Affinity Sites Research Articles

Zinc-based organophyllosilicate nanosheets as novel carbonic anhydrase-like nanozyme for efficient CO2 fixation

The unparalleled catalytic activity of carbonic anhydrases (CAs) towards CO2 hydrolysis can potentially contribute to carbon capture, utilization, and storage technology aimed at global decarbonization; however, the high cost and low stability of CAs significantly limit their practical applications. Recently, the development of robust and effective nanozymes with CA-mimicking activity has been proposed as a promising solution to this challenge. Herein, a novel zinc-based organophyllosilicate (ZnAC) nanozyme with CA-like catalytic activity was synthesized by a facile one-pot sol–gel method for the first time, leading to a Zn-O coordination structure that mimicked the catalytic center fragments of natural CAs, with surface amino groups acting as the affinity sites and proton shuttles. The synergistic effect of the two components on the open surface of the layered structure resulted in excellent CA-like activity with a Michaelis–Menten constant (Km) of 4.071 mM, maximum velocity (Vmax) of 0.220 mM/s, and turnover number (kcat) of 11.213 /s at 25 °C and pH=7.5, which were superior to those of most previously reported CA-like nanozymes. The catalytic performance of ZnAC significantly improved with increasing pH and temperature; furthermore, ZnAC exhibited excellent stability, maintaining high catalytic activity after the exposure to extreme temperatures or pH values, eight reaction cycles, and 30 d of storage. Additionally, ZnAC significantly increased the rate of CO2 hydration and the extent of CO2 mineralization and precipitation. This work proposes a new strategy for designing and constructing efficient CA-like nanozymes, providing valuable insights into the structure–activity relationship and new perspectives for the implementation of CO2 fixation.

Joint experimental-computational: Revealing the role of sodium poly(heptazineimide) in the selective separation of CO2 in PEBAX-based mixed matrix membranes

Mixed matrix membranes (MMMs) with the dual properties of porous materials and polymer matrix membranes have become one of the most optimal solutions to the gas separation problem. However, the adapted porous material and the explicit transport mechanism of gases within the mixed matrix membrane hinder its further commercialization. Thanks to their porous nature and tunable gas adsorption properties, heteroatom-doped porous materials, especially nitrogen-doped porous materials, are widely used to design and prepare advanced CO2 separation membranes. Herein, a layered two-dimensional nano-sheet with a nitrogen content of 44.7 % was successfully synthesized via thermal polymerization. And nano-sheets were homogeneously dispersed in a Pebax matrix to make a composite film. Since the CO2 affinity of the nitrogen functional site and the PEO flexible chain, realizing the synergistic improvement of CO2 permeability and selectivity. A comprehensive combined permeation experimental/simulation characterization reveals the CO2 transport pathway within the membrane, indicating the mechanism of action of nitrogen-doped porous packing on CO2 diffusion. Among these, the CO2 permeability of the mixed matrix film containing 5 wt% sodium poly(heptazine imide) (NaPHI) reached 153 Barrer at a feed gas pressure of 9 bar, nearly 149 % compared to the pure Pebax film. The selectivity of CO2/N2 was enhanced to 105, exhibiting a remarkable increase of 159 % and exceeding the 2019 Robeson limit. Importantly, the present work further reveals and demonstrates the gas separation mechanism of nitrogen-doped porous filler/polymer hybrid matrix membranes, which provides new ideas for the design and application of heteroatom-doped porous fillers in the field of gas separation.

Investigation of the Anti-Inflammatory Properties of Bioactive Compounds from Olea europaea: In Silico Evaluation of Cyclooxygenase Enzyme Inhibition and Pharmacokinetic Profiling.

