Broad-spectrum Drug Research Articles

Design, Synthesis, and Biological Evaluation of 1,3,4-Thiadiazole Derivatives as Novel Potent Peptide Deformylase Inhibitors for Combating Drug-Resistant Gram-Positive and -Negative Bacteria.

The prevalence of drug-resistant bacteria is a major challenge throughout the world, especially with respect to Gram-negative bacteria, such as drug-resistant Acinetobacter baumannii, which are regarded as the greatest bacterial threat to human health by the World Health Organization (WHO). In this work, 1,3,4-thiadiazole was introduced into the main skeleton of the classical peptidomimetic peptide deformylase (PDF) inhibitor in pursuit of highly efficient and broad-spectrum bacteriostatic drugs. Upon detailed structure-activity relationship study, PDF inhibitors that possess satisfactory activity against both Gram-positive and Gram-negative bacteria as well as a lower potential for methemoglobin toxicity were screened out. The mechanism of the empowered antibacterial activity against Gram-negative bacteria was also investigated. Finally, for the first time, remarkable protective efficacy against drug-resistant A. baumannii in a mouse model was achieved by a PDF inhibitor (compound 43). These findings can pave a way to new approaches to the development of novel broad-spectrum PDF inhibitors.

Nerol attenuates doxorubicin-induced heart failure by inhibiting cardiomyocyte apoptosis in rats.

As a broad-spectrum anti-tumour drug, the clinical application of DOX is often limited owing to its cardiotoxicity. Nerol is a naturally occurring compound with both anti-inflammatory and antioxidant properties. However, the ability of Nerol to improve DOX-induced heart failure and its underlying mechanisms remain unclear. Rat models of DOX-induced heart failure were established and rats were treated with various doses of Nerol. Apoptosis in cardiomyocytes was detected using TUNEL staining and the expression levels of apoptosis-related proteins were detected using western blotting and immunofluorescence. In addition, mitochondrial structure was observed using electron microscopy, mitochondrial membrane potential was detected using a JC-1 fluorescent probe, and superoxide dismutase were detected to comprehensively evaluate the regulatory effect of Nerol on mitochondrial function and oxidative stress. Analysis showed that the number of apoptotic cardiomyocytes was significantly reduced after Nerol treatment, accompanied by the downregulation of Bax protein expression and upregulation of Bcl-2 protein expression, suggesting that Nerol may inhibit the apoptotic process of cardiomyocytes by regulating the balance of Bcl-2 family proteins. In addition, the mitochondrial function of Nerol-treated rats was protected, as indicated by the stability of the mitochondrial membrane potential, integrity of mitochondrial morphology. These changes suggest that Nerol may reduce the severity of heart failure by improving mitochondrial function. Nerol plays a positive role in alleviating DOX-induced heart failure in rats, possibly by inhibiting cardiomyocyte apoptosis. These findings provide novel evidence and potential targets for developing new cardioprotective drugs.

Deep Learning-driven Identification and Evaluation of Potential Therapeutic Agents for COVID-19 Co-infection Diseases Targeting METTL3

Background: Patients with COVID-19 often have an impact on populations with underlying co-infection. METTL3 plays a crucial role in numerous infectious diseases, making the research on broad-spectrum anti-infective drugs targeting METTL3 both captivating and compelling. Objective: The study aims to identify anti-infection potential compounds targeting METTL3 using a comprehensive virtual screening approach. Method: The approach involves ensemble docking and molecular dynamics simulations to predict and validate the potential of natural compounds. This was complemented by the application of deep learning, specifically CNN binary classification and regression models, to predict the anti-infection capabilities of the compounds identified through docking. Experimental validation through Surface Plasmon Resonance (SPR) further confirmed the computational predictions. Network and gene ontology analyses provided insights into the compounds' biological targets and pathways. Results: Ensemble docking approach achieved high prediction accuracy. We identified 17 natural compounds as potential METTL3 inhibitors, with Rosavin and Eriocitrin showing strong antiinfection potential against HIV-1, SARS-CoV-2, Mycobacterium tuberculosis, and Influenza A. Rosavin's low toxicity and strong METTL3 binding affinity were confirmed by SPR experiments and MD simulations. Network and gene ontology analyses suggest Rosavin may enhance immune responses by disrupting interferon degradation. Conclusion: The multidimensional analysis identified Rosavin as a potent METTL3 inhibitor with significant anti-COVID-19 co-infection diseases potential.

