Pharmaceutical cocrystals have emerged as an effective strategy for enhancing the solubility and bioavailability of poorly water-soluble drugs. The successful design of cocrystals largely depends on the selection of suitable coformers, which requires an understanding of the molecular interactions governing cocrystal formation during the process. The objective of this study was to develop robust machine learning models for predicting Hansen solubility parameters of pharmaceutical cocrystal using molecular descriptors generated from COSMO-RS and group contribution methods. A dataset consisting of 181 samples and 86 input features was utilized, incorporating molecular descriptors related to hydrogen-bonding capability, van der Waals interactions, and structural functional groups. Data preprocessing included outlier detection using the Isolation Forest algorithm and feature selection through Sequential Floating Forward Selection (SFFS). Three tree-based ensemble learning models, namely Random Forest (RF), Extra Trees (ET), and Gradient Boosting Regression Trees (GBRT), were developed and optimized using the Dragonfly algorithm for hyperparameter tuning. The obtained results demonstrated that the Extra Trees model consistently outperformed the RF and GBRT models in predicting all three Hansen solubility parameters. For the three target outputs, the ET model achieved test-set R2 values of 0.9175, 0.8661, and 0.9815, respectively, while also exhibiting the lowest RMSE and MAE values. Feature importance analysis revealed that both group contribution descriptors and COSMO-RS-derived molecular properties play significant roles in determining the Hansen solubility parameters of coformers. Overall, the proposed machine learning framework provides an efficient and accurate approach for predicting Hansen solubility parameters and screening pharmaceutical cocrystals. The findings highlight the superior predictive capability of the Extra Trees model and demonstrate the potential of combining molecular thermodynamic descriptors with advanced machine learning techniques for pharmaceutical cocrystal design.
Antipsychotic use in people living with dementia has been linked to serious adverse outcomes. Guidelines recommend limiting antipsychotic treatment to short-term use (<12 weeks), followed by tapering and cessation. However, antipsychotic treatment in routine practice often exceeds the recommended duration. The effect of discontinuing antipsychotics on reducing adverse outcomes in practice remains unclear. We aimed to use linked primary and secondary care data in England to investigate whether tapering or abrupt discontinuation of antipsychotics, versus continuation, affected the risk of stroke, death, fracture, delirium, and pneumonia in people living with dementia. Using UK primary care data from the Clinical Practice Research Datalink, linked with hospital and mortality data from Jan 1, 1998, to March 31, 2021, we emulated two sets of target trials, one after 12 weeks of antipsychotic treatment and another after 24 weeks of antipsychotic treatment, to compare continuing treatment versus tapering treatment and continuing treatment versus abrupt discontinuation of treatment. Patients aged 65 years and older at incident dementia diagnosis with antipsychotic treatment duration of at least 12 weeks were included. Study outcomes were incidence of fracture, stroke, hospitalisation for delirium, hospitalisation for pneumonia, and all-cause mortality within 24 months. A negative control outcome of skin conditions was included to measure potential unmeasured confounding. A clone-censor-weight approach was used with a 6-month grace period allowed for discontinuing antipsychotics. Weighted pooled logistic regression models were used to estimate 24-month absolute risk differences (ARDs). 134 549 eligible patients had a new antipsychotic treatment after dementia diagnosis, of whom 24 822 (18·4%) had a treatment period of at least 12 weeks and 16 795 (12·5%) had a treatment period of at least 24 weeks and met the eligibility criteria. The mean age was 83·57 (SD 7·07) in the 12-week trial and 83·65 (SD 7·02) in the 24-week trial; 16 725 (67·4%) of 24 822 patients were female and 8097 were male in the 12-week trial and 11 523 (68·6%) of 16 795 patients were female and 5272 were male in the 24-week trial. Compared with continuation after 12 weeks of treatment, estimated risks of delirium and fracture under the tapering strategy corresponded to 24-month ARDs of -2·46% (95% CI -4·10 to -1·27) and -2·80% (-4·14 to -1·29), respectively. Estimated risks for stroke, pneumonia, and all-cause mortality were similar between strategies. No difference in risk was observed for the negative control outcome. Similar results were found after 24 weeks of treatment and in sensitivity analyses. Irrespective of method, antipsychotic discontinuation decreased the risk of delirium and fracture. Antipsychotics can be discontinued safely when treatment duration has exceeded guideline recommendations without increasing the risk of death, stroke, or pneumonia in those living with dementia. PharmAlliance Research Clusters for Doctoral Training.