In a landmark study, oleocanthal (OLC), a major phenolic in extra virgin olive oil (EVOO), was found to possess anti-inflammatory activity similar to ibuprofen, involving inhibition of cyclooxygenase (COX) enzymes. EVOO is a rich source of bioactive compounds including fatty acids and phenolics; however, the biological activities of only a small subset of compounds associated with Olea europaea have been explored. Here, the OliveNetTM library (consisting of over 600 compounds) was utilized to investigate olive-derived compounds as potential modulators of the arachidonic acid pathway. Our first aim was to perform enzymatic assays to evaluate the inhibitory activity of a selection of phenolic compounds and fatty acids against COX isoforms (COX-1 and COX-2) and 15-lipoxygenase (15-LOX). Olive compounds were found to inhibit COX isoforms, with minimal activity against 15-LOX. Subsequent molecular docking indicated that the olive compounds possess strong binding affinities for the active site of COX isoforms, and molecular dynamics (MD) simulations confirmed the stability of binding. Moreover, olive compounds were predicted to have favorable pharmacokinetic properties, including a readiness to cross biological membranes as highlighted by steered MD simulations and umbrella sampling. Importantly, olive compounds including OLC were identified as non-inhibitors of the human ether-à-go-go-related gene (hERG) channel based on patch clamp assays. Overall, this study extends our understanding of the bioactivity of Olea-europaea-derived compounds, many of which are now known to be, at least in part, accountable for the beneficial health effects of the Mediterranean diet.

Upstream CtrA-binding sites both induce and repress pilin gene expression in Caulobacter crescentus

Pili are bacterial surface structures important for surface adhesion. In the alphaproteobacterium Caulobacter crescentus, the global regulator CtrA activates transcription of roughly 100 genes, including pilA which codes for the pilin monomer that makes up the pilus filament. While most CtrA-activated promoters have a single CtrA-binding site at the − 35 position and are induced at the early to mid-predivisional cell stage, the pilA promoter has 3 additional upstream CtrA-binding sites and it is induced at the late predivisional cell stage. Reporter constructs where these additional sites were disrupted by deletion or mutation led to increased activity compared to the WT promoter. In synchronized cultures, these mutations caused pilA transcription to occur approximately 20 min earlier than WT. The results suggested that the site overlapping the − 35 position drives pilA gene expression while the other upstream CtrA-binding sites serve to reduce and delay expression. EMSA experiments showed that the − 35 Site has lower affinity for CtrA∼P compared to the other sites, suggesting binding site affinity may be involved in the delay mechanism. Mutating the upstream inhibitory CtrA-binding sites in the pilA promoter caused significantly higher numbers of pre-divisional cells to express pili, and phage survival assays showed this strain to be significantly more sensitive to pilitropic phage. These results suggest that pilA regulation evolved in C. crescentus to provide an ecological advantage within the context of phage infection.

Insecticidal Properties and Chemical Characterization of Laurus nobilis L. Essential Oils from Two Regions of Morocco against Callosobruchus maculatus (Coleoptera: Bruchinae)

Morocco is a significant botanical reservoir that boasts a wealth of raw materials with promising applications across various industrial sectors, notably in pharmaceuticals and food. The objective of this study was to assess the effectiveness of essential oils (EOs) derived from Laurus nobilis L. leaves originating from the Tanger (EOT) and Meknes (EOM) regions in combating Callosobruchus maculatus infection. The chemical compositions of these oils were examined using Fourier transform infrared (FTIR) spectroscopy and gas chromatography–mass spectrometry (GC-MS). The biological activity of the EOs was evaluated via repulsion and fumigation tests against C. maculatus at varying concentrations. FTIR analysis revealed distinct vibrational bands indicative of various chemical compounds. GC-MS analysis was used to delineate the major chemical constituents of the EOs. The three predominant compounds in the EOT were 1,8-cineole (37.64%), linalool (16.40%), and adamantane (12.00%), whereas 1,8-cineole (47.84%), toluene (17.60%), and α-phellandrene (8.44%) were the most abundant in the EOM. Notably, the EOs exhibited significant repellent activity against C. maculatus, with repulsion percentages ranging from 51.11 to 90.00% in Tanger and 67.78 to 93.33% in Meknes. Mortality rates varied from 0 to 100% depending on the treatment. However, the mean concentrations showed mortality rates ranging from 29.44 to 65.56% for the EOT and from 21.11 to 67.78% for the EOM, with LD50 values of 11.96 μL/L and 5.22 μL/L. Docking studies revealed that 1,8-cineole had the highest binding affinity for the active site of acetylcholinesterase, thus confirming its toxic activity against C. maculatus. The findings of this study highlight the ability of EOs extracted from L. nobilis in the Moroccan regions of Tanger and Meknes to act as effective insecticides and repellents against C. maculatus, thereby highlighting avenues for further exploration of pest management and agricultural practices.

