Lipoxygenases are enzymes found in plants, mammals, and other organisms that catalyse the hydroperoxidation of polyunsaturated fatty acids, such as arachidonic, linoleic, and linolenic acids. They have attracted a lot of attention as molecular targets for industrial and biomedical applications, due to their implication in key biological processes, such as plant development and defence, cell growth, as well as immune response and inflammation. Soybean (Glycine max) lipoxygenase (LOX) is a versatile biocatalyst used in biotechnology, pharmaceutical, and food industries. sLOX1, a soybean LOX isoform, is central in various industrial applications; thus, it is of particular interest to develop an efficient sLOX1 isolation process, control its activity, and leverage its potential as an effective industrial biocatalyst, tailoring it to a specific desired outcome. In this study, sLOX1 was extracted and purified from soybean seeds using an optimized protocol that yielded an enzyme preparation with higher activity compared to the commercially available lipoxygenase. Comprehensive biophysical characterization employing dynamic and electrophoretic light scattering, fluorescence, and Fourier-transform infrared spectroscopies revealed that sLOX1 exhibits remarkable structural and functional stability, particularly in sodium borate buffer (pH 9), where it retains activity and integrity up to at least 55 °C and displays minimal aggregation under thermal, ionic, and temporal stress. In contrast, sLOX1 in sodium phosphate buffer (pH 6.8) remained relatively stable against ionic strength and time but showed thermally induced aggregation above 55 °C, while in sodium acetate buffer (pH 4.6), the enzyme exhibited a pronounced aggregation tendency under all tested conditions. Overall, this study provides physicochemical and stability assessments of sLOX1. The combination of enhanced catalytic activity, high purity, and well-defined stability profile across diverse buffer systems highlights sLOX1 as a promising and adaptable biocatalyst for industrial applications, offering valuable insights into optimizing lipoxygenase-based bioprocesses.

We introduce the compound (E)-2-(2-(3,4-dihydroxystyryl)-4H-chromen-4-ylidene)malononitrile, termed Epirus-Yellow (EY), as the first dual mode optical sensor for the naked-eye colorimetric and near-infrared (NIR) fluorometric selective detection of Sn2+. Designed for point-of-care (POC) cellular imaging and food-safety monitoring, EY exhibits exceptional sensitivity for colorimetric (23.4 × 10-8mol/L) and NIR fluorimetric (25.7 × 10-8mol/L) sensing, with a near-instantaneous response and complete colorimetric transition within 400 s. 1D/2D119Sn,1H, and13C NMR, UV-Vis, FT-IR, thermogravimetric analysis, ICP-MS, and DFT calculations elucidated the underlying mechanism. DFT suggests EY's oxidation, forming a quinone that interacts with Sn(OH)₂ through hydrogen bonding, leading to fluorescence. EY's applicability was demonstrated through paper strip and office scanner-assisted color quantification, with a visual detection limit of 12.0 × 10-7mol/L. EY was successfully applied to the real-time estimation of Sn2+in food and water samples, underscoring its potential for on-site food safety monitoring. Finally, a smartphone-assisted analytical platform and mobile application (Epirus4Tin) were developed, enabling user-friendly, real-time, and digital quantification of Sn²⁺ in agro-food samples.

Gonadotropin-Releasing Hormone receptors (GnRHRs) are overexpressed in several hormone-dependent malignancies, making them attractive molecular targets for selective anticancer drug delivery. Peptide-drug conjugates (PDCs) are a promising therapy for cancer and autoimmune diseases with high specificity and reduced toxicity. In this study, the three-dimensional structures of two previously synthesized mitoxantrone-GnRH conjugates, con3 and con7, were elucidated using high-resolution NMR spectroscopy in combination with molecular dynamics (MD) simulations. Complete 1H and 13C resonance assignments were achieved in DMSO-d6 through two-dimensional NMR experiments. NOESY-derived distance restraints were subsequently used to refine the conformational ensembles obtained from MD simulations performed in water and DMSO. Both conjugates exhibited compact bent conformations with a U-shaped peptide backbone. The mitoxantrone moiety is positioned close to the peptide backbone in water simulations and NMR-refined structures, while it is positioned farther away in DMSO, without affecting the orientation of key residues involved in GnRH receptor binding. Importantly, His2, Trp3, and Arg8 remain solvent-exposed, whereas the disulfide bond is easily accessible to the solvent, consistent with the proposed drug release mechanism by the thioredoxin system. NMR-restrained molecular modeling confirmed the dominant conformational features predicted by the unconstrained theoretical simulations. Overall, these findings provide better structural understanding of the molecular organization of mitoxantrone-GnRH conjugates, highlighting key receptor-recognition residues and supporting both the proposed thioredoxin-mediated drug release mechanism and their previously reported biological properties. These insights may facilitate the rational design and optimization of improved GnRH peptide-drug conjugates for targeted therapy.

