Drug Target Insights最新文献

Introduction: Atypical hemolytic uremic syndrome (aHUS) is a potentially life-threatening condition associated with poor clinical outcomes if not treated adequately. Eculizumab has become the standard of care, whereas ravulizumab, a second-generation, high-affinity complement C5 inhibitor, demonstrates comparable efficacy in improving renal function, hematological markers, and dialysis rates. In addition, ravulizumab offers practical advantages, including a longer dosing interval and immediate, complete, and sustained inhibition of free C5, making it a valuable therapeutic option.

Methods: Given the limited real-world experience with ravulizumab, we present a case series of six treatment-naïve aHUS patients who received ravulizumab as first-line therapy.

Results: These cases include one pregnancy-related aHUS, one postpartum case, one related to a urinary tract infection, one associated with hypertension, one with a pneumonia-related trigger, and one kidney transplant patient with a prior verotoxin-producing E. coli infection. Altogether, these cases illustrate the challenges in diagnosing aHUS. The choice to administer ravulizumab as first-line treatment was sometimes made in the presence of a clear clinical suspicion, even when not all minor criteria seemed to confirm the diagnosis. In most patients, renal function improved rapidly after ravulizumab administration, followed by recovery of hematological parameters, which were stable in the longer term. As improvements remained sustained over time, the possibility of discontinuing ravulizumab can be evaluated on a case-by-case basis.

Conclusion: These cases highlight the importance of early diagnosis, prompt intervention, and multidisciplinary care in managing aHUS. Ravulizumab as first-line therapy proved effective and well-tolerated, with sustained clinical improvements observed across diverse real-world scenarios.

Introduction: This study aimed to detect the co-occurrence of carbapenem resistance genes along with aminoglycoside-modifying enzyme (AME) genes in clinical Enterobacterales isolates to understand the distribution of multiple resistance genes among clinical isolates.

Methods: This prospective study was conducted for six months (November 2024 to April 2025) in the department of microbiology of a tertiary care hospital. A total of 30 blood culture isolates were identified as resistant to both carbapenem and aminoglycoside antibiotics using the automated VITEK 2 compact system. The genes responsible for carbapenem resistance (bla_NDM , bla_OXA-48 , bla_KPC , bla_IMP , and bla_VIM ) were detected by multiplex real-time PCR, and the aminoglycoside-modifying enzyme genes [APH(3')-Ia, APH(2")-Ib, AAC(3)-IIc, AAC(6')-Ib, and ANT(3")-I] were detected by the conventional polymerase chain reaction method. All the clinical data, patient demographics, and molecular findings were entered in an MS Excel spreadsheet version 14.0.4734.1000 and analyzed using GraphPad/PRISM software version 10.5.0.

Results: Of the 30 Enterobacterales isolates, Klebsiella pneumoniae was the most common isolate (66.7%). Molecular detection revealed bla_NDM in 40% isolates and bla_OXA48 in 10% isolates. The majority of the AME genes were in combination. The most common combination of the AME gene was AAC(6')-Ib+ AAC(3)-IIc+ ANT(3")-I + APH(3')-I detected in 4 (13.3%) isolates. The most common combination of carbapenem and aminoglycoside resistant genes was bla_NDM + bla_OXA48 + AAC(6')-Ib+ AAC(3)-IIc+ ANT(3")-I+ APH(3')-I (13.3%). The bla_OXA-48 gene had a statistically significant association with AAC(6')-Ib, ANT(3")-I, and APH(3')-I (p <0.05).

Conclusion: The Co-occurence of carbapenem resistance and aminoglycoside-modifying enzyme genes in clinical Enterobacterales isolates limits the therapeutic option.

Introduction: Diabetes mellitus (DM), particularly type 2 DM (T2DM), is a chronic metabolic disorder requiring novel therapeutic approaches as the available therapies are not meeting the current challenges. This study investigates the anti-diabetic potential of Vigna unguiculata using a network pharmacology approach, supported by in vitro and in silico analyses.