ANTIBACTERIAL AND ANTIBIOFILM ACTIVITY OF ERYNGIUM FOETIDUM ESSENTIAL OIL

Eryngium aquaticum L. (Apiaceae), a culinary herb enjoyed in global cuisines, is also valued for its medicinal properties. The essential oil (EO) extracted from E. aquaticum leaves by hydro-distillation was subjected to antibacterial and antibiofilm activity using a microtiter plate-based in vitro assay against Staphylococcus aureus and Pseudomonas aeruginosa. Compounds identified by GC-MS analysis of EO were screened against the transcriptional regulatory proteins SarA of S. aureus and LasR of P. aeruginosa by molecular docking analysis. The minimum inhibitory concentration (MIC) was recorded as 250 μg/mL against both of these two pathogens. The EO of E. aquaticum also showed concentration-dependent antibiofilm activity against these pathogens, with a maximum inhibition of 50.9 and 48.03% against P. aeruginosa and S. aureus, respectively, at the highest concentration (500 μg/mL) tested. The GC-MS analysis identified 17 compounds and all of them showed moderate to weak binding affinity for the active sites of SarA and LasR, with pentanedioic acid (2,4-di-t-butylphenyl) mono-ester showing the best docking score against SarA (-5.7 kcal/mol) and LasR (-8.0 kcal/mol). This study suggests that E. aquaticum can be a good source of EO with antibacterial and antibiofilm activity against P. aeruginosa and S. aureus.

Baicalein loaded liposome with hyaluronic acid and Polyhexamethylene guanidine modification for anti methicillin-resistant Staphylococcus aureus infection

Targeting delivery to the infection site and good affinity of vehicle to the bacterial are two main concerns in therapy of bacterial infection, and on-demand release of drug is another important issue. In this work, a liposome drug delivery system (HA/P/BAI-lip) incorporated with baicalein and modified by PHMG and HA was prepared. Several characterizations were conducted to examine the physical properties of liposome. Then it was applied to treatments of MRSA induced dorsal subcutaneous abscess model and the thigh muscle infected model. The presence of guanidine group in HA/P/BAI-lip rendered the liposome satisfactory bacterial target ability and good pH sensitive properties. The lipase secreted by bacterial could promote the hydrolysis of soybean phosphatidylcholine (SPC) in liposome. The modification of HA in HA/P/BAI-lip could lead the drug system to the exact infected site where CD44 was abundant because of inflammation. The low pH microenvironment characteristic of bacterial infection could induce the swelling of liposome following by degradation. Taken together, baicalein could be released selectively at the infected site to exert antibacterial capacity. HA/P/BAI-lip showed impressive antibacterial ability and dramatically decrease the bacterial burden of infection site and alleviate the infiltration of inflammatory cells, facilitating the recovery of infection.

Aggregation of Apo/Glycated Human Serum Albumins and Aptamer-Saturated Graphene Quantum Dot: A Simulation Study.