Although third-generation epidermal growth-factor receptor (EGFR)-tyrosine-kinase inhibitors (TKIs) are standard first-line therapies for patients with advanced EGFR-mutated non-small-cell lung cancer (NSCLC), their effectiveness is often limited by the emergence of drug resistance and subsequent disease progression. Given the previously established clinical efficacy and adverse event profile of aumolertinib, we aimed to evaluate the efficacy and adverse event profile of aumolertinib in combination with platinum-based chemotherapy versus aumolertinib monotherapy as first-line treatment for patients with locally advanced or metastatic NSCLC patients with EGFR-sensitive mutations. The open-label, multicentre, randomised, controlled, phase 3 AENEAS2 trial was done across 60 hospitals in China. Patients aged at least 18 years with Eastern Cooperative Oncology Group performance status (ECOG PS) 0-1; treatment-naive; histologically or cytologically confirmed locally advanced or metastatic NSCLC harbouring EGFR-sensitive mutations (ex19del/L858R with or without other EGFR mutations) were eligible. Brain metastases were allowed if neurologically stable. Previous EGFR-TKI therapy was an exclusion criterion. Patients were randomly assigned (1:1) with block randomisation (block size of 6), stratified by EGFR mutation type and baseline brain metastasis, to receive aumolertinib monotherapy (110 mg orally once a day) or combination therapy (aumolertinib 110 mg orally once a day plus pemetrexed 500 mg/m2 intravenously with cisplatin [75 mg/m2] or carboplatin [area under the plasma concentration-time curve 5] intravenously on day 1 of 21-day cycles for 4-6 cycles), followed by maintenance therapy (aumolertinib 110 mg orally once a day and pemetrexed 500 mg/m2 intravenously once every 3 weeks). The primary endpoint was progression-free survival assessed by blinded independent central review (BICR; RECIST version 1.1). Efficacy was analysed in the full-analysis set, which included all randomly assigned patients, and safety was analysed in patients who received at least one dose of the actual trial treatment. The trial is registered at ClinicalTrials.gov, NCT04923906, and is ongoing, but closed to enrolment. Between Aug 4, 2021, to June 18, 2024, of 1011 patients assessed for eligibility, 624 randomly assigned patients (median age 59·0 years [IQR 52·0-66·0]; 337 [54%] were female, 287 [46%] were male) were randomly assigned. 310 (50%) patients received combination therapy and 314 (50%) received monotherapy. As of the data cutoff date (June 18, 2024), the median follow-up was 23·4 months (IQR 20·5-26·5), In the full-analysis set, median BICR-assessed progression-free survival was 28·9 months (95% CI 26.3, NA) in the combination therapy versus 18·9 months (17·8-21·1) in the monotherapy (hazard ratio [HR] 0·47, 95% CI 0·37-0·60; log-rank p<0·0001). The most common grade 3-4 adverse events (occurring in at least 20% in any group) were neutrophil count decreased (168 [55%] of 304 in the combination group versus four [1%] of 316 in monotherapy group), white blood cell count decreased (103 [34%] vs one [<1%]), and platelet count decreased (62 [20%] vs two [1%]). Serious adverse events occurred in 109 (36%) patients in the combination group and 53 (17%) in the monotherapy group, the most common of which were platelet count decreased (22 [7%] vs 0), neutrophil count decreased (17 [6%] vs 0), white blood cell count decreased (13 [4%] vs 0), and anaemia (ten [3%] vs two [1%]). Treatment-related deaths occurred in one (<1%) patient in the combination group (encephalopathy) and two (1%) in the monotherapy group (pulmonary embolism and respiratory failure with circulatory collapse). Aumolertinib in combination with chemotherapy significantly improved progression-free survival. Although this regimen was associated with increased toxicity, the side-effects were managed with dose adjustment and supportive treatment aligned with clinical practice. Long-term follow-up is required to assess overall survival. The AENEAS2 study provides evidence to guide clinical practice regarding EGFR-TKIs and their combination use in treating patients with advanced EGFR-mutated NSCLC. Jiangsu Hansoh Pharmaceutical Group, and the Collaborative Innovation Center for Clinical and Translational Science by Ministry of Education & Shanghai. For the Chinese translation of the abstract see Supplementary Materials section.
