👤 Castelli S

Drug-drug interactions (DDIs) are a significant source of morbidity and adverse drug events (ADEs), particularly in situations of polypharmacy and complex medication regimens. While rules-based software integrated in electronic health records (EHRs) has demonstrated proficiency in identifying DDIs present in medication regimens, large language model (LLM) based identification requires thorough benchmarking and performance evaluation using high-quality datasets for safe use. The purpose of this study was to develop a series of performance benchmarking experiments specifically for LLM performance in identification and management of DDIs using a specifically curated clinician-annotated dataset of clinically-relevant DDIs. Show less

Early in the evolution of life, a proto-metabolic network was encapsulated within a membrane compartment. The permeability characteristics of the membrane determined several key functions of this network by determining which compounds could enter the compartment and which compounds could not. One key feature of known life is the utilization of right-handed d-ribose and d-deoxyribose sugars and left-handed l-amino acid stereochemical isomers (enantiomers); however, it is not clear why life adopted this specific chirality. Generally, archaea have l-phospholipid membrane chemistries and bacteria and eukaryotes have d-phospholipid membrane chemistries. We previously demonstrated that an l-archaeal and a d-intermediate membrane mimic, bearing a mixture of bacterial and archaeal lipid characteristics (a 'hybrid' membrane), displayed increased permeability for several key compounds compared to bacterial-like membranes. Here, we investigate if these membranes can drive stereochemical selection on pentose sugars, hexose sugars, and amino acids. Using permeability assays of homogenous unilamellar vesicles, we demonstrate that both membranes select for d-ribose and d-deoxyribose sugars while the hybrid membrane uniquely selects for a reduced alphabet of l-amino acids. This repertoire includes alanine, the plausible first l-amino acid utilized. We conclude such compartments could provide stereochemical compound selection matching those used by the core metabolism of life. Show less

Geological structures known as alkaline hydrothermal vents (AHVs) likely displayed dynamic energy characteristics analogous to cellular chemiosmosis and contained iron-oxyhydroxide green rusts in the early Earth. Under specific conditions, those minerals could have acted as non-enzymatic catalysts in the development of early bioenergetic chemiosmotic energy systems while being integrated into the membrane of AHV-produced organic vesicles. Here, we show that the simultaneous addition of two probable AHV components, namely nickel and amino acids, impacts green rust's physico-chemical properties, especially those required for its incorporation in lipid vesicle's membranes, such as decreasing the mineral size to the nanometer scale and increasing its hydrophobicity. These results suggest that such hydrophobic nano green rusts could fit into lipid vesicle membranes and could have functioned as a primitive, inorganic precursor to modern chemiosmotic metalloenzymes, facilitating both electron and proton transport in early life-like systems. Show less

In the healthcare industry, the ever-increasing volume of clinical trial data presents challenges for ensuring drug safety and detecting adverse drug reactions (ADRs). This study aims to address the challenge of accurately detecting Serious Adverse Events (SAEs) in pharmacovigilance, a critical component in ensuring drug safety during and after clinical trials. The key problem lies in the underreporting and delayed detection of Adverse Drug Reactions (ADRs) due to the heterogeneous nature of medical data, class imbalance, and the limited scope of traditional monitoring techniques. This study proposes a hybrid AI-driven framework that integrates structured (e.g., patient demographics, lab results) and unstructured data (e.g., clinical notes) to detect ADRs using advanced deep learning and NLP methods. The objective is to outperform traditional signal detection methods and provide interpretable predictions to aid clinicians in real-time. By leveraging advanced Machine Learning (ML) and Deep Learning (DL) techniques, including Random Forests, Gradient Boosting Machines, and Convolutional Neural Networks (CNNs), our model aims to identify potential ADRs across different patient subgroups. Through meticulous feature engineering and the application of techniques to address data imbalance, our model demonstrates improved accuracy and interpretability in predicting ADRs. The CNN model achieved an accuracy of 85 %, outperforming traditional models, such as Logistic Regression (78 %) and Support Vector Machines (80 %). These findings suggest that specific demographic and clinical factors significantly influence the likelihood of adverse reactions, offering valuable insights for targeted monitoring and risk mitigation strategies[11]. This research underscores the potential of predictive modeling to enhance pharmacovigilance efforts and ensure safer clinical trial outcomes.•The research methodology includes a comparison of supervised learning algorithms, such as Logistic Regression, Random Forest, Gradient Boost, CNN, and genetic algorithms, to identify patterns and anomalies in clinical trial data. BERT and GPT, were also employed to provide the functionality of textual interactions over medical data.•Performance metrics such as accuracy, precision, recall, and F1-score were systematically applied to evaluate each model's performance. Among the models tested, the CNN model with BERT achieved the highest accuracy, providing valuable insights into the potential of deep learning for enhancing pharmacovigilance practices.•These findings suggest that an inclusion of diverse clinical data when supplied to advanced ML and NLP techniques can significantly improve the detection of ADRs, leading to better alignment with the fundamental principles of Good Clinical Practice (GCP). Show less