Human serum albumin (HSA) is a protein carrier that transports a wide range of drugs and nutrients. The amount of glycated HSA (GHSA) is used as a diabetes biomarker. To quantify the GHSA amount, the fluorescent graphene-based aptasensor has been a successful method. In aptasensors, the key mechanism is the adsorption/desorption of albumin from the aptamer-graphene complex. Recently, the graphene quantum dot (GQD) has been reported to be an aptamer sorbent. Due to its comparable size to aptamers, it is attractive enough to explore the possibility of GQD as a part of an albumin aptasensor. Therefore, molecular dynamics (MD) simulations were performed here to reveal the binding mechanism of albumin to an aptamer-GQD complex in molecular detail. GQD saturated by albumin-selective aptamers (GQDA) is studied, and GHSA and HSA are studied in comparison to understand the effect of glycation. Fast and spontaneous albumin-GQDA binding was observed. While no specific GQDA-binding site on both albumins was found, the residues used for binding were confined to domains I and III for HSA and domains II and III for GHSA. Albumins were found to bind preferably to aptamers rather than to GQD. Lysines and arginines were the main contributors to binding. We also found the dissociation of GLC from all GHSA trajectories, which highlights the role of GQDA in interfering with the ligand binding affinity in Sudlow site I. The binding of GQDA appears to impair albumin structure and function. The insights obtained here will be useful for the future design of diabetes aptasensors.

Novel Approach to Exploring Protease Activity and Targets in HIV-associated Obstructive Lung Disease using Combined Proteomic-Peptidomic Analysis.

Obstructive lung disease (OLD) is increasingly prevalent among persons living with HIV (PLWH). However, the role of proteases in HIV-associated OLD remains unclear. We combined proteomics and peptidomics to comprehensively characterize protease activities. We combined mass spectrometry (MS) analysis on bronchoalveolar lavage fluid (BALF) peptides and proteins from PLWH with OLD (n=25) and without OLD (n=26) with a targeted Somascan aptamer-based proteomic approach to quantify individual proteases and assess their correlation with lung function. Endogenous peptidomics mapped peptides to native proteins to identify substrates of protease activity. Using the MEROPS database, we identified candidate proteases linked to peptide generation based on binding site affinities which were assessed via z-scores. We used t-tests to compare average forced expiratory volume in 1 second per predicted value (FEV1pp) between samples with and without detection of each cleaved protein and adjusted for multiple comparisons by controlling the false discovery rate (FDR). We identified 101 proteases, of which 95 had functional network associations and 22 correlated with FEV1pp. These included cathepsins, metalloproteinases (MMP), caspases and neutrophil elastase. We discovered 31 proteins subject to proteolytic cleavage that associate with FEV1pp, with the top pathways involved in small ubiquitin-like modifier mediated modification (SUMOylation). Proteases linked to protein cleavage included neutrophil elastase, granzyme, and cathepsin D. In HIV-associated OLD, a significant number of proteases are up-regulated, many of which are involved in protein degradation. These proteases degrade proteins involved in cell cycle and protein stability, thereby disrupting critical biological functions.

Biomimetic designing MOFs nanosheet based membranes with self-recovery two-dimensional selective channels for specific molecules long-lasting precise separation

Two-dimensional (2D) materials-based membranes with artificial transfer channels have shown significant potential for selective separation. However, the challenges such as uncontrollable interlayer spacing and undesirable molecular sieving capabilities of 2D channels have impeded their further application in separation. Inspired by biological selectivity transport channels with proper steric and affinity sites, herein we have designed biomimetic 2D selective transport channels based on a ZIF-L nanosheet membrane. In this design, the UiO-66-NH2 nanoparticles tune the appropriate interlayer confinement and compensate for laminate framework defects of 2D selectivity transport channels, the artificial imprinting recognition sites establishes the essential chemical environment for specific separation. As a result, the obtained MOFs nanosheet based membranes with imprinted recognition sites (MN-IMs) exhibited enhanced permeation flux (J = 1.0847 × 10−3 and 1.0423 × 10−3 mg min−1 cm−2) and permselectivity (α = 3.77 and 4.10), outperforming state-of-the-art similar technologies. Besides, the composite MOFs demonstrated good photoinduced self-recovery ability, which also enables MN-IMs to have long-lasting selective separation performance (the separation efficiency is 90.76 %) in the continuous separation process. This study introduces a novel design strategy for developing sophisticated 2D materials-based membranes and offers new insights into the precise separation of specific molecules.

