5-LOX Inhibitor Research Articles

Protective effects of licofelone on scopolamine-induced spatial learning and memory impairment by enhancing parkin-dependent mitophagy and promotion of neural regeneration and in adult mice

Inhibition of COX and LOX could contribute to memory formation and prevention of neurodegeneration, by alleviation of neuroinflammation and improvement of mitochondrial homeostasis. We aimed to assess the effect of licofelone, a dual COX and 5-LOX inhibitor on memory formation, neural apoptosis, neural regeneration, and mitophagy in acute and chronic dosages, given that licofelone could regulate nitric oxide levels. Y-maze and Passive Avoidance tests were used to evaluate memory function in NMRI mice using the EthoVision setting, following scopolamine administration (1 mg/kg, i.p.) as an acute amnestic drug. Hippocampi were used to evaluate the levels of apoptosis via TUNEL assay, neural regeneration via immunohistochemistry method detecting doublecortin and nestin, and mitophagy via Western blot of mitophagy proteins Parkin and ATG5. While acute high-dose licofelone (20 mg/kg) could reverse amnestic effects of scopolamine in passive avoidance test (p = 0.0001), Chronic licofelone (10 mg/kg for 10 consecutive days) could improve performance in Y-maze (p = 0.0007). Molecular analysis revealed that the chronic form of the drug could enhance neural regeneration in CA1 and SGZ regions, reset mitophagy levels as much as the healthy state, and reduce apoptosis rate. Licofelone appears to show a desirable anti-amnestic profile in a low dose chronically; it is hence recommended for future clinical studies on the prevention of neuroinflammation and memory deficit.

In Vitro Evaluation of Pharmacokinetic Properties of Selected Dual COX-2 and 5-LOX Inhibitors.

Evaluation of pharmacokinetic properties is a significant step at the early stages of drug development. In this study, an in vitro evaluation of the pharmacokinetic properties of five newly synthesized compounds was performed. These compounds belong to N-hydroxyurea and hydroxamic acid derivatives and analogs of NSAIDs indomethacin, flurbiprofen, diclofenac, ibuprofen, and naproxen (compounds 1, 2, 3, 11, and 12, respectively) with dual COX-2 and 5-LOX inhibitory activity. Two in vitro methods (biopartitioning micellar chromatography (BMC) and PAMPA) were used to evaluate passive gastrointestinal absorption, while high-performance affinity chromatography (HPAC) and differential pulse voltammetry (DPV) were used to evaluate binding to human serum albumin (HSA). The introduction of N-hydroxyurea and hydroxamic acid groups into the structure of NSAIDs decreases both expected passive gastrointestinal absorption (BMC k values were from 3.02 to 9.50, while for NSAIDs were from 5.29 to 13.36; PAMPA -logPe values were between 3.81 and 4.76, while for NSAIDs were ≤3.46) and HSA binding (HPAC logk values were from 2.03 to 9.54, while for NSAIDs were ≥11.03; DPV peak potential shifts were between 7 and 34, while for NSAIDs were ≥54). Structural modifications of all tested compounds that increase lipophilicity could be considered to enhance their passive gastrointestinal absorption. Considering lower expected HSA binding and higher lipophilicity of tested compounds compared to corresponding NSAIDs, it can be expected that the volume of distribution of compounds 1, 2, 3, 11, and 12 will be higher. Reduced HSA binding may also decrease interactions with other drugs in comparison to corresponding NSAIDs. All tested compounds showed significant microsomal instability (25.07-58.44% decrease in concentration) in comparison to indomethacin (14.47%) and diclofenac (20.99%).

12-Lipoxygenase inhibition suppresses islet immune and inflammatory responses and delays autoimmune diabetes in human gene replacement mice.

Type 1 diabetes (T1D) is characterized by the autoimmune destruction of insulin-producing β cells and involves an interplay between β cells and cells of the innate and adaptive immune systems. We investigated the therapeutic potential of targeting 12-lipoxygenase (12-LOX), an enzyme implicated in inflammatory pathways in β cells and macrophages, using a mouse model in which the endogenous mouse Alox15 gene is replaced by the human ALOX12 gene. Our findings demonstrate that VLX-1005, a potent 12-LOX inhibitor, effectively delays the onset of autoimmune diabetes in human gene replacement non-obese diabetic mice. By spatial proteomics analysis, VLX-1005 treatment resulted in marked reductions in infiltrating T and B cells and macrophages with accompanying increases in immune checkpoint molecule PD-L1, suggesting a shift towards an immune-suppressive microenvironment. RNA sequencing analysis of isolated islets and polarized proinflammatory macrophages revealed significant alteration of cytokine-responsive pathways and a reduction in interferon response after VLX-1005 treatment. Our studies demonstrate that the ALOX12 human replacement gene mouse provides a platform for the preclinical evaluation of LOX inhibitors and supports VLX-1005 as an inhibitor of human 12-LOX that engages the enzymatic target and alters the inflammatory phenotypes of islets and macrophages to promote the delay of autoimmune diabetes.