Βackround/Objectives: The rising global burden of cardiometabolic disorders and chronic low-grade inflammation underscores the need for therapies capable of modulating multiple interconnected pathways. Methods: In this work, a ligand-based virtual screening campaign centered on a previously reported scaffold (compound 1a) was combined with molecular docking, 200 ns molecular dynamics simulations and ADMET prediction to identify and prioritize small-molecule multitarget candidates against PCSK9, GLP1R, FGFR1, GIPR, NF-κB and NLRP3. Results: Among the screened analogs, D4Z emerged as the most balanced lead, displaying consistently favorable binding profiles, stable interactions within functionally relevant pockets and a drug-like physicochemical and pharmacokinetic profile with high predicted oral absorption. Conclusions: Although these findings remain purely computational, they support D4Z as a prioritized multitarget lead for synthesis and experimental validation and illustrate the potential of rational multitarget design for addressing the cardiometabolic-inflammatory axis.

Objectives: Thiocarbohydrazones represent an important class of Schiff base derivatives with versatile chemical and biological properties. Methods: Herein, we present a combined in silico spectroscopic and molecular docking investigation of N'-benzylidenehydrazinecarbothiohydrazide (1). Results: Conformational docking studies were conducted against cathepsin B, acetylcholinesterase, HER2, protein kinase C, and protein kinase A. The compound displayed favorable binding affinities and key interactions within the catalytic sites of all targets, with the strongest predicted binding observed for acetylcholinesterase. Notably, all conformers exhibited higher affinity for protein kinase C than the reference inhibitor balanol, and hydroxylation led to an approximately 10% enhancement in docking performance. Density functional theory (DFT) calculations were employed to analyze vibrational properties, and IR and Raman spectra were computed to elucidate structural features and conformational behavior. Conclusions: The integrated spectroscopic and docking analyses provide mechanistic insights into ligand-target interactions and support rational drug design. These findings identify thiocarbohydrazone derivatives as promising multi-target candidates for the development of enzyme inhibitors relevant to neurodegenerative, oncological, and inflammatory diseases.

In the original publication [...].

The investigation of quinoxalinones, nitrogen-rich heterocyclic compounds, has attracted considerable attention in scientific research due to their versatile functionalization potential and their important biological properties. The present study reports the synthesis and structural characterization of novel 6,7-dihalogenated quinoxalin-2(1H)-one derivatives, focusing on their complex imine-enamine tautomeric equilibrium. Using a high-yield protocol in glacial acetic acid, dibromo, dichloro, and difluoro derivatives were prepared with yields ranging from 70% to 83%. Detailed 1D and 2D NMR spectroscopy in DMSO-d6, complemented by Density Functional Theory (DFT) calculations, was employed to decipher their structural behavior and electronic transition states. Results reveal a slow exchange on the NMR timescale, allowing the distinct identification of both tautomers. For the dibromo and dichloro analogs, the enaminoester is the predominant species (88%), stabilized by an intramolecular hydrogen bond between the enamine proton and the ester carbonyl group, forming an intramolecularly hydrogen-bonded six-membered pseudo-ring. Conversely, the difluoro derivative exists primarily as the iminoester (64%), a shift attributed to the high electronegativity of fluorine destabilizing the intramolecular hydrogen bond. Calculations, also reveal that for the fluoro compound both enamino and imino tautomer are almost energetically degenerate in agreement with the experimental findings. Furthermore, they suggest the formation of homodimers stabilized by hydrogen bonding between amide groups, i.e., the calculated 1H NMR spectra of the dimers are in good agreement with the experimental ones. Diffusion Ordered Spectroscopy (DOSY) confirmed the existence of tautomerism and dimerism. The dimers were very stable up to 333 K and the hydrogen bonds formed between the dimers were much stronger for the imino versus the enamino tautomer. This is showcased from the chemical shift changes of H-4 from the variable temperature 1H NMR spectra, as well as the hydrogen bond lengths calculated from the DFT studies. Interestingly, transition state calculations indicate that the tautomeric exchange is water-catalyzed; residual moisture in the solvent significantly lowers the activation energy (from ∼74–91 kcal/mol to 34 kcal/mol) compared to a non-catalyzed route. Ab initio MD calculations at 300 Κ show that both enamino and imino tautomers are stable, allowing the distinct identification of both tautomers via NMR. However, in the water catalyzed reaction of tautomerism, the enamino structures the N1-H1 distance can be enlarged from 1.01 Å to 1.40 Å in femtosecond scale assisting the tautomerism. These findings provide essential insights into the structural dynamics of quinoxalinones, facilitating future structure-activity relationship (SAR) studies for developing advanced therapeutics.