Phytochemical analysis, therapeutic and molecular docking studies for the compounds of wild type and Senkambu variants of curry leaves targeting HER2 Kinase domain a potential gastric cancer receptor

Gastric cancer is the fifth most notable health concern globally. In recent years, molecular docking, a computational technique, has emerged as tool in drug discovery. The present investigation aimed to identify the major bioactive compounds in the wild-type curry leaves found in the Shevaroy Hills and the local Senkambu variant from Karamadai. Virtual screening of 40 ligands from Curry leaves of wild type and Senkambu type was identified through GC-MS profiling. These compounds were targeted against HER2 Kinase domain which is a potential receptor for Gastric cancer. Information regarding the binding site residues for the receptor was predicted using CASTp server. Molecular docking was performed for HER2 kinase domain with the predicted compounds through GC-MS profiling. The top 3 hits reported with least binding affinity for the target protein were considered for further interaction analysis using Biovia Discovery studio visualizer. Upon analyzing the interacted compounds, the Piperine from Wild type curry leaves was found to have good interaction with HER2 Kinase domain by forming two hydrogen bonds and binding score of -8.3 k cal/mol. The current study might guide the designing of analogues of piperine in the evolution of effective broad spectrum drug development in cancer therapy.

Non-Polio Enterovirus Inhibitors: Scaffolds, Targets, and Potency─What's New?

Enterovirus (EV) is a genus that includes a large diversity of viruses spread around the world. They are the main cause of numerous diseases with seasonal clusters, like hand-foot-mouth disease (HFMD). A vaccine is marketed in China for the prevention of HFMD caused by EV-A71. Despite the need, no antiviral is marketed to date. Therefore, several compounds have been currently evaluated to inhibit non-polio Enterovirus (NPEV), namely direct antiviral agents and host target inhibitor. We propose to make a review of the latest molecules evaluated as NPEV inhibitors and to summarize structure-activity relationships between these inhibitors and their target. We provide access to all recent information on Enterovirus inhibitors, regardless of the species, to facilitate the design of future broad-spectrum drugs.

Tailoring moxifloxacin hydrochloride loaded oleic acid liposomes for the topical management of Methicillin-Resistant Staphylococcus aureus (MRSA)- Induced skin infection: In-vitro characterization and in-vivo assessment.

Oleic acid liposomes (OALs) are novel vesicular carriers ofunsaturated fatty acids and their corresponding ionized species, arranged within an enclosed lipid bilayer. This study aimed to encapsulate moxifloxacin HCl (MOX), a broad-spectrum antibacterial drug into OALs for effective treatment of Methicillin-resistant Staphylococcus aureus (MRSA) infection through topical application. Various OALs were formulatedby combining varied quantities of phosphatidylcholine (PC), oleic acid (OA), and cholesterol (CH) with 50 mg of MOX. The OALs were produced utilizing varying sonication durations. MOX-loaded OALs were formulated using the thin film hydration method by applying (24) a full factorial design utilizing the Design-Expert® software. The formula for MOX-loaded OALs was OAL13, which consisted of 200 mg of PC and 20 mg of OA. The mixture was sonicated for 5 min. The OAL13 exhibited spherical vesicles with a small diameter and a smooth outer surface. Additionally, the entrapment efficiency was measured to be 75.00 ± 1.41 %, the particle size was 234.65 ± 4.74 nm, the polydispersity index was 0.53 ± 0.01, and the zeta potential was -38.50 ± 0.42 mV. The OAL13 formula exhibited an extended release profile. Moreover, the antibiofilm activity of OAL13 gel and MOX-loaded liposomes gel against MRSA infection demonstrates greater activity than the MOX gel at the maximum concentration used (MIC/2). Furthermore, the in-vivo study showed that OAL13 improved MOX's antimicrobial and immunomodulatory effects against MRSA infection by increasing TLR-2 and IL-1β, as well as their downstream molecules NF-κB and TNF-α. Moreover, the histopathological examination conducted by a skin irritation test has verified the safety of OAL13. Overall, the results demonstrated the significant efficacy of MOX-loaded OALs in the treatment of MRSA infected wounds when applied topically.