Computational drug discovery is essential for screening potential treatments and reducing the costs and time associated with proposing or combining drugs for disease management. Despite the extensive research conducted in this field, it remains an emerging area, particularly with the advent of machine learning, deep learning, and large language models (LLMs). This systematic review examines the integration of machine learning and deep learning techniques in drug discovery, concentrating on three critical areas: drug-drug interactions (DDIs), drug-target interactions (DTIs), and adverse drug reactions (ADRs). The review analyzes over 100 papers published between 2020 and 2025, categorizing the methods into deep learning, machine learning, graph learning, and hybrid models. It highlights the transformative impact of natural language processing (NLP) and LLMs in extracting meaningful insights from biomedical literature and chemical data. Furthermore, this work introduces key databases and data sets widely utilized in drug discovery. Additionally, this review identifies gaps in the existing research, such as the lack of comprehensive studies that simultaneously address DDI, DTI, and ADR extraction, and it proposes a more holistic approach to fill these gaps. The paper concludes by thoroughly evaluating various models, underscoring their performance metrics. Show less

Claudin (CLDN) proteins are extensively studied due to their critical role in maintaining tissue barriers and cell polarity. However, significant gaps remain in understanding the functional mechanisms of their sequence motifs and the molecular mechanisms of their interactions with other tight junction proteins. This review systematically examines the multifunctional properties of the CLDN protein family from the perspectives of sequence and structure. During evolution, CLDN family members have developed highly conserved structural features, particularly key conserved sites within the first extracellular loop (ECL1) and the C-terminal PDZ-binding domain, which play a central role in regulating the barrier function of tight junctions, ion selectivity, and protein-protein interactions. Furthermore, the distribution pattern of acidic and basic amino acids in ECL1 has been shown to directly determine ion selectivity and paracellular permeability. Meanwhile, the assembly and functional stability of tight junctions are precisely regulated by the C-terminal PDZ-binding domain through its interactions with the ZO protein family. Additionally, the study further elucidates how CLDN proteins modulate critical signaling pathways governing cellular proliferation, survival, and permeability, thereby participating in diverse physiological and pathological processes. These insights have deepened the understanding of the functional diversity of CLDN proteins and provided a new theoretical basis for developing disease diagnostic markers and designing targeted treatment strategies based on CLDN proteins. Show less

N6-methyladenosine (m6A) is the most prevalent internal modification of eukaryotic mRNA and has emerged as a pivotal regulator of gene expression at the post-transcriptional level. In the tumor immune microenvironment, tumor-associated macrophages (TAMs) represent a highly plastic and heterogeneous population that profoundly influences cancer progression, immune evasion, and therapeutic response. Recent studies have uncovered that m6A modification, mediated by dynamic "writers," "erasers," and "readers," exerts critical regulatory effects on TAM differentiation, polarization, and functional reprogramming. By modulating the stability, translation, and decay of transcripts involved in inflammatory signaling, metabolic adaptation, and immune checkpoints, m6A shapes the balance between tumor-promoting (M2-like) and tumor-suppressive (M1-like) macrophage phenotypes. Moreover, dysregulation of m6A machinery in TAMs has been linked to the suppression of anti-tumor immunity and resistance to immunotherapy, highlighting its translational potential as a therapeutic target. This review summarizes current advances in understanding the roles and mechanisms of m6A modification in TAM biology, discusses its implications in tumor immunity, and outlines the challenges and opportunities of targeting the m6A-TAM axis for cancer treatment. Show less

Deep learning, a cornerstone of artificial intelligence, is driving rapid advancements in computational biology. Protein-protein interactions (PPIs) are fundamental regulators of biological functions. With the inclusion of deep learning in PPI research, the field is undergoing transformative changes. Therefore, there is an urgent need for a comprehensive review and assessment of recent developments to improve analytical methods and open up a wider range of biomedical applications. This review meticulously assesses deep learning progress in PPI prediction from 2021 to 2025. We evaluate core architectures (GNNs, CNNs, RNNs) and pioneering approaches-attention-driven Transformers, multi-task frameworks, multimodal integration of sequence and structural data, transfer learning via BERT and ESM, and autoencoders for interaction characterization. Moreover, we examined enhanced algorithms for dealing with data imbalances, variations, and high-dimensional feature sparsity, as well as industry challenges (including shifting protein interactions, interactions with non-model organisms, and rare or unannotated protein interactions), and offered perspectives on the future of the field. In summary, this review systematically summarizes the latest advances and existing challenges in deep learning in the field of protein interaction analysis, providing a valuable reference for researchers in the fields of computational biology and deep learning. Show less

