Authors: Yanshu Zhang, Jingjing Sun, Jing Zhang...

BACKGROUND This study aimed to analyze the factors associated with the development of antibiotic-associated diarrhea (AAD) in critically ill patients receiving enzyme inhibitor antibiotics. MATERIAL AND METHODS A retrospective study of patients with and without AAD admitted to the intensive care unit (ICU) of the First Teaching Hospital of Xi'an Jiaotong University from February 1, 2014, to January 31, 2016, was undertaken. Relevant clinical data underwent univariate or multivariate regression analysis. RESULTS Of 184 patients who received enzyme inhibitor antibiotic therapy, 70 patients (38.04%) developed AAD, with a mean duration of onset of 6.97±3.64 days. AAD was associated with the use of enzyme inhibitor antibiotic therapy alone (OR, 1.142; 95% CI, 1.038-1.256; P=0.007), and in combination with antifungal agents (OR, 2.449; 95% CI, 1.116-5.372; P=0.025), quinolones (OR, 5.219; 95% CI, 1.746-15.601; P=0.003), and oxazolidinones (OR 2.895; 95% CI, 1.183-7.083; P=0.020). The mean duration of ICU stay was significantly increased in patients with AAD (19.00±11.49 days vs. 9.60±6.76 days) (P<0.001). Mean duration of antibiotic therapy (14.09±8.82 days vs. 8.10±4.91 days) (P<0.001) and duration of enzyme inhibitor antibiotic therapy (9.26±5.06 days vs. 6.61±3.24 days) (P<0.001) were significantly increased in patients with AAD. CONCLUSIONS Duration of use of enzyme inhibitor antibiotic therapy and the combined use of antifungals, quinolones, and oxazolidinones increased the incidence and duration of AAD and increased the length of stay in ICU.

Topics: Adult; Aged; Anti-Bacterial Agents; Antifungal Agents; China; Critical Illness; Diarrhea; Enzyme Inhibitors; Female; Humans; Incidence; Intensive Care Units; Male; Middle Aged; Oxazolidinones; Quinolones; Retrospective Studies

PubMed: 30512009
DOI: 10.12659/MSM.913739

Authors: Hope Adamson, Modupe O Ajayi, Emma Campbell...

Robust technology is required to underpin rapid point-of-care and in-field diagnostics to improve timely decision making across broad sectors. An attractive strategy combines target recognition and signal generating elements into an "active" enzyme-switch that directly transduces target-binding into a signal. However, approaches that are broadly applicable to diverse targets remain elusive. Here, an enzyme-inhibitor switch sensor was developed by insertion of non-immunoglobulin Affimer binding proteins, between TEM1-β-lactamase and its inhibitor protein, such that target binding disrupts the enzyme-inhibitor complex. Design principles for a successful switch architecture are illustrated by the rapid (min), simple (wash-free), and sensitive (pM) quantification of multimeric target analytes in biological samples (serum, plasma, leaf extracts), across three application areas. A therapeutic antibody (Herceptin), protein biomarker (human C-reactive protein), and plant virus (cow pea mosaic virus) were targeted, demonstrating assays for therapeutic drug monitoring, health diagnostics, and plant pathogen detection, respectively. Batch-to-batch reproducibility, shelf-life stability, and consistency with validated enzyme-linked immunosorbent assay analysis confirm that the principle of an Affimer-enzyme-inhibitor switch provides a platform for point-of-care and in-field diagnostics.

Topics: Biosensing Techniques; Enzyme Inhibitors; Enzyme-Linked Immunosorbent Assay; Humans; beta-Lactamases

PubMed: 31578863
DOI: 10.1021/acssensors.9b01574

Authors: Kevin M Cottrell, Kimberly J Briggs, Douglas A Whittington...

It has been shown that PRMT5 inhibition by small molecules can selectively kill cancer cells with homozygous deletion of the gene if the inhibitors can leverage the consequence of deletion, namely, accumulation of the MTAP substrate MTA. Herein, we describe the discovery of TNG908, a potent inhibitor that binds the PRMT5·MTA complex, leading to 15-fold-selective killing of -deleted (MTAP-null) cells compared to intact (MTAP WT) cells. TNG908 shows selective antitumor activity when dosed orally in mouse xenograft models, and its physicochemical properties are amenable for crossing the blood-brain barrier (BBB), supporting clinical study for the treatment of both CNS and non-CNS tumors with loss.

Topics: Protein-Arginine N-Methyltransferases; Humans; Animals; Mice; Antineoplastic Agents; Drug Discovery; Enzyme Inhibitors; Cell Line, Tumor; Xenograft Model Antitumor Assays; Neoplasms; Brain; Structure-Activity Relationship

PubMed: 38595098
DOI: 10.1021/acs.jmedchem.4c00133

Authors: Shane M Buker, P Ann Boriack-Sjodin, Robert A Copeland...

