and excellent predicted blood-brain barrier penetration, supporting the potential of these derivatives as promising multifunctional candidates for the development of anti-Alzheimer agents.
The current study devised and synthesized a novel class of pyrazole derivatives based on indole as possible inhibitors of cyclin-dependent kinase-2 (CDK-2). 1H NMR, 13C NMR, NOESY, HMQC, and elemental analysis were used to confirm the structural integrity of the synthesized compounds. Promising CDK-2 inhibitory activity was observed in biological assays, and numerous compounds exhibited sub-micromolar IC50 values. Compound 5 d outperformed the reference inhibitor Roscovitine (IC50 = 0.716 µM) as the most potent inhibitor (IC50 = 0.536 µM), followed by compound 9 g (IC50 = 0.675 µM). SAR analysis showed that the observed activity was significantly influenced by the electronic nature of the added substituents as well as the orientation of the indole bond, with brominated derivatives exhibiting greater potency. The antiproliferative activity of the most potent compounds against the cancer cell lines HepG2, HCT-116, and MCF-7 was further assessed. In addition to having an enhanced selectivity index for normal MCF-10A cells (SI = 8.00 vs. 4.96 for Roscovitine), compound 5 d had the greatest activity against MCF-7 cells (IC50 = 6.78 µM), surpassing Roscovitine (IC50 = 8.11 µM). According to mechanistic investigations, compound 5 d significantly reduced the S-phase population, markedly promoted apoptosis, and caused G1 and G2/M cell-cycle arrest. Additionally, the consistent binding of compound 5 d within the ATP-binding pocket of CDK-2 was confirmed by molecular docking and molecular dynamics simulations, and attractive drug-like and pharmacokinetic features, similar to those of Roscovitine, were demonstrated by in silico ADMET predictions. All of these results point to compound 5 d as a promising lead scaffold for developing potent CDK-2-targeted anticancer agents.
Breast cancer is one of the leading causes of cancer-related deaths among women worldwide, with estrogen receptor-positive (ER+) breast cancer being the most common subtype. Cyclin-dependent kinase 4/6 inhibitors (CDK4/6i) have become a crucial therapeutic approach for this type of cancer. However, ER+ breast cancer frequently develops resistance to CDK4/6i, limiting therapeutic efficacy. Alternative splicing is a key post-transcriptional regulatory mechanism that may drive such resistance. In this review, we comprehensively analyzed the literature on the molecular mechanisms and key signaling pathways associated with CDK4/6i resistance in ER+ breast cancer and introduced the role of alternative splicing, with a particular focus on its function in tumor drug resistance. Available evidence suggests that splicing dysregulation may influence resistance through multiple pathways, including cell-cycle control, epithelial-mesenchymal transition, growth factor and RAS/MAPK signaling, and immune-related programs. Recent work has also suggested that reduced expression of the splicing regulator NSRP1 may be associated with CDK4/6i resistance through altered NSD2 splicing and activation of interferon signaling, although this mechanism currently requires further independent validation. Collectively, these findings support alternative splicing as a promising but still evolving area of investigation in CDK4/6i resistance. This work provides deeper insights into the role of alternative splicing in CDK4/6i resistance and offers a theoretical foundation for developing novel therapeutic approaches. Future research may focus on developing drugs that precisely modulate specific splicing events or combining CDK4/6i with splicing modulators to reverse or delay resistance.
Non-small cell lung cancer (NSCLC) exhibits stem-like characteristics that drive tumor aggressiveness and treatment resistance. The molecular chaperone Glucose-Regulated Protein 78 (GRP78) is substantially elevated in NSCLC compared to normal tissues and cell lines. In clinical samples, GRP78 protein levels correlated with advanced tumor stage and lymph node metastasis. Pharmacological inhibition of GRP78 using BAPTA-AM and GRP78-IN-3 significantly suppressed NSCLC cell viability while promoting apoptosis. The inhibitors effectively attenuated cell migration by downregulating matrix metalloproteinases and reversed epithelial-mesenchymal transition by modulating key cadherin expression profiles. Importantly, GRP78 inhibition substantially reduced cancer stem cell markers and impaired spheroid formation capability. Genetic knockdown of GRP78 in A549 and H1975 cells recapitulated the effects of pharmacological inhibition, reducing cell migration, reversing EMT, attenuating stemness, and suppressing β-catenin signaling. Conversely, GRP78 overexpression in A549 and H1975 cells induced opposite effects, promoting mesenchymal markers, enhancing stemness proteins, and increasing spheroid size. Mechanistic studies revealed that this anti-tumor activity operates through suppression of β-catenin signaling mediated by SIX1, as evidenced by SIX1-overexpression rescue experiments. These findings demonstrate that GRP78 acts as a key regulator of NSCLC stemness and metastatic potential by coordinating epithelial plasticity and stemness pathways, positioning it as a promising therapeutic target for NSCLC treatment.
Nanotechnology has emerged as a promising avenue for producing nanomedicines as alternatives to conventional drugs. Many organic nanoparticles, such as liposomes used in formulations like Doxil, Onivyde, and Marqibo, are approved by the Food and Drug Administration (FDA) and instrumental in treating various types of cancers. However, due to the many challenges associated with these formulations, alternative delivery systems have been explored, particularly those derived from inorganic sources. In particular, bioinorganic nano-based delivery systems (BNDSs) such as gold, silver, platinum, silicon-based, metal oxide and hybrid nanoparticles have been found to be useful in treating cancer and infectious and noncommunicable diseases. This review presents the most commonly used BNDSs, providing an in-depth discussion on their use as therapeutic and imaging agents, the challenges associated with their use and current trends and future perspectives in their development for enhancing efficacy.