Ferroptosis, the non-apoptotic, iron-dependent form of cell death is an unavoidable outcome and byproduct of cellular metabolism. Reactive oxygen species generation during metabolic activities transcends to Fe2+-induced lipid peroxidation, leading to ferroptosis. Cancer cells being highly metabolic are more prone to ferroptosis. However, their neoplastic nature enables them to bypass ferroptosis and become ferroptosis-resistant. The capability of cancer cells to reprogram its metabolic activities is one of its finest abilities to abort oxidative damage, and hence ferroptosis. Moreover, the reprogrammed metabolism of cancer cells, also associates with the radical trapping antioxidant systems to enhance the scavenging of ferroptosis and thereby tumor progression. Additionally, the TME, which is an inevitable part and regulator of carcinogenesis, presents an intricate cooperation with tumor metabolism to build an immuno-metabolic environment to regulate the sustenance of cell proliferation and survival. This review focuses on the current understanding of ferroptosis in carcinogenesis and its resistance acquired by cancer cells via several modulators including the radical trapping antioxidant systems, the reprogrammed metabolism, the TME, and intertwined role of cancer metabolism and tumor immunity. The reprogrammed metabolism section further comprehends the functional role of lipids, iron and glucose metabolism against ferroptosis defense separately. The affiliation of TME in ferroptosis regulation is further sectioned with reference to different immune cells present within the TME such as tumor-associated macrophages, tumor-infiltrating neutrophils, myeloid-derived suppressor cells, T-cells, natural killer cells, dendritic cells, and B-cells, modifying the TME in both pro and anti-tumorigenic manner. Subsequently, this review also discusses the convergence of immuno-metabolic environment in ferroptosis regulation, and eventually brings up research gaps in this context providing consequential and significant questions to explore for better understanding of the immuno-metabolic environment's role in driving ferroptosis resistance for anti-cancer treatment progress. Show less

Ferroptosis, a novel form of regulated cell death induced by the excessive accumulation of lipid peroxidation products, plays a pivotal role in the suppression of tumorigenesis. Two prominent mitochondrial ferroptosis defense systems are glutathione peroxidase 4 (GPX4) and dihydroorotate dehydrogenase (DHODH), both of which are localized within the mitochondria. However, the existence of supplementary cellular defense mechanisms against mitochondrial ferroptosis remains unclear. Our findings unequivocally demonstrate that inactivation of mitochondrial respiratory chain complex I (MCI) induces lipid peroxidation and consequently invokes ferroptosis across GPX4 low-expression cancer cells. However, in GPX4 high expression cancer cells, the MCI inhibitor did not induce ferroptosis, but increased cell sensitivity to ferroptosis induced by the GPX4 inhibitor. Overexpression of the MCI alternative protein yeast NADH-ubiquinone reductase (NDI1) not only quells ferroptosis induced by MCI inhibitors but also confers cellular protection against ferroptosis inducers. Mechanically, MCI inhibitors actuate an elevation in the NADH level while concomitantly diminishing the CoQH2 level. The manifestation of MCI inhibitor-induced ferroptosis can be reversed by supplementation with mitochondrial-specific analogues of CoQH2. Notably, MCI operates in parallel with mitochondrial-localized GPX4 and DHODH to inhibit mitochondrial ferroptosis, but independently of cytosolically localized GPX4 or ferroptosis suppressor protein 1(FSP1). The MCI inhibitor IACS-010759, is endowed with the ability to induce ferroptosis while concurrently impeding tumor proliferation in vivo. Our results identified a ferroptosis defense mechanism mediated by MCI within the mitochondria and suggested a therapeutic strategy for targeting ferroptosis in cancer treatment. Show less

Modern quantitative image analysis techniques have enabled high-throughput, high-content imaging experiments. Image-based profiling leverages the rich information in images to identify similarities or differences among biological samples, rather than measuring a few features, as in high-content screening. Here, we review a decade of advancements and applications of Cell Painting, a microscopy-based cell-labeling assay aiming to capture a cell's state, introduced in 2013 to optimize and standardize image-based profiling. Cell Painting's ability to capture cellular responses to various perturbations has expanded owing to improvements in the protocol, adaptations for different perturbations, and enhanced methodologies for feature extraction, quality control, and batch-effect correction. Cell Painting is a versatile tool that has been used in various applications, alone or with other -omics data, to decipher the mechanism of action of a compound, its toxicity profile, and other biological effects. Future advances will likely involve computational and experimental techniques, new publicly available datasets, and integration with other high-content data types. Show less

Ewing sarcoma (EwS) cell line culture largely relies on standard techniques, which do not recapitulate physiological conditions. Here, we report on a feasible and cost-efficient EwS cell culture technique with increased physiological relevance employing an advanced medium composition, reduced fetal calf serum, and spheroidal growth. Improved reflection of the transcriptional activity related to proliferation, hypoxia, and differentiation in EwS patient tumors was detected in EwS cells grown in this refined in vitro condition. Moreover, transcriptional signatures associated with the oncogenic activity of the EwS-specific FET::ETS fusion transcription factors in the refined culture condition were shifted from proliferative toward metabolic gene signatures. The herein-presented EwS cell culture technique with increased physiological relevance provides a broadly applicable approach for enhanced in vitro modeling relevant to advancing EwS research and the validity of experimental results. Show less