Contemporary chemical biology and drug discovery are increasingly focused on the discovery of inhibitory molecules that interact with enzyme targets in specific ways, such as allosteric or orthosteric binding. Hence, there is increasing interest in evaluating hit compounds from high-throughput diversity screening to determine their mode of interaction with the target. In this work, the common inhibition modalities are reviewed and clarified. The impact of substrate concentration, relative to substrate K, for each common inhibition modality is also reviewed. The pattern of changes in IC that accompany increasing substrate concentration are shown to be diagnostic of specific inhibition modalities. Thus, replots of IC as a function of the ratio [S]/K are recommended as a simple and rapid means of assessing inhibition modality. Finally, specific recommendations are offered for ideal experimental conditions for the determination of inhibition modality through the use of IC replots.

Topics: Drug Discovery; Drug Evaluation, Preclinical; Enzyme Inhibitors; Enzymes; High-Throughput Screening Assays; Humans; Kinetics; Substrate Specificity

PubMed: 30811960
DOI: 10.1177/2472555219829898

Authors: Gareth Davies, Hannah Semple, Megan McCandless...

Enzymes represent a significant proportion of the druggable genome and constitute a rich source of drug targets. Delivery of a successful program for developing a modulator of enzyme activity requires an understanding of the enzyme's mechanism and the mode of interaction of compounds. This allows an understanding of how physiological conditions in disease-relevant cells will affect inhibitor potency. As a result, there is increasing interest in evaluating hit compounds from high-throughput screens to determine their mode of interaction with the target. This work revisits the common inhibition modalities and illustrates the impact of substrate concentration relative to K upon the pattern of changes in IC that are expected for increasing substrate concentration. It proposes a new, high-throughput approach for assessing mode of inhibition, incorporating analyses based on a minimal descriptive model, to deliver a workflow that allows rapid and earlier compound classification immediately after high-throughput screening.

Topics: Drug Discovery; Enzyme Activation; Enzyme Inhibitors; Enzymes; High-Throughput Screening Assays; Humans; Substrate Specificity; Workflow

PubMed: 33482076
DOI: 10.1177/2472555220983809

Review

Authors: Si'Ana A Coggins, Bijan Mahboubi, Raymond F Schinazi...

Enzyme kinetic analysis reveals a dynamic relationship between enzymes and their substrates. Overall enzyme activity can be controlled by both protein expression and various cellular regulatory systems. Interestingly, the availability and concentrations of intracellular substrates can constantly change, depending on conditions and cell types. Here, we review previously reported enzyme kinetic parameters of cellular and viral DNA and RNA polymerases with respect to cellular levels of their nucleotide substrates. This broad perspective exposes a remarkable co-evolution scenario of DNA polymerase enzyme kinetics with dNTP levels that can vastly change, depending on cell proliferation profiles. Similarly, RNA polymerases display much higher values than DNA polymerases, possibly due to millimolar range rNTP concentrations found in cells (compared with micromolar range dNTP levels). Polymerases are commonly targeted by nucleotide analog inhibitors for the treatments of various human diseases, such as cancers and viral pathogens. Because these inhibitors compete against natural cellular nucleotides, the efficacy of each inhibitor can be affected by varying cellular nucleotide levels in their target cells. Overall, both kinetic discrepancy between DNA and RNA polymerases and cellular concentration discrepancy between dNTPs and rNTPs present pharmacological and mechanistic considerations for therapeutic discovery.

Topics: Animals; DNA-Directed DNA Polymerase; DNA-Directed RNA Polymerases; Drug Discovery; Enzyme Inhibitors; Humans; Kinetics; Nucleotides; Substrate Specificity

PubMed: 32737197
DOI: 10.1074/jbc.REV120.013746

Authors: Rina Baba, Satoru Matsuda, Yuuichi Arakawa...

Persistent epigenetic dysregulation may underlie the pathophysiology of neurodevelopmental disorders, such as autism spectrum disorder (ASD). Here, we show that the inhibition of lysine-specific demethylase 1 (LSD1) enzyme activity normalizes aberrant epigenetic control of gene expression in neurodevelopmental disorders. Maternal exposure to valproate or poly I:C caused sustained dysregulation of gene expression in the brain and ASD-like social and cognitive deficits after birth in rodents. Unexpectedly, a specific inhibitor of LSD1 enzyme activity, 5-((1,2)-2-((cyclopropylmethyl)amino)cyclopropyl)--(tetrahydro-2-pyran-4-yl)thiophene-3-carboxamide hydrochloride (TAK-418), almost completely normalized the dysregulated gene expression in the brain and ameliorated some ASD-like behaviors in these models. The genes modulated by TAK-418 were almost completely different across the models and their ages. These results suggest that LSD1 enzyme activity may stabilize the aberrant epigenetic machinery in neurodevelopmental disorders, and the inhibition of LSD1 enzyme activity may be the master key to recover gene expression homeostasis. TAK-418 may benefit patients with neurodevelopmental disorders.

