Deciphering the toxicological and computational assessment of Verbascum yemenese.: Integrating phytochemistry and bioinformatics tools.

This study explores the phytochemical composition and biological activities of Verbascum yemenense, a plant known for its medicinal properties. The plant extract revealed a rich presence of bioactive compounds that exhibited significant antioxidant properties against free radicals. The enzyme inhibition potential was particularly notable against cholinesterases (AChE: 2.56 mg GALAE/g; BChE: 1.98 mg GALAE/g), and tyrosinase (87.94 mg KAE/g), α-glucosidase suggesting potential therapeutic applications in neurodegenerative diseases, skin disorders and diabetes. Molecular docking studies and Molecular Dynamics simulations, providing insights into the interaction mechanisms of the identified compounds with the target proteins. Molecular docking studies revealed high binding affinities of the phytoconstituents, with compounds like VY4 and phyllanthusol-A (VY15) showing substantial docking scores against AChE (-9.840 kcal/mol) and BChE (-9.853 kcal/mol), respectively. For instance, the RMSD values during the MD simulations for compound VY17 in the AML complex showed a stable conformation, fluctuating within a range of 0.75 Å to 1.75 Å, indicating a strong and consistent interaction with the enzyme. MESP studies highlighted VY17's distinctive electrostatic features, notably a pronounced electronegative region, which might contribute to its binding efficiency. These findings suggest that V. yemenense is a promising candidate for developing novel therapeutic agents.

Exploring Phytochemical profiling, Antioxidant, Enzyme Inhibition, and Antibacterial Potential of Callistemon viminalis (Sol. ex Gaertn.) G.Don ex Loudon)by In vitro and In silico Approaches.

Callistemon viminalis (Sol. ex Gaertn.) G. Don ex Loudon) (Family: Myrtaceae) is used for its medicinal properties in treating various metabolic disorders. We investigated the chemical characterization and biological screening of n-hexane extract of C. viminalis. Total phenolic content was (37.45 ± 7.40 mg GA.E/g D.E ± S.D) and total flavonoid content was (18.43 ± 6.34 mg R.E/g D.E ± S.D). The GC-MS screening of n-hexane extract tentatively identified 70 bioactive phytochemicals. The maximum antioxidant potential (289.99 ± 9.01 mg T.E/g D.E±S.D) was observed by FRAP assay. The enzyme inhibition assays revealed α-glucosidase and α-amylase inhibition (6.9 ± 0.13 and 7.2 ± 0.56 mmol of ACA.E/g D.E±S.D), urease (4.95 ± 0.9 mg of TU.E/g D.E±S.D), acetylcholinesterase (2.9 ± 0.08 mg GALA.E/g D.E±S.D), lipoxygenase (4.93 ± 1.05 mg of Indo.E/g D.E±S.D) and tyrosinase (4.33 ± 0.62 mg of KA.E/g D.E±S.D) inhibition. The extract showed antibacterial potential against Salmonella typhi (53.90±5.05) Bacillus subtilis (68.55±2.70%), Salmonella aureus (71.30±4.44%) and Pseudomonas aeruginosa (57.86±6.02%). The docking studies revealed a good docking interaction of ligands against the studied enzymes while ADME analysis revelaed pharmacokinetic profile of the phytoconstituents. C. viminalis possesses promising therapeutic potential and can be further explored for drug development and drug designing.

Bioactive compounds and in vitro biological properties of Arthrospira platensis and Athrospira maxima: a comparative study.

