Fungal pathogens are an escalating global public health concern, particularly in the context of invasive and opportunistic infections. Cryptococcosis, primarily caused by Cryptococcus neoformans var. grubii, can affect multiple organs and often leads to life-threatening meningitis in immunocompromised individuals. Given limited antifungal therapies and emergence of resistance and toxicity-related constraints, the development of novel anti-cryptococcal agents remains an urgent priority. A library of innovative 3-hydroxypyridin-4(1H)-one-based hybrids (5a-f) was synthesized and evaluated for antimicrobial activity against clinically relevant Gram-positive and Gram-negative bacteria, as well as fungal species Candida albicans and C. neoformans var. grubbi. Safety was assessed through cytotoxicity studies in HEK293 and HepG2 cells and hemolytic evaluation, while iron-chelating capacity and lipophilicity were also investigated. All compounds formed stable iron(III) complexes and displayed no significant toxicity up to 25 μM. Series 1 compounds (5a-c) were less lipophilic than Series 2 (5d-f), mainly due to regioisomeric position of hydroxyl group on 2-methyl-4-pyridone scaffold, whereas fluorination increased lipophilicity in both series. Notably, compounds 5c-f emerged as potent, selective, and nontoxic antifungal agents against C. neoformans var. grubii (MIC < 16 µg/mL; CC50 > 32 µg/mL; HC10 > 32 µg/mL), highlighting the potential of 3-hydroxypyridin-4(1H)-one-based hybrids as a promising approach for cryptococcal meningitis therapy.
Bioluminescence resonance energy transfer (BRET) is a technique based on energy transfer between a luminescent enzyme and a fluorescent acceptor. Since its first description in 1999, BRET has been widely applied in chemical biology research and drug discovery. Today, it is regarded as one of the most versatile methods for investigating the dynamics of ligand-protein and protein-protein interactions, both in cell-free systems and living cells. Owing to several advantages, including compatibility with high-throughput screening, suitability for cellular target engagement studies, and the ability to monitor real-time kinetics, BRET-based binding assays have become indispensable tools, particularly in early-stage drug discovery. Consequently, over the past decade these approaches have been extensively used for major drug target classes such as G protein-coupled receptors (GPCRs), kinases, and proteases. Despite the high relevance of ion channels as drug targets-approximately one-fifth of all approved drugs act on them-BRET-based assays have so far played only a limited role in this field. Here, we summarize recent applications of BRET-based methods for ion channels with a particular focus on approaches for studying ligand-ion channel interactions, highlighting current challenges and, more importantly, the significant potential of this technology to advance ion channel-focused drug discovery.
In recent years, the role of traditional Chinese medicine (TCM) in comprehensive cancer treatment has become increasingly prominent, gradually emerging as an important adjunctive modality alongside conventional therapies such as surgery, chemotherapy, and radiotherapy, and attracting widespread attention. Berberine, an isoquinoline alkaloid extracted from natural plants such as Coptis chinensis, possesses broad-spectrum and potent pharmacological activities. A large body of evidence has demonstrated that berberine exerts definite antitumor effects in various cancer models, with its mechanisms of action spanning multiple dimensions, including cell-cycle arrest, induction of cell death, modulation of the tumor microenvironment (TME), inhibition of invasion and metastasis, and intervention in angiogenesis. Based on the latest research progress worldwide, this review focuses on key oncogenic signaling pathways and immune regulatory mechanisms to systematically elucidate the cellular and molecular mechanisms underlying the antitumor activity of berberine, and further discusses its clinical application potential and the challenges ahead. Although the clinical translation of berberine remains constrained by bottlenecks such as formulation optimization and bioavailability enhancement, its broad prospects in the field of cancer prevention and treatment establish it as a highly valuable TCM-derived antitumor candidate for further development.
