Enzyme Kinetic Studies Research Articles

Mechanistic insights into alkaloid-based inhibition of squalene epoxidase: A combined in silico and experimental approach for targeting cholesterol biosynthesis.

Squalene epoxidase (SQLE) is a key enzyme in the cholesterol biosynthesis pathway and an attractive therapeutic target for hypercholesterolemia and antifungal treatment. In this study, we investigated the inhibitory potential of six alkaloids-berberine, evodiamine, harmine, reserpine, matrine, and sanguinarine-against SQLE using a combined in silico and in vitro approach. Molecular docking revealed strong binding affinities ranging from -8.1 to -11.0 kcal/mol, with evodiamine demonstrating the highest affinity, followed by sanguinarine and berberine. 200 ns MD simulations confirmed stable interactions for all alkaloid-enzyme complexes, characterized by low RMSD values, robust hydrogen bonding, and favorable free energy landscapes, as supported by MM/PBSA analysis. Experimental validation through in vitro inhibition assays revealed that evodiamine (IC₅₀ = 2.87 ± 0.08 μM) exhibited potent inhibition comparable to the standard inhibitor TNSCPA (IC₅₀ = 2.65 ± 0.18 μM), while berberine (IC₅₀ = 3.57 ± 0.18 μM) and reserpine (IC₅₀ = 4.91 ± 0.34 μM) showed strong and moderate inhibition, respectively. Harmine, matrine, and sanguinarine were less effective. Enzyme kinetics studies demonstrated that berberine and reserpine act as noncompetitive inhibitors, binding to allosteric sites, whereas evodiamine exhibited competitive inhibition at the active site. ADMET analysis highlighted favorable pharmacokinetic properties for berberine, evodiamine, and sanguinarine, while reserpine and matrine exhibited limited bioavailability due to solubility and size constraints. The unique inhibitory mechanisms observed were consistent with the structural and physicochemical properties of the compounds. These findings establish berberine, evodiamine, and reserpine as promising SQLE inhibitors, with evodiamine emerging as a lead candidate.

Salvicsite A-C: Three Seco-Norabietane Diterpenoids Including an Unusual Tetracyclic Skeleton from Salvia castanea Diels f. tomentosa Stib.

Three seco-norabietane diterpenoids, salvicsites A-C (1-3), along with two known compounds, were isolated from the roots and rhizomes of Salvia castanea Diels f. tomentosa Stib. Salvicsite A (1) represents an unprecedented structural combination, featuring an eight-membered α-methyl-α,β-unsaturated lactone ring and a five-membered α,β-unsaturated lactone ring, based on a 6/6/5/8 ring system. Their structures were elucidated through comprehensive spectroscopic analyses, X-ray crystallography, and quantum chemical calculations. Putative biosynthetic pathways of 1-3 were proposed, with compound 5 being the plausible biogenetic precursor. The AChE inhibition assay demonstrated that salvicsite A (1) exhibited a notable inhibitory effect against AChE, with an IC50 value of 9.56 ± 1.05 μM. Enzymatic kinetic studies indicated that salvicsite A (1) acted as a mixed-type inhibitor, and its binding mode was explored through molecular docking. The results of the cytotoxic activity demonstrated that only compound 5 exhibited inhibitory activity.

Multi-pronged molecular insights into flavonoid-mediated inhibition of squalene epoxidase: a pathway to novel therapeutics.

