Journal of pharmacological and toxicological methods最新文献

{"title":"Chemometrics-assisted spectrophotometric method for the simultaneous determination of phenylephrine hydrochloride, chlorpheniramine maleate and paracetamol","authors":"Muhammad W. Sabr ,&nbsp;Diyar S. Ali ,&nbsp;Slim Smaoui ,&nbsp;Teresa D'Amore","doi":"10.1016/j.vascn.2025.107741","DOIUrl":"10.1016/j.vascn.2025.107741","url":null,"abstract":"<div><div>This study presents an eco-friendly, combined UV spectrophotometric method based on derivative ratio and second derivative for the simultaneous determination and quantification of phenylephrine HCl (PHE), chlorpheniramine maleate (CPM), and paracetamol (PAR) in pharmaceutical mixtures. In the derivative ratio method, sample spectra are divided by standardized spectra of the other components, followed by calculation of the first derivative. Analysis was performed at 226 nm for PHE, 271 nm for CPM, and 253 nm for PAR. The second derivative method uses second-order derivative absorption spectra, using zero-crossing points to selectively quantify each drug at 280.5 nm for PHE, 250.5 nm for CPM, and 245.5 nm for PAR. The linearity ranges were 0.1–30 μg/mL (PHE), 0.5–36 μg/mL (CPM), and 1–30 μg/mL (PAR) for the derivative ratio method; and 3–24 μg/mL (PHE), 1–15 μg/mL (CPM), and 1–14 μg/mL (PAR) for the second derivative method. The relative standard deviations were below 0.2 % for the derivative ratio method and below 3 % for the second derivative method. Method performances were benchmarked against a conventional HPLC-DAD technique and the eco-friendliness and greenness were evaluated using advanced assessment tools. The proposed method was effectively applied to commercial pharmaceutical formulations, confirming its reliability and environmental compatibility.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107741"},"PeriodicalIF":1.3,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143873571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The statistical and pharmacological sensitivity of nonclinical QTc analysis comparing jacketed telemetry and multilead snapshot recording in the same dogs","authors":"Matthew M. Abernathy,&nbsp;Derek D. Best,&nbsp;Derek J. Leishman","doi":"10.1016/j.vascn.2025.107742","DOIUrl":"10.1016/j.vascn.2025.107742","url":null,"abstract":"<div><div>Quantitative QTc assessment in toxicology studies can use one of two techniques, either continuous telemetry recording in ambulatory animals or short snapshots of electrocardiograms recorded using multilead electrodes in restrained animals. QTc assessments can augment clinical QTc data in regulatory submissions provided both the statistical and pharmacological sensitivity can be demonstrated.</div><div>Analyses in dogs (<em>N</em> = 24) used both jacketed external telemetry (ET) and short segment or ‘snapshot’ (SS) multilead recordings in separate phases. Three groups (moxifloxacin, and two doses of ondansetron) were compared to a vehicle group. In phases 1 and 2, one group received moxifloxacin (30 mg/kg), two received doses of ondansetron (3 and 10 mg/kg), and the last received vehicle (0.5 % (<em>w</em>/<em>v</em>) methylcellulose in deionized water) in a parallel group study design. The third phase of the study involved the original 12 male dogs receiving all treatments in a randomized triple Latin square design.</div><div>ET (<em>n</em> = 6/group) detected a significant effect on QTc of moxifloxacin for all but one hour of the postdose period and detected an effect at fewer timepoints with ondansetron in a dose- and exposure-dependent manner. QTc prolongation at a single timepoint was significant for both the lower dose of ondansetron and moxifloxacin using SS. SS did not detect any significant QTc effect at the higher dose of ondansetron. In the cross-over phase (ET only), moxifloxacin significantly prolonged the QTc interval through all but one hour of the post dose period. Low dose ondansetron did not significantly prolong the QTc interval while there was a small dose-dependent effect with isolated time points being significant with the higher dose. The least significant differences detectable were approximately 9 (ET), 17 (SS) and 5 ms (ET with Latin square) for the three phases, respectively.</div><div>These analyses demonstrate the statistical and pharmacological sensitivity of the techniques commonly used in large animal toxicology studies. Overall, QTc assessment using telemetry is the more sensitive and consistent technique and is a model sufficiently sensitive to supplement clinical QTc assessment.