Methods: The plant was collected from Khyber Pakhtunkhwa, Pakistan, and subjected to hydroalcoholic extraction and fractionation. In vitro assays included α-amylase, α-glucosidase, and aldose reductase. Target prediction using STITCH and SwissTargetPrediction identified 88 common genes linked to T2DM. Protein-protein interaction (PPI) network analysis highlighted key genes like EGFR, PTGS2, and TLR4 as central nodes in diabetes-related pathways. Molecular docking was used to study the binding affinities of compounds.

Results: IC50 values were determined using IBM SPSS Statistics 21 software. The data underwent analysis using one-way ANOVA followed by Dunnett's multiple comparison test. Significance value was determined at *p   0.05, **p   0.01 and ***p   0.001. In-vitro assays demonstrated significant α-amylase, α-glucosidase, and aldose reductase inhibitory activities. Phytochemical screening identified several bioactive compounds. Functional annotation and KEGG pathway analysis confirmed these genes' roles in crucial metabolic pathways. Virtual screening revealed strong binding affinities of compounds like Stigmasterol, Luteoline, and Quercetin with GSK3B, PTGS2, and TLR4. The Molecular Dynamics (MD) simulation, binding free energy calculations (MM-PBSA and MM-GBSA), confirmed the results of Virtual screening.

Conclusion: In short, these findings underscore V. unguiculata as a promising source for anti-diabetic agents, supporting further clinical trials for T2DM management.

Introduction: Bushenhuoxue formula is a traditional Chinese medicine formula with relatively safe clinical effects, but its mechanism in recurrent spontaneous abortion (RSA) is still unclear. Our present study used Network pharmacology an experimental validation to discuss how Bushenhuoxue formula improves prethrombotic state in RSA.

Materials and methods: The active ingredients of Bushenhuoxue formula (Drug) were acquired from our previous study. The putative targets of ZYP relevant to AS were obtained from TCMSP, Swiss Target Prediction, STITCH, DisGeNET, and Gene Cards databases. Protein-protein interactions (PPI) network, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted using the Cytoscape software. Furthermore, in vivo experiments were carried out for target validation in BALB/c mice, collecting placental tissue from different groups, the cell apoptosis by TUNEL assay; the pathology by HE staining; relative mRNA expression by qRT-PCR assay; relative protein expression by IHC and WB assay.

Results: Animal experiments, compared with the NC group, the AT-III, PROG, HCG, APC and t-PA concentrations were significantly depressed (P˂0.05, respectively), Apoptosis cell numbers were significantly up-regulated with PI3K/AKT/HIF-1α VEGF significantly depressing (P˂0.001, respectively). With Bushenhuoxue formula supplement, AT-III, PROG, HCG, APC and t-PA concentrations were significantly improved in RSA model mice; and improved pathological changes and apoptosis cell number in placenta tissues (P˂0.05, respectively). However, with LY294002 supplement, the drug treatment effects were disappeared.

Conclusion: Bushenhuoxue formula improves prethrombotic state in RSA via stimulating PI3K/AKT/HIF-1α/VEGF pathway in vivo.

Introduction: Keloid scars are a kind of skin disorder in which the scar grows beyond the boundaries of the original wound. Baicalein, a flavonoid, may treat keloids by targeting fibrosis, inflammation, and possible viral factors.

Methods: In silico studies were conducted to evaluate the potential anti-keloid effects of baicalein by predicting its interactions with three key proteins of the transforming growth factor-β (TGF-β) family (PDB IDs: 1VJY, 3TZM, and 7DV6). POM analysis was also used to understand the conditions that could enhance baicalein's efficacy.

Results: The results indicated that baicalein binds effectively to TGF-β family proteins via hydrogen bonds, showing strong affinities (1VJY: -9.9 kcal/mol, 3TZM and 7DV6: -9.3 kcal/mol), indicating its potential as a TGF-β receptor ligand. Osiris analysis gave a drug score of 75% for baicalein, while Molinspiration indicated good bioavailability with a cLogP of 2.84. Atomic charge distribution and pharmacophore site mapping through POM analysis indicate that baicalein exhibits an antiviral pharmacophoric moiety akin to known antiviral agents. This indicates that baicalein may act as a pro-drug, undergoing metabolic transformation to form a bis-bidentate ligand. Such ligands are crucial for forming bimetallic complexes that can function as efficient biocatalysts against various biological targets.