Calcium-dependent antibiotics (CDAs) represent an important class of antimicrobial agents active against Gram-positive pathogens. Olikomycin A, produced by Streptomyces ghanaensis ΔwblA, displays potent activity against multidrug-resistant bacteria; however, its molecular mode of action remained unclear. In this study, we investigated the interaction of olikomycin A with bacterial membrane lipids. Phospholipid antagonization assays demonstrated a calcium-dependent interaction with phosphatidylglycerol (PG), a major anionic phospholipid of Gram-positive membranes. Cryo-electron microscopy and fluorescence microscopy using model membranes revealed rapid membrane disruption induced by olikomycin A, resulting in vesicle deformation and fragmentation. Compared with the clinically used lipopeptide daptomycin, olikomycin A caused markedly faster and more extensive membrane damage. Furthermore, treatment of Staphylococcus aureus with olikomycin A in the presence of calcium resulted in rapid cellular aggregation consistent with severe membrane perturbation. These findings indicate that olikomycin A targets bacterial membranes via calcium-dependent binding to PG and disrupts membrane integrity through a mechanism distinct from daptomycin.
Among the human neuropeptide Y (NPY) receptors, the Y5 receptor (Y5R) represents a potential therapeutic target for the treatment of obesity and stress-related diseases. As the Y5R is expressed in various malignant tumors, it is also considered a potential target for cancer diagnosis and therapy. Although numerous non-peptidic Y5R antagonists with high binding affinity have been reported, only labeled peptidic agonists are used in routine Y5R competition binding assays, which are needed for the development and characterization of Y5R ligands. In the present study, three radiolabeled derivatives of the high-affinity Y5R antagonist CGP71683A (1) were synthesized ([3H]26, [3H]32, [3H]42) and studied in saturation binding assays using adherent or suspended HEC-1B-hY5R cells or membrane preparations thereof. Their Kd values (1.5-13 nM) were consistent with the Ki values (1.3-11 nM) of the non-labeled analogs. However, all synthesized radioligands exhibited very high non-specific binding, primarily caused by binding of the radioligands to cellular membranes, which limits their use as tool compounds in radiochemical Y5R binding assays. Nevertheless, a competition binding assay with [3H]32 and the endogenous Y5R agonist hNPY afforded a Ki that was in agreement with literature data.
Breast cancer remains the most prevalent malignancy among women worldwide and continues to present major therapeutic challenges, including drug resistance, systemic toxicity, and limited long-term efficacy despite advances in multimodal treatment strategies. These limitations have intensified interest in venom-derived peptides as structurally diverse bioactive scaffolds with anticancer potential. This review evaluates venom-derived peptides in breast cancer from a medicinal chemistry and translational drug development perspective. A structured narrative synthesis of the literature was conducted, covering preclinical and early translational studies of peptides derived from snakes, spiders, scorpions, and bees, with emphasis on structure-function relationships, mechanisms of action, and optimization strategies. Unlike previous reviews that primarily focus on individual venom sources, this work integrates mechanistic pharmacology with peptide engineering and translational constraints to provide a unified drug-development framework across breast cancer subtypes. Venom-derived peptides exert anticancer effects through ion channel modulation, apoptosis induction, cell cycle arrest, angiogenesis inhibition, membrane disruption, and immune modulation, with reported selectivity toward malignant cells in triple-negative, HER2-positive, and hormone receptor-positive models. Despite encouraging preclinical evidence, clinical translation is limited by instability, immunogenicity, poor pharmacokinetics, delivery barriers, and manufacturing challenges. Emerging advances in peptide engineering, nanodelivery systems, and artificial intelligence-assisted design offer promising strategies to improve pharmacological performance and drug-like properties. Overall, this review provides a medicinal chemistry-guided framework that identifies the key steps and remaining challenges required to support the future development of venom-derived peptides as anticancer lead scaffolds for breast cancer.