Некоторые цианозамещённые спиросочленённые пиразолины и пирролидины, обладающие антимикробной активностью

This work summarizes one-pot synthetic approaches to spiro-fused (di)azoles, which are promising in terms of studying biological activity. Proposed approaches involve condensation reactions of (hetero)cyclic ketones, methylene active moieties, and N-nucleophiles, occurring under mild conditions by both stepwise and concerted mechanisms. The antimicrobial activity of signals from the proposed approaches of cyanosubstituted spiro-membered pyrazolines and pyrrolidines against gram-negative (P. aeruginosa, E. coli) and gram-positive (S. aureus) losses has been studied (EC50 determination (μg/ml)).The well-known component of broad-spectrum antimicrobial drugs, ciprofloxacin, has been used as a control. It has been found that 3-amino-2-(4-nitrophenyl)-1,2-diazaspiro[4.5]dec-3-ene-4-carbonitrile exhibits the greatest inhibitory effect against selected gram-negative bacteria. In this case, the phenyl substituent at the second nitrogen atom of the pyrazoline ring plays an important role, since its replacement with an aroyl fragment reduces the studied effect by several tens of times, regardless of the type of the second ring. Substituted spiroindolinopyrrolizidinedicarbonitriles, containing cyano groups at the 2nd carbon atom of the pyrrolizidine ring and a phenyl substituent at the first, also exhibit a moderate and high inhibitory effect. For pairs with the same type of substituent position, compounds containing a 3,4 Cl2 substituent in the phenyl fragment have the greatest antimicrobial activity.

Physiologically based pharmacokinetic modeling and simulation of topiramate in populations with renal and hepatic impairment and considerations for drug-drug interactions.

Topiramate (TPM) is a broad-spectrum antiepileptic drug (AED) commonly prescribed for approved and off-label uses. Routine monitoring is suggested for clinical usage of TPM in special population due to its broad side effect profile. Therefore, it is crucial to further explore its pharmacokinetic characteristics. Physio-chemical properties of TPM were initially determined from online database and further optimized while establishing the PBPK model for healthy adults using the PK-Sim software. The model was then extrapolated to patients with renal impairment and patients who were hepatically impaired. A drug-drug interaction (DDI) model was also built to simulate plasma TPM concentrations while concomitantly used with carbamazepine (CBZ). The goodness-of-fit method and average fold error (AFE) method were used to compare the differences between predicted and observed values to assess the accuracy of the PBPK model. Almost all of the predicted concentration fell within twofold error range of corresponding observed concentrations. The AFE ratio of predicted to observed values of Cmax and AUC0-inf was all within 0.5 and 2. It is recommended that the doses be reduced to 70%, 50%, and 40% of the healthy adult dose for the chronic kidney disease (CKD) stage 3, stage 4, and stage 5 patients, respectively, and reduced to ~70%, and 35% for the Child-Pugh-B, and Child-Pugh C scored patient with hepatic impairment, respectively. If TPM is co-administered with CBZ, increasing TPM doses to 150%-175% of the monotherapy dose is recommended according to model simulation.

Formulation and in-vitro permeation kinetic studies of β-cyclodextrin complexed ketoconazole drug capped silver nanoparticles

Background: Invasive fungal infections are a growing global crisis due to environmental shifts and growing vulnerable populations. Pathogenic fungi that infect humans are developing resistance against all approved systemic antifungal drugs. Topical drug delivery systems provide a non-invasive way to distribute therapeutic substances directly to the site of action. Ketoconazole (KZ) is a BCS Class II broad-spectrum drug that is used to treat and prevent fungal infections in which Trichophyton rubrum is the main causative agent.Materials and Methods: The present study utilizes an in-vitro model in which KZ is complexed with β-CD and then capped with AgNPs to formulate a cream and evaluated for the underlying mechanisms and amount of improved passive penetration. The formulated cream is characterized for physiochemical evaluation, Fourier transform infrared spectroscopy (FTIR), kinetic studies indicating the effective stability and encapsulation of the composite.Results: The prepared sample was evaluated in terms of its appearance, drug content, spreadability, and viscosity. Drug diffusion through the dialysis membrane, and the findings showed the β-CD-KZ-AgNPs formulation had a significantly higher permeability than KZ alone and followed first-order kinetics with an anomalous release pattern. The higher solubility and stability of β-CD and nano-sized AgNPs resulted in improved penetration. This study investigates that β-CD-capped AgNPs can significantly enhance KZ passive penetration and diffusion, providing a potentially useful nanocarrier technology for antifungal treatment.Conclusion: Enhancing the passive permeation and diffusion characteristics of KZ on β-Cyclodextrin (β-CD)-capped silver nanoparticles (AgNPs) presents a viable approach for enhancing drug delivery efficiency.