Squalene epoxidase (SQLE) is a crucial enzyme in the sterol biosynthesis pathway and a promising target for therapeutic intervention in hypercholesterolemia and fungal infections. This study evaluates the inhibitory potential of six flavonoids namely silibinin, baicalin, naringenin, chrysin, apigenin-7-O-glucoside, and isorhamnetin against SQLE using an integrative approach combining in silico and experimental methods. Molecular docking revealed that apigenin-7-O-glucoside, silibinin, and baicalin displayed the highest binding affinities (-10.7, -10.2, and -10.0 kcal mol-1, respectively) and robust interactions with the SQLE binding site. These findings were corroborated by 200 ns molecular dynamics (MD) simulations, which demonstrated stable binding trajectories, minimal structural fluctuations, a thermodynamically favored potential energy landscape (PEL) and favorable MM/PBSA binding free energies for three flavonoids. Experimental validation via in vitro inhibition assays confirmed the computational predictions, with apigenin-7-O-glucoside emerging as the most potent inhibitor (IC50 = 1.74 ± 0.05 μM), followed by silibinin (IC50 = 1.88 ± 0.28 μM) and baicalin (IC50 = 2.50 ± 0.46 μM). Enzyme kinetics studies revealed distinct mechanisms of action: apigenin-7-O-glucoside exhibited competitive inhibition, while silibinin and baicalin showed mixed inhibition. Furthermore, in silico ADMET analysis indicated favorable pharmacokinetic and pharmacodynamic profiles for these flavonoids, with silibinin demonstrating particularly high bioavailability and lipophilicity. This study highlights apigenin-7-O-glucoside, silibinin, and baicalin as potent SQLE inhibitors with promising therapeutic potential. The congruence between in silico predictions and experimental results underscores the reliability of computational approaches in drug discovery, paving the way for future preclinical development of these compounds as novel SQLE-targeted therapeutics.

Quinazoline derivatives as novel bacterial sphingomyelinase enzyme inhibitors.

Bacillus cereus sphingomyelinase C (B. cereus SMase), which plays a crucial role in bacterial virulence, has emerged as a new therapeutic target for treating opportunistic infections caused by this pathogen. It also shares catalytic domain similarity with human neutral sphingomyelinase 2 (nSMase2), which is implicated in Alzheimer's disease. In this study, a series of quinazoline derivatives were synthesized and evaluated for their inhibition of B. cereus SMase, electric eel acetylcholinesterase (EeAChE), and equine butyrylcholinesterase (eqBuChE). Moreover, the antibacterial, anti-hemolytic and metal chelation properties of the selected compounds were determined. Among the synthesized compounds, 6-chloro-2-thioxo-2,3-dihydroquinazolin-4(1H)-one (compound 4) and 6-fluoro-2-thioxo-2,3-dihydroquinazolin-4(1H)-one (compound 5) exhibited promising inhibition of B. cereus SMase, with IC50 values of 6.43 and 6.50µM, respectively. The mode of inhibition of compound 4 was determined as mixed-type inhibition by enzyme kinetic study. In addition, compounds 4 and 5 showed 59.50% and 51.66% eqBuChE inhibition at 50µM concentration, respectively. Furthermore, compound 4 reduced B. cereus-induced hemolysis on sheep erythrocytes and able to form a complex with Cu2+ in ligand:metal ratio of 2:1. Additionally, cambinol, an inhibitor of both nSMase2 and B. cereus SMase, was found to exhibit inhibitory activity against eqBuChE, with IC50 value of 7.40µM. The biological data were also supported by the results of molecular docking studies and in-silico physicochemical properties/ADME predictions of the selected compounds were determined.

A method for HDAC activity screening in postmortem human brain. A proof-of-concept study with antipsychotics.

Histone deacetylase (HDAC) density and activity are altered in different brain disorders. Antipsychotic drugs (APs) might modulate HDAC activity in brains of schizophrenia subjects. HDAC activity assay amenable for enzyme kinetics and HDAC inhibitor (HDACi) screening studies in postmortem human brain samples. The optimization and characterization work involved several steps. The nucleosolic subcellular fraction and total protein amount needed for an optimal HDAC activity on Boc-Lys(Ac)-AMC substrate were characterized. Signal-to-noise ratio (1.8) and Z-score values (0.82) were indicators of the assay quality. Inhibition studies with non-selective (belinostat, vorinostat, valproic acid) and selective (apicidin, MS275, romidepsin, tacedinaline and EX527) HDACis showed that the optimized assay detected class I HDAC activity. The obtained IC50 values were similar to those previously reported, proving the assay reliability. We used the optimized assay to study the effect of APs on HDAC activity. Inhibition studies with APs in postmortem human brain, together with enzyme kinetic studies in brains of rats chronically treated with APs observed no modulation of class I HDAC activity. This study describes the optimization of a reliable and cost efficient HDAC activity assay for its use in postmortem human brain samples. The assay does not depend on antibody specificity and it is valid for enzyme kinetic studies and for the screening of new potential class I HDACis. We optimized and characterized an assay to measure HDAC activity in postmortem human brain samples. We did not observe any modulatory effect of APs on HDAC activity.