Cyanobacteria, especially Arthrospira, are valuable resources of nutrients and natural pigments with many beneficial health-related properties. This study optimized the extraction conditions of Arthrospira to achieve high phenolic contents and antioxidant activities. Under optimized extraction conditions, the bioactive compounds (phenolics and pigment components), antioxidant activities, and inhibitions of the key enzymes relevant to some non-communicable diseases were compared between Arthrospira platensis and Arthrospira maxima. Optimized extraction conditions were determined as 2 h shaking time, 50 °C extraction temperature, and 1% (w/v) solid-to-liquid ratio, giving effective phenolic and phycocyanin contents using aqueous extraction, while 80% (v/v) aqueous ethanolic extraction provided high total chlorophyll content. Most antioxidant activities were higher using 80% (v/v) aqueous ethanolic extracts. Both Arthrospira species inhibited the key enzymes involved in controlling non-communicable diseases including hyperlipidemia (lipase), diabetes (α-amylase, α-glucosidase, and dipeptidyl peptidase-IV), Alzheimer's disease (acetylcholinesterase, butyrylcholinesterase and ß-secretase), and hypertension (angiotensin-converting enzyme). High inhibitory activities were detected against ß-secretase (BACE-1), the enzyme responsible for ß-amyloid plaque formation in the brain that acts as a significant hallmark of Alzheimer's disease. Arthrospira extract and donepezil (Alzheimer's disease drug) synergistically inhibited BACE-1, suggesting the potential of Arthrospira extracts as effective BACE-1 inhibitors. Interestingly, A. maxima exhibited higher bioactive compound contents, antioxidant activities, and key enzyme inhibitions than A. platensis, indicating high potential for future food and medicinal applications.

Inhibition of Key Virulence Enzymes of Botrytis cinerea by Flavonoids from the Antarctic Plant Colobanthus quitensis.

This study investigates the antifungal efficacy of flavonoids derived from Colobanthus quitensis against key virulence-related enzymes implicated in the pathogenic mechanisms of Botrytis cinerea. The flavonoids swertiajaponin, schaftoside, vitexin, and saponarin significantly inhibited pectinase, cellulase, and laccase activity. Specifically, swertiajaponin showed mixed inhibition of pectinase and cellulase, characterized by high affinity (low inhibition constant -Ki-) for enzyme-substrate complexes. Schaftoside showed mixed inhibition of pectinase and competitive inhibition of laccase, effectively reducing enzymatic activity by competing directly with the substrate. In contrast, vitexin showed competitive inhibition of pectinase and non-competitive inhibition of laccase, suggesting it induces conformational changes within the enzyme. Finally, saponarin uniquely showed uncompetitive inhibition of laccase, stabilizing the enzyme-substrate complex and thereby markedly reducing catalytic turnover. Supported by kinetic parameters (maximum velocity -Vmax-, Michaelis constant -Km-, and Ki), these findings highlight the potential of flavonoids from C. quitensis as natural fungicides, offering a sustainable and eco-friendly alternative to synthetic fungicides for managing agricultural diseases.

Cheminformatic Identification of Tyrosyl-DNA Phosphodiesterase 1 (Tdp1) Inhibitors: A Comparative Study of SMILES-Based Supervised Machine Learning Models.

BACKGROUND: Tyrosyl-DNA phosphodiesterase 1 (Tdp1) repairs damages in DNA induced by abortive topoisomerase 1 activity; however, maintenance of genetic integrity may sustain cellular division of neoplastic cells. It follows that Tdp1-targeting chemical inhibitors could synergize well with existing chemotherapy drugs to deny cancer growth; therefore, identification of Tdp1 inhibitors may advance precision medicine in oncology. OBJECTIVE: Current computational research efforts focus primarily on molecular docking simulations, though datasets involving three-dimensional molecular structures are often hard to curate and computationally expensive to store and process. We propose the use of simplified molecular input line entry system (SMILES) chemical representations to train supervised machine learning (ML) models, aiming to predict potential Tdp1 inhibitors. METHODS: An open-sourced consensus dataset containing the inhibitory activity of numerous chemicals against Tdp1 was obtained from Kaggle. Various ML algorithms were trained, ranging from simple algorithms to ensemble methods and deep neural networks. For algorithms requiring numerical data, SMILES were converted to chemical descriptors using RDKit, an open-sourced Python cheminformatics library. RESULTS: Out of 13 optimized ML models with rigorously tuned hyperparameters, the random forest model gave the best results, yielding a receiver operating characteristics-area under curve of 0.7421, testing accuracy of 0.6815, sensitivity of 0.6444, specificity of 0.7156, precision of 0.6753, and F1 score of 0.6595. CONCLUSIONS: Ensemble methods, especially the bootstrap aggregation mechanism adopted by random forest, outperformed other ML algorithms in classifying Tdp1 inhibitors from non-inhibitors using SMILES. The discovery of Tdp1 inhibitors could unlock more treatment regimens for cancer patients, allowing for therapies tailored to the patient's condition.