ANTIBIOGRAM OF PRE-PROCESSED RAW CHICKEN MEAT FROM DIFFERENT SUPERSHOPS OF DHAKA CITY, BANGLADESH

Pathogenic strains of salmonella, S. aureus, S. epidermidis, shigella, enterobacter and citrobacter are serious health threat for human being. Present investigation was conducted to assess the antibiogram of pre-processed raw chicken meat collected from different chain-shops of Dhaka city. Pre-processed chicken meat samples were collected from three different super-shops (S1, S2, and S3) in Dhaka. The Samples are serially diluted to 10-4, and after growth on MSA, MAC, and SS agars, the total counts showed The most growth S2 (360 × 10.4  18.33), (235 × 10.4), (25 × 10.14.4  3.6) followed by S3 (353.3 × 10.4  42.39), (151.67 × 10.4  22.19), 15 × 10-4  3.0) and finally S1 (3.26.67 × 10.4  29.48), (64.5 × 10.4  8.62) (14 × 10.4  5.13) respectively. Salmonella S aureus, S. epidermidis, Shigella, Enterobacter and Citrobacter were identified after a number of morphological and biochemical tests; Gram staining, MIU, KIA, oxidase and catalase tests, and citrate utilization test following aseptic techniques. Six antibiotics; Vancomycin, and five broad spectrum drugs; Streptomycin, Norfloxacin, Novobiocin, Chloramphenicol, and Cefotaxime were used to test the sensitivity or resistance of the organisms. All the pathogens were susceptible to chloramphenicol. Staphylococcusepidermidis were novobiocin-resistant. Citrobacter spp. and Enterobacter spp. were sensitive to all except novobiocin. Staphylococcusepidermidis was susceptible and intermediate to norfloxacin. Staphylococcusaureus was resistant to norfloxacin and novobiocin; intermediate to cefotaxime, and sensitive to the rest. Salmonella spp. was found to be resistant to novobiocin, vancomycin, and streptomycin; intermediate to norfloxacin, but sensitive to chloramphenicol and cefotaxime. Shigella.spp was found to be sensitive to all the antibiotics but resistant to novobiocin and vancomycin. Fecal coliform E. coli was absent showing some degree of sanitation in the chicken, and all the pathogens were found susceptible to at least two antibiotics

Cryo-EM structure of Nipah virus L-P polymerase complex

Nipah virus (NiV) is a non-segmented, negative-strand (NNS) RNA virus, belonging to Paramyxoviridae. The RNA polymerase complex, composed of large (L) protein and tetrameric phosphoprotein (P), is responsible for genome transcription and replication by catalyzing NTP polymerization, mRNA capping and cap methylation. Here, we determine the cryo-electron microscopy (cryo-EM) structure of fully bioactive NiV L-P polymerase complex at a resolution of 3.19 Å. The L-P complex displays a conserved architecture like other NNS RNA virus polymerases and L interacts with the oligomerization domain and the extreme C-terminus region of P tetramer. Moreover, we elucidate that NiV is naturally resistant to the allosteric L-targeting inhibitor GHP-88309 due to the conformational change in the drug binding site. We also find that the non-nucleotide drug suramin can inhibit the NiV L-P polymerase activity at both the enzymatic and cellular levels. Our findings have greatly enhanced the molecular understanding of NiV genome replication and transcription and provided the rationale for broad-spectrum polymerase-targeted drug design.

Human essential gene identification based on feature fusion and feature screening.