Topics: Autism Spectrum Disorder; Autistic Disorder; Enzyme Inhibitors; Epigenesis, Genetic; Female; Histone Demethylases; Humans

PubMed: 33712455
DOI: 10.1126/sciadv.aba1187

Authors: Jayaprakasam Bolleddula, Hao Chen, Lawrence Cohen...

Metabolism and disposition of pevonedistat, an investigational, first-in-class inhibitor of the NEDD8-activating enzyme (NAE), were characterized in patients with advanced solid tumors after intravenous infusion of [C]pevonedistat at 25 mg/m (∼60-85 Ci radioactive dose). More than 94% of the administered dose was recovered, with ∼41% and ∼53% of drug-related material eliminated in urine and feces, respectively. The metabolite profiles of [C]pevonedistat were established in plasma using an accelerator mass spectrometer and excreta with traditional radiometric analysis. In plasma, unchanged parent drug accounted for approximately 49% of the total drug-related material. Metabolites M1 and M2 were major (>10% of the total drug-related material) circulating metabolites and accounted for approximately 15% and 22% of the drug-related material, respectively. Unchanged [C]pevonedistat accounted for approximately 4% and 17% of the dose in urine and feces, respectively. Oxidative metabolites M1, M2, and M3 appeared as the most abundant drug-related components in the excreta and represented approximately 27%, 26%, and 15% of the administered dose, respectively. Based on the unbound plasma exposure in cancer patients and in vitro NAE inhibition, the contribution of metabolites M1 and M2 to overall in vivo pharmacological activity is anticipated to be minimal. The exposure to these metabolites was higher at safe and well tolerated doses in rat and dog (the two preclinical species used in toxicology evaluation) plasma than that observed in human plasma. Reaction phenotyping studies revealed that CYP3A4/5 are primary enzymes responsible for the metabolic clearance of pevonedistat. SIGNIFICANCE STATEMENT: This study details the metabolism and clearance mechanisms of pevonedistat, a first-in-class NEDD8-activating enzyme inhibitor, after intravenous administration to patients with cancer. Pevonedistat is biotransformed to two major circulating metabolites with higher exposure in nonclinical toxicological species than in humans. The pharmacological activity contribution of these metabolites is minimal compared to the overall target pharmacological effect of pevonedistat. Renal clearance was not an important route of excretion of unchanged pevonedistat (∼4% of the dose).

Topics: Administration, Oral; Animals; Cyclopentanes; Dogs; Enzyme Inhibitors; Feces; Infusions, Intravenous; Neoplasms; Pyrimidines; Rats

PubMed: 35504658
DOI: 10.1124/dmd.122.000842

Authors: Ashley P Dudey, Jake M Rigby, Gregory R Hughes...

The HECT E3 ubiquitin ligases 1 (WWP1) and 2 (WWP2) are responsible for the ubiquitin-mediated degradation of key tumour suppressor proteins and are dysregulated in various cancers and diseases. Here we expand their limited inhibitor space by identification of NSC-217913 displaying a WWP1 IC of 158.3 µM (95% CI = 128.7, 195.1 µM). A structure-activity relationship by synthesis approach aided by molecular docking led to compound which displayed increased potency with an IC of 32.7 µM (95% CI = 24.6, 44.3 µM) for WWP1 and 269.2 µM (95% CI = 209.4, 347.9 µM) for WWP2. Molecular docking yielded active site-bound poses suggesting that the heterocyclic imidazo[4,5-]pyrazine scaffold undertakes a π-stacking interaction with the phenolic group of tyrosine, and the ethyl ester enables strong ion-dipole interactions. Given the therapeutic potential of WWP1 and WWP2, we propose that compound 11 may provide a basis for future lead compound development.

Topics: Ubiquitin-Protein Ligases; Humans; Structure-Activity Relationship; Molecular Structure; Dose-Response Relationship, Drug; Molecular Docking Simulation; Enzyme Inhibitors

PubMed: 39223706
DOI: 10.1080/14756366.2024.2394895

Review

Authors: Philine Kirsch, Alwin M Hartman, Anna K H Hirsch...

In this review, a general introduction to fragment-based drug design and the underlying concepts is given. General considerations and methodologies ranging from library selection/construction over biophysical screening and evaluation methods to in-depth hit qualification and subsequent optimization strategies are discussed. These principles can be generally applied to most classes of drug targets. The examples given for fragment growing, merging, and linking strategies at the end of the review are set in the fields of enzyme-inhibitor design and macromolecule-macromolecule interaction inhibition. Building upon the foundation of fragment-based drug discovery (FBDD) and its methodologies, we also highlight a few new trends in FBDD.

Topics: Drug Design; Drug Discovery; Enzyme Inhibitors; Humans; Macromolecular Substances

PubMed: 31779114
DOI: 10.3390/molecules24234309