Characterising the Metabolomic Diversity and Biological Potentials of Extracts from Different Parts of Two Cistus Species Using UHPLC-MS/MS and In Vitro Techniques.

This study investigates the biochemical composition and biological properties of different parts (leaves, roots, and twigs) of two Cistus species (Cistus monspeliasis and Cistus parviflorus). The extracts were analysed using UHPLC-MS/MS to determine their chemical profiling. A range of antioxidant assays were performed to evaluate the extract's antioxidant capabilities. The enzyme inhibition studies focused on acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-amylase, and α-glucosidase and tyrosinase. In addition, the study examined the antimicrobial effects on different bacteria and yeasts and evaluated the toxicity using the MTT assay. Quinic acid, citric acid, gallic acid, catechin, quercetin derivatives, kaempferol, myricetin, ellagic acid, prodelphinidins, procyanidins, scopoletin, and flavogallonic acid dilactone are the main bioactive compounds found in both species. In enzyme inhibition assays, C. monspeliasis roots exhibited significant activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with the values of 2.58 ± 0.02 mg GALAE/g and 11.37 ± 1.93 mg GALAE/g, respectively. Cytotoxicity studies showed mostly weak toxicity, with some samples moderately reducing viability in RAW and HepG2 cells. These findings underscore the diverse biochemical profiles and bioactive potential of Cistus species, suggesting their utility as natural sources of antioxidants and enzyme inhibitors for pharmaceutical and nutraceutical development.

Synthesis of Novel Soritin Sulfonamide Derivatives as Potential α-Glucosidase Inhibitor and Their Molecular Docking Studies.

Diabetes Mellitus (DM) is linked to various factors causing cardiovascular diseases, with uncontrolled postprandial hyperglycemia being a direct contributor. α-Glucosidase inhibitors (AGIs) aid in reducing postprandial hyperglycemia, potentially mitigating cardiovascular risks. In order to synthesize novel chemical scaffolds with possible α-glucosidase inhibition activity, a series of novel soritin sulfonamide derivatives were synthesized. The soritin hydrazide was treated with various aryl sulfonyl chlorides to obtain targeted compounds (1-16). Findings suggested that all compounds have better α-glucosidase inhibition compared to standard drugs, acarbose (2187.00 ± 1.25 µM) and 1-deoxynojirimycin (334.90 ± 1.10 µM), with IC50 values ranging from 3.81 ± 1.67 µM to 265.40 ± 1.58 µM. The most potent analog was Compound 13, a trichloro phenyl substituted compound, with IC50 value of 3.81 ± 1.67 µM. Structure-activity relationship (SAR) showed that introducing an additional chlorine group into the parent nucleus increases the potency. The docking studies validated that Compound 13 established hydrogen bonds with the active site residues Asp214, Glu276, and Asp349, while being further stabilized by hydrophobic interactions, providing an explanation for its high potency.

Exploring biological activities of novel Schiff bases derived from amlodipine and in silico molecular modeling studies.

Aim: Calcium channel antagonists are of considerable interest in treating elevated blood pressure and its pathologies.Materials & methods: Schiff base derivatives of amlodipine were produced to check its urease inhibition potentials as well antibacterial and antioxidant activities. Structural illustration along with chemical characterization were achieved by spectral techniques (1H NMR, FTIR, 13C NMR) and docking studies also performed.Results & conclusion: 3g displayed remarkable anti-hypertensive activity compared with parent drug. 3b, 3f and 3g showed urease inhibition potentials. These compounds can aid as lead for further investigations since they exhibited comparable or superior interactions.

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The Z isomer of pyridoxilidenerhodanine 5'-phosphate is an efficient inhibitor of human pyridoxine 5'-phosphate oxidase, a crucial enzyme in vitamin B6 salvage pathway and a potential chemotherapeutic target.