Essential genes are necessary to sustain the life of a species under adequate nutritional conditions. These genes have attracted significant attention for their potential as drug targets, especially in developing broad-spectrum antibacterial drugs. However, studying essential genes remains challenging due to their variability in specific environmental conditions. In this study, the authors aim to develop a powerful prediction model for identifying essential genes in humans. The authors first obtained the essential gene data from human cancer cell lines and characterised gene sequences using 7 feature encoding methods such as Kmer, the Composition of K-spaced Nucleic Acid Pairs, and Z-curve. Subsequently, feature fusion and feature optimisation strategies were employed to select the impactful features. Finally, machine learning algorithms were applied to construct the prediction models and evaluate their performance. The single-feature-based model achieved the highest area under the Receiver Operating Characteristic curve (AUC) of 0.830. After fusing and filtering these features, the classical machine learning models achieved the highest AUC at 0.823 while the deep learning model reached 0.860. Results obtained by the authors show that compared to using individual features, feature fusion and feature optimisation strategies significantly improved model performance. Moreover, the study provided an advantageous method for essential gene identification compared to other methods.

Enzymes of ADP-Heptose Biosynthesis As Targets for the Creation of Broad-Spectrum Antibacterial Drugs

Enzymes of ADP-Heptose Biosynthesis As Targets for the Creation of Broad-Spectrum Antibacterial Drugs

Quantitative fluorescent detection of tetracycline in animal-derived foods using quantum dots

Tetracycline (Tc) antibiotics, a class of synthetically produced broad-spectrum antimicrobial drugs, have been widely used in animal husbandry, leading to their widespread presence in animal-derived foods. However, misuse, overuse, and non-compliance with withdrawal periods in animal farming have resulted in excessive Tc residues in these foods, which can cause various adverse reactions in humans, induce bacterial resistance, and pose a significant threat to public health. Consequently, the detection of Tc antibiotic residues in animal-derived food has become a critical issue. This study aims to establish a novel method for quantifying Tc residues in animal-derived food using quantum dots (QDs) fluorescence immunoassay (FLISA). The developed method was optimized to achieve a detection limit of 0.69 ng/mL and a quantitative detection range of 1.30 ~ 59.22 ng/mL. The applicability of the method was demonstrated by successfully determining Tc residues in pork, chicken, fish, milk, eggs, and honey samples spiked with Tc standard solutions, yielding recoveries ranging from 94.01% to 110.19% and relative standard deviations between 1.10% and 11.39%. The significance of this study lies in its potential to provide a rapid and reliable approach for monitoring Tc residues in animal-derived food products, thereby contributing to the enhancement of food safety monitoring practices.Key points• Screen out tetracycline-specific blocking monoclonal antibodies• The quantitative detection has high specificity and sensitivity• This method can be a useful tool for laboratories or testing facilities

Ivermectin inhibits the growth of ESCC by activating the ATF4-mediated endoplasmic reticulum stress-autophagy pathway.

Esophageal squamous cell carcinoma (ESCC) is one of the most common forms of malignancy worldwide. However, there is currently a lack of effective chemotherapeutic drugs for ESCC. Ivermectin is a broad-spectrum antiparasitic drug with notable antitumor activity. However, the cellular and molecular mechanisms by which ivermectin inhibits cancer growth remain unclear. In this study, we elucidate the role of ivermectin in ESCC suppression by activating the endoplasmic reticulum (ER) stress and autophagy pathways. In transcriptome analyses, we find that activating transcription factor 4 (ATF4) and DNA damage inducible transcript 3 (DDIT3) are involved in the activation of ER stress by ivermectin. Moreover, ivermectin treatment suppresses the growth of ESCC xenograft tumors in nude mice. Taken together, our results establish the antitumor molecular role of ivermectin in targeting the ER stress-autophagy pathway and suggest that ivermectin is a potential drug candidate for the treatment of ESCC.