The interaction between the transcription factor c-Myb and the CBP/p300 KIX domain is a critical regulatory node in hematopoietic gene expression and an attractive target in MYB-dependent leukemia. Herein, we aimed to identify new small-molecule disruptors of the c-Myb-CBP/p300 KIX protein-protein interaction using integrated in silico and experimental workflow. A focused library of 1143 biaryl hydroxy-naphthamides (naphthanilides) was subjected virtual screening filters yielding five prioritized candidates (C1-C5). Docking showed that all selected compounds occupied the c-Myb-facing groove of the KIX domain and reproduced key interfacial contacts. Subsequent 1000 ns molecular dynamics simulations revealed distinct stability and disruption profiles, with C1 and C4 showing the most favorable combination of stable groove occupancy, protein-protein interface perturbation, and ligand-binding energetics. Free-energy landscape analysis further supported these compounds as the most conformationally stable interfacial binders. Differential MM/PBSA analysis of the protein-protein interface showed that C1 produced the largest predicted weakening of the c-Myb-KIX interaction (ΔΔGPPI = +38.81 kcal/mol), followed by C4 (+29.90), C5 (+27.78), and C3 (+25.81 kcal/mol), whereas C2 was predicted to stabilize the complex (-5.41 kcal/mol). In silico ADMET profiling indicated that the series was drug-like by Lipinski criteria but carried liabilities related to solubility, metabolism, and predicted toxicity. Experimental validation by microscale thermophoresis confirmed direct binding of the assay proteins (Kd = 26.72 ± 0.82 µM) and demonstrated that the naphthanilides disrupt the interaction in vitro. C1 was the most potent disruptor (IC50 = 9.50 ± 0.22 µM), outperforming Naphthol AS-E phosphate (IC50 = 32.84 ± 9.46 µM). These findings establish 3-hydroxy-2-naphthamides as promising scaffolds for targeting the c-Myb-CBP/p300 KIX interface.
Angiogenesis, regulated by vascular endothelial growth factor (VEGF), is crucial in tumor growth, metastasis, and inflammation. Leonurus japonicus Houtt., a traditional Chinese herb, was investigated for its anti-angiogenic potential. DNA sequencing confirmed its identity, distinguishing it from the common morphological misidentification with Leonurus sibiricus by the public. Using UHPLC-MS/MS and GNPS (Global Natural Products Social) molecular networking on the active methanol partition, over 6000 nodes were grouped into 273 clusters. Diterpenoids and flavonoids were targeted. Six compounds were successfully isolated from L. japonicus, including four labdane-type diterpenoids (1-4) and two flavonoids (5, 6) using column chromatography. Compound 1, (-)-8S-acetoxy-15,16-epoxy-8,9-seco-13(16),14-labdadiene, was a new stereoisomer, while Compound 6, nevadensin, was reported for the first time from this species. Compounds were characterized by NMR, ESI-MS, and ECD. At 50 μM, Compounds 1, 4, and 5 exhibited inhibitory effects in endothelial progenitor cell (EPC) tube formation. Molecular docking and dynamics simulations supported this anti-angiogenic activity. Compound 1 showed the highest VEGFR-2 binding affinity (-9.24 kcal/mol), comparable to sunitinib (-9.52 kcal/mol), further verified by an immunoblotting assay. This study demonstrated the successful integration of GNPS and bioactivity-guided isolation for identifying natural VEGFR-2 inhibitors. Our findings highlight Compounds 1 and 4 as promising lead candidates for anti-angiogenic therapy.
In this study, 20 novel biphenyl-sulfonamide-indole-based thiosemicarbazone derivatives (1-20) were synthesized and evaluated for their inhibitory activities against AChE, hCA I, and hCA II. Among the synthesized compounds, compound 20 (4-nitrophenyl substituted) exhibited the highest inhibitory activity against all three enzymes (AChE Ki = 54.24 nM; hCA I Ki = 12.75 nM; hCA II Ki = 8.72 nM) and behaved as a competitive inhibitor in enzyme kinetic studies. To further explore the experimentally observed AChE inhibition, induced fit docking (IFD), MM-GBSA calculations, and molecular dynamics simulations were performed for compound 20, suggesting a plausible binding mode within the AChE active site. In silico ADME analysis indicated generally favorable drug-like properties for the synthesized compounds. These findings identify biphenyl-sulfonamide-indole-based thiosemicarbazones as promising AChE and carbonic anhydrase inhibitors for further biological investigation.