Viral infections continue to pose a significant health problem to all parts of the world because of a high rate of mutation and the ability to resist traditional single-target antivirals. Multi-target inhibition of core viral proteins (main protease, RNA-dependent RNA polymerase, and spike glycoprotein) with plant metabolites as the target of drug discovery is a promising alternative. This review presented a systematic review of the antiviral potential and mechanism of plantderived compounds published from 2020 to 2025. The systematic search of the literature was performed with the help of PubMed, Scopus, Web of Science, Science Direct, and EMBASE in accordance with PRISMA 2020. Peer-reviewed articles that reported precise molecular targets, quantifiable antiviral activity (IC50, EC50 or %inhibition), and mechanistic information were included. Qualitative synthesis of data was done because of heterogeneity in methodology. Sixty studies were eligible. Flavonoids (quercetin, kaempferol) had an inhibitory effect on viral protease and polymerase with an IC50 value of 2-12 μM. The terpenoids (glycyrrhizin, betulinic acid) inhibited spike-ACE2 binding and viral fusion with up to 80% infectivity in vitro. The polymerases of hepatitis and influenza viruses were inhibited by polyphenols (EGCG, resveratrol). The bioavailability increased 150-200% with nano formulations, which enhanced therapeutic potency. Metabolites of plants have direct antiviral and host immunomodulatory properties. Nevertheless, a lack of clinical validation and inconsistency in the standardization of phytochemicals are critical issues. Plant-based metabolites are highly promising, multi-target antiviral leads. Nanotechnology, together with standardized pharmacological assessment and enhanced regulatory validation, has the potential to hasten clinical translation of plant metabolites into safe, broad-spectrum antiviral therapies.
The HD21 trial demonstrated efficacy and safety of brentuximab vedotin, etoposide, cyclophosphamide, doxorubicin, dacarbazine, and dexamethasone (BrECADD) vs. bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone (eBEACOPP) as frontline therapy for advanced-stage, classical Hodgkin lymphoma. This analysis evaluated the cost-effectiveness of BrECADD from a US healthcare payer perspective. Building upon a multicenter, randomized, open-label phase 3 HD21 trial (NCT02661503) that evaluated BrECADD versus eBEACOPP in advanced-stage, classical Hodgkin lymphoma patients, we constructed a Markov model with 3-week cycles over a 50-year horizon. The model's primary outcomes encompassed total costs, quality-adjusted life years (QALYs), and incremental cost-effectiveness ratios, with all economic parameters discounted at 3.0% annually. Cost-utility analyses employed a willingness-to-pay threshold of $100,000 per QALY, supplemented by comprehensive sensitivity and scenario analyses to verify model robustness. Economic evaluation demonstrated that compared to eBEACOPP, the BrECADD regimen yielded an additional 2.24 QALYs at an incremental cost of $113,134.17, producing an ICER of $50,411.69/QALY, substantially below the $100,000 WTP threshold. One-way sensitivity analysis identified BrECADD drug acquisition costs and health state utility values as the predominant model drivers; all ICERs remained below the $100,000 WTP threshold across the plausible ranges of all parameters. Probabilistic sensitivity analysis indicated that at a willingness-to-pay threshold of $100,000 per QALY, BrECADD was cost-effective compared to eBEACOPP in 100% of 1000 Monte Carlo iterations. The cost-effectiveness acceptability curve demonstrated that BrECADD achieved a greater than 50% probability of being cost-effective at WTP thresholds above approximately $55,000 per QALY. BrECADD can be considered a cost-effective treatment versus eBEACOPP in treating advanced-stage, classical Hodgkin lymphoma in America. Not applicable.
Counterions and the solvent environment are critical factors determining the electrical properties of DNA. Previous studies have reported the effects of Na+ ion concentration variations and solvent environments on the electrical conductivity of B-DNA. However, the mechanism by which they exert their influence on the drug-DNA complex remains unclear. This model integrates density functional theory and the nonequilibrium Green's function formalism to explore the impact of Na+ ions on the electrical properties of three drug-DNA complexes in both aqueous and vacuum environments. By minimizing the energy, Na+ ions are added to the drug-DNA complexes to render them electrically neutral. Subsequently, the electrical properties of the three DNA molecules are calculated under neutral conditions and after randomly removing one Na+ ion in different solvent environments to investigate the effects of the solvent environment and Na+ ions. The results show that the presence of Na+ hinders charge transport at the HOMO level both in aqueous solvent and vacuum. The removal of one Na+ ion from each of the three complexes leads to an increase in molecular conductivity and charge transfer efficiency, among which the Noga-DNA complex is particularly significantly affected by Na+ ions in the aqueous environment.