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107742"},"PeriodicalIF":1.3,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143882987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A comparative study of kidney lithogenic agents in Wistar rats by invasive and noninvasive methods","authors":"S. Sakthikarthikeyan ,&nbsp;S. Ramesh ,&nbsp;P. Senthilkumar ,&nbsp;K. Priya ,&nbsp;M.R. Srinivasan ,&nbsp;M. Balagangatharathilagar ,&nbsp;R.B. Vishnurahav ,&nbsp;R. Thangathurai","doi":"10.1016/j.vascn.2025.107743","DOIUrl":"10.1016/j.vascn.2025.107743","url":null,"abstract":"<div><div>Urolithiasis has been globally reported in humans and animals due to multiple factors. Research on the management and treatment of urolithiasis would benefit if suitable lithogenic agents replicate various degrees of severity as seen in humans. Based on the literature, ethylene glycol with ammonium chloride and sodium oxalate are the commonly used agents for the induction of kidney stones in rats, but the duration of the induction period varies between the researchers. Hence, in the present study, ethylene glycol with 1 % ammonium chloride (EG1AC), ethylene glycol with 2 % ammonium chloride (EG2AC) in drinking water for 14 days, and sodium oxalate (SO) at 70 mg/kg body weight administered through intraperitoneal route for 14 days and post monitored up to 50 days to know the kidney damage. In the studies on urolithiasis model, the severity of kidney damage is determined based on renal tissue analysis, urine, and serum parameters. For the first time, instantaneous noninvasive ultrasound scorecard technique has been used to assess the severity of kidney damage in urolithiasis. Based on the urine, serum, renal tissue, and ultrasound examinations, one day is enough for sodium oxalate for kidney stone formation, whereas seven days for EG1AC or EG2AC and the degree of kidney severity were in the order of sodium oxalate &gt; EG2AC &gt; EG1AC from days 1 to 50. Ultrasound scorecard can be effectively used to evaluate kidney stone formation and damage without sacrifice of animal from day 1 onwards.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107743"},"PeriodicalIF":1.3,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143869358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Surgical instrumentation of cardiovascular telemetry device has minimal impact on the general toxicology data interpretation in Cynomolgus monkeys","authors":"Xiefan Fang,&nbsp;Jui-Tung (Ray) Liu,&nbsp;Stephen D. Tichenor","doi":"10.1016/j.vascn.2025.107740","DOIUrl":"10.1016/j.vascn.2025.107740","url":null,"abstract":"<div><div>Increasing numbers of general toxicology studies with integrated implanted cardiovascular (CV) telemetry are being conducted due to the constrained supply of nonhuman primates and demands for higher method sensitivities as per ICH S7B Q&amp;A. However, limited information is available for the potential impact of surgical instrumentation on the toxicology data. In this study, we reviewed the reports of implanted CV telemetry studies conducted in cynomolgus monkeys from 2017 to 2022 at our facility. The instrumentation had no impact on food consumption, respiration, pulse oximetry, neurological parameters, ophthalmology, troponin I, urinalysis, immunophenotyping, or organ weights. There was a low probability of needing to exclude animals from studies due to instrumentation-related technical (4.8 %) or health issues (0.8 %). Incidences were low (0.3 % to 2.6 %) to have instrumentation-related in-life or laboratory findings. Pathology findings were regional, rarely associated with clinical pathology changes, non-adverse, and clearly distinguishable from the test article effects in the studies examined. They were limited to localized effects (fibrosis and low-grade inflammation) associated with a foreign body reaction in the area immediately in contact with the device. Also, thrombus formation (0.2 %) was noted when the femoral artery was used for blood pressure catheter placement. Changing from intramuscular to intra-peritoneal placement greatly reduced the incidence of unusable ECG data and abnormal clinical signs. Use of the internal iliac artery for catheter placement also eliminated thrombus formation and the subsequent health impact. In conclusion, implanted telemetry integrated into a nonhuman primate toxicology study does not compromise the study objectives, study execution, or toxicology data interpretation.