Protein kinase CK2 is the subject of numerous studies in medicinal chemistry due to its involvement in the development of several diseases, primarily cancers. Its overexpression in tumor cells is related to key processes such as tumor immune evasion and cell proliferation. The scientific approach of this study aims to investigate the thermal shift assay (TSA) as a pre-screening tool and to complement it with a co-crystallization approach in post-screening. Therefore, the synthesis of seven small-molecule CK2 inhibitors derived from indeno[1,2-b]indoles was supplemented by 18 related derivatives from our in-house compound library. The 25 molecules belong to four sub-scaffolds, namely 4b,9b-dihydroxy-4b,5,6,7,8,9b-hexahydroindeno[1,2-b]indole-9,10-dione (D-0), 5,6,7,8-tetrahydroindeno[1,2-b]indole-9,10-dione (D-1), 9-hydroxy-5H-indeno[1,2-b]indol-10-one (D-2), and 5H-indeno[1,2-b]indole-6,9,10-trione (D-3). The most active CK2 inhibitors identified by capillary electrophoresis (CE)-based assay belong to the D-1 sub-scaffold. In the TSA, these compounds also generate significant shifts of the melting temperature (Tm) of CK2, indicating a clear correlation between the results of the CE-based assay and those of the TSA. The contribution of co-crystallization in post-screening also demonstrated the effectiveness of D-1 sub-scaffold compared with D-0 sub-scaffold.
ABSTRACT Chalcones are versatile scaffolds with broad pharmacological relevance, particularly as anti‐inflammatory and anti‐malarial agents. In the current investigation, a series of (3‐arylediene‐1‐phenyl‐1 H ‐pyrazol‐4‐yl)acryloyl‐4‐hydroxy‐6‐methyl‐2 H ‐pyran‐2‐one analogs/1,3‐diarylprop‐2‐en‐1‐ones were successfully synthesized, leveraging dehydroacetic acid (DHA) as the precursor molecule. The choice of DHA is strategic as it is a bioactive and highly versatile scaffold bearing a 4‐hydroxy‐6‐methyl‐2 H ‐pyran‐2‐one moiety, which enhances structural diversity and provides multiple reactive sites for functionalization. Its inherent anti‐microbial, anti‐inflammatory, and anti‐malarial potential offers an added advantage over conventional precursors, thereby increasing the likelihood of generating pharmacologically active chalcone hybrids. The synthetic protocol employed in this study involved the implementation of a Claisen condensation reaction, wherein DHA was reacted with formyl pyrazole to afford the corresponding chalcones, with yields ranging from moderate to good. The synthesized compounds were subjected to a rigorous characterization protocol, involving a battery of analytical techniques, including 1 H‐NMR, 13 C‐NMR, FT‐IR, and HRMS analyses. Biological evaluation focused on anti‐malarial and anti‐inflammatory activities, while molecular docking (PyRx 0.8) was performed to explore binding interactions with cyclooxygenase‐2 (COX‐2; PDB ID: 3LN1) and enoyl‐acyl‐carrier‐protein reductase (EACPR; PDB ID: 1NHG). Notably, compound 3c emerged as the most potent one within the synthesized series, exhibiting remarkable anti‐inflammatory and anti‐malarial potency, as evidenced by its IC 50 values of 7.05 ± 0.17 µM and 0.95 ± 0.06 µM, respectively. Docking studies revealed strong binding affinities and favorable molecular interactions, supporting the observed bioactivities. This study highlights chalcone–pyrazole hybrids as promising therapeutic scaffolds. In particular, compound 3c emerges as a compelling lead candidate for further optimization and preclinical investigation as a dual‐action agent against malaria and inflammation.
