Non-competitive Research Articles

Features of biliary tract diseases in ketamine abusers: a systematic review of case reports

Background and aimsAnesthesiologists prefer ketamine for certain surgeries due to its effectiveness as a non-competitive inhibitor of the N-methyl-D-aspartate receptor in the brain. Recently, this agent has also shown promise as an antidepressant. However, ketamine can cause hallucinogenic effects and is sometimes abused as an illicit drug. Ketamine abuse has been associated with liver and bile duct complications. This systematic study aims to better understand cholangiopathy in ketamine abusers by reviewing case reports.Methods and materialIn this systematic review, a comprehensive literature search was conducted with the terms “biliary tract diseases” and “ketamine”. Case reports and case series of adult patients with documented ketamine abuse and reported cholangiopathy or biliary tract disease were included. We extracted the data of relevant information and the results were reported through narrative synthesis and descriptive statistics.ResultsA total of 48 studies were initially identified, and 11 studies were finally included in the review. The mean age of the patients was 25.88 years. Of the 17 patients, 64.7% were men. Symptoms often included abdominal pain, nausea, and vomiting. Most patients were discharged with improved symptoms and liver function. Common bile duct dilation and other findings were observed in imaging results and other diagnostic studies.ConclusionThis review highlights the diverse presentations and diagnostic modalities used in ketamine-induced cholangiography. These patients tend to be young men with deranged liver function tests and abdominal pain, which should be taken into consideration. These patients often require a multidisciplinary approach in their management.

Rehmannioside A inhibits the activity of CYP3A4, 2C9 and 2D6 in vitro

To assess the effect of Rehmannioside A on CYP450s activity and to estimate its inhibitory properties. The effect of Rehmannioside A on the activity of major CYP450s in human liver microsomes (HLMs) was assessed with the corresponding substrates and marker reactions, and compared with a blank control and the respective inhibitors. Suppression of CYP3A4, 2C9 and 2D6 was assessed by the dose-dependent assay and fitted with non-competitive or competitive inhibition models. The inhibition of CYP3A4 was determined in a time-dependent manner. Rehmannioside A suppressed the activity of CYP3A4, 2C9, and 2D6 with IC50 values of 10.08, 12.62, and 16.43 μM, respectively. Suppression of CYP3A4 was fitted to a non-competitive model with Ki value of 5.08 μM, whereas CYP2C9 and 2D6 were fitted to a competitive model with Ki values of 6.25 and 8.14 μM. Additionally, the inhibitory effect on CYP3A4 was time-dependent with KI value of 8.47 μM−1 and a Kinact of 0.048 min−1. In vitro suppression of CYP3A, 2C9 and 2D6 by Rehmannioside A indicated that Rehmannioside A or its source herbs may interact with drugs metabolised by these CYP450s, which could guide the clinical application.

The Impact of Baohuoside I on the Metabolism of Tofacitinib in Rats.

To study the potential drug-drug interactions between tofacitinib and baohuoside I and to provide the scientific basis for rational use of them in clinical practice. A total of eighteen Sprague-Dawley rats were randomly divided into three groups: control group, single-dose group (receiving a single dose of 20 mg/kg of baohuoside I), and multi-dose group (receiving multiple doses of baohuoside I for 7 days). On the seventh day, each rat was orally administered with 10 mg/kg of tofacitinib 30 minutes after giving baohuoside I or vehicle. Blood samples were collected and determined using UPLC-MS/MS. In vitro effects of baohuoside I on tofacitinib was investigated in rat liver microsomes (RLMs), as well as the underlying mechanism of inhibition. The semi-inhibitory concentration value (IC50) of baohuoside I was subsequently determined and its inhibitory mechanism against tofacitinib was analyzed. Furthermore, the interactions between baohuoside I, tofacitinib and CYP3A4 were explored using Pymol molecular docking simulation. The administration of baohuoside I orally has been observed to enhance the area under the concentration-time curve (AUC) of tofacitinib and decrease the clearance (CL). The observed disparity between the single-dose and multi-dose groups was statistically significant. Furthermore, our findings suggest that the impact of baohuoside I on tofacitinib metabolism may be a mixture of non-competitive and competitive inhibition. Baohuoside I exhibit an interaction with arginine (ARG) at position 106 of the CYP3A4 enzyme through hydrogen bonding, positioning itself closer to the site of action compared to tofacitinib. Our study has demonstrated the presence of drug-drug interactions between baohuoside I and tofacitinib, which may arise upon pre-administration of tofacitinib. Altogether, our data indicated that an interaction existed between tofacitinib and baohuoside I and additional cares might be taken when they were co-administrated in clinic.