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"133 ","pages":"Article 107740"},"PeriodicalIF":1.3,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143733828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intra-laboratory validation of yeast-based reporter gene assays for human thyroid hormone receptors","authors":"Masahiro Ogawa ,&nbsp;Junya Kitamoto ,&nbsp;Mayuko Nakashima ,&nbsp;Yuto Hanaichi ,&nbsp;Sayoko Ito-Harashima ,&nbsp;Itaru Takeda ,&nbsp;Takashi Yagi ,&nbsp;Masanobu Kawanishi ,&nbsp;Taku Tanaka","doi":"10.1016/j.vascn.2025.107593","DOIUrl":"10.1016/j.vascn.2025.107593","url":null,"abstract":"<div><div>Thyroid hormones (THs) function by activating TH receptors (THRα and THRβ) on the target cells. Several chemicals adversely affect human health and ecosystems by disrupting TH signaling. Multiple assays for thyroid disruption have been reviewed for validation, especially with respect to assay reliability, sensitivity, efficiency, and technical criteria in the OECD Test Guidelines framework. The reporter gene assay is a sensitive method used to observe cellular events associated with signal transduction and gene expression. Some mammalian cell-based THR reporter gene assays have been developed optimized, and verified. In contrast, yeast-based reporter gene assays offer a cost-effective and rapid approach compared to mammalian-based assays and are considered advantageous for detecting direct or indirect interactions between test chemicals and the receptor of interest. In this study, the previously developed yeast-based reporter gene assays for human-derived THRα and THRβ were validated using several test substances, including THs and TH-disrupting chemicals, to establish scientific confidence. Our results showed that the yeast-based THRs reporter gene assays had high repeatability and the use of a fluorescent substrate improved the detection of TH disruption caused by several chemicals. Although inter-laboratory studies are needed to verify the acceptance and data interpretation criteria, our assays can provide information on the potential of chemicals to interfere with THR transactivation as animal-free <em>in vitro</em> assays.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"132 ","pages":"Article 107593"},"PeriodicalIF":1.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143485106","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The application of Bayesian forecasting to explore the effects of sex and high-fat diet on the pharmacokinetics of ropivacaine in the rat","authors":"Shamima Parvin,&nbsp;Hamdah M. Al Nebaihi,&nbsp;John R. Ussher,&nbsp;Dion R. Brocks","doi":"10.1016/j.vascn.2025.107592","DOIUrl":"10.1016/j.vascn.2025.107592","url":null,"abstract":"<div><div>Bayesian forecasting is commonly applied as part of therapeutic drug monitoring to obtain individual estimates of pharmacokinetic parameters in patients. Here its utility was explored in a preclinical study involving ropivacaine, in which sparse blood sampling data was available in the rat. Initially sample-population estimates of parameters were obtained by injecting male cannulated male Sprague-Dawley rats subcutaneously with ropivacaine HCl. Blood samples were serially drawn from each rat for 12 h after the dose (rich sampling); the concentrations were used with compartmental analysis to optimize model selection and obtain mean and variances of pharmacokinetic parameters. Two additional single doses, spaced by 5 days, were injected, each followed by 1 to 3 sparse blood draws. Other sparsely sampled age-matched groups of male and female rats given standard diet, and a group of males given high-fat diet, were dosed. Bayesian forecasting was conducted for each of these sparsely sampled rats to estimate pharmacokinetic parameters. Plasma was assayed using liquid-chromatographic method using mass spectrometry. For validation, the Bayesian parameter forecasts were compared to those using nonlinear mixed-effects modelling (NLMEM). Ropivacaine had a high clearance compared to hepatic blood flow, and a large volume of distribution. Excellent correlations were present between observed and estimated plasma concentrations using Bayesian forecasting, as was the relationship between those estimates and those obtained from NLMEM. The male rats given high-fat diet had a significant decrease in the weight-normalized clearance of ropivacaine, and female rats had a slower absorption rate. The effects were also identified using NLMEM. Bayesian forecasting has applicability in estimating pharmacokinetic properties of drugs in preclinical studies.