Cloning of the Thermus thermophilus methionine aminopeptidase gene and confirmation of the enzyme functional activity

Background. Methionine aminopeptidases (MAPs) are a class of enzymes that catalyze the removal of the N-terminal initiator methionine from a polypeptide chain. Bacterial MAPs are considered as targets for the development of broad-spectrum antibacterial drugs, and using MAPs in biotechnology necessitates the search for new MAPs and the study of their functioning and inhibition mechanisms.The aim of the study. To identify methionine aminopeptidase in the Thermus thermophilus genome (Tt-MAP) and to confirm its functional activity.Materials and methods. To identify Tt-MAP, we analyzed the Thermus thermophilus genome in the GeneBank database. Modern genetic engineering techniques (polymerase chain reaction, restriction, transformation, heterologous expression) were used to clone the putative open reading frame (ORF) encoding Tt-MAP in the pHUE vector. Various chromatography techniques (affinity, ion exchange, and size-exclusion) were used to obtain a purified enzyme preparation. The fluorogenic substrate L-methionine 7-amino-4-methylcoumarin (Met-AMC) was used to confirm the specific functional aminopeptidase activity of the enzyme.Results. An ORF encoding MAP was identified in the Thermus thermophilus bacterium genome. Oligonucleotide primers were designed based on the nucleotide sequence. The ORF was cloned in the vector, and the recombinant enzyme was produced in E. coli cells. A method for purifying the enzyme to a homogeneous state was developed using a series of sequential chromatographies, allowing up to 30 mg to be obtained from 1 liter of culture. Using the fluorogenic substrate Met-AMC, the specific functional activity of the enzyme was demonstrated (the enzyme cleaves methionine from the substrate).Conclusion. We have identified the Thermus thermophilus MAP and tested its functional activity. It has been shown that the ORF product TTHA1670 encodes a methionine-specific aminopeptidase, i. e. methionine aminopeptidase. The enzyme can be used in various fields of biotechnology and scientific research.

Effect of UGT1A6 and UGT2B7 polymorphisms on the valproic acid serum concentration and drug-induced liver injury.

Aim: Valproic acid (VPA) is a classic broad-spectrum antiepileptic drug, with significant pharmacokinetic variability. Genetic polymorphisms contribute to this variability, influencing both VPA trough serum concentration (VPA concentration) and VPA-induced liver injury. Our study aims to investigate the association between polymorphisms of uridine diphosphate glucuronyl transferase (UGT) 1A6, UGT2B7 and VPA concentration and screen for potential genetic loci affecting VPA-induced liver injury.Methods: This study included epilepsy patients treated with VPA. PCR-RFLP method was used to determine the genotypes of UGT1A6 and UGT2B7. Chemiluminescent microparticle immunoassay was used to measure VPA concentration. Multiple linear regression and logistic regression were employed to analyze factors influencing VPA concentration and VPA-induced liver injury, respectively.Results: The correlation between UGT polymorphism and VPA concentration was analyzed in 133 samples. For VPA-induced liver injury, 105 patients were analyzed, with 29 in the liver injury group and 76 in the control group. Our finding showed patients with the UGT1A6-T19G variant had significantly lower VPA concentrations compared with wild-type patients and UGT1A6-T19G, A541G, A552C and UGT2B7-C802T, G211T, A268G polymorphisms showed no impact on VPA-induced liver injury.Conclusion: This study demonstrated UGT1A6-T19G polymorphisms affected the VPA concentration, providing a theoretical basis for the individualized clinical use of VPA.

Understanding Voriconazole Metabolism: A Middle-Out Physiologically-Based Pharmacokinetic Modelling Framework Integrating In Vitro and Clinical Insights.