The Mechanism of Inhibition of Mycobacterial (p)ppGpp Synthetases by a Synthetic Analog of Erogorgiaene.

The synthesis of (p)ppGpp alarmones plays a vital role in the regulation of metabolism suppression, growth rate control, virulence, bacterial persistence, and biofilm formation. The (p)ppGpp alarmones are synthesized by proteins of the RelA/SpoT homolog (RSH) superfamily, including long bifunctional RSH proteins and small alarmone synthetases. Here, we investigated enzyme kinetics and dose-dependent enzyme inhibition to elucidate the mechanism of 4-(4,7-dimethyl-1,2,3,4-tetrahydronaphthalen-1-yl)pentanoic acid (DMNP) action on the (p)ppGpp synthetases RelMsm and RelZ from Mycolicibacterium smegmatis and RelMtb from Mycobacterium tuberculosis. DMNP was found to inhibit the activity of RelMtb. According to the enzyme kinetics analysis, DMNP acts as a noncompetitive inhibitor of RelMsm and RelZ. Based on the results of molecular docking, the DMNP-binding site is located in the proximity of the synthetase domain active site. This study might help in the development of alarmone synthetase inhibitors, which includes relacin and its derivatives, as well as DMNP - a synthetic analog of the marine coral metabolite erogorgiaene. Unlike conventional antibiotics, alarmone synthetase inhibitors target metabolic pathways linked to the bacterial stringent response. Although these pathways are not essential for bacteria, they regulate the development of adaptation mechanisms. Combining conventional antibiotics that target actively growing cells with compounds that impede bacterial adaptation may address challenges associated with antimicrobial resistance and bacterial persistence.

Insecticidal, antifeedant and acetylcholinesterase inhibitory activity of sesquiterpenoids derived from eudesmane, their molecular docking and QSAR

This work evaluated the insecticidal, antifeedant and AChE inhibitory activity of compounds with eudesmane skeleton. The insecticidal activity was tested against larvae of Drosophila melanogaster and Cydia pomonella, the compounds 3 and 4 were the most active (LC50 of 104.2 and 106.7 μM; 82.0 and 84.4 μM, respectively). Likewise, the mentioned compounds were those that showed the highest acetylcholinesterase inhibitory activity, with IC50 of 0.26 ± 0.016 and 0.77 ± 0.016 μM, respectively. Enzyme kinetic studies, as well as molecular docking, show that the compounds would be non-competitive inhibitors of the enzyme. The antifeedant activity on Plodia interpunctella larvae showed an antifeedant index (AI) of 99% at 72 h for compounds 16, 27 and 20. The QSAR studies show that the properties associated with the polarity of the compounds would be responsible for the biological activities found.

2-Styrylchromones as inhibitors of α-amylase and α-glucosidase enzymes for the management of type 2 diabetes mellitus

α-amylase and α-glucosidase are key enzymes implicated in carbohydrate digestion and their inhibition has been suggested as a powerful approach for regulating blood glucose levels. The present work describes for the first time their inhibition by a group of twelve hydroxylated 2-styrylchromones (2-SC). Our findings revealed that 2-SC display strong systematic inhibition of α-glucosidase rather than α-amylase activity. The number and position of the hydroxy groups in the chromone moiety further modulate the inhibitory profile of the studied compounds, and the derivatives bearing one catechol unit are efficient inhibitors of both enzymes. Enzyme kinetic studies indicate that all active compounds act as competitive inhibitors of α-amylase while most of them behave as non-competitive inhibitors of α-glucosidase. The results are promising and pave the way to further deciphering the potential of this class of compounds as a suitable alternative for the management of type 2 diabetes and its complications.