Pyridoxal 5'-phosphate (PLP), the catalytically active form of vitamin B6, acts as a cofactor in many metabolic processes. In humans, PLP is produced in the reactions catalysed by pyridox(am)ine 5'-phosphate oxidase (PNPO) and pyridoxal kinase (PDXK). Both PNPO and PDXK are involved in cancer progression of many tumours. The silencing of PNPO and PDXK encoding genes determines a strong reduction in tumour size and neoplastic cell invasiveness in models of acute myeloid leukaemia (in the case of PDXK) and ovarian and breast cancer (in the case of PNPO). In the present work, we demonstrate that pyridoxilidenerhodanine 5'-phosphate (PLP-R), a PLP analogue that has been tested by other authors on malignant cell lines reporting a reduction in proliferation, inhibits PNPO in vitro following a mixed competitive and allosteric mechanism. We also show that the unphosphorylated precursor of this inhibitor (PL-R), which has more favourable pharmacokinetic properties according to our predictions, is phosphorylated by PDXK and therefore transformed into PLP-R. On this ground, we propose the prototype of a novel prodrug-drug system as a useful starting point for the development of new, potential, antineoplastic agents.

Studies on the Bioactivity (Antioxidant, Enzyme Inhibitory, Antimicrobial and Cytotoxic) and Chemical Composition of Aqueous Extracts of Some Plant Species Collected from Turkmenistan.

Alhagi persarum (AP), Alhagi persarum flowers (APF), Ephedra equisetina (EE) and Glycyrrhiza glabra root (GGR) have been utilized in Central Asian folk medicine for centuries. In the present study, antioxidant, enzyme inhibition, antimicrobial and cytotoxic effects the aqueous extracts of these plants were studied. In addition, 23 compounds were identified in the extracts using UHPLC-DAD-QqQ-MS/MS analysis. The four plant samples demonstrated promising antioxidant activities in vitro for scavenging of 2, 20-azinobis-(3-ethylbenz-thiazoline-6-sulphonate) radical (ABTS), 1, 1-diphenyl-2-picrylhydrazyl radical (DPPH), and total reducing power. AP (DPPH: 47.39 mg TE/g; ABTS:108.58 mg TE/g; CUPRAC: 117.41 mg TE/g; FRAP: 82.22 mg TE/g) and EE (DPPH: 46.75 mg TE/g; ABTS: 110.51 mg TE/g; CUPRAC: 140.77 mg TE/g; FRAP: 91.00 mg TE/g) displayed the strongest antiradical and reduction abilities in the test system. In terms of enzyme inhibition efficacy, AP and EE showed higher bioactivity. Aqueous extracts demonstrated the most potent efficacy against eight different bacterial and fungal strains in antibacterial (MIC values: 1.5-4 mg/ml) and antifungal (MIC values: 1-8 mg/ml) testing. Contrary to other activities, these aqueous extracts did not show cytotoxic effects against HaCaT cells. All bioactivities suggested that the plants could be utilized as candidate resources in the food and pharmaceutical industries.

Valorization of grape pomace extracts against cranberry, elderberry, rose hip berry, goji berry and raisin extracts: Phytochemical profile and in vitro biological activity.

The circular economy is gaining attention around the world as a sustainable approach to tackling environmental problems, promoting more responsible management of resources. The aim of this work is the valorization of grape pomace as a waste product of agrifood chain. We prepared decoction (DC), ultrasound-assisted and microwave-assisted extracts (UAE and MAE respectively) of grape pomace, determining their phytochemical profile (using HPLC-ESI-Q-TOF-MS), antioxidant activity and enzyme inhibitory effects. Then, the results were compared with those of raisins and several edible berries already present in the market. Grape pomace extracts presented the highest total phenolic content (62-68 mg gallic acid equivalents/g; mg GAE/g), whereas the concentrations in the other berries were 4-43 mg GAE/g. These results were in agreement with the higher antioxidant activity and tyrosinase inhibition observed in grape pomace compared with the other berries, except for the metal chelating activity. The main compounds in grape pomace extracts were flavonoids (particularly quercetin glycosides), followed by organic acids (citric, isocitric and gallic acids). These results open new perspectives in the development of food supplements and nutraceuticals based on grape pomace extracts.

Hydrazinecarboxamides: Comprehensive review of their anticancer, anticonvulsive, anti-inflammatory, enzyme inhibition, antioxidant and other activities.