Neurodegenerative diseases, notably Alzheimer's Disease (AD) and Parkinson's Disease (PD), represent a significant and growing global health burden, characterized by progressive cognitive and motor dysfunction. Despite advances in understanding their multifactorial pathogenesis, current pharmacotherapies primarily provide symptomatic relief and fail to modify disease progression. This review critically evaluates the emerging role of bioactive phytocompounds as multi-target neuroprotective agents in the management of AD and PD, emphasizing mechanistic insights and translational challenges. A comprehensive literature search was conducted in PubMed, Scopus, and ScienceDirect, focusing on both preclinical and clinical studies that investigated the neuroprotective potential of key phytoconstituents, including luteolin, catechin, apigenin, and quercetin, derived from botanicals such as Commiphora wightii (Guggul), Curcuma longa, and Salvia officinalis. Accumulating evidence demonstrates that these phytoconstituents exert potent antioxidant, anti-inflammatory, anti-amyloidogenic, and anti-apoptotic effects. Mechanistically, they modulate critical signalling pathways implicated in neurodegeneration, including attenuation of oxidative stress, suppression of pro-inflammatory cytokines (e.g., TNF-α, IL-6), mitochondrial stabilization, inhibition of acetylcholinesterase activity, and prevention of amyloid-β aggregation. Their pleiotropic actions position them as promising adjuncts or alternatives to current mono-targeted therapies. However, clinical translation remains constrained by poor bioavailability, lack of standardisation, and limited pharmacokinetic profiling. Phytocompounds show pleiotropic mechanisms (antioxidant, anti-inflammatory, anti-amyloidogenic) but clinical translation is limited by pharmacokinetic and standardisation barriers; novel delivery systems and rigorous clinical trials are required. Phytocompounds represent a compelling, multi-target strategy for AD and PD management, yet significant barriers to clinical application persist. Future research should focus on the development of advanced drug delivery platforms (e.g., nanoformulations), robust clinical trials, and standardized phytopharmaceutical preparations to validate efficacy and safety. Integrative therapeutic frameworks combining phytochemicals with existing pharmacotherapies may offer a viable path toward disease modification in neurodegeneration.
LJF (Lonicerae japonicae flos) is a high-value plant that is both edible and medicinal. It has multiple functions such as clearing heat, detoxifying and reducing inflammation, and is widely used in the field of food and traditional Chinese medicine. Due to the low yeiled and high value of LJF, LF (Lonicerae flos), which has a similar appearance and lower price, is often used as LJF. This not only affects the medicinal quality but also poses a risk to clinical medication. Therefore, establishing rapid and reliable detection methods is crucial for market supervision and ensuring the safe use of drugs. This paper collects the characteristic mass spectra of two medicinal materials based on miniaturized direct ionization mass spectrometry. Through the integration of chemometrics and systematic comparison of pretreatment methods and prediction models, the combination of original data and random forest was ultimately chosen, enabling accurate prediction of unknown samples. Meanwhile, based on variable importance analysis from the random forest model, swertimarin was identified as the characteristic discriminating component in LJF. Finally, the TLC-MS method verified that the feature component mined by machine learning was consistent with that recorded in the pharmacopoeia. Miniaturized direct ionization mass spectrometry is unrestricted by environment or location and offers a high degree of flexibility. The integration with machine learning methods offers a promising proof-of-concept for rapid and accurate quality assessment of traditional Chinese medicine.