Synthesis, structure analysis, Hirshfeld surface studies, molecular docking studies against cyclin-dependent kinases (CDKs) of sulfonamide decorated N6-benzyl aminopurines for cancer treatment

Cyclin-dependent kinases (CDKs) are recognized as the primary regulators of the cell cycle. CDKs are over-expressed in various types of cancer. Inhibiting CDKs with small molecules can reduce tumor growth and benefit cancer patients. In the current study, sulfonamide-decorated N6-benzyl aminopurines such as ligand 1: (α- alpha (Purin-6-ylamino)-p-toluenesulfonamide) and ligand 2: (α-(2-Amino purin-6-ylamino)-p-toluenesulfonamide)) purine were synthesized and explored for their anticancer efficacy. Crystallographic analysis revealed that ligand 1 crystallizes in P-1 of the triclinic and ligand 2 molecule in C2/c or P21/c of the monoclinic systems. Hirshfeld surface and X-ray crystallographic studies discovered that ligand 2 possesses stronger noncovalent interaction ability than roscovitine. In silico analysis showed that ligand 2 had a higher binding affinity for ATP binding sites of CDK1, CDK2, and CDK4 receptors than roscovitine (a known CDK inhibitor). The enhanced binding affinity of ligand 2 with CDKs was found to be associated with strong noncovalent interactions between ligand 2 and specific amino acids in CDKs. Cytotoxicity studies were conducted on the glioblastoma cell line (U251) by incubating cells with ligands 0, 1, and 2, roscovitine, and the established anticancer drug Temozolomide (TMZ). Ligand 2 (66.12 ± 1.09 µM) demonstrated better cytotoxicity compared to ligand 1 (81.22 ± 0.30 µM), roscovitine (127.10 ± 0.47 µM), and TMZ (165.11 ± 1 µM). It was well known that CDK inhibition can lead to cell cycle arrest. Intriguingly, ligand 2 induced G2/M phase cell cycle arrest by increasing the percentage of the G2/M phase cell population from 21.08 % to 47.79 % in U251 cells. Based on these results, ligand 2 showed anticancer properties by binding to the ATP binding site of CDKs. Hence, ligand 2 can serve as a potential lead molecule in the development of effective anticancer agents.

Mixed matrix membranes consisting of PDMS and IL@MoS2 for enhancing 2-phenylethanol pervaporation separation

The MoS2-based mixed matrix membranes (MMMs) exhibited excellent pervaporation separation for aromatic compounds, but the narrow interlayer spacing and few affinity sites in MoS2 limited the simultaneous improvement in the total flux and separation factor. Therefore, constructing 2-PE transport channels and affinity sites in MMMs is of significant importance to improve the pervaporation separation performance. Herein, 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (IL) was intercalated into the interlayer of the molybdenum disulfide (MoS2) under electrostatic and concentration diffusion interactions to obtain IL@MoS2. Then, IL@MoS2 was incorporated into polydimethylsiloxane (PDMS) to prepare MMMs for efficient 2-PE separation, which had good miscibility with polymer matrix. The total flux, separation factor and pervaporation separation index of as-prepared MMMs reached 1466 g m−2 h−1, 33 and 47,571 g m−2 h−1, separately, which increased by 26 %, 31 % and 44 % compared with that of PDMS embedding with MoS2. This is because IL@MoS2 in MMMs not only can afford 2-PE transport channels, but also provide 2-PE affinity sites due to the hydrogen bonds and π-π interactions, which promotes the solubility of 2-PE in MMMs. When the concentration of maltol increased to 1500 ppm in 2-PE aqueous solution, the 2-PE flux and separation factor of 2-PE/water only decreased by 23 % and 26 %, revealing that MMMs had good 2-PE selectivity in 2-PE/maltol/water mixture. In addition, it displayed exceptional durability during 168 h testing, demonstrating that this membrane had a good application prospect for 2-PE separation.