This review comprehensively summarizes recent advances in the field of hydrazinecarboxamide (semicarbazide) derivatives, highlighting their significant therapeutic potential and a broad spectrum of biological activities. As a promising and privileged scaffold in medicinal chemistry, hydrazinecarboxamides have emerged as a versatile class of compounds with significant bioactive properties. Based on their substitutions, their structural diversity permits extensive chemical modifications to enhance their interactions with various biological targets to combat multiple disorders. Notable, this group of compounds has shown significant efficacy against numerous cancer cell lines through diverse mechanisms of action and potent inhibition of enzymes, including cholinesterases, carbonic anhydrases, cyclooxygenases, lipoxygenases, etc. Beyond these, they have also been investigated for their anticonvulsive, analgesic/anti-inflammatory, and antioxidant properties, with detailed structure-activity relationships. For many applications, the hybridization of hydrazinecarboxamides with other bioactive scaffolds, such as primaquine, is of particular interest and offers advantages. Despite their promises, challenges such as suboptimal physicochemical properties and selectivity issues of certain derivatives require further effort. The review aims to inspire future innovation in the design and development of new potential hydrazinecarboxamide-based drugs, addressing existing challenges and expanding their therapeutic applications.

Phytochemical composition, simulated digestive bioaccessibility and cytotoxicity of Ficus auriculata Lour. fruits: In vitro and in silico insights.

Ficus auriculata Lour. (Moraceae) is an underutilized wild edible fruit widely consumed for its nutritional properties. The present study aimed to determine the phytochemical composition and in vitro antioxidant, enzyme inhibitory, anti-inflammatory and anti-cancerous properties of the F. auriculata fruit extracts through in vitro digestion (oral, gastric and intestinal phases). The extracts were obtained by hot extraction and cold maceration methods using aqueous and methanolic solvents. Major phytoconstituents identified through LC-MS was subjected to molecular docking against the target proteins. The elemental analysis shows the presence of major elements; high levels of total phenolics (124.61 ± 0.82 mg gallic acid equivalent/g), flavonoids (76.38 ± 0.82 mg quercetin equivalent/g), vitamin E (32.48 ± 0.09 mg alpha-tocopherol equivalent/g), and carbohydrate (34.59 ± 0.45 mg glucose equivalent/g) in hot extracted methanolic undigested extract (HEM UD) and high level of total protein (124.71 ± 0.34 mg bovine serum albumin equivalent/g) in cold extracted methanolic undigested fruit extract were found. HEM UD showed high antioxidant activity in 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid), 2,2-diphenyl-1-picryl-hydrazyl, and superoxide radical scavenging assays with IC50 of 53.30 ± 0.57, 80.69 ± 0.12, and 65.47 ± 1.13 µg/mL, respectively. The HEM UD extract also potentially inhibited the enzyme activity of α-amylase, α-glucosidase, tyrosinase, and protein denaturation (IC50 of 67.76 ± 1.22, 83.18 ± 1.23, 87.24 ± 1.15, and 65.76 ± 0.60 µg/mL). The most potent extract (HEM UD) was studied for its anticancer effects by MTT assay against the MCF-7 and HeLa cell lines and showed the IC50 of 89.80 ± 0.56 and 60.76 ± 0.04 µg/mL, respectively. The LC-MS analysis elucidated ten phytoconstituents. Based on the molecular docking study, querciturone could potentially be an effective constituent in treating diabetes and inflammation-related issues. The findings indicated the ability of F. auriculata fruits as a promising functional food.

Elucidation of the Microwave-Assisted Synthesis and Characterization of Heteronuclear Complexes of Bisbenzimidazole Derivatives and Their Biological Activities by In Vitro and In Silico Assays.