Molecular insights into the aspartate protease Plasmepsin II activity inhibition by fluoroquinolones: A pathway to antimalarial drug development

Plasmepsin II (PlmII) belongs to the aspartate proteases and is involved in hemoglobin degradation in Plasmodium falciparum. Due to its critical role in the survival of the Plasmodium, PlmII is considered as a potent drug target for antimalarial therapy. We have done recombinant protein production of pro-plasmepsin II (Pro-plmII). Pro-PlmII was further activated to mature form (mPlmII) and its activity was confirmed by enzyme kinetics studies. The fluorescence spectroscopic and isothermal titration calorimetric studies show that fluoroquinolone-based antibiotic drugs norfloxacin, ciprofloxacin, and sparfloxacin bind with mPlmII. Molecular docking results show that only norfloxacin and ciprofloxacin are able to bind at the active site of mPlmII via hydrogen binding and hydrophobic interactions. Enzyme kinetics analysis reveals that norfloxacin and ciprofloxacin effectively inhibit mPlmII activity, while sparfloxacin does not exhibit any inhibitory effect on the enzyme's catalytic function. The two methyl groups on the 3rd and 5th carbon atoms of the piperazine ring make sparfloxacin unable to go inside mPlmII and bind at its active site. Overall, the results here suggested that fluoroquinolone-based antibiotic drugs norfloxacin, and ciprofloxacin, can be repurposed as antimalarial inhibitors targeting aspartic proteases. These findings contribute to pave the way for potential therapeutic interventions targeted at malaria.

Acquiring stereospecific new pseudosugars: Obtaining rac-decahydro-1,4-epoxynaphthalene-2,3,5,6,7,8-hexaols from the Diels-Alder reaction and investigating their biological effects.

In this study, Diels-Alder reaction was performed to sulfolene and endo/exo-diacetate compounds. After a series of reactions, new conduritol A and F analogs containing oxo-bridge and naphthalene rings in their structures were synthesized. To the starting compound, bromination, elimination, singlet oxygen reaction, acetylation, selective oxidation with osmium tetroxide (OsO4), and m-chloroperbenzoic acid (m-CPBA), re-acetylation, and finally hydrolysis of the compounds by NH3(g)/MeOH reactions were carried out. 1H NMR, 13C NMR, IR, and elemental analysis elucidated the structures of all synthesized compounds. The α, β-glucosidase, and α-amylase inhibitory potentials of the new polycyclitols, conduritol A and F analogs, were examined for biological activity. Also, enzyme kinetic studies of well-active compounds were carried out. Compound 30 showed the best inhibition activity against α, β-glucosidase, and α-amylase enzymes. Compound 28a showed the best activity against L929 and Capan-1 cell lines, and compound 22 showed the best activity against the A549 fibroblast cell line. Moreover, hemolysis (ASTM F756 standard) and genotoxicity test results were recorded.