Synthesis, structural characterization, fukui functions, DFT calculations, molecular docking and biological efficiency of some novel heterocyclic systems

The electron deficient centers in α,β-unsaturated ketone 3 are C-2 and C=O of chromone moieties as well as C=O and H-β of α,β-unsaturated ketone segment. To investigate the reactivity of various electron deficient centers and to synthesize some heterocyclic systems, compound 3 was allowed to react with some binucleophilic reagents. The antimicrobial activity of some compounds showed a notable inhibitory effect on the microorganisms that were being studied. The synthesized compounds were verified by spectral and analytical data. The structure of the prepared compounds were optimized at the DFT/B3LYP level of theory employing 6-311G(d,p). Density Functional Theory (DFT) was used to compute the chemical reactivity descriptors. Compound 9 is more stable and less reactive than other compounds. MEP mapping was applied at the same computational level to identify the favored locations for electrophilic and nucleophilic assault, providing additional insight into the regioselectivity in the desired reaction. Moreover, the most reactive sites of compound 3 were investigated by Fukui function descriptor using Mulliken charges. The 13C and 1H-NMR spectra of the prepared compound were measured experimentally in DMSO, and correlated with the theoretical calculations that performed by 6-311G(d,p) of DFT-B3LYP method. All compounds met Veber's and Lipinski's requirements according to in silico predictions made with the SwissADME online server, suggesting the possibility for usage as oral active drugs. The synthesized compounds were also subjected to molecular docking studies to investigate their binding pattern and affinity for the CDKs active site and correlated with antimicrobial data.

Transient interactions modulate the affinity of NF-κB transcription factors for DNA

The dimeric nuclear factor kappa B (NF-κB) transcription factors (TFs) regulate gene expression by binding to a variety of κB DNA elements with conserved G:C-rich flanking sequences enclosing a degenerate central region. Toward defining mechanistic principles of affinity regulated by degeneracy, we observed an unusual dependence of the affinity of RelA on the identity of the central base pair, which appears to be noncontacted in the complex crystal structures. The affinity of κB sites with A or T at the central position is ~10-fold higher than with G or C. The crystal structures of neither the complexes nor the free κB DNAs could explain the differences in affinity. Interestingly, differential dynamics of several residues were revealed in molecular dynamics simulation studies, where simulation replicates totaling 148 μs were performed on NF-κB:DNA complexes and free κB DNAs. Notably, Arg187 and Arg124 exhibited selectivity in transient interactions that orchestrated a complex interplay among several DNA-interacting residues in the central region. Binding and simulation studies with mutants supported these observations of transient interactions dictating specificity. In combination with published reports, this work provides insights into the nuanced mechanisms governing the discriminatory binding of NF-κB family TFs to κB DNA elements and sheds light on cancer pathogenesis of cRel, a close homolog of RelA.

The Identification of Peptide Inhibitors of the Coronavirus 3CL Protease from a Fucus ceranoides L. Hydroalcoholic Extract Using a Ligand-Fishing Strategy.