Dual COX/ 5-LOX inhibitory potential and antioxidant activities by extracts isolated from Melia Azedarach L. leaves

This study was aimed at evaluating the dual cyclooxygenase (COX) and 5-Lipoxygenase (5-LOX) enzyme inhibition and antioxidant activity of the crude extract (CE) and various solvent fractions from Melia azedarach leaves. Multiple solvents were used to fractionate the 80% ethanolic extract of the leaves, which was then examined for enzyme inhibitory effectiveness against COX and 5-LOX enzymes. All fractions and CE were examined for antioxidant activity, and phenolic acids and flavonoids were identified using Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC). The ethyl acetate fraction (EaF) had the greatest COX/5-LOX inhibitory impact among tested fractions.In vitro, COX activity studies showed that EaF and butanol fraction (BF) possess a higher COX-2 selectivity index than Indomethacin. Besides, the EaF exhibited the highest reduction potential among the control and the CE when tested for antioxidant activities. These findings support the traditional use of Melia azedarachas an anti-inflammatory drug, as the plant inhibited both COX/5-LOX and COX-2 with high selectivity.1.Introduction The most frequent chemical species that cause oxidative damage in the body are reactive oxygen species (ROS) like peroxides, superoxide, singlet oxygen, hydroxyl radical, hydrogen peroxides, and so on. These species produced in the body can cause various cardiovascular and other chronic diseases [1-3], neurodegenerative diseases [4,5], and inflammatory diseases [6,7]. Different inflammatory diseases are linked to oxidative stress (an imbalance between ROS and their removal by protective mechanisms such as antioxidants) [7-9]. Inflammatory mediators such as prostaglandins produced by the arachidonic acid pathway need to be reduced and inhibited as they mainly cause inflammation, pain, and fever in the body [10]. Currently, the symptomatic treatment of inflammation is mainly done by non-steroidal anti-inflammatory drugs (NSAIDs). However, the traditional NSAIDs used in COX reduction are non-selective, lead to gastrointestinal side effects, heart strokes and renal failure. On the other hand, the currently used selective COX-2 inhibitors like ‘coxibs' are reported for cardiovascular side effects [11]. In addition to COX-2, the increased level of leukotriene from the 5-LOX enzyme can result in bronchospasm [11]. In this regard, medicinal preparations that inhibit both Cyclooxygenase (COX-2) and 5-Lipoxygenase (5-LOX) pathways are superior to commonly used NSAIDs since they could produce a synergetic effect to achieve optimal anti-inflammatory activities [10,11]. This can be done by blocking the production of both leukotrienes and prostaglandins. Hence, indeed, there is a need for new, effective, and safe anti-inflammatory drugs with dual COX-2/5-LOX inhibition capacities, which may obliterate the side effects caused by the inhibition of COX-2 alone [12,13]. Melia azedarachL. (known commonly as “Chinaberry, bead tree, Bakain and Persian lilac") is a deciduous tree belonging to the Meliaceae family that is native to tropical Asia and Australia but has now spread throughout the world. [14-16]. The antioxidantand anti-inflammatory potential of Melia azedarachL. from various regions werereported by other groups [14,15, 17-21]. The plant with different species and some species of different cultivars markedly differin their phenolics, flavonoids and antioxidant activities [18,20]. The antioxidant and anti-inflammatory activities of plant extracts are closely linked to phytoconstituents like total phenolics and flavonoids [22-25]. There is no scientific report on the anti-inflammatory action and antioxidant potential of Taiwanese-originMelia azedarach. Therefore, the purpose of the current study is to investigate the dual COX-2 and 5-LOX inhibitor potential, antioxidant potential, quantification of key flavonoids and phenolic acids, as well as total phenolic and flavonoid content of Melia azedarach.

Exploration of 4-tolyl-5-(p-tolyloxymethyl)-4H-1,2,4-triazole thioethers as potent 15-LOX inhibitors supported by in vitro, in silico, MD simulation and DNA binding studies