The aim of the UniProt Knowledgebase (UniProtKB; https://www.uniprot.org/) is to provide users with a comprehensive, high-quality and freely accessible set of protein sequences annotated with functional information. In this publication, we describe ongoing changes to our production pipeline to limit the sequences available in UniProtKB to high-quality, non-redundant reference proteomes. We continue to manually curate the scientific literature to add the latest functional data and use machine learning techniques. We also encourage community curation to ensure key publications are not missed. We provide an update on the automatic annotation methods used by UniProtKB to predict information for unreviewed entries describing unstudied proteins. Finally, updates to the UniProt website are described, including a new tab linking protein to genomic information. In recognition of its value to the scientific community, the UniProt database has been awarded Global Core Biodata Resource status. Show less

Rare diseases affect over 300 million people worldwide and are often caused by genetic variants. While variant detection has become cost-effective, interpreting these variants-particularly collecting literature-based evidence like ACMG/AMP PM3-remains complex and time-consuming. Show less

LitSense 2.0 (https://www.ncbi.nlm.nih.gov/research/litsense2/) is an advanced biomedical search system enhanced with dense vector semantic retrieval, designed for accessing literature on sentence and paragraph levels. It provides unified access to 38 million PubMed abstracts and 6.6 million full-length articles in the PubMed Central (PMC) Open Access subset, encompassing 1.4 billion sentences and ∼300 million paragraphs, and is updated weekly. Compared to PubMed and PMC, the primary platforms for biomedical information search, LitSense offers cross-platform functionality by searching seamlessly across both PubMed and PMC and returning relevant results at a more granular level. Building on the success of the original LitSense launched in 2018, LitSense 2.0 introduces two major enhancements. The first is the addition of paragraph-level search: users can now choose to search either against sentences or against paragraphs. The second is improved retrieval accuracy via a state-of-the-art biomedical text encoder, ensuring more reliable identification of relevant results across the entire biomedical literature. Show less

Lung cancer is most terrible cause of cancer-related death throughout the world. This study focused on the synthesis and characterisation of novel flavokawain A ruthenium-p-cymene complex and to investigate the chemotherapeutic activity against lung carcinoma via in silico, in vitro and in vivo approaches. The complex was characterised via several spectroscopic techniques. In vitro study including cell viability, transwell migration, Western blot and flow cytometric analysis have been executed on both A549 and NCI-H460 cells. The toxicological assessment was performed and subsequently anticancer activity of complex was evaluated in benzo[α]pyrene persuaded lung carcinoma in mice. The molecular docking study demonstrated the compound has greater binding ability with β-catenin, Akt, HER2 and PARP. Followed by the complex treatment, the downregulation of β-catenin, PI3K, Akt, HER2 and PARP were investigated by Western blot analysis and cell cycle arrest was determined through flow cytometry. The outcomes of in vivo experimentation represented fruitful restoration of typical lung architecture after complex treatment. Immunohistochemical analysis demonstrated the downstream of β-catenin/m-TOR/Akt and upstream of caspase-3 and p53 expression, thereby initiating apoptosis. The complex exhibited a potent chemotherapeutic activity via the alteration of tumour microenvironment by modulating PI3K/Akt/β-catenin/HER2/PARP transduction in correlates with apoptotic events in lung carcinoma. Show less

Ruthenium-based metallodrugs have garnered attention as a promising alternative for anticancer therapy, aiming to overcome chemoresistance and severe side effects linked to platinum-based drugs. However, ruthenium complexes tested in clinical trials to date have yielded unsatisfactory results. This study synthesized a positively charged ruthenium complex (Ru-2) that effectively penetrated cancer cells and exhibited superior cytotoxicity to cisplatin in vitro against cancer cell lines and organoids. Ru-2 selectively targeted mitochondria, disrupting their function by depolarizing mitochondrial membrane potential, elevating reactive oxygen species production, and impairing both oxidative phosphorylation and the tricarboxylic acid cycle. Furthermore, Ru-2 triggered endoplasmic reticulum (ER) stress and apoptosis. Integrative transcriptomic and proteomic analyses, performed using RNA sequencing and mass spectrometry, identified key molecular changes in cancer cells treated with Ru-2. For enhanced in vivo application, we developed a transferrin-based nanomedicine formulation, TF/Ru-2, incorporating Ru-2 into transferrin. In vivo studies demonstrated that both Ru-2 and TF/Ru-2 exhibited superior antitumor efficacy and improved biosafety compared to cisplatin. This study presents a novel ruthenium complex and a transferrin-based drug delivery platform with significant potential for future cancer therapies. Show less