Inhibition of Insulin-Regulated Aminopeptidase by Imidazo [1,5-α]pyridines-Synthesis and Evaluation.

Inhibition of insulin-regulated aminopeptidase (IRAP) has been shown to improve cognitive functions in several animal models. Recently, we performed a screening campaign of approximately 10,000 compounds, identifying novel small-molecule-based compounds acting as inhibitors of the enzymatic activity of IRAP. Here we report on the chemical synthesis, structure-activity relationships (SAR) and initial characterization of physicochemical properties of a series of 48 imidazo [1,5-α]pyridine-based inhibitors, including delineation of their mode of action as non-competitive inhibitors with a small L-leucine-based IRAP substrate. The best compound displays an IC50 value of 1.0 µM. We elucidate the importance of two chiral sites in these molecules and find they have little impact on the compound's metabolic stability or physicochemical properties. The carbonyl group of a central urea moiety was initially believed to mimic substrate binding to a catalytically important Zn2+ ion in the active site, although the plausibility of this binding hypothesis is challenged by observation of excellent selectivity versus the closely related aminopeptidase N (APN). Taken together with the non-competitive inhibition pattern, we also consider an alternative model of allosteric binding.

Anti-diabetic and anti-inflammatory indole diterpenes from the marine-derived fungus Penicillium sp. ZYX-Z-143

Seven new indole-diterpenoids, penpaxilloids A–E (1–5), 7-methoxypaxilline-13-ene (6), and 10-hydroxy-paspaline (7), along with 20 known ones (8–27), were isolated from the marine-derived fungus Penicillium sp. ZYX-Z-143. Among them, compound 1 was a spiro indole-diterpenoid bearing a 2,3,3a,5-tetrahydro-1H-benzo[d]pyrrolo[2,1–b][1,3]oxazin-1-one motif. Compound 2 was characterized by a unique heptacyclic system featuring a rare 3,6,8-trioxabicyclo[3.2.1]octane unit. The structures of the new compounds were established by extensive spectroscopic analyses, NMR calculations coupled with the DP4 + analysis, and ECD calculations. The plausible biogenetic pathway of two unprecedented indole diterpenoids, penpaxilloids A and B (1 and 2), was postulated. Compound 1 acted as a noncompetitive inhibitor against protein tyrosine phosphatase 1B (PTP1B) with IC50 value of 8.60 ± 0.53 μM. Compound 17 showed significant α-glucosidase inhibitory activity with IC50 value of 19.96 ± 0.32 μM. Moreover, compounds 4, 8, and 22 potently suppressed nitric oxide production on lipopolysaccharide-stimulated RAW264.7 macrophages.

Quercetin analogs as α-glucosidase inhibitors with antidiabetic activity

This study investigated the inhibitory effects of synthesized quercetin analogs (Q1–Q23) on α-glucosidase, particularly Q7. Q7, a synthetic analog of quercetin, exhibits superior α-glucosidase inhibitory properties compared with quercetin and the standard drug acarbose. This effect is attributed to the unique –CF3 group on the benzene ring of Q7, which enhances its inhibitory effect. As a non-competitive inhibitor, Q7 shows strong binding and stability within the α-glucosidase-Q7 complex. In vivo experiments showed that Q7 significantly reduced fasting blood sugar levels in diabetic mice, surpassing the effects of metformin. In addition, it affects the gut microbiota, particularly the microbes associated with anti-inflammatory responses and short-chain fatty acid production, which are crucial for the treatment of type 2 diabetes. However, the current Q7 dosage led to carbohydrate accumulation in the colon of mice, impairing the function of the intestinal microbiota. This study highlights the potential of Q7 for treating type 2 diabetes and emphasizes the need to determine the optimal dosages that balance therapeutic effects and gut health.