A novel and efficient protocol for the microwave-assisted synthesis of diversely substituted 2,2'-bisbenzimidazol-5,6'-dicarboxylic acid (BIMCA) from the reaction of 3,4-diaminobenzoic acid with oxalic acid has been developed, which proceeds through sequential nucleophilic addition and electrophilic substitution in accordance with the Philips method. The synthetic utility of this strategy was demonstrated by the concise, one-pot synthesis of (BIMCA) and metal complexes. (BIMCA) with a [{Fe(salen)}2O] Schiff base ligand complex and new benzimidazole coordination compounds with double oxygen [(BIMCA){Fe(salen)}2] ligand complexes were obtained. The resulting [(BIMCA){Fe(salen)}2] ligand complex was then synthesized from Co(CH3COO)2.4H2O, Ni(CH3COO)2.4H2O and Cu(CH3COO)2.H2O heteronuclear complexes. The condensations proceed with good yield to give products that, in certain instances, are not readily attainable by conventional condensation techniques. The structures of the compounds were identified by Fourier-transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance (1H NMR), elemental analysis and magnetic susceptibility. The mutagenic potential of the synthesized chemicals was evaluated by the Ames test towards mutant Salmonella typhimurium strains TA98 and TA100. It was recorded that these chemicals had no mutagenic action. Also, antimicrobial activities were screened by broth microdilution test. It was seen that the minimum inhibitory concentration (MIC) against Klebsiella pneumoniae, Staphylococcus aureus and Staphylococcus epidermidis was 0.195 mg/mL, followed by a MIC value of 0.390 mg/mL against Escherichia coli and Salmonella typhimurium. [(BIMCA){Fe(salen)}2Co(II)] demonstrated significant antimicrobial activity against Proteus mirabilis and Staphylococcus aureus, with an MIC of 0.195 mg/mL, followed by an MIC of 0.390 mg/mL against Pseudomonas aeruginosa, K. pneumonia and Salmonella typhimurium. The antioxidant properties were examined using various chemical assays, and [(BIMCA){Fe(salen)}2O] and (BIMCA) exhibited greater 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability, when compared with other compounds. Enzyme inhibitory effects were tested against acetylcholinesterase (AChE), amylase, butyrylcholinesterase (BChE) and tyrosinase. [(BIMCA){Fe(salen)}2Cu(II)] displayed the best AChE (IC50 0.51 mg/mL), BChE (IC50 0.51 mg/mL) and tyrosinase (IC50 1.52 mg/mL) inhibitory effects. Furthermore, molecular docking calculations were performed to gain insights into the interaction between [(BIMCA){Fe(salen)}2] and AChE, and between [(BIMCA){Fe(salen)}2Cu(II)] and amylase. Both compounds showed the potential inhibition of the protein targets.

Exploring Acyl Thiotriazinoindole Based Pharmacophores: Design, Synthesis, and SAR Studies with Molecular Docking and Biological Activity Profiling against Urease, α-amylase, α-glucosidase, Antimicrobial, and Antioxidant Targets.

A diminutive chemical library of acyl thiotriazinoindole (ATTI) based bioactive scaffolds was synthesized, instigated by taking the economical starting material Isatin, through a series of five steps. Isatin was first nitrated followed by the attachment of pentyl moiety via nucleophilic substitution reaction. The obtained compound was reacted with thiosemicarbazide to obtain thiosemicarbazone derivative, which was eventually cyclized using basic conditions in water as solvent. Finally, the reported series was obtained through reaction of nitrated thiotriazinoindole moiety with differently substituted phenacyl bromides. The synthesized compounds were characterized using NMR spectroscopy and elemental analysis. Finally, the synthesized motifs were scrutinized for their potential to impede urease, α-glucosidase, DPPH, and α-amylase. Compound 5 h with para cyano group manifested the most pivotal biological activity among all, displaying IC50 values of 29.7 ± 0.8, 20.5 ± 0.5 and 36.8 ± 3.9 µM against urease, α-glucosidase, and DPPH assay, respectively. Simultaneously, for α-amylase compound 5 g possessing a p-CH3 at phenyl ring unfolded as most active, with calculated IC50 values 90.3 ± 1.1 µM. The scaffolds were additionally gauged for their antifungal and antibacterial activity. Among the tested strains, 5d having bromo as substituent exhibited the most potent antibacterial activity, while it also demonstrated the highest potency against Aspergillus fumigatus. Other derivatives 5b, 5e, 5i, and 5j also exhibited dual inhibition against both antibacterial and antifungal strains. The interaction pattern of derivatives clearly displayed their SAR, and their docking scores were correlated with their IC50 values. In molecular docking studies, the importance of interactions like hydrogen bonding was further asserted. The electronic factors of various substituents engendered variety of interactions between the ligands and targets implying their importance in the structures of the synthesized heterocyclic scaffolds. To conclude, the synthesized compounds had satisfactory biological activity against various important targets. Further studies are therefore encouraged by attachment of different substitutions in the structure at various positions to enhance the activity of these compounds.