Hypoxia, a hallmark of many solid tumors, is linked to increased cancer aggressiveness, metastasis, and resistance to conventional therapies, leading to poor patient outcomes. This challenges the efficiency of photodynamic therapy (PDT), which relies on the generation of cytotoxic reactive oxygen species (ROS) through the irradiation of a photosensitizer (PS), a process partially dependent on oxygen levels. In this work, we introduce a novel family of potent PSs based on ruthenium(II) polypyridyl complexes with 2,2'-bipyridyl ligands derived from COUPY coumarins, termed COUBPYs. Ru-COUBPY complexes exhibit outstanding in vitro cytotoxicity against CT-26 cancer cells when irradiated with light within the phototherapeutic window, achieving nanomolar potency in both normoxic and hypoxic conditions while remaining nontoxic in the dark, leading to impressive phototoxic indices (>30,000). Their ability to generate both Type I and Type II ROS underpins their exceptional PDT efficiency. The lead compound of this study, SCV49, shows a favorable in vivo pharmacokinetic profile, excellent toxicological tolerability, and potent tumor growth inhibition in mice bearing subcutaneous CT-26 tumors at doses as low as 3 mg/kg upon irradiation with deep-red light (660 nm). These results allow us to propose SCV49 as a strong candidate for further preclinical development, particularly for treating large hypoxic solid tumors. Show less

In this work, we have carefully designed and synthesized two Ru(II) metal complexes: [Ru(phen)₂(HMPIP)](PF₆)₂ (6a, where phen = 1,10-phenanthroline, HMPIP = 2-(2-hydroxy-3-methylphenyl-1H-imidazo[4,5-f][1,10]phenanthroline) and [Ru(bpy)₂(HMPIP)](PF₆)₂ (6b, where bpy = 2,2'-bipyridine). Using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to explore the cytotoxicity of 6a and 6b towards HepG2, B16, A549, SGC-7901, HCT116 and non-cancer LO2. The complexes exhibited cytotoxicity activity against HepG2 cells. The capacity of 6a and 6b to impede the proliferation and dissemination of cancer cells was evaluated by conducting proliferation and migration experiments and 3D model. The anticancer mechanism was investigated in detail. The utilization of cycle blocking assays revealed that 6a and 6b induced a G0/G1 phase arrest in HepG2 cells. The cellular uptake experiments show that the complexes enter the cell nuclei, then escape from the cell nuclei into the cytoplasm, finally accumulate in the mitochondria. Apoptosis assays and the examination of proteins indicated that the complexes were capable of efficiently inducing apoptosis in HepG2 cells. Additionally, the potential induction of autophagy-mediated cell death was explored. The observed reduction in glutathione (GSH) levels and glutathione peroxidase 4 (GPX4) expression suggested a disruption of redox homeostasis within cancer cells, an increment in malondialdehyde (MDA) amount, together with BODIPY staining experiment, confirm that 6a and 6b can induce ferroptosis. Interestingly, in a nude mouse model, 6a showed a significant suppression of tumor growth with an inhibition rate of 63.4 %, without causing any weight loss of mice. The studies on the mechanism show that 6a causes immune cell death, increase the amount of TNF-α and IFN-γ, reduce IL-10 content, which further activates immune response to increase CD8⁺ T cells to prevent tumor growth. Therefore, 6a inhibits the tumor growth through stimulating the immune response to increase CD8⁺ T cells. In addition, the experiments in vitro show that the complexes through inhibition of PI3K/AKT/mTOR signaling pathway and intrinsic mitochondria pathway to cause cell apoptosis. These results demonstrate that Ru(II) complexes may be potent anticancer candidates for HepG2 tumor. Show less

In this work, we studied six Ruthenium(II)-diphosphine compounds containing different mercapto ligands (N-S), with general formula [Ru(N-S)(dppm)₂]Cl (dppm=1,1-bis(diphenylphosphino)methane). These compounds were characterized by several techniques (NMR [¹H, ³¹P(¹H), and ¹³C], HRMS, IR, UV-Vis and XRD) and their purity confirmed by elemental analysis. DLS experiments revealed low diameters and polydispersity indexes, and positive log P values in n-octanol/PBS indicated their preference for the organic phase. In general, these compounds are stable in different media over 48 h. Cytotoxicity experiments revealed promising IC₅₀ values on A549 breast cancer cells, 0.48 μM and 0.80 μM for [Ru(mtz)(dppm)₂]Cl (1) and [Ru(mmi)(dppm)₂]Cl (2), respectively (mtz and mmi are 2-mercapto-2-thiazoline and mercapto-1-methylimidazole in their deprotonated form, respectively). Clonogenic and migration experiments indicated their antiproliferative and anti-migratory capacity. ICP-MS results indicated their cellular accumulation in the nucleus, with little amounts in mitochondria. No covalent DNA binding was observed by ICP-MS. JC-1 and cell Mito Stress test confirmed mitochondrial dysfunction, which was verified by mitochondrial membrane potential uncoupling and drastic alterations in the oxygen consumption rate. Taken together, our results provide crucial insights regarding the anticancer potential of ruthenium(II)-phosphine compounds. Show less