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"132 ","pages":"Article 107592"},"PeriodicalIF":1.3,"publicationDate":"2025-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143488375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Predicting clinical outcomes from off-target receptor interactions using Secondary Intelligence™","authors":"Will S. Redfern ,&nbsp;Chris E. Pollard ,&nbsp;Mark Holbrook ,&nbsp;Barira Islam ,&nbsp;Mitra Abbasi ,&nbsp;Joanne Mahmud ,&nbsp;Katie Lambert ,&nbsp;Augustus Haslam ,&nbsp;Heeseung Jo ,&nbsp;Hiba Khalidi ,&nbsp;Zofia Bielecka ,&nbsp;Josh Starkey ,&nbsp;Thomas Ellinger ,&nbsp;Simon Bryan ,&nbsp;Angeli Savas ,&nbsp;Steve Andrews ,&nbsp;Rob Aspbury ,&nbsp;Lyn Rosenbrier Ribeiro ,&nbsp;Kim A. Henderson Park ,&nbsp;Hugo M. Vargas ,&nbsp;Clare R. Gilmer","doi":"10.1016/j.vascn.2024.107570","DOIUrl":"10.1016/j.vascn.2024.107570","url":null,"abstract":"<div><div>Adverse effects due to off-target activity can be predicted by careful comparison of the relationship between expected plasma concentration and off-target activity of the test compound with that of reference drugs targeting that receptor for their therapeutic efficacy. The ratio between plasma concentration (unbound) and the K_i at the receptor is a surrogate measure reflecting receptor occupancy. Where data are available for reference drugs, we have curated and evaluated this at 100 receptors, 72 of which can involve both negative and positive modulations by drugs: a total of 172 ‘receptor modulations’. This provides a quantitative framework upon which to achieve consistent risk assessment of off-target interactions across receptors, across compounds and between assessors. It therefore represents a significant departure from an opinion-based to an evidence-based approach to secondary pharmacology. Demonstration of proof-of-principle was achieved for one of the receptor interactions (α_1A-adrenoceptor antagonism leading to postural hypotension in clinical use) due to the availability of high-quality off-target K_i data for &gt;30 drugs at this receptor.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"131 ","pages":"Article 107570"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142693935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Drug combination assays using Caenorhabditis elegans as a model system","authors":"Guillermina Hernando,&nbsp;Cecilia Bouzat","doi":"10.1016/j.vascn.2025.107583","DOIUrl":"10.1016/j.vascn.2025.107583","url":null,"abstract":"<div><div>The <em>C. elegans</em> drug combination assay evaluates the effects of drug combinations in the nematode <em>Caenorhabditis elegans</em>, serving as a valuable tool to assess the efficacy of pharmaceutical agents and natural compounds. Using <em>C. elegans</em> as a model organism, this method allows for the efficient screening of the combined effects of different drugs and evaluation of synergistic effects in drug combinations, which reduces the risk of developing drug resistance.</div><div>Combination therapy, involving commercial drugs, new agents, or natural products, broadens treatment effectiveness by targeting multiple pathways, effectively managing complex diseases with minimized side effects. The method focuses on discovering effective drug combinations, such as anthelmintic drugs, streamlining early-stage drug discovery to save time and resources. Additionally, its versatility allows for application across most areas of pharmacology and toxicology, extending its usefulness beyond anthelmintic treatments.</div><div>In the experiments, synchronized worms are exposed to different drug concentrations to evaluate behavioral changes, mostly alterations in worm locomotion. Concentration-response curves for changes in behavior are generated and EC₅₀ or IC₅₀ values determined for the individual drugs. To determine whether the effects of a drug combination are synergistic, additive, or antagonistic, at least three different concentration ratios must be tested. These combinations are then analyzed using specialized drug combination analysis software. This methodology ensures consistent and precise outcomes and evaluates drug impacts on worm behavior parameters crucial for effective pharmacological activity.