Exploration of inhibitor effect of Gly-Pro (GP), Arg-Gly-Asp-Ser (RGDS) and Ser-Asp-Gly-Arg-Gly (SDGRG) bioactive peptides on angiotensin-converting enzyme activity purified from human serum.

Bioactive peptides (BPs) are a natural and important alternative to synthetic angiotensin-converting enzyme (ACE) inhibitors used in the treatment of hypertension. In this study, ACE was 3575-fold purified from human serum with the affinity chromatography process in one step. The molecular weight and purity of ACE were identified using the SDS-PAGE process and seen in two bands at around 60 kDa and 70 kDa on the gel. Vmax and KM values from the Lineweaver-Burk graphic were determined as 96.15 (µmol/min) mL-1 and 0.2 mM, respectively. The effects of Gly-Pro (GP), Arg-Gly-Asp-Ser (RGDS) and Ser-Asp-Gly-Arg-Gly (SDGRG) BPs on purified ACE were researched. Also, lisinopril was used as a reference inhibitor. GP, RGDS and SDGRG on purified ACE demonstrated an inhibitory effect. IC50 values for these peptides were found as 184.71, 107.16 and 32.54 µM, respectively. Ki values and type of inhibitory for GP, RGDS and SDGRG by the Lineweaver-Burk chart were found. The type of inhibitory for these peptides was calculated as reversible-competitive inhibitory. Ki values for GP, RGDS and SDGRG were calculated to be 260.02, 63.44 and 11.42 µM, respectively. Also, the SDGRG indicated a higher inhibition effect on ACE activity than the GP and RGDS. The IC50 value of lisinopril was designated as 0.35 nM. The inhibition type of lisinopril was designated as reversible noncompetitive inhibition from the Lineweaver-Burk chart and the Ki value was 0.15 nM. Herein, it was concluded that GP, RGDS and SDGRG have ACE inhibitor potential.Communicated by Ramaswamy H. Sarma.

Mechanisms of Allosteric Inhibition of Insulin-Regulated Aminopeptidase

Inhibition of Insulin-Regulated Aminopeptidase is being actively explored for the treatment of several human diseases and several classes of inhibitors have been developed although no clinical applications have been reported yet. Here, we combine enzymological analysis with x-ray crystallography to investigate the mechanism employed by two of the most studied inhibitors of IRAP, an aryl sulfonamide and a 2-amino-4H-benzopyran named HFI-419. Although both compounds have been hypothesized to target the enzyme's active site by competitive mechanisms, we discovered that they instead target previously unidentified proximal allosteric sites and utilize non-competitive inhibition mechanisms. X-ray crystallographic analysis demonstrated that the aryl sulfonamide stabilizes the closed, more active, conformation of the enzyme whereas HFI-419 locks the enzyme in a semi-open, and likely less active, conformation. HFI-419 potency is substrate-dependent and fails to effectively block the degradation of the physiological substrate cyclic peptide oxytocin. Our findings demonstrate alternative mechanisms for inhibiting IRAP through allosteric sites and conformational restricting and suggest that the pharmacology of HFI-419 may be more complicated than initially considered. Such conformation-specific interactions between IRAP and small molecules can be exploited for the design of more effective second-generation allosteric inhibitors.