Chronic obstructive pulmonary disease (COPD) remains a significant global health challenge, which urges the discovery of novel drugs. In this article, we investigated the therapeutic potential and action mechanism of a new benzoxazolone derivative, 4-(5'-dimethylamino)-naphthalenesulfonyl-2(3H)-benzoxazolone (W3D), synthesized by our research team, against COPD both in vivo and in vitro. The results demonstrated that W3D could down-regulate inflammatory cytokines such as interleukin-6 (IL-6), interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), and MMP-9, thereby reducing airway inflammation and improving lung function, which together alleviated lung injury in COPD. Meanwhile, W3D increased the expression of tight junction proteins claudin-1 and occludin and attenuated the activation of the Toll-like receptor 4/nuclear factor kappa B (TLR4)/NF-κB) signaling pathway to maintain the integrity of bronchial epithelial cells. Additionally, W3D restored the expression of glycolytic enzymes such as LDHA, PKM2, and HK2 to modulate lactate levels, thereby correcting glycolytic pathway dysregulation. W3D decreased intracellular lactate content, down-regulated global Kla levels and H3K18la expression, and regulated macrophage polarization in cigarette smoke extract (CSE)-induced macrophages. Furthermore, these therapeutic effects of W3D were compromised in the presence of the glycolytic inhibitor 2-deoxy-d-glucose (2-DG), indicating that W3D regulated macrophage polarization by inhibiting glycolysis. Our results demonstrated that glycolysis was activated in macrophages exposed to CSE and served as a key role in the macrophage polarization process. Inhibiting glycolysis in macrophages might be a potential therapeutic direction for COPD. In addition, given the confirmed protective effect against COPD, W3D could serve as a promising lead compound for further structural modifications of innovative drugs.
New series of thiazole analogues were designed, synthesized, and tested for their potential anticancer activity. All the synthesized compounds were biologically tested against HCT-116 and MCF-7 cell lines. Compounds 14d, 14h, and 14i were determined to be the most active members in this series against HCT-116 cancer cell lines with a considerable safety profile. The three lead compounds were further investigated for their inhibitory activities against EGFR and CDK-2. Compound 14i exhibited the most potent inhibition, with IC50 values of 0.056 and 0.215 µM against EGFR and CDK-2, respectively, suggesting a possible association between kinase inhibition and the observed cellular activity. Compounds 14d, 14h, and 14i induced early apoptosis (14.40%-18.09%) and G2/M arrest (51.55%-66.15% population) in HCT-116 cells, with minimal necrosis. Additionally, the screened compounds upregulated pro-apoptotic factors (Bax, cytochrome c, and cleaved caspase-3), while downregulating pro-survival/anti-apoptotic markers (p-AKT1, Bcl-2) and the angiogenic factor VEGF, offering preliminary insight into the potential mechanism of action. EGFR and CDK-2 were among the most prominent genes identified in the network pharmacology analysis of the tested compounds. Molecular docking and molecular dynamics simulations suggested favorable binding modes of compound 14i within the active sites of EGFR and CDK-2, with stable interaction patterns observed during the simulation period. Furthermore, compounds 14d, 14h, and 14i showed encouraging in silico ADMET and drug-likeness profiles. Overall, these findings highlight the thiazole series as promising lead compounds with potential dual inhibitory activity, warranting further optimization and mechanistic validation.