The enzyme kinetic studies, DNA protection and antioxidant activities of furan/ thiophene-2-carboxamide derivatives

Three furan and/or thiophene-2-carboxamide compounds, namely N-(furan-2-ylmethyl)thiophene-2-carboxamide (1), N-(furan-2-carbonyl)furan-2-carboxamide (2), and N-(Thiophene-2-ylmethyl)furan-2-carboxamide (3) were investigated the enzyme kinetic studies by urease, acetylcholinesterase (AChE), and butyrylcholinesterase (BChE). The inhibition constant (Ki) of Compound (CPD)3 by AChE was determined as 0.10 mM, and the Ki value by BChE was determined as 0.07 mM. In comparison, the Ki value of CPD1 by urease was determined as 0.10 mM. These CPDs were examined for antioxidant activity by the DPPH˙ scavenging method. CPD3 exhibited 98.93% DPPH scavenging activity compared to ascorbic acid, the positive control group. Furthermore, the DNA-protective activities of the compounds were investigated, and the DNA protection activity of CPD1 was observed to be 78%. The findings suggest that thiophene/furan carboxy amide-containing CPD1 and CPD3 might be exploited as potential structures for evaluating pharmaceuticals with greater potency.

Temperature-Dependent Parameters in Enzyme Kinetics: Impacts on Enzyme Denaturation

Enzymes are vital proteins in biological systems, responsible for regulating and coordinating numerous essential processes. The incorporation of denaturation rate accounts for the gradual loss of enzyme activity over time, which is particularly significant under experimental conditions where enzymes are susceptible to denaturation. It is noteworthy that adverse environmental conditions, such as high temperature or pH imbalance, can induce enzyme denaturation, leading to a loss of functionality over time. This structural disruption renders enzymes inactive, posing a crucial consideration in long-term enzyme kinetics studies. Furthermore, enzymes typically exhibit reduced catalytic activity at lower temperatures, which is pivotal for understanding their stability and efficacy in biological systems and industrial applications. Accordingly, we developed a mathematical model to investigate enzyme kinetics under varying temperature, aiming to analyse their respective impacts on both enzyme behaviour and product formation.

Phytocompounds hesperidin, rebaudioside a and rutin as drug leads for the treatment of tuberculosis targeting mycobacterial phosphoribosyl pyrophosphate synthetase

The main aim of this study is to address the global health crisis posed by tuberculosis (TB) through the exploration of novel therapeutic strategies targeting Mycobacterial phosphoribosyl pyrophosphate synthetase (MtPrsA), an untried enzyme involved in essential metabolic pathways of Mycobacterium tuberculosis. This enzyme plays a crucial role in cell wall synthesis, nucleotide biosynthesis and amino acid synthesis in M tb. Any hindrance to these may affect the growth and survival of the organism. Phytochemicals were systematically screened for potential inhibitors to MtPrsA. Subsequently, based on molecular docking studies, three compounds, namely, hesperidin, rebaudiosideA and rutin were selected. The binding stabilities of these compounds were analyzed using molecular dynamics simulation. Based on the RMSD score obtained, the binding stability of the compounds was confirmed. To validate the findings, an enzyme inhibition assay was done using recombinant MtPrsA. Ligation Independent Cloning (LIC cloning) method was used to produce recombinant His-tagged MtPrsA, followed by purification using Histrap columns. Enzyme kinetic studies unveiled the distinct modes of inhibition exhibited by each compound towards MtPrsA. RebaudiosideA and rutin emerged as competitive inhibitors, while hesperidin showcased a mixed inhibition profile. In conclusion, the study contributes valuable insights into potential therapeutic strategies for TB, through the exploration of alternative enzyme targets and the identification of phytochemical inhibitors. Notably, todate, no effective plant compounds have been reported as inhibitors to MtPrsA.

Assessment of the Effect of Berberine on Metalloprotease Enzymes Inhibition and Antioxidant Activity: Possible Application in Skin Aging