Researchers are increasingly focusing on developing target-specific, highly cytoselective, lipophilic, water-soluble Ru(II) arene complexes to mitigate the side effects of commercially available platinum-based anticancer drugs. In this context, we present novel Ru(II) arene complexes, (Ru1 and Ru1a-f), which are based on a 1,10-phenanthroline-substituted imidazolium core derivatized with alkyl (butyl(a), octyl(b), dodecyl(c)) or benzyl ((benzyl(d), 2,4,6-trimethylbenzyl(e), pentamethylbenzyl(f)) groups. The structures of these complexes were characterized using ¹H, ¹³C, ¹⁹F and, ³¹P nuclear magnetic resonance (NMR) spectroscopy, Fourier transform infrared spectroscopy, mass spectrometry and elemental analysis. The cytotoxic activities of Ru1 and Ru1a-f complexes were tested against the cancer cell lines MCF-7 and MDA-MB-231 and normal cell lines, such as MCF-10 A. The cell cycle distribution in the MCF-7 and MDA-MB-231 breast cancer cell lines after 72 h of incubation with IC₅₀ concentration of the complex Ru1c can validly inhibit cell growth in the G2/M phase. Flow cytometry analysis showed that the complex Ru1c induced apoptosis in MCF-7 and MDA-MB-231 breast cancer cells. Additionally, the binding mode of the complex Ru1c with Fish-Salmon DNA was examined using ultraviolet-visible spectroscopy. Interaction of Ru1c complex with bovine serum albumin was analyzed by absorption study. The stability of all complexes in the solvent was assessed using ¹H NMR spectroscopy. Additionally, quantitative determination of the total ruthenium level within the cells was performed by inductively coupled plasma mass spectrometry (ICP-MS). Molecular docking was performed to evaluate the interaction residues and docking scores of Ru1c and the reference drug cis‑platinum against CDK1, cyclin B1, Bcl-xL and Bcl-2 proteins. Show less

Title: The role of ancillary ligands on benzodipyridophenazine-based Ru(II)/Ir(III) complexes in dark and light toxicity against TNBC cells. Abstract: In this study, we investigated the impact of ancillary ligands on the anticancer activity of benzodipyridophenazine-based Ru(II) and Ir(III) complexes (Ru1, Ru2, Ir1, and Ir2). These metal complexes displayed three significant absorption bands attributed to the ligand-centered (LC) transitions, ligand-to-ligand charge transfer (LLCT), and metal-to-ligand charge transfer (MLCT). Binding studies of biomolecules were performed with the complexes along with the ligand, and it was found that after binding with Ru(II)/Ir(III), the properties of the ligands were enhanced. In vitro screening revealed that complex [(η5-Cp*)IrIIICl(κ2-N,N-benzo[i]dipyrido[3,2-a:2',3'-c])phenazine] (Ir1) exhibited the highest potency and selectivity (IC50 ∼ 2.14 μM, PI > 13) under yellow light irradiation. The photo-toxicity trend was Ir1 > Ru1 > Ir2 ≫ Ru2, which was found to be directly correlated with the singlet oxygen quantum yield (1O2). Chloro-substituted complexes (Ir1 and Ru1) were effective for hypoxic tumor treatment, particularly Ir1, which could generate high amounts of reactive oxygen species (ROS, type I PDT) in cells under photo irradiation. The high value of fluorescence quantum yield (fφ = 0.26) and significant emission at λ = 571 nm of Ir1 were certainly useful for bio-imaging applications. Colocalisation and DCFDA studies of Ir1 revealed that it can accumulate in the mitochondria, leading to depolarization of the mitochondrial membrane. These studies confirm that the complex Ir1 is a promising candidate for TNBC treatment in hypoxic tumors, with efficacy comparable to the current PDT drug Photofrin. Show less

In recent years, photodynamic therapy (PDT) and gas therapy (GT) have emerged as research hotspots due to their excellent cancer treatment efficacy. By combining the advantages of both, the simultaneous and controllable release of reactive oxygen species (ROS) and nitric oxide (NO) has become a possibility. This paper describes the design of two Ru(II) complexes, [Ru(bpy)₂(NFIP)](PF₆)₂ (Ru1, bpy = 2,2'-bipyridine, NFIP = 4-nitro-3-trifluoromethylaniline-1H-imidazo[4,5-f][1,10]phenanthroline) and [Ru(phen)₂(NFIP)](PF₆)₂ (Ru2, phen = 1,10-phenanthroline), through the integration of the polypyridyl ruthenium structure and a photoresponsive NO donor. The structures and purity of the complexes were confirmed by several methods, including ¹H NMR, mass spectrometry, elemental analysis, high performance liquid chromatography (HPLC) and UV-Vis absorption spectra. Both complexes were demonstrated to efficiently generate singlet oxygen (¹O₂) (Φ_Δ = 0.40 and 0.44 in phosphate buffered saline (PBS) for Ru1 and Ru2, respectively) and release NO under visible light irradiation. Upon light exposure, Ru2 exhibited significant phototoxicity against human cervical cancer HeLa cells. In vitro experiments indicated that Ru2 elevated the levels of ROS and NO in HeLa cells when exposed to light, resulting in mitochondrial impairment and caspase-mediated cell death. Overall, Ru2 proves to be a potent phototherapeutic compound, capable of producing ROS and NO, thus providing precision in cancer phototherapy. Show less