Inflammation is an intricate process exacerbated by the sequential release of lipid mediators during arachidonic acid metabolism through the LOX and COX pathways. The inhibition of enzymes accountable for the arachidonic acid metabolism is attributed to a synergistic anti-inflammatory impact with a broader spectrum of activity. The present study was concerned with the search for lead 15-LOX inhibitors. A new series of alkyl/aralkyl substituted 4-tolyl-5-(p-tolyloxymethyl)-4H-1,2,4-triazole (6a-h) was designed, synthesized, and characterized to determine their 15-LOX inhibitory potential. The analogue 3-(phenethylthio)-4-tolyl-5-(p-tolyloxymethyl)-4H-1,2,4-triazole (6h) was the most potent with IC50 2.73 ± 0.15 µM while 6g, 6d, 6f, 6c exhibited excellent inhibitory activities with IC50 values between 11.39 ± 0.13 and 14.63 ± 0.17 µM. All analogues maintained > 72 % blood mononuclear cells viability at 0.25 mM concentration during MTT assay. In silico ADME profiles predicted druggability potential. The molecular docking studies disclosed binding free energies of -9.78 to -7.94 kcal/mol and His373 and His378 were majorly involved in π-interactions in all molecules. Van der Waals interactions dominated hydrogen bonding in the stabilization of ligand-receptor complexes. MD simulations and free energy calculations revealed the significance of complexes in stabilizing the biomolecular system and helped in identifying the key active site residues that contributed towards the energetics. Studies against DNA dodecamer displayed minor groove binding and intercalation, suggesting a possible role in modulating DNA-related processes. The results approve the active analogues possessing promising druglike properties and work is in progress on the synthesis of more analogues in search for leads as 15-LOX inhibitors.

Evaluation of Antioxidant and Anti-inflammatory Activities of a Medicinal Plant Recipe from Burkina Faso

<i>Background:</i> Oxidative stress and chronic inflammation are crucial factors in the development of various diseases, such as hepatitis and cancer. In the case of hepatitis, persistent inflammation of the liver contributes to cellular damage and progression to more severe stages of the disease. <i>Objective:</i> The aim of this study is to evaluate the antioxidant and anti-inflammatory activity of a recipe, <i>Hepatib tiben,</i> traditionally used in the treatment of viral hepatitis B, and often associated with <i>Momordica charantia</i> in the treatment of liver cancer in Burkina Faso. <i>Material and Methods:</i> The recipe and associated plants were infused, macerated and the yields were evaluated and being freeze-dried. The extracts obtained were used to determine the total content of phenolic and flavonoid compounds by measuring the antioxidant activity through the FRAP, DPPH and ABTS methods and then the evaluation of the anti-inflammatory properties by inhibition of 15-LOX and COX 1 & 2. <i>Results: </i>The aqueous infusion of <i>Hepatib tiben</i> had the best yield, the highest content of phenolic compounds and flavonoids, an antioxidant activity comparable to that of Rutin, by the FRAP method, DPPH and ABTS method. But the best antioxidant activity by FRAP method was observed with the DCM fraction. It is on the other hand the hydroalcoholic extract which had the best anti-inflammatory activity by the inhibition of the 15-LOX. Both Aqueous and Hydroalcoholic Extracts inhibited COX 1 & 2. <i>Momortica charantia</i> had high phenolic content and significant antioxidant activity by FRAP, DPPH and ABTS methods. <i>Conclusion:</i> These observations suggest that the recipe of traditional medicinal plants used in Burkina Faso could be beneficial in the treatment of inflammatory conditions and oxidative stress. Further studies are needed to identify the active compounds responsible for these activities and to assess their safety and clinical effectiveness.

Novel Dual COX-2/5-LOX Inhibitory Activity by Chalcone Derivatives: A Safe and Efficacious Anti-inflammatory Agent

Background: Non-communicable diseases are chronic systemic inflammation in humans that occurs because of enhanced inflammatory mediators of the arachidonic acid cas-cade. We aimed to explore whether the lead chalcone compounds could exhibit anti-inflam-matory activity via dual blockage of COX-2/5-LOX enzymes and their regulatory mechanism. Methods: RAW 264.7 macrophages were collected from NCC, Pune, for in-vitro experiments. The IC50 values of chalcone compounds C45 and C64 were calculated. RAW 264.7 macro-phages were treated with C45 and C64 (10%, 5%, 2.5%, 0.125%, and 0.0625% concentration). The cell viability was carried out with an MTT assay. The COX-1, COX-2, 5-LOX, PGE2, and LTB4 levels were detected by ELISA-based kits. The in-vivo evaluation was carried out in Male Wistar rats (250-300 g, 7-8 weeks old) with acute and chronic anti-inflammatory models and histopathological studies on the stomach, liver, and kidney. Results: The present study described the in-vitro and in-vivo biological evaluation of dual COX-2/5-LOX inhibitors in chalcone derivatives (C45 and C64) compounds showed the most effective COX-2 and 5-LOX inhibition with IC50 values 0.092 and 0.136μM respectively. Simultaneously, compound C64 showed comparable selectivity towards COX-2 with a Selec-tivity Index (SI) of 68.43 compared to etoricoxib, with an SI of 89.32. In-vivo carrageenan-induced rat paw oedema activity, the compound C64 showed a significant reduction in oedema with 78.28% compared to indomethacin with 88.07% inhibition. Furthermore, cotton pellet-induced granuloma activity revealed that compound C64 significantly reduced 32.85% com-pared with standard 40.13% granuloma inhibition. Conclusion: The chalcone compound C64, (E)-1-(4-Amino-2-hydroxyphenyl)-3-(3,4,5-tri-methoxyphenyl)-prop-2-en-1-one was proved to be a potent and novel Dual COX-2/5-LOX inhibitor with improved gastric safety profiling.