Skin aging has been defined to enclose both intrinsic and extrinsic aging. Phytochemicals are frequently used for developing skin care formulations and could protect the skin’s epidermal and dermal layers, consisting mainly of elastin and collagen, from UV radiation. Berberine is an isoquinoline alkaloid and a biologically active component from plant sources. Our objective was to assess Berberine’s anti-aging capabilities by conducting elastase and collagenase enzyme inhibition and kinetic studies and to also evaluating its antioxidant capacity with three different methods. Furthermore, heat stability, pH and sun protection factor (SPF) of the formulated cream containing 1.5% berberine was evaluated. The elastase and collagenase IC50 values of berberine were estimated to be 47.54 and, 22.16 µg/mL respectively. Berberine was determined as an un-competitive inhibitor of elastase and collagenase. It scavenged DPPH and ABTS free radicals with IC50 values of 66.81 and 180.5 µg/mL respectively. 210.387 mg/L of berberine was equivalent in reducing power of 176 mg/L of ascorbic acid. SPF and pH value of cream containing berberine was found to be 12.3 and 5.62 respectively. In conclusion, these findings suggest that Berberine is a promising candidate for use as an active ingredient in cosmeceuticals, offering a natural approach to enhance skin health and reduce the visible signs of aging.

Green preparation and isolation of carrageenan oligosaccharides and assessment of their ability to inhibit melanogenesis

Green preparation and isolation of carrageenan oligosaccharides and assessment of their ability to inhibit melanogenesis

Isoindolinedione-Benzamide Pyridinium Derivatives for Targeting Alzheimer's Disease.

An Isoindolinedione-benzamide pyridinium derivatives were designed through a structure-based strategy and synthesized as novel multifunctional anti-Alzheimer agents. The inhibitory activities of all 17 derivatives against acetylcholinesterase and butyrylcholinesterase were evaluated. Results exhibited that compound 7j displayed promising AChE inhibitory activity with an IC50 value of 0.26 ± 0.07 μM, and compound 7c exhibited an IC50 value of 0.08 ± 0.01 μM against BChE with 132-fold better inhibitory activity in comparison with positive control. Next, the enzyme kinetics studies and detailed binding mode via molecular docking were performed for the most potent compounds. Additionally, molecular dynamics simulations were accomplished to further investigate the potent compound's interaction, orientation, and conformation over the related enzymes. The neurotoxicity of the most potent derivative was executed against SH-SY5Y, and the mRNA levels of GSK-3α and GSK-3β after treatment with 7c on SH-SY5Y were evaluated. Results exhibited the mRNA levels of GSK-3β were decreased compared to the control group. All these results indicate that 7c is a good starting point for developing a multifunctional anti-Alzheimer compound.

Metal catalyst-free β-amination of branched rac-C8N-type such as C7N carbasugars via intramolecular aza-michael addition: Biological evolution, DFT studies and ADME properties

Metal catalyst-free β-amination of branched rac-C8N-type such as C7N carbasugars via intramolecular aza-michael addition: Biological evolution, DFT studies and ADME properties

Benzylidenehydrazine Derivatives: Synthesis, Antidiabetic Evaluation, Antioxidation, Mode of Inhibition, DFT and Molecular Docking Studies.

Diabetes mellitus (DM) is a metabolic condition that is a profound health concern across the globe due to its contribution to the increased mortality rate. It affects millions of people around the world and is associated with severe complications among people diagnosed with it. Among the array of approaches used for the management of DM, inhibition of enzymes viz. α-amylase and α-glucosidase which are responsible for sugars hydrolysis is regarded as a feasible therapeutic protocol for the management of DM. Herein, we report the synthesis of benzylidenehydrazine derivatives as well as their evaluation as α-glucosidase and α-amylase inhibitors including their antioxidant testing. Generally, all the synthesized derivatives were more potent inhibitors of α-amylase than of α-glucosidase. Specifically, 2,4 fluoro substituted analogue 9 (IC50 =116.19 μM) emerged as the strongest α-amylase inhibitor with ~5-fold superior activity in comparison to the standard drug, acarbose (IC50 =600 μM). Compounds 18 (IC50 =240.59) and 19 (IC50 =198.32 μM) displayed the strongest NO scavenging activity compared to Trolox (IC50 =272.36 μM). In addition, the enzyme kinetic studies indicated that compound 9 acts as a non-competitive inhibitor of α-amylase. Finally, density functional theory and molecular docking studies for compound 9 were conducted to explore its structural and electronic properties as well as to determine protein-ligand interactions, respectively to decipher its observed activity. The obtained findings present promising possibilities for developing new drug candidates to control postprandial glucose levels in persons with diabetes.