Imparting aromaticity to 2-pyridone derivatives by O-alkylation resulted in new competitive and non-competitive PIM-1 kinase inhibitors with caspase-activated apoptosis

New aromatic O-alkyl pyridine derivatives were designed and synthesised as Proviral Integration Moloney (PIM)-1 kinase inhibitors. 4c and 4f showed potent in vitro anticancer activity against NFS-60, HepG-2, PC-3, and Caco-2 cell lines and low toxicity against normal human lung fibroblast Wi-38 cell line. Moreover, 4c and 4f induced apoptosis in the four tested cancer cell lines with high percentage. In addition, 4c and 4f significantly induced caspase 3/7 activation in HepG-2 cell line. Furthermore, 4c and 4f showed potent PIM-1 kinase inhibitory activity with IC50 = 0.110, 0.095 µM, respectively. Kinetic studies indicated that 4c and 4f were both competitive and non-competitive inhibitors for PIM-1 kinase enzyme. In addition, in silico prediction of physiochemical properties, pharmacokinetic profile, ligand efficiency, ligand lipophilic efficiency, and induced fit docking studies were consistent with the biological and kinetic studies, and predicted that 4c and 4f could act as PIM-1 kinase competitive non-adenosine triphosphate (ATP) mimetics with drug like properties.

Mechanism of ferrous ions reversibly inhibiting of oxidase-Like activity of Co3O4 nanowires and its application in horticultural fertilizer analysis

Nanozymes are becoming substitutes of some natural enzymes due to their excellent properties, however, little attention has been paid to the catalytic mechanisms and especially inhibition mechanism of nanozymes, which impeded advancement in nanozymes and their wide application including bioanalysis. Herein, non-toxic filamentous virus, M13 phage templated Co3O4 nanowires (NWs) was facilely prepared, which exhibits excellent oxidase (OXD)-like activity and catalytic efficiency. The catalytic mechanism of OXD-like is oxygen vacancies, confirmed by high resolution X-ray photoelectron spectrum of O 1 s, electron paramagnetic resonance spectra and reactive oxygen species scavenging experiments. The substrate of 3,3′,5,5′-tetramethylbenzidine (TMB) for OXD-like is oxidized into blue ox-TMB, and then turned to yellow diimine, depending on TMB concentration and incubation time. Then the mechanism of ferrous ions (Fe2+) inhibition of the oxidase-like activity of Co3O4 NWs was uncovered that Fe2+ competes with TMB for oxygen vacancies. The inhibition is reversible with the type of hybrid of competitive inhibition and non-competitive inhibition with the inhibition constants (KI and KI') of 0.134 mM and 1.012 mM, respectively. Finally, on the basis of the inhibitory effect, a fast, selective and sensitive Fe2+ colorimetric analysis was proposed with two linear parts and a limit of detection of 0.21 μM Fe2+ (S/N = 3), which is applicable to detect Fe2+ in horticultural fertilizers. This study may offer a new strategy for the in-depth study of inhibition mechanism of other nanozymes, which would extend the scope of nanozyme-based biosensing.

Monastrol disrupts KIFC1-ATP dynamics: Towards newer anticancer mechanism

Mitotic kinesins are eukaryotic proteins that play a vital role in cellular mitosis. Their over-expression in malignant rather than normal cells and their specific cellular role made kinesin inhibitors a promising cancer therapeutics. The realization of in-vitro antitumor activity of the investigational molecule-monastrol-was traced to its allosteric inhibition of the ATPase activity of the motor domain of Human Kensin-5 (Eg5) and prompted extensive efforts towards kinesin inhibitors. Many reports pointed to the prospect of Kinesin-like protein (KIFC1) as a novel anticancer target. Nevertheless, no verified KIFC1-inhibitor crystallized structure has been reported so far, confirming the scarcity of molecular studies devoted to deciphering claimed potentiality. The significant structural resemblance across the kinesins superfamily sparked our interest in investigating monastrol as a KIFC1 inhibitor.Time-scale findings of molecular dynamics simulation and molecular mechanics/generalized-born surface area (MM/GBSA) methods revealed that monastrol binds with high affinity to an allosteric pocket, inducing notable dynamical alterations evidenced by increased structural stability, rigidity, compactness, and overall folding tendency of KIFC1. Further, these conformational events negatively impacted the affinity of ATP to its site disrupting the essential ATP-KIFC1 dynamics leading to consequential loss of functionality. We concluded that monastrol can act as non-competitive inhibitor of KIFC1 and these findings speculate new anticancer mechanism contributing to the design of highly selective and novel cancer therapeutics.