Evaluation of the Interaction between Carvacrol and Thymol, Major Compounds of Ptychotis verticillata Essential Oil: Antioxidant, Anti-Inflammatory and Anticancer Activities against Breast Cancer Lines.

The objective of this study was to evaluate the antioxidant, anti-inflammatory, and anticancer properties of thymol, carvacrol, and their equimolar mixture. Antioxidant activities were assessed using the DPPH, ABTS, and ORAC methods. The thymol/carvacrol mixture exhibited significant synergism, surpassing the individual compounds and ascorbic acid in DPPH (IC50 = 43.82 ± 2.41 µg/mL) and ABTS (IC50 = 23.29 ± 0.71 µg/mL) assays. Anti-inflammatory activity was evaluated by inhibiting the 5-LOX, COX-1, and COX-2 enzymes. The equimolar mixture showed the strongest inhibition of 5-LOX (IC50 = 8.46 ± 0.92 µg/mL) and substantial inhibition of COX-1 (IC50 = 15.23 ± 2.34 µg/mL) and COX-2 (IC50 = 14.53 ± 2.42 µg/mL), indicating a synergistic effect. Anticancer activity was tested on MCF-7, MDA-MB-231, and MDA-MB-436 breast cancer cell lines using the MTT assay. The thymol/carvacrol mixture demonstrated superior cytotoxicity (IC50 = 0.92-1.70 µg/mL) and increased selectivity compared to cisplatin, with high selectivity indices (144.88-267.71). These results underscore the promising therapeutic potential of the thymol/carvacrol combination, particularly for its synergistic antioxidant, anti-inflammatory, and anticancer properties against breast cancer. This study paves the way for developing natural therapies against breast cancer and other conditions associated with oxidative stress and inflammation, leveraging the synergistic effects of natural compounds like thymol and carvacrol.

5-Lipoxygenase: The Therapeutically Unexplored Target for Bone Health

Bone fracture is a common orthopedic condition that represents a significant health concern. Bone fracture repair process is a complex process that involves the harmonic and synchronized activity of bone cells. 5-lipoxygenase (5-LOX) is an enzyme responsible for the conversion of arachidonic acid to form leukotrienes. While leukotrienes have an empirical inflammatory role, various evidence suggests that these 5-LOX mediators, by switching the inflammatory environment and altering the activity of bone cells, have a detrimental effect on the bone fracture healing process. Additionally, various evidence suggests that 5-LOX inhibition shows improvement in the overall bone healing process and improves overall bone health. Despite this evidence, the clinical use of 5-LOX inhibitors in bone fracture healing is largely unexplored. The current review aimed to summarize the available evidences and pave the way for future large scale pre-clinical and clinical studies to evaluate the effectiveness of selective 5-LOX inhibitors in bone fracture healing. A comprehensive literature search was conducted in PubMed, Google Scholar, and ScienceDirect to identify relevant articles related to the effect of 5-LOX in bone health. The summary of available scientific evidence highlights that the selective 5-LOX inhibition can modulate the functioning of osteoclasts and osteoblasts, favoring faster bone fracture healing. The observations of this study support the selective 5-LOX inhibition as a potential treatment option that can improve bone fracture healing rate and therefore future clinical studies can be designed to confirm the observations of the current review study.

Pharmacochemical Study of Multitarget Amino Acids' Hybrids: Design, Synthesis, In vitro, and In silico Studies.

Neuro-inflammation is a complex phenomenon resulting in several disorders. ALOX-5, COX-2, pro-inflammatory enzymes, and amino acid neurotransmitters are tightly correlated to neuro-inflammatory pathologies. Developing drugs that interfere with these targets will offer treatment for various diseases. Herein, we extend our previous research by synthesizing a series of multitarget hybrids of cinnamic acids with amino acids recognized as neurotransmitters. The synthesis was based on an In silico study of a library of cinnamic amide hybrids with glycine, γ- aminobutyric, and L - glutamic acids. Drug-likeness and ADMET properties were subjected to In silico analysis. Cinnamic acids were derived from the corresponding aldehydes by Knoevenagel condensation. The synthesis of the amides followed a two-step reaction with 1- hydroxybenzotriazole monohydrate and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride in dry dichloromethane and the corresponding amino acid ester hydrochloride salt in the presence of N,N,-diisopropyl-Nethylamine. The structure of the synthesized compounds was confirmed spectrophotometrically. The new compounds, such as lipoxygenase, cyclooxygenase-2, lipid peroxidation inhibitors, and antiinflammatories, were tested in vitro. The compounds exhibited LOX inhibition with IC50 values in the low μM region). Compounds 18a, 23b, and 11c are strong lipid peroxidation inhibitors (99%, 78%, and 92%). Compound 28c inhibits SLOX-1 with IC50 =8.5 μM whereas 11a and 22a highly inhibit COX-2 (IC50 6 and 5 μM Hybrids 14c and 17c inhibit both enzymes. Compound 29c showed the highest anti-inflammatory activity (75%). The In silico ADMET properties of 14c and 11a support their drug-likeness.