The effect of cold atmospheric plasma (CAP) (a pin-to-liquid DBD) (28-32 kV, 1-10 min) on virgin olive oil (VOO) lipid oxidation was kinetically investigated. Quality assessment was performed (bioactive compound concentrations and fatty acid profiles) while the samples were further characterized by Fourier Transform Infrared (FTIR) spectroscopy and proton Nuclear Magnetic Resonance (H-1 NMR). Intense processing (>5 min and voltages > 31 kV) significantly affected the quality of VOO, enhancing the oxidative reactions. CAP treatment led to an eight-fold increase in peroxide values and to a decrease in total antioxidants by up to 80% compared to untreated VOO. Carbonyl compounds (aldehydes, carboxylic acids) and hydroperoxide intermediates were the main oxidation products, while polyunsaturated fatty acids (PUFAs) dropped from 81.17% to 76.51%. The double bonds in the acyl chains were also highly reactive and facilitate the oxidation and subsequent fragmentation of the VOO.

Accurate and reliable quantification of protein-ligand energetics at the screening stage is often complicated by ligand aggregation, hydrophobicity-driven artifacts, and the need for cosolvents. Here, differential scanning calorimetry (DSC) as a quantitative, label-free screening method is evaluated using BCL-2 as a model oncogenic target. Nine inhibitors (i.e., venetoclax, navitoclax; and seven previously prioritized BCL-2 hit inhibitors by our research group) are profiled across solvent systems, including neat DMSO, 10% DMSO, and a ternary matrix (S3: 10% DMSO, 90% sulfobutylether-β-cyclodextrin (SBE-β-CD) in saline). DSC yielded thermal transition temperatures and thermodynamic parameters (ΔH, ΔG) that enabled ranking of binding strength. Solubility challenges are addressed by S3, which improved thermal signal quality. Comparisons with time-resolved fluorescence energy transfer (TR-FRET) analysis, in vitro assays, and MM/GBSA binding free energy results confirmed DSC's accuracy in detecting binding energetics. Collectively, these results position DSC as a robust, material-efficient tool for thermodynamic screening of BCL-2 ligands and other poorly soluble compounds, and as a practical complement to isothermal titration calorimetry when solubility or kinetic limitations prevail.

Renin, a key aspartic protease central to the renin-angiotensin-aldosterone system (RAAS), remains a therapeutic target for hypertension despite the withdrawal of the only approved direct renin inhibitor, Aliskiren, due to unfavorable drug-drug interactions and safety concerns. Here, we report a computational protein design-driven evaluation of (S)-3-((3-(1H-imidazol-1-yl)propyl)amino)-2-(((S)-1-carboxy-2-(cyclooctanecarboxamido)ethyl)amino)-3-oxopropanoic acid (N-CDAH), a novel lipophilic cyclooctanoyl- derivative, as a next-generation renin inhibitor scaffold. This scaffold was designed based on the rationale of leveraging the carnosine like backbone while optimizing lipophilicity and metabolic stability. Pharmacokinetic, ADME, and toxicity predictions (SwissADME, pkCSM) revealed greater predicted aqueous solubility, enhanced metabolic stability, and significantly reduced off-target liabilities compared with Aliskiren (specifically, non-inhibition of major CYP isoforms). Molecular docking (AutoDock Vina binding affinity: -8.08 kcal/mol; Maestro Induced Fit Docking score: -11.149 kcal/mol) and molecular dynamics simulations confirmed favorable binding interactions, conformational adaptability, and complex stability within the renin active site. To contextualize its performance within the broader chemical space, the diastereomeric analog of N-CDAH as well as structurally related compounds identified through SwissSimilarity were also examined using computational workflow. The MD analysis (200 ns) demonstrated that the inhibitor is anchored via a dual stabilization mechanism: hydrophobic enclosure coupled with persistent ionic interactions. These integrative in silico results highlight the potential of this derivative to overcome Aliskiren's pharmacological shortcomings, providing a strong computational rationale for experimental validation and underscoring the role of structure-based drug design in antihypertensive drug discovery.