Brown seaweeds of the Fucus genus represent a rich source of natural antiviral products. In this study, a Fucus ceranoides hydroalcoholic extract (FCHE) was found to inhibit 74.2 ± 1.3% of the proteolytic activity of the free SARS-CoV-2 3CL protease (3CLpro), an enzyme that plays a pivotal role in polyprotein processing during coronavirus replication and has been identified as a relevant drug discovery target for SARS- and MERS-CoVs infections. To purify and identify 3CLpro ligands with potential inhibitory activity using a one-step approach, we immobilized the enzyme onto magnetic microbeads (3CLpro-MPs), checked that the enzymatic activity was maintained after grafting, and used this bait for a ligand-fishing strategy followed by a high-resolution mass spectrometry analysis of the fished-out molecules. Proof of concept for the ligand-fishing capacity of the 3CLpro-MPs was demonstrated by doping the FCHE extract with the substrate peptide TSAVLQ-pNA, resulting in the preferential capture of this high-affinity peptide within the macroalgal complex matrix. Ligand fishing in the FCHE alone led to the purification and identification via high-resolution mass spectrometry (HRMS) of seven hepta-, octa-, and decapeptides in an eluate mix that significantly inhibited the free 3CLpro more than the starting FCHE (82.7 ± 2.2% inhibition). Molecular docking simulations of the interaction between each of the seven peptides and the 3CLpro demonstrated a high affinity for the enzyme's proteolytic active site surpassing that of the most affine peptide ligand identified so far (a co-crystallographic peptide). Testing of the corresponding synthetic peptides demonstrated that four out of seven significantly inhibited the free 3CLpro (from 46.9 ± 6.4 to 76.8 ± 3.6% inhibition at 10 µM). This study is the first report identifying peptides from Fucus ceranoides with high inhibitory activity against the SARS-CoV-2 3CLprotease which bind with high affinity to the protease's active site. It also confirms the effectiveness of the ligand-fishing strategy for the single-step purification of enzyme inhibitors from complex seaweed matrices.

Exploration of morpholine-thiophene hybrid thiosemicarbazones for the treatment of ureolytic bacterial infections via targeting urease enzyme: Synthesis, biochemical screening and computational analysis

An important component of the pathogenicity of potentially pathogenic bacteria in humans is the urease enzyme. In order to avoid the detrimental impact of ureolytic bacterial infections, the inhibition of urease enzyme appears to be an appealing approach. Therefore, in the current study, morpholine-thiophene hybrid thiosemicarbazone derivatives (5a-i) were designed, synthesized and characterized through FTIR, 1H NMR, 13C NMR spectroscopy and mass spectrometry. A range of substituents including electron-rich, electron-deficient and inductively electron-withdrawing groups on the thiophene ring was successfully tolerated. The synthesized derivatives were evaluated in vitro for their potential to inhibit urease enzyme using the indophenol method. The majority of compounds were noticeably more potent than the conventional inhibitor, thiourea. The lead inhibitor, 2-(1-(5-chlorothiophen-2-yl)ethylidene)-N-(2-morpholinoethyl)hydrazinecarbothioamide (5g) inhibited the urease in an uncompetitive manner with an IC50 value of 3.80 ± 1.9 µM. The findings of the docking studies demonstrated that compound 5g has a strong affinity for the urease active site. Significant docking scores and efficient binding free energies were displayed by the lead inhibitor. Finally, the ADME properties of lead inhibitor (5g) suggested the druglikeness behavior with zero violation.

New oxadiazol‐5‐ones derivatives and their performance as angiotensin‐converting enzyme (ACE) inhibitors

AbstractAngiotensin‐converting enzyme inhibitors are widely used in treating arterial hypertension, acting on the renin‐angiotensin‐aldosterone system and controlling blood pressure. We present a novel, greener, and faster methodology to assess the 1,2,4‐oxadiazol‐5‐one ring and perform molecular modifications to obtain angiotensin‐converting enzyme (ACE) inhibitors using this heterocyclic core. Molecular docking simulations indicate that the tested compounds exhibited an affinity for the ACE binding site, with scores comparable to the commercial inhibitor lisinopril. However, in vitro assays revealed that the compounds were ineffective in inhibiting ACE activity. The lack of inhibition may be related to the compounds' more apolar nature. These results emphasize the importance of integrating computational and experimental approaches in developing new drugs, providing valuable insights for planning future studies to optimize the activity of synthesized compounds.

The effects of intensive trapping on invasive round goby densities.