To overcome the undesirable side effects and acquired resistance associated with platinum-based chemotherapeutics, scientists are searching for alternative strategies involving novel metal-based compounds with improved pharmacological properties. Ruthenium complexes have emerged as prospective candidates to combat side effects and improve the selectivity of anticancer agents. In this work, a benzimidazole-based chelating ligand, HL (4-(1H-naphth[2,3-d]imidazol-2-yl)-1,3-benzenediol) with O and N donor atoms, was synthesized and used for complexation with ruthenium to obtain three Ru(II) arene complexes represented by [Ru(η⁶-p-cym)(L)Cl], [Ru(η⁶-p-cym)(L)(PPh₃)]⁺ and [Ru(η⁶-p-cym)(L)(PTA)]⁺ (where p-cym = p-cymene, PPh₃ = triphenylphosphine and PTA = 1,3,5-triaza-7-phosphaadamantane). The synthesized complexes were characterized using spectroscopic techniques. UV-Vis absorption spectroscopy and LC-MS were used to study the stability of the complexes in biological medium. Their lipophilicity was studied by calculating the partition coefficient in n-octanol and water. The complexes showed significant binding with biomolecules like albumin proteins and nucleic acids. All the complexes were found to be cytotoxic, with complex [Ru(η⁶-p-cym)(L)PPh₃]PF₆ exhibiting the highest anticancer activity. The mechanism of anticancer activity was attributed to the ability of the complexes to induce apoptosis and generate reactive oxygen species (ROS). The complexes also exhibited antimetastatic properties. Furthermore, complex [Ru(η⁶-p-cym)(L)PPh₃]PF₆ was loaded onto amine-functionalized mesoporous silica nanoparticles which led to an increase in its cytotoxic activity. Show less

In recent years, photodynamic therapy (PDT) has emerged as a promising alternative to classical chemotherapy for treating cancer. PDT is based on a nontoxic prodrug called photosensitizer (PS) activated by light at the desired location. Upon irradiation, the PS reacts with the oxygen present in the tumor, producing cytotoxic reactive oxygen species (ROS). Compounds with highly conjugated π-bond systems, such as porphyrins and chlorins, have proven to be excellent light scavengers, and introducing a metal atom in their structure improved the generation of ROS. In this work, a series of tetrapyrrole-ruthenium(II) complexes derived from protoporphyrin IX and the commercial drug verteporfin were designed as photosensitizers for PDT. The complexes were almost nontoxic on human gastric cancer cells under dark conditions, revealing remarkable cytotoxicity upon irradiation with light. The ruthenium atom in the central cavity of the chlorin ligand allowed combined mechanisms in photodynamic therapy, as both singlet oxygen and superoxide radicals were detected. Additionally, one complex produced large amounts of singlet oxygen under hypoxic conditions. Biological assays demonstrated that the ruthenium derivatives caused cell death through a caspase 3 mediated apoptotic pathway and via CHOP, an endoplasmic reticulum stress-inducible transcription factor involved in apoptosis and growth arrest. Show less

Title: Necrosis inducing tetranuclear Ru(II)-Re(I) metal complex for anticancer therapy. Abstract: Chemotherapy is one of the most widely used anticancer treatments worldwide. However, despite its clinical effectiveness, most chemotherapeutic agents are associated with severe side effects. To address this limitation, there is an urgent need for the development of novel anticancer agents. Among the promising alternatives, Ruthenium and Rhenium complexes have garnered significant attention in the scientific literature. This study proposes combining these two metal moieties into a single tetranuclear complex, bridged by a 2,2'-bipyrimidine ligand. Cytotoxicity tests revealed broad activity of the novel metal complex against multiple cancer cell lines. Mechanistic studies suggested that the complex induces cell death by necrosis. Further analyses demonstrated its ability to eradicate colon carcinoma tumor spheroids at micromolar concentrations. To the best of our knowledge, this represents the first example of a Ru(II)-Re(I) tetranuclear metal complex as an anticancer agent. Show less