</div><div>In conclusion, the <em>C. elegans</em> drug combination assay provides critical insights for developing successful market formulations applicable across a wide range of pharmacological treatments. Its ability to efficiently screen for synergistic, additive, or antagonistic effects makes it a valuable tool for identifying effective therapeutic strategies, potentially reducing drug resistance and improving treatment outcomes in various medical and toxicological fields.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"131 ","pages":"Article 107583"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143043949","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and validation of a sensitive LC-MS/MS assay for determination of upadacitinib in human plasma and its application in patients with inflammatory bowel disease","authors":"Jing-jing Wu ,&nbsp;Lang Lin ,&nbsp;Jia-jia Yan ,&nbsp;Jia-he Kong ,&nbsp;Qiao-lan Xuan ,&nbsp;Xiang Gao ,&nbsp;Kang Chao ,&nbsp;Xia Zhu","doi":"10.1016/j.vascn.2025.107581","DOIUrl":"10.1016/j.vascn.2025.107581","url":null,"abstract":"<div><h3>Background</h3><div>Upadacitinib is a selective Janus kinase (JAK) 1 inhibitor approved by the Food and Drug Administration for the treatment of moderate-to-severe inflammatory bowel disease (IBD). We aimed to establish and validate a method for determining Upadacitinib in patients with IBD by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method.</div></div><div><h3>Methods</h3><div>The mobile phase was 0.1 % formic acid: acetonitrile (35:65, <em>v</em>/v) at a flow rate of 0.40 mL/min. Upadacitinib and its internal standard Upadacitinib <img>¹⁵N, d₂ were separated by a Waters Xbridge BEH C18 column (4.6 × 100 mm, 2.5 μm) and subjected to mass analysis using positive electrospray ionization (ESI).</div></div><div><h3>Results</h3><div>The calibration range of Upadacitinib was 0.5–200 ng/mL with the correlation coefficient r² ≥ 0.99. Accuracies ranged from −9.48 % ∼ 8.27 % and the inter- and intra-day precisions were less than 15 % for all analytes in quality control samples. There was no significant matrix effect. The range of extraction recoveries was 87.53–93.47 % for all analytes. Twenty-one plasma samples were obtained from the sixth affiliated hospital of Sun Yat-sen University. The median plasma concentration of Upadacitinib was 7.32 (0.56–26.78) ng/mL.</div></div><div><h3>Conclusion</h3><div>This newly developed method is sensitive, simple, and successfully applied in determining Upadacitinib in IBD patients to provide reference for safe and effective drug administration in clinical practice.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"131 ","pages":"Article 107581"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143043947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Compromising the immunogenicity of diphtheria toxin-based immunotoxins through epitope engineering: An in silico approach","authors":"Behrouz Golichenari ,&nbsp;Mohammad Heiat ,&nbsp;Ehsan Rezaei ,&nbsp;Amirreza Ramshini ,&nbsp;Amirhossein Sahebkar ,&nbsp;Nazila Gholipour","doi":"10.1016/j.vascn.2024.107571","DOIUrl":"10.1016/j.vascn.2024.107571","url":null,"abstract":"<div><div>Immunotoxins are genetically engineered recombinant proteins consisting of a targeting moiety, such as an antibody, and a cytotoxic toxin moiety of microbial origin. <em>Pseudomonas</em> exotoxin A and diphtheria toxin (DT) have been abundantly used in immunotoxins, with the latter applied as the toxin moiety of the FDA-approved drug Denileukin diftitox (ONTAK®). However, the use of immunotoxins provokes an adverse immune response in the host body against the toxin moiety, limiting their efficacy. <em>In silico</em> approaches have received increasing attention in protein engineering. In this study, the epitopes responsible for immunogenicity were identified through multiple platforms. By subtracting conserved and ligand-binding residues, K33, T111, and E112 were identified as common epitopes across all platforms. Substitution analysis evaluated alternative residues regarding their impact on protein stability, considering 19 different amino acid substitutions. Among the mutants explored, the T111A-E112G mutant exhibited the most destabilizing substitution for DT, thereby reducing immunogenicity. Finally, a 3D model of the mutant was generated and verified. The model was then docked with its native ligand NADH, and the complex's molecular behavior was simulated using molecular dynamics.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"131 ","pages":"Article 107571"},"PeriodicalIF":1.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}