Conclusion: In-silico analysis suggests that baicalein may influence TGF-β receptors and exhibit anti-keloid activity. Additionally, POM analysis recommends that baicalein may serve as a lead compound with the potential to modulate TGF-β signalling and exhibit antiviral properties, indicating it as a dual-action agent against keloids and viral infections.

Introduction: Wound management presents significant challenges, requiring effective treatments. Herbal extracts have been traditionally used to support healing due to their anti-inflammatory, antimicrobial, and cell-regenerative properties.

Methods: This study aimed to evaluate the therapeutic efficacy of Pau D'Arco (Tabebuia), Yarrow (Achillea millefolium), Gotu Kola (Centella asiatica), Figwort (Scrophularia nodosa), and Broadleaf (Plantago major) extracts, both individually and combined, on wound healing in vitro and in vivo in equine models. In vitro tests using human macrophages and keratinocyte cell lines to assess cellular responses such as cytokine secretion and phagocytic activity under simulated inflammatory conditions. Additionally, pilot case studies on equines with open wounds provided practical insights into the extracts' healing capabilities.

Results: MTT assay was used to assess cytotoxicity. The extracts did not significantly affect the viability of HaCaT or THP-1 cells. The herbal extracts reduced IL-8 levels and increased phagocytic activity in macrophages, indicating an ability to modulate inflammatory responses. In vivo, the extracts were well tolerated and associated with supported healing in equines. These effects were suggested to be attributed to the synergistic actions of the herbal components.

Conclusion: These findings suggest that the herbal extracts may be useful for supporting wound healing. Their natural anti-inflammatory and healing properties could provide an additional option alongside traditional wound management approaches.

Introduction: The current scenario of tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) has presented an almost insurmountable challenge to hospitals with high patient numbers. Delayed diagnosis of TB is a major hurdle in preventing the employment of efficient therapeutics, leading to the development of drug resistance. Hence, an easily accessible diagnostic method, particularly for resource for resource-limited settings, is pertinent for the rapid identification of MTB-infected patients. In pursuit of developing such an assay, the present study offers a CLAP-TB (CRISPR-Cas coupled RT-LAMP Amplification Protocol for Tuberculosis) assay, which will allow us to diagnose TB rapidly and visually.

Methods and results: Herein, the visual MTB detection consists of a method utilizing 232 different samples (sputum, urine, serum) from 82 patients for reverse transcription loop-mediated isothermal amplification (RT-LAMP). Additionally, the assay also utilizes the integration of a CRISPR-Cas12-based system using different guide RNAs of IS6110 and an internal control POP7 (human RNase P) genes along with visual detection via lateral flow readout-based dipsticks with the unaided eye (~134 min). Overall, the limit of detection for CLAP-TB assay was up to 1 ag of RNA, while the clinical sensitivity and specificity were 98.27% and 100%, respectively, on the pilot scale.

Conclusion: Together, our CLAP-TB assay offers proof of concept for a rapid, sensitive, and specific method with the minimum technical expertise required for TB diagnosis in developing and resource-limited settings.

Background: Withania somnifera is among the most widely prescribed medicinal plants in traditional Indian medicine. Hydroalcoholic extract of the roots of this plant was investigated for its effects on the overall health and lifespan of the model worm Caenorhabditis elegans.

Methods: The extract's effect on worm lifespan and fertility was observed microscopically. Worm motility was quantified through an automated worm tracker. The metabolic activity of the worms was captured using the Alamar Blue® assay. Differential gene expression in extract-treated worms was revealed through a whole transcriptome approach.