Biochemometric-guided isolation of new Isosteroidal alkaloids from Fritillaria cirrhosa D.Don (Liliaceae, syn. Fritillaria roylei Hook) as acetylcholinesterase inhibitors

Globally, Alzheimer's disease is an urgent public health concern with the ageing population in developing nations. Recent studies have identified isosteroidal alkaloids as promising therapeutic agents for Alzheimer's treatment. Fritillaria species are well-known rich sources of steroidal and isosteroidal alkaloids. In this context, the current study focuses on the biochemometric-guided isolation of three previously undescribed and two known isosteroidal alkaloids as acetylcholinesterase (AChE) inhibitors from the bulbs of Fritillaria cirrhosa D.Don. The isolated molecules were characterized by NMR, HR-ESI-MS, FT-IR, and DP4+ analysis. Subsequently, all isolates were evaluated for AChE inhibitory activity using Ellman's method. Among the evaluated molecules, 1 (IC50: 33.0±4.4μM) and 5 (IC50: 24.7±4.5μM) showed promising AChE inhibition in vitro. Enzyme kinetic studies of isolated molecules revealed mixed inhibition kinetics with Ki varying from 1.3 to 24.4μM. Moreover, the in silico studies showed excellent binding affinities of isolated molecules with the target protein and good drug-like ADMET properties. The present study identified new isosteroidal alkaloids as promising AChE inhibitors from F. cirrhosa bulbs via a biochemometric approach and advocated their further exploration for treating neurodegenerative disorders.

Synthesis, Urease Inhibition, Molecular Docking, and Optical Analysis of a Symmetrical Schiff Base and Its Selected Metal Complexes.

Designing and developing small organic molecules for use as urease inhibitors is challenging due to the need for ecosystem sustainability and the requirement to prevent health risks related to the human stomach and urinary tract. Moreover, imaging analysis is widely utilized for tracking infections in intracellular and in vivo systems, which requires drug molecules with emissive potential, specifically in the low-energy region. This study comprises the synthesis of a Schiff base ligand and its selected transition metals to evaluate their UV/fluorescence properties, inhibitory activity against urease, and molecular docking. Screening of the symmetrical cage-like ligand and its metal complexes with various eco-friendly transition metals revealed significant urease inhibition potential. The IC50 value of the ligand for urease inhibition was 21.80 ± 1.88 µM, comparable to that of thiourea. Notably, upon coordination with transition metals, the ligand-nickel and ligand-copper complexes exhibited even greater potency than the reference compound, with IC50 values of 11.8 ± 1.14 and 9.31 ± 1.31 µM, respectively. The ligand-cobalt complex exhibited an enzyme inhibitory potential comparable with thiourea, while the zinc and iron complexes demonstrated the least activity, which might be due to weaker interactions with the investigated protein. Meanwhile, all the metal complexes demonstrated a pronounced optical response, which could be utilized for fluorescence-guided targeted drug delivery applications in the future. Molecular docking analysis and IC50 values from in vitro urease inhibition screening showed a trend of increasing activity from compounds 7d to 7c to 7b. Enzyme kinetics studies using the Lineweaver-Burk plot indicated mixed-type inhibition against 7c and non-competitive inhibition against 7d.