It is well known that monoamine oxidase (MAO) plays a pivotal role in neurodegeneration and the inhibition of this enzyme can manifest anti-depressant properties as well as have a positive impact in Alzheimer’s and Parkinson’s diseases. Specifically, the MAO enzyme catalyzes the oxidative deamination of a variety of monoamines. This reaction leads to the formation of aldehydes, together with H2O2 and ammonia. Hydrogen peroxide can generate additional reactive oxygen species (ROS), this way leading to neurotoxicity. When MAO is activated, it induces the amyloid-beta (Aβ) deposition via abnormal cleavage of the amyloid precursor protein (APP) and contributes to the generation of neurofibrillary tangles and cognitive impairment due to neuronal loss. MAO has two isoforms: MAO-A and MAO-B. The main hMAO-B inhibitors used for the treatment of Alzheimer’s and Parkinson’s diseases, encompass a terminal triple bond in their structure, which provides their potency. Recently, a new class of inhibitors has emerged, bearing the carbon-carbon triple bond not necessarily at the end of the chain. In this review, the structure and physiological function of the MAO enzymes is discussed, as well as their mechanism of inhibition via terminal propargylamines. Moreover, it is highlighted the current development and discovery of potential hMAO-B inhibitors from propargylamine scaffolds and docking studies are performed to four of them by our group, in order to assess their binding energy with the enzyme. Finally, molecules which do not contain a propargylamine moiety in their structure were studied and compared against a known hMAO-B inhibitor, deprenyl. From the superimposition results of these molecules with deprenyl, as well as the interactions of the molecules with the amino acids of the active site of hMAO-B, it appears that these compounds have several similarities with deprenyl, opening new paths for the creation of novel molecules against Alzheimer’s disease.

The corticotropin-releasing factor (CRF) and its type 1 receptor (CRF1R) play a key role in the regulation of the hypothalamic-pituitary-adrenal (HPA) axis. Dysregulation of the HPA axis is associated with congenital adrenal hyperplasia (CAH) and depression. Non-peptide CRF1R-selective antagonists displayed antidepressant effects on animal models and are used for the management of CAH. To develop novel non-peptide CRF1R antagonists, we have previously designed and synthesized a series of substituted pyrimidines. Among these analogs, molecule 43 (M43) binds to CRF1R with the highest affinity. Based on this finding, we selected M43 for further pharmacological characterization in the present study. The results suggest that M43 is a potent CRF1R antagonist, blocking the ability of the CRF-related agonist, Tyr0-sauvagine, to stimulate (1) cAMP accumulation in HEK 293 cells expressing CRF1R and (2) the proliferation rate of RAW 264.7 macrophages. Computational studies suggest that the antagonist properties of M43 are mostly attributed to its ability to interact with residues in the allosteric pocket of CRF1R, comprised of the third, fifth, and sixth transmembrane domain residues, which block activation-associated structural rearrangements of the receptor. Our data will be used to design novel non-peptide CRF1R antagonists for clinical use.