Thymidine kinase 2 deficiency (TK2d) (MIM 609560) is an ultra-rare, autosomal recessive mitochondrial myopathy caused by TK2 variants, leading to mitochondrial DNA depletion and/or multiple deletions. People with thymidine kinase 2 deficiency experience progressive myopathy, bulbar weakness and respiratory insufficiency, often losing the ability to walk, eat and breathe independently. Doxecitine and doxribtimine represents the first approved treatment for patients with thymidine kinase 2 deficiency with age of symptom onset ≤12 years by the US Food and Drug Administration and the European Medicines Agency; previously, disease management was limited to supportive care. We investigated the efficacy and safety of pyrimidine nucleos(t)ide therapy in thymidine kinase 2 deficiency. Patients treated with pyrimidine nucleos(t)ides were pooled from retrospective (NCT03701568, NCT05017818) and prospective (NCT03845712) studies and company-supported Expanded Access Programs. Untreated patients were pooled from literature reviews and a retrospective chart review study (NCT05017818). Patient subgroups were stratified by age of thymidine kinase 2 deficiency symptom onset (≤12 years and >12 years). The primary outcome was survival in 50th-percentile matched pairs of treated and untreated patients. Other outcomes included status of developmental motor milestones, ventilatory and feeding tube support, and safety. In total, 218 patients were included (treated: 104; untreated: 114). Baseline demographics and characteristics were comparable between subgroups. Most patients had an age of symptom onset ≤12 years [treated: 82/104 (78.8%); untreated: 93/114 (81.6%)]. In the age-of-symptom-onset-≤12-years subgroup, restricted mean survival time (95% confidence interval) was 29.2 (28.2, 30.3) years over the 30 years after symptom onset for treated patients and 14.4 (11.1, 17.6) years for untreated patients. Loss of ≥1 acquired motor milestone was more frequent before treatment start than after. Substantially more patients regained ≥1 lost motor milestone after treatment start than before. Ventilatory and feeding support were used across all age-of-symptom-onset subgroups, but some patients reduced or discontinued support after starting treatment and fewer patients initiated support after treatment start than before. Most treatment-emergent adverse events (TEAEs) did not lead to discontinuation. The most frequent TEAE was diarrhoea [43/50 patients (86.0%)], which was generally mild or moderate and resolved with dose reduction. Serious TEAEs occurred in 28/50 patients (56.0%); few were considered to be drug related [4/50 (8.0%)]. In total, 3/67 patients (4.5%) experienced a fatal serious TEAE, which were not considered to be drug related. These findings indicate that pyrimidine nucleos(t)ide therapy improves survival and functional outcomes in people with thymidine kinase 2 deficiency, especially those with age of symptom onset ≤12 years, and has an acceptable safety profile.
Glioma progression is shaped by molecular heterogeneity, therapy resistance, and an immunosuppressive tumor microenvironment. Lysosomal remodeling has emerged as a hallmark of glioma adaptation; however, the regulation of individual lysosomal enzymes to malignant progression remains poorly understood. Lysosomal acid phosphatase 2 (ACP2) has been implicated in developmental and metabolic disorders, but its role in glioma has not been systematically investigated. In this study, we conducted an integrative multi-omics analysis to define the transcriptional, clinical, functional, and cellular correlates of ACP2 in glioma. Bulk RNA-seq datasets from TCGA and CGGA were used to analyze gene expressions, survival modeling, and machine-learning-based prognostic classification to evaluate the predictive contribution of ACP2 across glioma grades. Immune infiltration was quantified using TIMER2.0. Functional pathways were assessed using MetaCore, KEGG, GO, and Hallmark GSEA. Single-cell RNA-seq and single-nucleus RNA-seq analyses provided cell-type and subtype-specific validation. Protein-protein interactions were examined using STRING and GeneMANIA. Further pharmacogenomic associations were examined using GDSC/CTRP, and molecular docking was performed to simulate the drug ability of ACP2. Findings of this study indicated that ACP2 was significantly overexpressed in glioma relative to normal tissues and demonstrated the strongest prognostic impact among ACP family members. Elevated ACP2 expression correlated with reduced overall survival across multiple independent cohorts and was associated with increased infiltration of macrophages, neutrophils, and dendritic cells. Enrichment analyses revealed consistent activation of PI3K/AKT, KRAS, E2F and extracellular matrix remodeling pathways. MetaCore identified APP processing and cytoskeletal remodeling as top ACP2-associated modules. Single-cell and single-nucleus analyses localized ACP2 expression mainly to malignant glioma and myeloid populations, with higher expression in recurrent and advanced malignant states. This multi-omics framework identified ACP2 as a lysosomal regulator of glioma aggressiveness and immune remodeling. ACP2 functions as a robust biomarker of malignancy and may represent a candidate target for therapeutic exploration in glioma.