Potential skincare benefits and self-healing properties of Lignosus rhinocerotis polysaccharides as affected by ultrasound-assisted H2O2/Vc treatment

Natural polysaccharides have been recognized as major bioactive components in skincare and wound care products. In this study, the skincare benefits and self-healing properties of Lignosus rhinocerotis polysaccharides (LRP) and its degraded products (DLRP-1, DLRP-2 and DLRP-3) by ultrasound assisted H2O2/Vc treatment (U-H/V) at different ultrasonic intensity (28.14, 70.35, and 112.56W/cm2) were investigated. U-H/V altered the internal crystalline structure and microstructure of LRP, and enhanced the thermal stability. Due to the breakage of molecular chains after U-H/V, the moisture absorption of LRP was enhanced but the moisturizing property showed a different degree of reduction. U-H/V significantly improved the antioxidant, anti-tyrosinase and anti-inflammatory activities of LRP. Furthermore, the results of enzyme kinetic studies showed a mixed competitive-noncompetitive inhibition of tyrosinase activity by DLRP-3 and the inhibition constant of DLRP-3 on tyrosinase was 2.97mg/mL. The apparent viscosity of LRP dispersions showed a first increasing followed by decreasing trend as ultrasonic intensity rose. U-H/V enhanced the viscoelastic properties of LRP gels without destroying their self-healing properties. This findings reveal that U-H/V is beneficial for improving the skincare efficacy of LRP, providing a theoretical foundation for the applicability of LRP in wound dressings.

Activity Variations of CYP2B6 Determine the Metabolic Stratification of Efavirenz.

To investigate the effects of hepatic enzyme activity variations and CYP2B6 gene polymorphisms on the in vivo and in vitro metabolism of efavirenz. In vitro enzyme systems using rat and human liver microsomes (RLM/HLM) were established, with in vivo studies conducted on Sprague-Dawley rats. Metabolite detection was performed via LC-MS/MS. Human recombinant CYP2B6 microsomes were prepared using a baculovirus-insect cell system and ultracentrifugation, with efavirenz serving as the substrate to study enzyme kinetics. Isavuconazole exhibited an IC50 of 21.14 ± 0.57 μM in RLM, indicating a mixed competitive and noncompetitive mechanism, and an IC50 of 40.44 ± 4.23 μM in HLM, suggesting an anticompetitive mechanism. In rats, coadministration of efavirenz and isavuconazole significantly increased the AUC, Tmax, and Cmax of efavirenz. Co-administration of efavirenz and rifampicin significantly elevated the AUC, Tmax, and Cmax of 8-OH-efavirenz. The activity of CYP2B6.4, 6, and 7 increased significantly compared to CYP2B6.1, with relative clearance ranging from 158.34% to 212.72%. Conversely, the activity of CYP2B6.3, 8, 10, 11, 13-15, 18-21, 23-27, 31-33, and 37 was markedly reduced, ranging from 4.30% to 79.89%. Variations in liver enzyme activity and CYP2B6 genetic polymorphisms can significantly alter the metabolism of efavirenz. It provides laboratory-based data for the precise application of efavirenz and other CYP2B6 substrate drugs.

Potent α-glucosidase inhibitors with benzimidazole-propionitrile hybridization; synthesis, bioassay and docking study.

Background: Diabetes is characterized by a lack of insulin and insensitivity to insulin. In 2013, the global diabetes population was 382million, with 90% of them having type 2 diabetes (non-insulin-dependent). It is predicted that this number will increase to 592million by 2035.Aim: Here, we aimed to synthesize a series of benzimidazole-based derivatives B1-B32 with α-glucosidase inhibition potential as antidiabetic agents.Methods: Compounds B1-B32 were prepared in three three-step reactions, and the structures were elucidated using spectroscopic methods, namely 1H NMR, 13C NMR, MS and IR. Enzyme inhibition and kinetic study were done using commercial assay kits, and molecular docking study using autodock4.Results: Bioassay data showed that twenty-four out of the thirty-two tested compounds exhibited IC50 values ranging from 44 to 745μM, surpassing the standard molecule, acarbose (IC50: 750μM). it was determined that the best compound, B10, functions as a competitive inhibitor. Additionally, a molecular docking study provided insights into the interactions between the four most promising compounds (B5, B6, B10 and B28) and the active site residues within the enzyme.Conclusion: The tested compounds are interesting α-glucosidase inhibitors, which indicates the benefit of more bioassay studies, especially in vivo studies.