Interactions between Inhibitors and 5-Lipoxygenase: Insights from Gaussian Accelerated Molecular Dynamics and Markov State Models.

Inflammation is a protective stress response triggered by external stimuli, with 5-lipoxygenase (5LOX) playing a pivotal role as a potent mediator of the leukotriene (Lts) inflammatory pathway. Nordihydroguaiaretic acid (NDGA) functions as a natural orthosteric inhibitor of 5LOX, while 3-acetyl-11-keto-β-boswellic acid (AKBA) acts as a natural allosteric inhibitor targeting 5LOX. However, the precise mechanisms of inhibition have remained unclear. In this study, Gaussian accelerated molecular dynamics (GaMD) simulation was employed to elucidate the inhibitory mechanisms of NDGA and AKBA on 5LOX. It was found that the orthosteric inhibitor NDGA was tightly bound in the protein's active pocket, occupying the active site and inhibiting the catalytic activity of the 5LOX enzyme through competitive inhibition. The binding of the allosteric inhibitor AKBA induced significant changes at the distal active site, leading to a conformational shift of residues 168-173 from a loop to an α-helix and significant negative correlated motions between residues 285-290 and 375-400, reducing the distance between these segments. In the simulation, the volume of the active cavity in the stable conformation of the protein was reduced, hindering the substrate's entry into the active cavity and, thereby, inhibiting protein activity through allosteric effects. Ultimately, Markov state models (MSM) were used to identify and classify the metastable states of proteins, revealing the transition times between different conformational states. In summary, this study provides theoretical insights into the inhibition mechanisms of 5LOX by AKBA and NDGA, offering new perspectives for the development of novel inhibitors specifically targeting 5LOX, with potential implications for anti-inflammatory drug development.

Probing N-substituted 4-(5-mercapto-4-ethyl-4H-1,2,4-triazol-3-yl)-N-phenylpiperdine-1-carboxamides as potent 15-LOX inhibitors supported with ADME, DFT calculations and molecular docking studies

In our continuous efforts to find out leads against the enzyme 15-lipoxygenase (15-LOX), the current study deals with the synthesis of a series of new N-alkyl/aralkyl/aryl derivatives of 2-(4-ethyl-5-(1-phenylcarbamoyl)piperidine-4H-1,2,4-triazol-3-ylthio)methylacetamide (7a-n) with anti-LOX activities. The synthesis was started by reacting phenylisocyanate with isonipecotate that sequentially converted into N-substituted ester (1), hydrazide (2), semicarbazide (3) and N-ethylated 5-(1-phenylcarbamoyl)piperidine-1,2,4-triazole (4). The final compounds, 7a-n, were obtained by reacting 4 with various N-alkyl/aralkyl/aryl electrophiles. Both the intermediates and target compounds were characterized by FTIR, 1H, 13C NMR spectroscopy, EI-MS and HR-EI-MS spectrometry and screened against soybean 15-LOX by chemiluminescence method. The eight compounds 7e, 7j, 7h, 7a, 7g, 7b, 7n, 7c showed potent inhibitory activities against 15-LOX with values ranging from IC50 0.36 ± 0.15 μM (7e) to IC50 6.75 ± 0.17 μM (7c) compared with the reference quercetin (IC50 4.86 ± 0.14 μM) and baicalein (IC50 2.24 ± 0.13 μM). Two analogues (7l, 7f) had significantly outstanding inhibitory potential with IC50 values 12.15 ± 0.23 μM and 15.54 ± 0.26 μM, whereas, the derivatives 7i, and 7d displayed IC50 values of 21.56 ± 0.27 μM, 23.59 ± 0.24 μM and the compounds 7k, 7m were found inactive. All analogues exhibited blood mononuclear cells (MNCs) viability >75 % at 0.25 mM concentration as determined by MTT method. Calculated pharmacokinetic properties projected good lipophilicity, bioavailability and drug-likeness properties and did not violate Lipinski's/Veber rule. Molecular docking studies revealed lower binding free energies of all the derivatives than the reference compounds. The binding free energies were −9.8 kcal/mol, −9.70 k/mol and −9.20 kcal/mol for 7j, 7h and 7e, respectively, compared with the standard quercetin (−8.47 kcal/mol) and baicalein (−8.98 kcal/mol). The docked ligands formed hydrogen bonds with the amino acid residues Gln598 (7e), Arg260, Val 126 (7h), Gln762, Gln574, Thr443, Arg580 (7j) while other hydrophobic interactions observed therein further stabilized the complexes. The results of density functional theory (DFT) revealed that analogues with more stabilized lower unoccupied molecular orbital (LUMO) had significant enzyme inhibitory activity. The data collectively supports these molecules as leads against 15-LOX and demand further investigations as anti-inflammatory agents.