Inhibition of staphylococcal nuclease by benzimidazole-based Ligand: Implications in DNA-Mediated entrapment and uptake of MRSA by Macrophage-like cells

The staphylococcal nuclease also referred as micrococcal nuclease (MNase) is a key drug target as the enzyme degrades the neutrophil extracellular trap (NET) and empowers the pathogen to subvert the host innate immune system. To this end, the current study presents a critical evaluation of MNase inhibition rendered by benzimidazole-based ligands (C1 and C2) and probes its therapeutic implications. A nuclease assay indicated that MNase inhibition rendered by C1 and C2 was ∼ 55 % and ∼ 72 %, respectively, at the highest tested concentration of 10 µM. Studies on enzyme kinetics revealed that C2 rendered non-competitive inhibition and significantly reduced MNase turnover number (Kcat) and catalytic efficiency (Kcat/Km) with an IC50 value of ∼ 1122 nM. In CD spectroscopy, a notable perturbation in the β-sheet content of MNase was observed in presence of C2. Fluorescence-microscope analysis indicated that MNase inhibition by C2 could restore entrapment of methicillin-resistant Staphylococcus aureus (MRSA) in calf-thymus DNA (CT-DNA). Flow cytometry and confocal microscope analysis revealed that uptake of DNA-entrapped MRSA by activated THP-1 cells was reinstated by MNase inhibition rendered by C2. Inhibition of nuclease by the non-toxic ligand C2 holds therapeutic prospect as it has the potential to bolster the DNA-mediated entrapment machinery and mitigate MRSA infections.

Effect of tubeimoside I on the activity of cytochrome P450 enzymes in human liver microsomes

This study assessed the effect of tubeimoside I on CYP1A2, 2A6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4 to reveal the potential of tubeimoside I to induce drug-drug interaction. The evaluation of cytochromes P450 enzyme (CYP) activity was performed in pooled human liver microsomes with probing substrates of CYP1A2, 2A6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4. Typical inhibitors were employed as positive controls and the effect of 0, 2.5, 5, 10, 25, 50, and 100 μM tubeimoside I was investigated. The activity of CYP2D6, 2E1, and 3A4 was significantly inhibited by tubeimoside I with the IC50 values of 10.34, 11.58, and 9.74 μM, respectively. The inhibition of CYP2D6 and 2E1 was competitive with the Ki value of 5.66 and 5.29 μM, respectively. While the inhibition of CYP3A4 was non-competitive with the Ki value of 4.87 μM. Moreover, the inhibition of CYP3A4 was time-dependent with the KI and K inact values of 0.635 μM−1 and 0.0373 min−1, respectively. Tubeimoside I served as a competitive inhibitor of CYP2D6 and 2E1 exerting weak inhibition and a non-competitive inhibitor of CYP3A4 exerting moderate inhibition.

Application of natural fungi in bioconversion of lignocellulosic waste to second-generation ethanol