Drug-resistant bacteria such as Methicillin-resistant Staphylococcus aureus (MRSA) and Quinolone-resistant S. aureus (QRSA), are a growing problem, creating a need for the development of novel antimicrobial agents. In this study, we designed and synthesized two novel 1,4-naphthoquinone derivatives, LHN-1034 and LHN-1035, and evaluated their antibacterial efficacy. Both compounds exhibited great antimicrobial activity against a group of Gram-positive bacteria, including MRSA and QRSA, with Minimum Inhibitory Concentrations (MICs) ranging from 50 to 100 µM. Mechanism studies revealed that the antibacterial effect is oxygen-dependent. The compounds act as redox-cycling agents that induce intracellular reactive oxygen species (ROS), which disrupts cell membrane integrity and leads to cell lysis. In silico assessments predict low intestinal absorption and skin permeability, suggesting these compounds are suited for topical application. In conclusion, LHN-1034 and LHN-1035 are candidates for developing topical agents against resistant Gram-positive infections.
A focused library of 19 donepezil-linked chalcones (DLCs) was efficiently synthesised through microwave-assisted Claisen-Schmidt condensation and subsequently profiled for their inhibitory activities against cholinesterases (AChE and BuChE) as well as monoamine oxidases (MAO-A and MAO-B). The DLCs exhibited potent and selective inhibition of MAO-B, with IC50 values ranging from 0.019 to 18.98 μM, whereas activity toward MAO-A was moderate to low (IC50 = 0.81 to > 20 μM). Among the tested DLCs, DLC9 and DLC14 showed the highest MAO-B inhibitory potential with IC50 values of 0.054 ± 0.004 μM and 0.019 ± 0.0015 μM, respectively, and high selectivity indexes (> 370 and > 1052, respectively), whereas DLC12 displayed notable MAO-A inhibition (IC50 = 0.81 ± 0.035 μM). Kinetic and reversibility studies revealed that the selected two lead DLCs (DLC9 and DLC14) acted as mixed-type reversible MAO-B inhibitors, with Kᵢ values of 20.0 ± 2.83 nM and 10.0 ± 2.82 nM, respectively. Furthermore, IC50 values of AChE inhibitory activities ranged from 5.40 to > 40 µM, whereas those of BuChE inhibitory activity range from 4.30 to > 40 µM. DLC6 showed the best AChE inhibitory potential with IC50 values of 5.40 ± 0.29 µM, while DLC13 revealed effective BuChE inhibitory potential with an IC50 value of 4.30 ± 0.89 µM. Molecular docking studies performed on hMAO-A and hMAO-B revealed that DLC14 establishes favourable π-π stacking within the aromatic cage of hMAO-B and maintains complementary hydrophobic contacts along the substrate cavity, whereas DLC6 lacks this key interaction due to steric interference of the ethoxy substituent. Among the three most potent MAO-B inhibitors (DLC2, DLC9, and DLC14), DLC2 exhibited the most favourable microsomal stability with the longest half-life and lowest intrinsic clearance, whereas DLC14 showed comparable metabolic profiles in rat and human liver microsomes. Experimental BBB permeability assays were hindered by compound-membrane interactions; however, in silico predictions indicated satisfactory oral bioavailability and brain penetration for all three candidates. In the MPTP-induced rat model of parkinsonism, the selective MAO-B inhibitor DLC14 and the non-selective inhibitor DLC6 significantly improved motor deficits and behavioural impairments across open field, pole, bar, rotarod, and forced swim tests, with progressive improvements observed up to Day 28. Notably, DLC14 consistently outperformed DLC6 and demonstrated an efficacy profile comparable to that of Selegiline, highlighting its therapeutic potential as an antiparkinsonian agent. These results indicate that DLC14 is potent and selective MAO-B inhibitor and could serve as promising candidate for the treatment of neurodegenerative disorders, such as Parkinson's disease.
Vascular endothelial growth factor receptor-2 (VEGFR-2) plays a central role in tumor angiogenesis and remains a well-established therapeutic target in cancer treatment. In this study, twenty-six benzimidazole-based derivatives (3a-k, 5a-d, 8a-g, and 10a-d) were rationally designed to incorporate the essential structural and pharmacophoric features required for effective VEGFR-2 inhibition, followed by their synthesis and biological evaluation. In vitro kinase assays demonstrated that the synthesized derivatives inhibited VEGFR-2 with IC50 values ranging from 0.061 to 1.895 µM. Based on their enzymatic potency, the most active compounds (3f, 3h, 3j, 8d, and 8g) were further investigated for antiproliferative activity against HUVEC, MCF-7, and HepG2 cell lines. Among them, Compound 3f exhibited IC50 values of 13.34, 5.32, and 5.02 µM, respectively, showing activity comparable to sorafenib. Evaluation against normal MCF-10A and Vero cell lines indicated favorable biocompatibility and a reasonable selectivity profile. Mechanistic studies revealed that 3f induced G2/M-phase arrest, promoted apoptosis, and suppressed cell migration, accompanied by a reduction in VEGFR-2 protein expression levels. Molecular docking suggested that the synthesized derivatives adopt a binding mode consistent with Type II VEGFR-2 inhibitors, while in silico ADME and toxicity predictions supported an acceptable toxicity profile and drug-like characteristics.