15-lipoxygenase and serine protease inhibitory activities of Heritiera fomes (buch. -ham), a mangrove plant of the Sundarbans

15-Lipoxygenase (15-Lox) oxidizes fatty acid, thereby producing inflammatory mediators which cause diverse inflammatory diseases such as atherosclerosis, asthma, cancer etc. Serine proteases are enzymes that activate tissue factor VII by proteolytic cleavage, eventually activating blood coagulation pathway. Antioxidants play role in 15-Lox and serine protease inhibition to dampen elevation of cytokine levels, thereby function as anti-inflammation and anticoagulation agent. In this study, we examined 15-Lox and serine protease inhibitory activities of the ethanolic bark and leaf crude extract of Heritiera fomes. The crude bark extract (HFB) exhibited 15-Lox and serine protease inhibition with IC50 117.14 and 409.61 μg/ml, respectively. Chromatographically separated active fraction HFBPF1 demonstrated highest 15-Lox and serine protease inhibitory activity with IC50 62.68 and 72.62 μg/ml, respectively. The antioxidant activity and total flavonoid content were substantially increased in the chromatographic fractions compared to the crude extracts. The LC-MS/MS analysis of bark and leaf active fractions revealed the presence of 2,4,6-octatrienal, apigenin 7 (4˝-Z-p-coumerylglucoside), brosimacutin B, gambrin B2, kandelin A1 and tetraneurin A which we report for the first time in H. fomes. Therefore, we conclude the significant augmentation in the 15-Lox and serine protease inhibition activity of the fractions which could be exerted by the enriched flavonoid content.

Metabolomic profiling of barley extracts obtained via different solvents and evaluation of their anti-inflammatory efficacy

Barley (Hordeum vulgare L.) stands out for its special nutritional value compared to other cereals, garnering significant attention in recent years. In this study, a comprehensive chemical and biological evaluation of different barley cultivars extracted with different solvents was performed. Ultra-high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC-ESI-QTOF-MS/MS) was used in the phytochemical study. The molecular docking with absorption, distribution, metabolism, excretion, and toxicity (ADMET) study and multivariate analysis alongside the quantification of total phenolic contents (TPC) were performed. Furthermore, the anti-inflammatory action was in vitro assessed by measuring COX-1, COX-2, and 5-LOX inhibition. This study seeks to establish a suitable solvent for extracting anti-inflammatory properties from diverse barley cultivars. The methanol extract obtained from the G.131 cultivar, followed by the 80% methanol extract from the G.136 cultivar, exhibited the most favorable IC50 values in comparison to the reference drugs indomethacin and celecoxib. The phytochemical profile revealed more than 60 compounds that belonged to nitrogen-containing compounds, phenolic compounds, organic acids, flavonoids, and their glycosides, hordatines, and other classes. The TPC and COX-2 inhibition was positively correlated with isoscoparin, coumaroyle–OH–eagmatine, isoorientin, isovitexin 7-O-rhamnosylglucoside, isoscoparin 7-O-glucoside, and isoorientin-7-O-[6-feruloyl]-glucoside. These findings along with their pharmacodynamics study suggested a potential synergistic and significant contribution of these compounds with the anti-inflammatory activity.

Hybrids of 4-aminosalicylic acid with dual anti-mycobacterial and anti-inflammatory activities: Synthesis, biological evaluation, in silico investigation and structure-activity relationships exploration