Dogs are a critical non-rodent species used in preclinical safety studies, particularly, in the pharmaceutical field, due to their physiological, metabolic, and immunological similarities to humans. As such, immunophenotyping of canine peripheral blood mononuclear cells (PBMCs) plays a crucial role in translational research, immune monitoring, and safety evaluations in drug development. However, the limited availability of canine-specific antibodies restricts detailed and accurate immune profiling, which is essential for advancing safety evaluations in drug development. To address this challenge, we developed a 15-marker panel for comprehensive mass cytometry-based immunophenotyping of cryopreserved canine PBMCs. This panel encompasses major leukocyte subsets, including B cells, CD4+ T helper cells, regulatory T cells, CD8+ cytotoxic T cells, memory T cell subsets, natural killer T cells, natural killer cells, dendritic cells, CD4+ monocytes, classical monocytes, and neutrophils. We utilized both extracellular and intracellular markers to facilitate in-depth immune profiling, despite the limited availability of canine-specific antibodies. The panel was thoroughly optimized in terms of marker selection, antibody clone validation, and metal isotope pairing. Additionally, by the use of mass cytometry, several channels remain unoccupied, providing flexibility for future panel expansion.
The pathological process of androgenic alopecia (AGA) is closely associated with oxidative stress in the perifollicular microenvironment. Existing antioxidant small-molecule drugs are often constrained by low permeability, necessitating the urgent development of active therapeutic agents that can effectively penetrate the skin barrier and precisely deliver antioxidants to hair follicles. This study reported a taurine-based active pharmaceutical ingredient ionic liquid (CTIL) synthesized via a one-step method, which not only acted as a delivery carrier but also served as a therapeutic material capable of modulating the perifollicular microenvironment by scavenging reactive oxygen species (ROS). An in vitro study demonstrated that the hydrogen-bond network structure of CTIL facilitated its penetration through the stratum corneum barrier, thus improving the targeted accumulation of taurine within deep hair follicle structures. More importantly, in the AGA animal model, CTIL effectively eliminated excessive ROS in the perifollicular microenvironment, alleviated tissue oxidative damage and inflammatory infiltration, thereby inhibiting the miniaturization process of hair follicles and promoting hair regrowth. This study provided an innovative strategy integrating delivery and treatment for AGA therapy and revealed the potential of bioactive ionic liquids in regulating the microenvironment of skin diseases.
Cathepsin L (Cat L), a lysosomal endopeptidase, is overexpressed in human renal clear carcinoma (RCC); however, its impact on apoptosis remains unclear. Results revealed that inhibition (SID, a specific inhibitor), knockdown (siRNA), or knockout of Cat L sensitizes human carcinoma cells to anticancer drugs-mediated apoptosis through the downregulation of Bcl-xL and Survivin at the post-translational level. Moreover, combined treatment with SID and sorafenib reduced the tumor growth in a xenograft model. Deletion of Cat L induced Parkin stabilization by increasing DUB3 expression at the transcriptional level. Parkin knockdown significantly prevented SID-induced Bcl-xL downregulation. Conversely, ectopic expression of Parkin suppressed Bcl-xL expression. Furthermore, SID-mediated Parkin upregulation inhibited USP53 deubiquitinase protein expression, leading to the degradation of Survivin expression through its ubiquitination. Parkin directly ubiquitinated Bcl-xL and USP53, but not Survivin. These results demonstrate that inhibition of Cat L enhances anticancer drugs-induced apoptosis through Parkin-mediated ubiquitination of Bcl-xL and USP53. Moreover, the downregulation of USP53 suppressed the expression of Survivin. Therefore, Cat L can be considered a potential candidate molecular target for the treatment of RCC.
The global outbreak of COVID-19, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has resulted in over 774 million cases and 7 million deaths worldwide since January 2020. Although widespread vaccination has been instrumental in reducing transmission and disease severity, there is a persistent and urgent need for effective treatments. This has made the virus's main protease (Mpro), a key enzyme in its replication cycle, a leading target for drug development. In the current research work, a schematic strategy was utilized to design, synthesize and screen the biological activity of newly prepared thiadiazole derivatives (5a-f, 9a-d and 12a-j) against Mpro as SARS-CoV-2 inhibitors. The research plan was organized into three schemes, where every step advanced from the outcome of the former one to create a clear structure activity relationship (SAR) and successively optimize biological activity. Specifically compounds 12c (IC50 = 0.03077 μM), and 12a (IC50 = 0.03356 μM) exhibited excellent inhibitory potency as compared to reference drug Nirmatrelvir (IC50 = 0.05855 μM). The Molecular docking study further validated the experimental results revealing that the synthesized molecules interacted strongly with the SARS-CoV-2 Mpro active site via multiple hydrogen bonds and hydrophobic interactions with important catalytic residues, resulting in stable enzyme inhibitor complexes consistent with the in-vitro results. Nevertheless, further cell-based antiviral studies are required to validate the ability of these compounds to suppress SARS-CoV-2 replication under physiological conditions. However, further selectivity studies against human proteases are necessary to evaluate potential off-target effects and safety profiles.