The round goby (Neogobius melanostomus) is an invasive benthic fish first introduced to the Laurentian Great Lakes in 1990 that has negatively impacted native fishes through increased competition for food and habitat, aggressive interactions, and egg predation. While complete eradication of the round goby is currently not possible, intensive trapping in designated areas during spawning seasons could potentially protect critical native fish spawning habitats. Baited minnow traps were spaced 10 meters apart in shallow water along a 100-meter stretch of shoreline within the Duluth-Superior Harbor during the round goby breeding period (June to October) with captured round gobies removed from interior traps (N = 10) every 48 hours. These traps were bracketed by two pairs of reference traps deployed weekly for 48 hours, from which round gobies were also tagged and released. The number of round gobies captured in the interior traps declined by 67% compared to reference traps over the course of the study, with extended periods of no captures. The tagged round gobies showed high site affinity, with 82.8% of tagged fish recaptured at the previous release site. The results indicate that even at open water sites, which allow natural migration of round gobies into the area, extensive trapping could reduce local population numbers.

Copaifera spp. oleoresins and two isolated compounds (ent-kaurenoic and ent-polyalthic acid) inhibit Toxoplasma gondii growth in vitro

Toxoplasmosis affects about one-third of the world's population. The disease treatment methods pose several side effects and do not efficiently eliminate the parasite, making the search for new therapeutic approaches necessary. We aimed to assess the anti-Toxoplasma gondii activity of four Copaifera oleoresins (ORs) and two isolated diterpene acids, named ent-kaurenoic and ent-polyalthic acid. We used HeLa cells as an experimental model of toxoplasmosis. Uninfected and infected HeLa cells were submitted to the treatments, and the parasite intracellular proliferation, cytokine levels and ROS production were measured. Also, tachyzoites were pre-treated and the parasite invasion was determined. Finally, an in silico analysis was performed to identify potential parasite targets. Our data show that the non-cytotoxic concentrations of ORs and diterpene acids controlled the invasion and proliferation of T. gondii in HeLa cells, thus highlighting the possible direct action on parasites. In addition, some compounds tested controlled parasite proliferation in an irreversible manner. An additional and non-exclusive mechanism of action involves the modulation of host cell components, by affecting the upregulation of the IL-6. Additionally, molecular docking suggested that ent-polyalthic acid has a high affinity for the active site of the TgCDPK1 protein. Copaifera ORs have great antiparasitic activity against T. gondii, and this effect can be partially explained by the presence of the isolated compounds ent-kaurenoic and ent-polyalthic acid.

Synthesis, In Silico Study, and In Vitro Antifungal Activity of New 5-(1,3-Diphenyl-1H-Pyrazol-4-yl)-4-Tosyl-4,5-Dihydrooxazoles.

The increase in multi-drug resistant Candida strains has caused a sharp rise in life-threatening fungal infections in immunosuppressed patients, including those with SARS-CoV-2. Novel antifungal drugs are needed to combat multi-drug-resistant yeasts. This study aimed to synthesize a new series of 2-oxazolines and evaluate the ligands in vitro for the inhibition of six Candida species and in silico for affinity to the CYP51 enzymes (obtained with molecular modeling and protein homology) of the same species. The 5-(1,3-diphenyl-1H-pyrazol-4-yl)-4-tosyl-4,5-dihydrooxazoles 6a-j were synthesized using the Van Leusen reaction between 1,3-diphenyl-4-formylpyrazoles 4a-j and TosMIC 5 in the presence of K2CO3 or KOH without heating, resulting in short reaction times, high compound purity, and high yields. The docking studies revealed good affinity for the active site of the CYP51 enzymes of the Candida species in the following order: 6a-j > 4a-j > fluconazole (the reference drug). The in vitro testing of the compounds against the Candida species showed lower MIC values for 6a-j than 4a-j, and for 4a-j than fluconazole, thus correlating well with the in silico findings. According to growth rescue assays, 6a-j and 4a-j (like fluconazole) inhibit ergosterol synthesis. The in silico toxicity assessment evidenced the safety of compounds 6a-j, which merit further research as possible antifungal drugs.