Studies point toward a beneficial effect of NSAIDs as an adjunctive tuberculosis treatment. To discover new antitubercular compounds with anti-inflammatory activity and favorable safety profiles, three new series of non-carboxylic 4-aminosalicylic acid hybrids bearing various hydrazides (3 and 5), heterocyclic scaffolds (6–15), hydrazones (16a–d) and carboxylic analogues bearing 1,2,3-triazoles (19 and 20), were synthesized. The synthesized hybrids were evaluated for their antimycobacterial activity against H37Rv using MABA assay. Among them, compounds 8, 16a–c, 19 and 20 exhibited MIC values (9.90–37.62 µM) with good selectivity index. 16b and 20 were the most potent with MIC 9.90 and 11.40 µM, respectively. Based on an in silico docking study at COX-2 active site, 11 compounds 13a, 13b, 14a–g, 16a and 16b were selected for testing their COX-2 inhibition. The most active compounds, 13a, 16a and 16b were further tested for COX-1 and 5-LOX inhibition. Also, their COX-2 selectivity indices (SI) were determined. 16a and 16b showed the highest potency and selectivity against COX-2 IC50 = 0.31, 1.26 µg/mL, respectively, and 5-LOX with IC50 = 5.49, 25.44 µg/mL, respectively. Computational docking simulations and in Silico ADME/pharmacokinetic studies for the most active hybrids were performed and discussed. Thus structure-activity relationships were explored. In conclusion, compounds 16a and 16b represent a good starting point for the development of new potential drugs of dual antitubercular and anti-inflammatory activities.

Mango peel extracts and mangiferin chromatographic Fourier-transform infrared correlation with antioxidant, antidiabetic, and advanced glycation end product inhibitory potentials using in silico modeling and in vitro assays.

Mangifera indica peels are a rich source of diverse flavonoids and xanthonoids; however, generally these are discarded. Computational studies revealed that mangiferin significantly interacts with amino acid residues of transcriptional regulators 1IK3, 3TOP, and 4f5S. The methanolic extract of Langra variety of mangoes contained the least phenol concentrations (22.6 ± 0.32 mg/gGAE [gallic acid equivalent]) compared to the chloroform (214.8 ± 0.12 mg/gGAE) and ethyl acetate fractions (195.6 ± 0.14 mg/gGAE). Similarly, the methanolic extract of Sindhri variety contained lower phenol concentrations (42.3 ± 0.13 mg/gRUE [relative utilization efficiency]) compared with the chloroform (85.6 ± 0.15 mg/gGAE) and ethyl acetate (76.1 ± 0.32 mg/gGAE) fractions. Langra extract exhibited significant α-glucosidase inhibition (IC50 0.06 mg/mL), whereas the ethyl acetate fraction was highly active (IC50 0.12 mg/mL) in Sindhri variety. Mangiferin exhibited significant inhibition (IC50 0.026 mg/mL). A moderate inhibition of 15-LOX was observed in all samples, whereas mangiferin was least active. In advanced glycation end product inhibition assay, the chloroform fraction of Langra variety exhibited significant inhibition in nonoxidative (IC50 64.4μg/mL) and oxidative modes (IC50 54.7μg/mL). It was concluded that both Langra and Sindhri peel extracts and fractions possess significant antidiabetic activities. The results suggest the potential use of peel waste in the management and complications of diabetes.

Phytochemical Constituents and Evaluation of Lipoxygenase Activity of Tetracera Macrophylla Twigs Wall.Ex Hook.f.& Thoms

Lipoxygenase is an enzyme that catalyzes the formation of leukotriene, which is involved in various diseases such as asthma, osteoporosis and atherosclerosis. Inhibition of this enzyme can overcome inflammatory diseases. Tetracera macrophylla Wall.ex Hook. f.& Thoms plant has shown various biological activities, especially as an anti-inflammatory. This study aims to analyze phytochemical constituents and evaluate the lipoxygenase activity of Tetracera macrophylla twig extract. The sample was gradually macerated using n-hexane, ethyl acetate, and methanol. Phytochemical analysis of the extract was carried out using the LC-MS/MS method. Lipoxygenase assay was carried out in vitro using RAW 264.7 macrophage cells induced with lipopolysaccharide using spectrofluorometry. As a comparison, zileuton, which has been established as a 5-LOX inhibitor and quercetin, which has the potential as a 5-LOX inhibitor. The results showed that methanol extract of Tetracera macrophylla twigs had lower specific LOX activity than standard zileuton and quercetin. This plant contains phenol and flavonoid compounds, which can be developed as lipoxygenase inhibitors to prevent inflammation.

Unveiling Anticancer Potential of COX-2 and 5-LOX Inhibitors: Cytotoxicity, Radiosensitization Potential and Antimigratory Activity against Colorectal and Pancreatic Carcinoma.