The second generation (2G) or cellulosic ethanol can help with diversification of the use of fossil energy sources. However, as bioconversion of plant waste into 2G bioethanol requires expensive additional steps of pre-treatment/hydrolysis of lignocellulosic materials, and this technology has not yet reached the technological readiness level which would allow it to be scaled-up, this process needs more interdisciplinary and comprehensive studies. This work was aimed at experimental study of a full cycle of successive processes of pre-treatment/saccharification using cellulolytic enzymes of filamentous fungi and fermentation of obtained syrups by xylose-fermenting yeast, using selected natural microorganisms for the fungal-based bioconversion of lignocellulosic agricultural waste to 2G ethanol. Using the Plackett-Burman and Box-Behnken methods of mathematical statistics, the optimal conditions for pre-treatment and enzymatic hydrolysis of wheat straw by a hemi- and cellulolytic multi-enzyme complex of the selected fungal strain Talaromyces funiculosus UCM F-16795 were established: microwave-assisted alkali pre-treatment with sodium hydroxide (NaOH) solutions (concentration range 4.6–4.8%), and saccharification conditions of medium pH 4, temperature 40 °С, hydrolysis duration 18 hours, and dilution of culture liquid with a buffer solution 1:1. The total energy of microwave irradiation 1.2 kJ and the ratio of substrate/enzyme solution 100 mg/1 mL were used. Under optimized conditions, wheat straw hydrolysates contained 5.0–7.5 g/L of reducing sugars, which, according to HPLC assessment, contained 0.7–1.0 g/L of glucose, 2.2–2.9 g/L of xylose and 0.7–0.8 g/L cellobiose. We used the selected strain of xylose-fermenting yeast in fermentation of mixtures of the most important monosaccharides in hydrolysates, xylose and glucose, in the concentration range relevant for syrups obtained by us during the optimized saccharification of lignocellulosic substrates with T. funiculosus enzymes. Based on sequencing and phylogenetic analysis, strain UCM Y-2810 was confirmed as Scheffersomyces stipitis; its nucleotide sequences of ITS region and 28S gene rDNA were deposited in GenBank under the accession numbers OP931914 and OP931915, respectively. The ethanologenic process for S. stipitis UCM Y-2810 was studied according to Box-Behnken design, assessing ethanol concentration by gas chromatography-mass spectrometry. Yeast fermentation under static microaerophilic conditions showed a 1.5 times higher rate of bioethanol production and 1.7 times greater efficiency of ethanologenesis per yeast biomass than for submerged cultivation. Optimization of the process of ethanologenesis resulted in the maximum rate of fermentation mixture of sugars, being 11.30 ± 0.36 g/L of ethanol, with optimal values of factors: 30 g/L of xylose, 5.5 g/L of glucose and cultivation for 5.5 days. It was revealed that the tested glucose concentrations did not significantly affect the process of xylose-fermentation by yeast, and non-competitive inhibition of xylose transport by glucose into yeast cells did not occur. This study demonstrated the potential of a full cycle bioconversion of lignocellulosic waste to 2G ethanol based on use of natural fungal strains and optimization of conditions for all steps.

Synthesis, in vitro potency of inhibition, enzyme kinetics and in silico studies of quinoline-based α-glucosidase inhibitors

Diabetes mellitus is a multifactorial global health disorder that is rising at an alarming rate. One effective therapeutic approach for controlling hyperglycemia associated with type-2 diabetes is to target α-glucosidase, which catalyzes starch hydrolysis in the intestine. In an attempt to find potential α-glucosidase inhibitors, a series of twenty new quinoline linked benzothiazole hybrids (8a–t) were synthesized in good yields from suitable reaction procedures and their chemical structures were analyzed by 1HNMR, 13CNMR, IR, and ESI–MS analysis. The synthesized derivatives further screened for their activity against α-glucosidase. Among them, compounds 8b, 8h, 8n and 8o exhibited remarkable α-glucosidase inhibitory activity with IC50 values ranging from 38.2 ± 0.3 to 79.9 ± 1.2 µM compared with standard drug acarbose (IC50 = 750.0 ± 2.0 µM). Enzyme kinetic studies of the most active compound (8h) indicated a non-competitive inhibition with Ki value of 38.2 µM. Moreover, the homology modeling, molecular docking and molecular dynamics simulation studies were conducted to reveal key interactions between the most active compound 8h and the targeted enzyme. These results are complementary to the experimental observations. In order to predict the druggability of the novel derivatives, the pharmacokinetic properties were also applied. These findings could be useful for the design and development of new α-glucosidase inhibitors.