K2P18.1 (TRESK) displays one of the most unusual two pore domain potassium channels, that is associated with different neurological and immune-related diseases emphasizing the therapeutic potential of K2P18.1 activators. Previous studies identified cloxyquin and nitroxoline as lead compounds that do not alter the function of other K2P channels. Although their 8-OH group is crucial for channel activation, 8-hydroxyquinolines are known for various effects including metal chelation and antibacterial properties increasing the risk of side effects. Therefore, we synthesized 14 quinoline derivatives to search for other tolerated substitutions in 8-position. Activity determination using two-electrode voltage clamp (TEVC) revealed that four derivatives achieved moderate inhibitory or stimulatory activity. Among these, benzaldimine 3d represents a new extended scaffold that unexpectedly increases K2P18.1 channel activity by about 60% at 100 µM. Docking studies suggest that despite the enlarged structure, 3d is still able to achieve in silico interactions with important amino acids from the cloxyquin binding site. However, its usability in vitro is limited by hydrolysis leading to a compound half-life of 39 min in aqueous solutions. Nevertheless, the identification of a possible scaffold extension for K2P18.1 channel activators opens a new opportunity for future compound diversification.
A new approach for the synthesis of marine natural product Sunshinamide has been developed, featuring macrolactonization as a key step. The important synthetic strategy is to first construct the unique 8-membered cyclic dithiazocane in the earlier stage and finally carry out macrolactonization according to Shiina's method. Compared with the reported macrolactamization of a linear precursor, the macrolactonization bearing the cyclic dithiazocane scaffold resulted in a lower macrocyclization yield, implying that the cyclic dithiazocane might not offer a more favorable conformational constraint for macrocyclization.
Benzosuberone is a promising scaffold with a remarkably varied pharmacological profile that has been attracting significant interest in the potential development of anticancer agents. This review focuses on the historical interest, diversity of chemical structures, and different medicinal applications of benzosuberone derivatives, particularly as agents with potential anticancer activity. We investigated the relationship between structure modification and biological activity by focusing on the molecular hybrids of the present hit with various functionalities or heterocycles, which led to improved cytotoxicity against several cancer cell lines, and miscellaneous biological activities such as antimicrobial, antiviral, neurological, antiparasitic, anti-diabetic, and antitubercular activities. Detailed in vitro analyses showed promising cytotoxicity against breast, lung, and colon cancers, presenting IC50 values in the low micromolar and submicromolar ranges and low toxicity profiles concerning normal cells. Novel approaches, including the hybrid molecule, combinatory therapy, and nano-formulation, are suggested to go beyond the current obstacles and expand the applications of benzosuberone-based derivatives toward clinical applications. This combined study demonstrates that the benzosuberone scaffold is a privileged structure in the search for miscellaneous biological drugs with high therapeutic potential.
The KCa2.2 and KCa3.1 channels are fundamental regulators of membrane potential and calcium signalling and promising targets to treat diseases such as spinocerebellar ataxia and cancer. To fully exploit their therapeutic potential, and to continue studying their pathophysiological role, it is crucial to develop selective modulators for each of these two channels. Here, we present a computational study to identify the molecular determinants behind the selectivity of two recently reported KCa2.2 modulators, namely, N-(2,1,3-benzoxadiazol-4-yl)-3-(4-methoxybenzene-1-sulfonamido)benzamide and N-(2,1,3-benzoxadiazol-4-yl)-4-(trifluoromethyl)benzamide. We leveraged a protocol combining in silico mutagenesis, molecular dynamics simulations, and protein-ligand docking to analyse the pockets targeted by these ligands. We identified the Ser353/Pro245 substitution to be the main driver of the distinct pocket shapes in KCa2.2 and KCa3.1 channels, ultimately defining modulator selectivity. This approach provides novel insights into the structural differences of this binding site across potassium channel subtypes, proposing potential selectivity determinants of the modulators targeting this pocket.