The six mononuclear Schiff's base Ru(III) complexes viz., [Ru(BZP)(LA)₂].2NO₃ (MRA), [Ru(BZP)(LB)₂].2NO₃ (MRB), [Ru(BZP))(LC)₂].2NO₃ (MRC), [Ru(BZP))(LD)₂].2NO₃ (MRD), [Ru(BZP)(LE)₂].2NO₃ (MRE) and [Ru(BZP)(LF)₂].2NO₃ (MRF), were synthesized using of (BZP=2,6-bis(2-benzimidazolyl)pyridine and p-sub-benzylthiosemicarbazones (BTS) [(Sub=4-NO₂ (LA), 4-N(CH₃)₂ (LB), 4-Cl (LC), 4-OCH₃ (LD), 4-OCH₂Ph (LE), and 4-OH (LF)] as an ancillary ligands. The thiosemicarbazones ligands (LA-LF) were obtained by the condensation of p-substituted benzaldehyde and thiosemicarbazide. These complexes were characterized by elemental analysis, IR, ESR, ESI-MS, electronic absorption spectroscopy. The geometry was optimized by theoretical calculation using DFT and structure reveals that MRA-MRF adopt octahedral geometry. Further, the complexes were examined for anti-cancer against Leukemia cancer cell line K562 and shown significant responses to these cells. Moreover, DNA binding studies were conducted with all complexes MRA-MRF and the binding constant (K_b) were found i.e., 1.10×10⁴, 1.54×10⁴, 2.87×10⁴, 1.67×10⁴, 1.98×10⁴ and 1.59×10⁴, respectively. It was found that DNA binds in intercalation mode which is also validated by the docking studies. Show less

Title: Systematic Investigation of Coordination Chemistry in Iridium(III) and Ruthenium(II) Complexes Derived from Pyridyl-Amine Ligands and Their Anticancer Evaluation. Abstract: A systematic investigation of the coordination chemistry of iridium(III) and ruthenium(II) complexes synthesized from pyridyl-amine ligands was performed, focusing on how ligand steric hindrance and metal centers affect oxidation behavior, coordination modes, and biological activities. The study revealed that steric hindrance at the ligand's bridge carbon strongly influenced both oxidation behavior and coordination modes. Smaller substituents (e.g., H and Me) facilitated oxidation to form pyridyl-imine species under adventitious oxygen, whereas bulky substituents (e.g., i-Bu and mesityl) suppressed oxidation, yielding stable pyridyl-amine or 16-electron pyridyl-amido complexes. Moreover, iridium(III) complexes were more prone to oxidation than the corresponding ruthenium(II) complexes under similar conditions. The aqueous stability of the newly synthesized complexes was confirmed. Cytotoxicity assays demonstrated that most of the complexes exhibited notable anticancer potency against A549, HeLa and cisplatin-resistant A549/DDP cancer cells. Mechanistic studies suggested a redox-driven pathway involving the catalytic oxidation of NADH to NAD+, the elevation of ROS levels and depolarization of the mitochondrial membrane. Notably, pyridyl-amine complexes induced apoptosis, while 16-electron pyridyl-amido complexes did not, though both caused S phase cell cycle arrest. Additionally these complexes can inhibit A549 cell migration, suggesting their potential to reduce cancer metastasis. Show less

Bioenergetic therapy targeting mitochondrial bioenergy is a promising therapeutic strategy for cancer. However, its clinical efficacy is limited by the metabolic adaptability of tumor cells, as they can switch between glycolytic and oxidative phosphorylation metabolic phenotypes to maintain energy homeostasis. In this study, we discovered 1,8-naphthyridine-piperazine-dithiocarbamate ruthenium(II) polypyridyl complexes (RuL1) that enhanced energy deprivation by inhibiting the activity of mitochondrial complex I and III, thereby disrupting oxidative phosphorylation. Simultaneously, RuL1 inhibits glycolysis while unexpectedly activating antitumor immunity. This dual metabolic-immunological targeting resulted in enhanced anticancer activity against MGC-803 cells. To the best of our knowledge, RuL1 is the first ruthenium polypyridyl complex reported to achieve high anticancer activity through dual metabolic inhibition. Show less

A series of half-sandwich ruthenium^II and iridium^III complexes bearing hybrid sp³-N/sp²-N amine-imine bidentate chelating ligands were strategically designed and synthesized. Their structures were fully characterized by ¹H and ¹³C NMR spectroscopy, mass spectrometry, and single-crystal X-ray diffraction, revealing nonplanar five-membered metallacycles in representative complexes. The complexes exhibited potent cytotoxicity against A549 lung, HeLa cervical, and HepG2 liver cancer cell lines, with IC₅₀ values ranging from 0.88 to 4.98 μM, significantly lower than that of cisplatin. Notably, the amine-imine complexes displayed superior cytotoxicity compared to their α-diimine analogues. Mechanistic studies indicated that DNA binding is not the primary mode of action. Instead, these complexes selectively target mitochondria, induce mitochondrial membrane depolarization, elevate intracellular reactive oxygen species (ROS) levels, and trigger apoptosis. Additionally, they enter A549 cells through an energy-dependent pathway and effectively inhibit cancer cell migration in vitro. Show less