Functional validation of host factors in whole-animal models is a major bottleneck in virology; it hinders the translation of data from in vitro studies into a deeper understanding of the viral life cycle and pathogenesis. To address this challenge, we developed a systematic in vivo screening platform for influenza A virus. This platform comprises a library of 84 CRISPR-Cas9-generated gene-modified mouse lines targeting host factors prioritized from the literature and in vitro small interfering RNA (siRNA) screening studies. Using this resource, we identified 17 host factors whose genetic ablation conferred resistance to influenza A virus infection. Further studies of two of these factors, Arhgef28 and Lasp1, revealed distinct protective mechanisms against influenza A virus. We offer this mouse library to the research community as a powerful platform for studying virus-host interactions in a physiologically relevant context.
Adenoviridae family members routinely infect humans, exhibit significant genetic diversity, and are associated with a variety of illnesses. Types 4 and 7 frequently circulate in the United States and are major causes of respiratory disease. Infections can result in hospitalization and, in severe cases, death. Although a live wild-type-virus vaccine targeting these two types exists, its use is restricted to military personnel due to concerns about viral-shedding and potential for genetic recombination. To overcome these limitations, we recently developed a virus-like particle (VLP) platform as an alternative vaccination strategy. These VLPs are stable, lack genomic material, and elicit a potent humoral immune response in mice, effectively neutralizing adenoviral infection. Here, we describe the cryo-EM structure of adenovirus 7 (AdV-7) VLPs. Structural insights are essential to ensure that neutralizing antigens displayed on the VLPs accurately mimic those of the virion, guide the design of particles with improved stability and efficacy, and enable engineering of VLPs with antigenic properties targeting multiple adenovirus types. The structure shows that hexon, penton, pIIIa, pVI, pVIII, and IX assemble comparable to AdV-5, hexon and penton neutralizing epitopes are appropriately displayed for antibody recognition, penton insertion into the hexon shell promotes cement protein pIIIa to increase its interaction with the peripentonal hexons, and presence of the core-genome is associated with increased interaction between cement protein pVIII and hexon. Finally, limited proteolysis and mass spectrometry demonstrate that VLP incorporated hexons digest more readily than virion incorporated hexons, indicating the greater dynamic nature of the VLP.
Scutellaria baicalensis, a perennial herb belonging to the genus Scutellaria (Lamiaceae), has dried roots that are widely used as a traditional Chinese medicinal herb known as "Huangqin". As one of the major chemical constituents of S. baicalensis, its polysaccharides exhibit diverse biological activities, including immunomodulatory effects, regulation of gut microbiota, antioxidant activity, hypoglycemic effects, anti-inflammatory activity, and antimicrobial properties. Modern pharmacological studies have further confirmed the intrinsic relationship between these activities and traditional therapeutic uses at the molecular level, while also revealing their potential applications in functional foods, dietary supplements, and food additives, thereby providing scientific support for the modernization and application of traditional medicine. To systematically summarize the latest research progress on the extraction, isolation, purification, structural characterization, and biological activities of Scutellaria baicalensis polysaccharides (SBP), and to explore their structure-activity relationships and potential research directions, with the aim of supporting their further development and application in the pharmaceutical and functional food industries. Following the PRISMA guidelines, relevant studies were systematically retrieved from PubMed, Web of Science, and CNKI databases, and the latest advances in the extraction, purification, structural characterization, and biological activities of SBP were comprehensively reviewed. Various extraction methods have been established, including hot water extraction, ultrasound-assisted extraction, and microwave-assisted extraction, among which the Sevag method is commonly employed for deproteinization during purification. Structural studies have primarily focused on monosaccharide composition, molecular weight, and glycosidic linkage patterns. Pharmacological studies have demonstrated that these polysaccharides possess multiple biological activities, including immunomodulatory, antioxidant, hypoglycemic, anti-inflammatory, antimicrobial, and gut microbiota-regulating effects. Structure-activity relationship studies indicate that the biological activities of SBP are collectively determined by their multi-level fine structures, including molecular weight, monosaccharide composition, types and patterns of glycosidic linkages, and higher-order spatial conformations, all of which play critical roles in their biological functions. Considerable progress has been achieved in the extraction, purification, structural characterization, and biological evaluation of SBP, demonstrating promising pharmacological potential in immunomodulation, anti-inflammatory activity, and metabolic regulation. However, their higher-order structural characteristics, the structure-activity relationships between specific structural motifs and biological activities, as well as in vivo processes such as bioavailability and metabolic fate, still require further investigation.