Heterotetrameric KV7.2/3 potassium channels were retrospectively identified as the target of the analgesic flupirtine and the antiepileptic retigabine (ezogabine). Clinical utility of these agents ended after decades or 6 years, respectively, before their full scope was explored. Market withdrawals in 2017 and 2018 hampered research in other medical fields and left a gap for researchers using these compounds off-label in experimental indications and for patients with KV7.2/3 malfunctions. Failure of these drugs might be regarded as a reason to abandon this class of compounds, due to the notion that toxicity might be a class effect and to competitive markets. Yet, evidence accumulated that the toxicity of both compounds is not an inherent property of modulators of KV7.2/3 channels, but rather the result of an oxidation-sensitive metabophore/toxicophore. Second, epilepsy associated with KV7.2/3 channelopathies might rationally be addressed best with modulators of this validated drug target. Employing retrometabolic drug design to remodel the enzyme-labile carbamate structure and the central highly substituted ring, azetukalner (formerly encukalner, XEN1101) emerged from the unlucky forerunners. This structural analog of retigabine exhibits improved metabolic stability, increased potency, and enhanced blood-brain barrier penetration compared with its predecessor. This review details the synthesis, physicochemical properties, and clinical results of azetukalner.
A novel thiopyrano[2,3-d]thiazole derivative Les-3384 (rel-N-(4-chlorophenyl)-3-[(5aR,11bR)-2-oxo-5a,11b-dihydro-2H,5H-chromeno[4',3':4,5]thiopyrano[2,3-d]thiazol-3(6H)-yl]propanamide) was evaluated for its anticonvulsant action. Anticonvulsant activity in mice was assessed using seizure models induced by picrotoxin, thiosemicarbazide, strychnine, caffeine, camphor action, or maximal electroshock (MES). Les-3384 was administered orally at 100 mg/kg. For comparison, valproate, carbamazepine, and inosine were applied as standard AEDs. Seizure parameters and lethality were recorded in treated animals. Significantly increased latency to seizure onset (insert time), reduced seizure frequency, severity, and lethality in animal models induced by picrotoxin, strychnine, caffeine, and MES. Its efficacy was comparable to the action of reference drugs or exceeded their effects. However, no protective effect of Les-3384 was observed in thiosemicarbazide and camphor models. Molecular docking studies revealed high binding affinity of Les-3384 to the GABAA-receptor, suggesting that GABAergic modulation is a possible mechanism of action. The high effectiveness of Les-3384 in the models of strychnine-induced and caffeine-induced seizures is confirmed by the results of molecular modeling, which indicate the affinity of the studied compound toward glycine and adenosine receptors. The LD50 of Les-3384 for mice is 1208 mg/kg by intragastric administration. The compound's efficacy and safety profile support its further preclinical study as a potential antiepileptic agent with the polytropic mechanisms of action.
Understanding the complex connections between cellular mechanisms is crucial for developing effective cancer treatments. In this study, new Schiff base derivative triazine compounds (5-8) were synthesized and structurally characterized using various spectroscopic techniques to investigate their cellular and gene-protein-level effects in lung cancer and to develop anticancer activity strategies. The cytotoxic activities of compounds were evaluated against A549 and MRC-5 cell lines using the WST-8 assay. Moreover, cellular death mechanisms in lung cancer were investigated using methods such as qRT-PCR, ELISA, membrane array, and Flow cytometry. In lung cancer cells, compounds 5 (58.24%) and 6 (59.08%) were mildly effective in terms of cell viability, and these two compounds exhibited cytotoxic activity at a dose of 50 µM. Also, while an increase in the expressions of the p21, p27, and p53 genes was observed across all compounds in A549 cells, a decrease in the expressions of the GRP78, GRP94, AKT, RIPK1, CDK1, CDK2, HSP27, HSP40, HSP60, and HSP90 genes was observed. Besides, GRP78 and Caspase-3 were considerably increased by compounds 5 and 6. While Caspase-3, CHK1, P38, P53, and TRAILR-2 pro-apoptotic protein expressions increased, BCL-2, BCL-W, IGF-II, and NFKB anti-apoptotic protein expressions decreased for all compounds. The G0/G1 phase was increased by compounds 5 and 6. Although these compounds attenuated both the S and G2/M phases. Overall, the new triazines target dysregulated cell-cycle, ER stress, and apoptotic pathways, demonstrating strong therapeutic potential in lung cancer. Notably, compounds 5 and 6 exhibited cytotoxic and pathway-modulating activities, highlighting their promise as clinically relevant lead candidates.