The effects of Cannabis sativa and cannabinoids on the inhibition of pancreatic lipase - An enzyme involved in obesity.

INTRODUCTION: Obesity is a chronic noncommunicable disease characterized by excessive body fat that can have negative health consequences. Obesity is a complex disease caused by a combination of genetic, environmental, and lifestyle factors. It is characterized by a discrepancy between caloric intake and expenditure. Obesity increases the risk of acquiring major chronic diseases, including heart disease, stroke, cancer, and Type 2 diabetes mellitus (T2DM). Currently, the inhibition of pancreatic lipases (PL) is a promising pharmacological therapy for obesity and weight management. In this study, the inhibition of pancreatic lipase by Cannabis sativa (C. sativa) plant extract and cannabinoids was investigated. METHODS: The inhibitory effect was assessed using p-nitrophenyl butyrate (pNPB), and the results were obtained by calculating the percentage relative activity and assessed using one-way analysis of variance (ANOVA). Kinetic studies and spectroscopy techniques were used to evaluate the mode of inhibition. Diet-induced; and diabetic rat models were studied to evaluate the direct effects of C. sativa extract on PL activity. RESULTS: Kinetic analyses showed that the plant extracts inhibited pancreatic lipase, with tetrahydrocannabinol (THC) and cannabinol (CBN) being the potential cause of the inhibition noted for the C. sativa plant extract. CBN and THC inhibited the pancreatic lipase activity in a competitive manner, with the lowest residual enzyme activity of 52 % observed at a 10 µg/mL concentration of CBN and 39 % inhibition at a 25 µg/mL concentration of THC. Circular dichroism (CD) spectroscopy revealed that the inhibitors caused a change in the enzyme's secondary structure. At low concentrations, THC showed potential for synergistic inhibition with orlistat. C.sativa treatment in an in vivo rat model confirmed its inhibitory effects on pancreatic lipase activity. CONCLUSION: The findings in this study provided insight into the use of cannabinoids as pancreatic lipase inhibitors and the possibility of using these compounds to develop new pharmacological treatments for obesity.

Unraveling the mechanism of carbonic anhydrase IX inhibition by alkaloids from Ruta chalepensis: A synergistic analysis of in vitro and in silico data.

Due to the pivotal role of carbonic anhydrase IX (CA IX) in pathological conditions, there's a pressing need for novel inhibitors to improve patient outcomes and clinical management. Herein, we investigated the inhibitory efficacy of six alkaloids from Ruta chalepensis against CA IX through in vitro inhibition assay and computational modeling. Skimmianine and maculosidine displayed significant inhibitory activity in vitro, with low IC50 values of 105.2 ± 3.2 and 295.7 ± 14.1 nM, respectively. Enzyme kinetics analyses revealed that skimmianine exhibited a mixed inhibition mode, contrasting with the noncompetitive inhibition mechanism observed for the reference drug (acetazolamide), as indicated by intersecting lines in the Lineweaver-Burk plots. The findings of docking calculations revealed that skimmianine and maculosidine exhibited extensive polar interactions with the enzyme. These alkaloids demonstrate substantial binding interactions and occupy identical binding site as acetazolamide, thereby enhancing their efficacy as inhibitors of CA IX. Utilizing a 100 ns molecular dynamics (MD) simulation, the dynamic interactions between isolated alkaloids and CA IX were intensively assessed. Analysis of diverse MD parameters revealed that skimmianine and maculosidine displayed consistent trajectories and notable energy stabilization during their interaction with CA IX. The findings of MM/PBSA analysis depicted the minimum binding free energy for skimmianine and maculosidine. In addition, the Potential Energy Landscape (PEL) analysis revealed distinct and stable conformational states for the CA IX-ligand complexes, with Skimmianine showing the most stable and lowest energy configuration. These computational findings align with experimental results, emphasizing the potential efficacy of skimmianine and maculosidine as inhibitors of CA IX.