To date, there is no reliable and valid screening tool for paraphilic disorders (PDs) based on the diagnostic criteria of the International Classification of Diseases (ICD-11). The present study aimed to develop and cross-culturally examine the cross-cultural applicability of a new screening tool for PDs following the ICD-11 guidelines. Top-down item development was conducted with expert practitioners in the field. Data were collected in 26 languages across 42 countries (N = 82,243; 39.5% men; 57.0% women; 3.4% gender-diverse; Mage = 32.39 years, SD = 12.52). Cut-off scores were established per ICD-11 guidelines, and at-risk groups were compared with low-risk groups along theoretically relevant correlates (e.g., depression, anxiety). The Paraphilic Disorders Short Screen (PDSS) assesses seven paraphilias via 21 items. Voyeurism was most common paraphilic interest (PI) (19.42% of the total sample; n = 15,970), which also had the highest number of participants meeting the ICD-11 diagnostic cut-off (3.24%; n = 2,662). PI and PD groups consistently demonstrated more severe depression and anxiety symptoms than others. The PDSS is a reliable and valid screening tool that follows the latest ICD-11 guidelines; therefore, it has the capacity to enhance clinical practice and research by identifying individuals at risk.
Brain infections, caused by various pathogens (such as viruses, bacteria, fungi, or parasites), have proven challenging to treat due to limited drug diffusion through the blood-brain barrier and the presence of intracellular reservoirs. As biologically derived nanocarriers, exosomes have emerged as viable candidates for crossing physiological barriers and effectively delivering target molecules into the central nervous system. This review aims to summarize what is currently known about exosome biogenesis, cargo sorting, and immunological function in relation to infectious disease. In addition, it provides information on how different pathogens have taken advantage of exosomal pathways to increase their virulence and modulate the immune response, while also suggesting options for the therapeutic engineering of exosomes. It critically evaluates technological advances made in exosome engineering, such as CRISPR/Cas9-based cargo loading, ligand-directed surface modification of exosomes, targeted delivery of nucleic acids, and creation of stimuli-responsive release systems for exosome cargo for their potential application as precision therapies against pathogens that infect the brain. Pharmacokinetic data and biodistribution studies, along with studies examining how route of administration, inflammatory status, and receptor mediated uptake affect CNS targeting efficacy reflect that exosome engineering offers a novel platform for creating precision therapeutics against pathogens that infect the brain.
Activation of tumor suppressors represents an attractive strategy for cancer treatment. Hepatocyte nuclear factor 4 alpha (HNF4A) functions as a tumor suppressor in the liver by inhibiting hepatocyte proliferation; however, no effective agonists have been identified. Here, we aimed to identify novel ligands for HNF4A and evaluate their role in hepatocarcinogenesis. We identified polyprenoic acid (PA, peretinoin) as the first ligand capable of directly binding HNF4A. Reanalysis of prior clinical trial data revealed that PA suppressed the progression of dysplastic nodules (DNs) to hepatocellular carcinoma (HCC), while showing no effect on local HCC recurrence after initial treatment. Consistently, PA inhibited DN growth but not established HCC in Pdgf-C transgenic mice. Mechanistically, PA selectively bound to the HNF4A P1 isoform, enhancing its transcriptional activity and upregulating hepatocyte maturation markers (ALB, TTR, and SLCO1B3), while suppressing alpha-fetoprotein expression driven by the HNF4A P2 isoform. Importantly, hepatocyte-specific Hnf4a knockdown or lipid-nanoparticle-mediated Hnf4a siRNA abrogated the protective effects of PA. These findings establish HNF4A as a pharmacologically controllable tumor suppressor and highlight PA-like compounds as promising agents for preventing liver carcinogenesis.