Results: Extract-exposed gnotobiotic worms, in the absence of any bacterial food, registered longer lifespan, higher fertility, better motility, and metabolic activity. Whole transcriptome analysis of the extract-treated worms revealed the differential expression of the genes associated with lifespan extension, eggshell assembly and integrity, progeny formation, yolk lipoproteins, collagen synthesis, cuticle molting, etc. This extract seems to exert its beneficial effect on C. elegans partly by triggering the remodeling of the developmentally programmed apical extracellular matrix (aECM). Differential expression of certain important genes (cpg-2, cpg-3, sqt-1, dpy-4, dpy-13, and col -17) was confirmed through PCR assay too. Some of the differently expressed genes (gfat-2, unc-68, dpy-4, dpy -13, col-109, col-169, and rmd-1) in worms experiencing pro-health effect of the extract were found through co-occurrence analysis to have their homologous counterpart in humans.

Conclusions: Our results validate the suitability of W. somnifera extract as a nutraceutical for healthy aging.

Introduction: Adequate hyperglycemic control is still a huge challenge with the clinically used therapeutics. New, more effective anti-diabetic agents are on the top list of drug discovery projects.

Methods: This article deals with the in vitro anti-diabetic potential of 2, 3 dichloroIndolinone (C1) and 2, 6-dichloroIndolinone (C2) on α-glucosidase and α-amylase followed by in silico analysis.

Results: Both compounds, C-1 and C-2, caused significant inhibition of α-glucosidase at various test concentrations with IC₅₀ of 35.266 μM and 38. 379 μM, respectively. Similarly, compounds C-1 and C-2 elicited significant anti-α-amylase action with IC₅₀ values of 42.449 μM and 46.708 μM, respectively. The molecular docking investigation regarding the α-glucosidase and α-amylase binding site was implemented to attain better comprehension with respect to the pattern in which binding mechanics occur between the C1 and C2 molecules and the active sites, which illustrated a higher binding efficacy in appraisal with reference inhibitor and acarbose. The interactions between the active compounds C1 and C2 with the active site residues were mainly polar bonds, hydrogen bonding, π-π, and π-H interactions, which contributed to a strong alignment with the enzyme backbone. Similarly, effective binding is frequently indicated by a strong and stable hydrogen-bonding pattern, which is suggested by the minimal fluctuation in MM-PBSA values.

Conclusion: In short, this study will contribute to providing these compounds with an improved anti-diabetic profile and decreased toxicity.

Introduction: Candida albicans biofilm formation is a significant contributor to antifungal resistance, necessitating new treatment strategies. Annona muricata Lin., a traditional herbal remedy, has shown promise in combating microbial infections. The purpose of this study was to assess the antibiofilm activity of the methanol extract of A. muricata leaves alone or with the addition of fluconazole against C. albicans.

Methods: Phytochemicals from the methanol extract were analyzed by LC-MS, the XTT assay was used for metabolic activity, and morphological characteristics were examined using scanning electron microscopy (SEM). Molecular docking screening of identified compounds in A. muricata methanol leaves extract against a Sap3 receptor (PDB: 2H6T) was also performed.

Results: The LC-MS analysis detected 17 possible phytochemicals. The methanol extract showed a dose-dependent inhibition of biofilm formation, with maximum inhibition of ~60% observed at 240 μg/ml, and inhibition by fluconazole increased from 32% to 76% as the concentration increased from 15 to 240 μg/ml. The combination of A. muricata and fluconazole increased the inhibition significantly, from 74% to 78% at 15 μg/ml to 240 μg/mL, respectively. SEM of control and treated C. albicans biofilms showed an altered morphology and loss of cell integrity by the combination, corroborating the findings. Plant phytochemicals also possess high binding affinity (-9.7 to 8.0 kcal/mol, respectively) for the Sap3 enzyme and may therefore have therapeutic potential against C. albicans.

Conclusion: Consequently, the findings indicate that compounds in the A. muricata methanol extract may function in concert with fluconazole at sub-inhibitory concentrations to suppress C. albicans biofilm formation. This finding paves the way for the formulation and development of antifungal treatment regimens that may limit the development of fluconazole resistance employing this plant part.