Distinct transcriptional profiles in rat thyroid glands after developmental exposure to three in vitro thyroperoxidase inhibiting chemicals.

Thyroperoxidase (TPO) is central in thyroid hormone (TH) synthesis and inhibition can lead to TH deficiency. Many chemicals can inhibit TPO activity in vitro, but how this may manifest in the developing thyroid gland at the molecular level is unclear. Here, we characterized the thyroid gland transcriptome of male rats developmentally exposed to the in vitro TPO-inhibitors amitrole, 2-mercaptobenzimidazole (MBI), or cyanamide by use of Bulk-RNA-Barcoding (BRB) and sequencing. Amitrole exposure caused TH deficiency and 149 differentially expressed genes in the thyroid gland. The effects indicated an activated and growing thyroid gland. MBI caused intermittent changes to serum TH concentrations in a previous study and this was accompanied by 60 differentially expressed genes in the present study. More than half of these were also affected by amitrole, indicating that they could be early effect biomarkers of developmental TH system disruption due to TPO inhibition. Further work to validate the signature is needed, including assessment of substance independency and applicability domain.

NMR Metabolite Profiling and Antioxidant Properties of Spartan, Jewels, Misty, and Camelia Blueberries.

The health-promoting properties of blueberries are widely recognized and are mainly attributed to anthocyanins. However, fruit's chemical composition includes also other components and strongly depends on varieties and climatic conditions. Here, 1H NMR metabolite profiling and biological activity of four blueberry cultivars (Spartan, Jewels, Misty, Camelia) grown in Central Italy over two years were reported. Untargeted and targeted NMR analyses allowed the quantification of sugars, organic acids, amino acids, anthocyanins, lipids, and other compounds. Spectrophotometric assays evaluated total phenolic and flavonoid content, antioxidant activity, and enzyme inhibitory activity toward cholinesterase, α-amylase, α-glucosidase, and tyrosinase. Statistical analysis showed a correlation between chemical composition and biological activity, revealing markers specific to blueberry cultivars (quinic acid, quercitrin, myo-inositol, myrtillin, and petunidin-3-O-glucoside). Almost all antioxidant assays were correlated with the chlorogenic acid levels. A strong effect of harvesting on chemical composition and biological activities was observed, with Misty cultivar having the highest antioxidant activity.

4-Phenyl-thiazole-based dual inhibitors of fatty acid amide hydrolase and soluble epoxide hydrolase do not alleviate orofacial inflammatory pain in female rats.

Pain arising from trigeminal systems such as headache is common, debilitating, and current treatments (e.g., sumatriptan) are limited. New treatments that target novel mechanisms of action may be required to innovate both short- and long-term pain therapy. Fatty acid amide hydrolase and soluble epoxide hydrolase are two pain-related enzymes that regulate pain and inflammation via independent pathways. We have previously demonstrated that simultaneous inhibition of these enzymes using a novel dual inhibitor alleviates acute inflammatory pain in the hindpaw and does not depress wheel running in rats. Here, we expanded on these findings and performed structure-activity relationships of our lead compound, the 4-phenyl-thiazole-based dual inhibitor SW-17, to generate 18 analogs and tested them for their inhibition at both enzymes. Conversion of the sulfonamide group to a tertiary amine led to a general decrease in the potency for the sEH enzyme, while this change was well-tolerated at the FAAH enzyme yielding several strong inhibitors. Six selected inhibitors were evaluated in mouse and rat sEH inhibition assays and results showed a species difference, i.e. 4-phenyl-thiazole-based analogs are significantly less or not active in mouse sEH compared to human and rat enzymes. The most potent inhibitor, SW-17, was evaluated in a plasma stability assay in human and rat plasma and showed moderate stability. However, SW-17 did not alleviate orofacial inflammatory pain in female rats compared to the traditional anti-migraine agent sumatriptan. Although modification of 4-phenyl-thiazole-based dual inhibitor SW-17 changes potencies at both FAAH and sEH, these approaches may not produce antinociception against trigeminal pain. Key Words: polypharmacology, formalin, inflammation, enzyme inhibition, structure-activity relationship studies.