Insights into the inhibition mechanisms of MERS-CoV and SARS-CoV2 papain-like proteases by inhibitors from Crinum distichum: In vitro and in silico analysis

Highly pathogenic coronaviruses, including the Middle East Respiratory Syndrome Coronavirus (Mers-CoV) and Severe Acute Respiratory Syndrome Coronavirus-2 (Sars-CoV2), have recently emerged and represent a serious threat to global health. Accordingly, a search for potent inhibitors against coronavirus diseases is warranted. Therefore, this study aimed at identifying potential inhibitors from Crinum distichum against the papain-like protease (PLpro), an important antiviral target due to its essential role in viral replication. A bacterial expression system was used for the recombinant expression of Mers-CoV-PLpro and Sars-CoV2-PLpro which was used to assess the inhibitory effect of compounds isolated from C. distichum against their proteolytic and deubiquitinating (DUB) activities. Then, the inhibition mechanism of hit compounds was determined through enzyme kinetic studies. Protein-ligand stability was assessed by Differential Scanning Fluorometry and Molecular docking was used to predict the possible molecular interactions. Five compounds, namely (2S)-4´,7-dimethoxyflavan (CDS1), (2S)-4´‑hydroxy-7-methoxyflavan (CDS2), Hippadine (CDS3), (2R)-4´‑hydroxy-5,7-dimethoxyflavan (CDS4) and Hippacine (CDS5) were tested in this study. They markedly inhibited both the proteolytic and deubiquitinating activities of CoV-PLpro. Their IC50 values were in the low micromolar range. The most potent inhibitors were CDS3 and CDS4, with IC50 values of 9.28 and 6.45 µM, and 11.63 and 6.10 µM against the proteolytic and DUB activities of Mers-CoV-PLpro and Sars-CoV2-PLpro, respectively. They exert their inhibitory effect through competitive and non-competitive inhibition mechanisms. These compounds induced the instability of the enzymes by reducing their melting temperature and molecular docking analysis revealed several molecular interactions between the compounds and various amino acids of the enzymes. Overall, our findings suggest C. distichum active constituents to be promising candidates for development into potential inhibitors against coronavirus.

Development of chemometric-assisted supercritical fluid extraction of effective and natural tyrosinase inhibitor from Syzygium aqueum leaves

Tyrosinase is a key enzyme in enzymatic browning, causing quality losses in food through the oxidation process. Thus, the discovery of an effective and natural tyrosinase inhibitor via green technology is of great interest to the global food market due to food security and climate change issues. In this study, Syzygium aqueum (S. aqueum) leaves, which are known to be rich in phenolic compounds (PC), were chosen as a natural source of tyrosinase inhibitor, and the effect of the sustainable, supercritical fluid extraction (SFE) process was evaluated. Response surface methodology-assisted supercritical fluid extraction (RSM-assisted SFE) was utilized to optimize the PCs extracted from S. aqueum. The highest amount of PC was obtained at the optimum conditions (55 °C, 3350 psi, and 70 min). The IC50 (661.815 μg/mL) of the optimized extract was evaluated, and its antioxidant activity (96.8 %) was determined. Gas chromatography-mass spectrometry (GC–MS) results reveal that 2′,6′-dihydroxy-4′-methoxychalcone (2,6-D4MC) (82.65 %) was the major PC in S. aqueum. Chemometric analysis indicated that 2,6-D4MC has similar chemical properties to the tyrosinase inhibitor control (kaempferol). The toxicity and physiochemical properties of the novel 2,6-D4MC from S. aqueum revealed that the 2,6-D4MC is safer than kaempferol as predicted via absorption, distribution, metabolism, and excretion (ADME) evaluation. Enzyme kinetic analysis shows that the type of inhibition of the optimized extract is non-competitive inhibition with Km = 1.55 mM and Vmax = 0.017 μM/s. High-performance liquid chromatography (HPLC) analysis shows the effectiveness of S. aqueum as a tyrosinase inhibitor. The mechanistic insight of the tyrosinase inhibition using 2,6-D4MC was successfully calculated using density functional theory (DFT) and molecular docking approaches. The findings could have a significant impact on food security development by devising a sustainable and effective tyrosinase inhibitor from waste by-products that is aligned with the United Nation's SDG 2, zero hunger.