Voriconazole (VRC), a broad-spectrum antifungal drug, exhibits nonlinear pharmacokinetics (PK) due to saturable metabolic processes, autoinhibition and metabolite-mediated inhibition on their own formation. VRC PK is also characterised by high inter- and intraindividual variability, primarily associated with cytochrome P450 (CYP)2C19 genetic polymorphism. Additionally, recent in vitro findings indicate that VRC main metabolites, voriconazole N-oxide (NO) and hydroxyvoriconazole (OHVRC), inhibit CYP enzymes responsible for VRC metabolism, adding to its PK variability. This variability poses a significant risk of therapeutic failure or adverse events, which are major challenges in VRC therapy. Understanding the underlying processes and sources of these variabilities is essentialfor safe and effective therapy. This work aimed to develop a whole-body physiologically-based pharmacokinetic (PBPK) modelling framework that elucidates the complex metabolism of VRC and the impact of its metabolites, NO and OHVRC, on the PK of the parent, leveraging both in vitro and in vivo clinical data in a middle-out approach. A coupled parent-metabolite PBPK model for VRC, NO and OHVRC was developed in a stepwise manner using PK-Sim® and MoBi®. Based on available in vitro data, NO formation was assumed to be mediated by CYP2C19, CYP3A4, and CYP2C9, while OHVRC formation was attributed solely to CYP3A4. Both metabolites were assumed to be excreted via renal clearance, with hepatic elimination also considered for NO. Inhibition functions were implemented to describe the complex interaction network of VRC autoinhibition and metabolite-mediated inhibition on each CYP enzyme. Using a combined bottom-up and middle-out approach, incorporating data from multiple clinical studies and existing literature, the model accurately predicted plasma concentration-time profiles across various intravenous dosing regimens in healthy adults, of different CYP2C19 genotype-predicted phenotypes. All (100%) of the predicted area under the concentration-time curve (AUC) and 94% of maximum concentration (Cmax) values of VRC met the 1.25-fold acceptance criterion, with overall absolute average fold errors of 1.12 and 1.14, respectively. Furthermore, all predicted AUC and Cmax values of NO and OHVRC met the twofold acceptance criterion. This comprehensive parent-metabolite PBPK model of VRC quantitatively elucidated the complex metabolism of the drug and emphasised the substantial impact of the primary metabolites on VRC PK. The comprehensive approachcombining bottom-up and middle-out modelling, therebyaccounting for VRC autoinhibition, metabolite-mediated inhibition, and the impact of CYP2C19 genetic polymorphisms, enhances our understanding of VRC PK. Moreover, the model can be pivotal in designing further in vitro experiments, ultimately allowing for extrapolation to paediatric populations, enhance treatment individualisation and improve clinical outcomes.

Ultrasound-assisted extraction of itraconazole from pellets: an efficient approach for the recovery of active pharmaceutical compounds from solid dosage forms

IntroductionItraconazole is a widely used broad-spectrum antifungal drug on a global scale. However, its poor water solubility necessitates the development of advanced formulations for its effective use. Currently, itraconazole is available commercially in capsule form, with pellets containing the active drug. The extraction of itraconazole from these granules is often required when developing new formulations. Extracting active pharmaceutical ingredients (APIs) from pellets is crucial in ensuring precise and consistent dosing when producing pharmaceutical dosage forms. Surprisingly, no existing methods for extracting itraconazole from pellets have been reported thus far.ObjectiveTherefore, this study aimed to develop and evaluate an efficient method for extracting this API from pellets.MethodsTwo extraction methods were assessed: conventional Soxhlet extraction and ultrasound-assisted extraction.Results and DiscussionThe results demonstrated that ultrasound-assisted extraction significantly outperformed the conventional method, yielding higher amounts of itraconazole with a high purity level of 96.8%.

The S2 Pocket Governs the Genus-Specific Substrate Selectivity of Coronavirus 3C-Like Protease.

Coronavirus 3C-like protease (CoV 3CLpro) is essential for viral replication, providing an attractive target for monitoring the evolution of CoV and developing anti-CoV drugs. Here, the substrate-binding modes of 3CLpros from four CoV genera are analyzed and found that the S2 pocket in 3CLpro is highly conserved within each genus but differs between genera. Functionally, the S2 pocket, in conjunction with S4 and S1' pockets, governs the genus-specific substrate selectivity of 3CLpro. Resurrected ancestral 3CLpros from four CoV genera validate the genus-specific divergence of S2 pocket. Drawing upon the genus-specific S2 pocket as evolutionary marker, eight newly identified 3CLpros uncover the ancestral state of modern 3CLpro and elucidate the possible evolutionary process for CoV. It is also demonstrated that the S2 pocket is highly correlated with the genus-specific inhibitory potency of PF-07321332 (an FDA-approved drug against COVID-19) on different CoV 3CLpros. This study on 3CLpro provides novel insights to inform evolutionary mechanisms for CoV and develop genera-specific or broad-spectrum drugs against CoVs.