Apart from cytotoxicity, inhibitors of the COX-2 enzyme have demonstrated additional effects important for cancer treatment (such as radiosensitization of tumor cells and cell antimigratory effects); however, the relationship between the inhibition of other inflammation-related enzyme 5-LOX inhibitors and anticancer activity is still not well understood. In our study, the cytotoxicity of thirteen COX-2 and 5-LOX inhibitors previously presented by our group (1-13) was tested on three cancer cell lines (HCT 116, HT-29 and BxPC-3) and one healthy cell line (MRC-5). Compounds 3, 5, 6 and 7 showed moderate cytotoxicity, but good selectivity towards cancer cell lines. IC50 values were in the range of 22.99-51.66 µM (HCT 116 cell line), 8.63-41.20 µM (BxPC-3 cell line) and 24.78-81.60 µM (HT-29 cell line; compound 7 > 100 µM). In comparison to tested, commercially available COX-2 and 5-LOX inhibitors, both cytotoxicity and selectivity were increased. The addition of compounds 6 and 7 to irradiation treatment showed the most significant decrease in cell proliferation of the HT-29 cell line (p < 0.001). The antimigratory potential of the best dual COX-2 and 5-LOX inhibitors (compounds 1, 2, 3 and 5) was tested by a wound-healing assay using the SW620 cell line. Compounds 1 and 3 were singled out as compounds with the most potent effect (relative wound closure was 3.20% (24 h), 5,08% (48 h) for compound 1 and 3.86% (24 h), 7.68% (48 h) for compound 3). Considering all these results, compound 3 stood out as the compound with the most optimal biological activity, with the best dual COX-2 and 5-LOX inhibitory activity, good selectivity towards tested cancer cell lines, significant cell antimigratory potential and a lack of toxic effects at therapeutic doses.

Phytochemical profiling and therapeutic potential of Baliospermum montanum: A promising inhibitor of COX-2/15-LOX enzymes and NO production in LPS-stimulated RAW 264.7 macrophages

IntroductionBaliospermum montanum (BM) is traditionally used to treat various ailments such as inflammation, rheumatoid arthritis, pain, etc. The root part of BM is well documented for its anti-inflammatory activity. However, the present study evaluates the multitargeted anti-inflammatory activity of the less explored BM leaf part. This is the first study investigating the COX-2, 15-LOX dual inhibiting potential and NO production inhibition of BM leaf extracts, fractions and compounds. MethodsThe BM leaf's water, ethanol, and hydroethanolic extracts were evaluated for anti-inflammatory activity by assessing the inhibition of COX-2, 15-LOX, and NO production. Additionally, the antioxidant potential of the extracts was determined through DPPH, ABTS, and FRAP assays. The most active ethanol extract was fractionated into 12 fractions and tested for anti-inflammatory, antioxidant, phenol and flavonoid content. Compounds were isolated from the most active fractions and identified using NMR techniques. The isolated compounds were checked for their anti-inflammatory activity. In addition, HPLC analysis of ethanol extract, fractions, and isolated compounds was carried out. ResultsThe ethanol extract exhibited higher inhibition of COX-2, 15-LOX enzymes and NO production (IC50 35.75 ± 1.95 μg/mL) in LPS-treated RAW 264.7 cells. It also showed higher antioxidant activity. Following the fractionation of the ethanol extract using different solvents, only the ethyl acetate fractions demonstrated inhibition of COX-2, 15-LOX enzymes, and NO production. The ethyl acetate and acetone fractions are excellent sources of antioxidants, phenolic and flavonoid compounds. An effort to find the active principles from these ethyl acetate fractions resulted in the isolation and identification of four potent anti-inflammatory agents, namely, (-)-epiafzelechin, afzelechin-(4α→8)-afzelechin, afzelechin-(4α→8)-epiafzelechin and β-sitosterol-d-glycoside. The first three compounds were reported for the first time from this plant and afzelechin-(4α→8)-afzelechin was found to be a potent inhibitor of the COX-2/15 LOX enzyme as well as NO production (IC50 = 5.9 ± 1.09 µM). ConclusionBased on these findings, we conclude that ethanol extract, fractions, and isolated compounds exert anti-inflammatory activity through multiple therapeutic mechanisms. The present study scientifically supports the traditional use of plant BM for treating inflammation and other related disorders.

UPLC-ESI-TQD-MS/MS Identification and Antioxidant, Anti-Inflammatory, Anti-Diabetic, Anti-Obesity and Anticancer Properties of Polyphenolic Compounds of Hawthorn Seeds

Hawthorn seeds are a by-product of fruit processing and due to the scale of processing of this raw material, they can be an important source of bioactive compounds. This work is the first report on the phenolic composition of hawthorn seeds and their antioxidant, anti-inflammatory, antidiabetic, antiobesity and anticancer activities. In the isolated phenolic fraction of six seed species, 23 phenolic compounds were identified using the UPLC-ESI-TQD-MS/MS method, the key ones of which included the B-type procyanidin dimer. The seeds of the tested species showed high antioxidant activity (mainly by scavenging O2•− and OH• radicals), anti-inflammatory (mainly through LOX inhibition), anti-diabetic, anti-obesity and anti-cancer, with the highest activity against colon cancer cells (Dld-1 line), showing no activity against healthy colon epithelial cells (CCD841CoN). This activity was significantly dependent on the analyzed hawthorn species and, according to PCA analysis, on the content of flavan-3-ols. These discoveries provided the theoretical basis for the possibility of industrial use of hawthorn seeds.Graphical