A frequent challenge that graduate and undergraduate students in chemistry, biology, and pharmacy laboratories face is accurately assigning proton and carbon peaks in the 1D and 2D Nuclear Magnetic Resonance (NMR) spectra of organic and pharmaceutical molecules. We propose a consistent, step-by-step approach to effectively assist students in simultaneously interpreting a variety of 1D and 2D NMR spectra. According to this approach, the anticipated NMR peaks are initially predicted based on the compound's molecular structure, followed by the interpretation of the actual spectra. The novelty of this teaching approach lies in the simultaneous use of experimental data from 1D and 2D homonuclear and heteronuclear spectra to validate or enhance the theoretical approach derived solely from the molecule's two-dimensional structure. The integrations obtained from 1D 1H NMR spectrum and bond and spatial correlations from homonuclear and heteronuclear 2D NMR spectra are the experimental data used for every structure elucidation. A special notation using arrows is adduced in order to indicate the bond and spatial correlations and to schematically present the proposed teaching approach in an easy and comprehensive way. The proposed notation could offer a new way to visualize and conceptualize the theoretical approach, potentially making it easier for students to apply. This methodology begins with predicting proton, carbon, and proton-carbon correlations based solely on the molecular structure, followed by listing the expected signals for all available spectra. The approach then emphasizes comparing these predicted correlations and signals with those observed in 1D and 2D NMR spectra. This methodology is only applicable when the synthesis of a particular product is anticipated, which is a typical scenario for students engaged in organic synthesis laboratories or conducting research in organic and medicinal chemistry.

Thiosemicarbazones and thiocarbohydrazones are key sulfur-containing organic compounds known for their diverse biological, pharmaceutical, and industrial applications. Beyond their well-established therapeutic potential, their strong chelating ability allows them to form stable complexes with transition metals, enabling uses in catalysis, corrosion inhibition, and dyeing processes. Their structural characteristics and dynamic conformations critically influence both biological activity and industrial performance, making nuclear magnetic resonance (NMR) spectroscopy an indispensable tool for their analysis. This review provides a comprehensive overview of the conformational and functional properties of bioactive thiosemicarbazones and thiocarbohydrazones, with a focus on how experimental NMR techniques are used to investigate their conformational behavior. In addition to experimental findings, available computational data are discussed, offering complementary insights into their structural dynamics. The integration of experimental and theoretical approaches offers a robust framework for predicting the behavior and interactions of these compounds, thereby informing the rational design of novel derivatives with improved functionality. By highlighting key structural features and application contexts, this work addresses a critical gap in the current understanding of these promising agents across both biomedical and industrial domains.

In this article, we report the design, synthesis, and biological evaluation of a new series of nitroheterocyclic aromatic adamantane amides targeting trypanosomes. These compounds feature diverse substituents on the adamantane scaffold, variations in side chain linker length, and a range of nitroheterocyclic moieties. This work represents a continuation of our previous efforts, with a particular focus on elucidating the structural and functional role of the linker connecting the phenyladamantane core to the nitroheterocyclic ring. The structure-activity relationship data underscore the importance of strategic modifications in enhancing the pharmacological profile of these compounds against trypanosome parasites. Further modifications are recommended to optimize the physicochemical properties of the current derivatives to improve intracellular targeting of trypanosomatids, an important clinical stage in their life cycle.

The renin–angiotensin–aldosterone system (RAAS) is essential for controlling blood pressure and maintaining fluid balance, driving significant structural changes throughout the cardiovascular system, including the heart and blood vessels. As a result, the RAAS is a key therapeutic target for various chronic cardiovascular diseases, ranging from arterial hypertension (AH) to heart failure (HF). In this review, one of our objectives is to describe the new evidence over the last 4 years regarding the RAAS. Moreover, we pay attention to the structure and function of the angiotensin II type 1 receptor (AT1R) and its role in hypertension, as well as define its active site. Later, we discuss the most potent, selective inhibitors of AT1 receptors, based on in vitro and in vivo experiments, from 2020 to 2024. Large peptide molecules, small non-peptide-like molecules, and sartan derivatives are analyzed. The low IC50 values of the entities that do not resemble sartans showcase the vast chemical space that can be explored for the creation of more potent antihypertensive medications. We have also employed computational chemistry tools in order to identify key molecular interactions between the compounds of the literature studied in order to elucidate the underlying reasons why these different molecules exhibit variations in their binding energies and overall potency.

Carob syrup presents significant commercial potential, providing unique nutritional along with significant pharmacological activity. The objective of this study was to comprehensively characterize the carob syrup to ascertain its potential health benefits and to estimate the levels of important compounds present in commercially available carob syrup, with a view to ensuring the quality and consistency of the manufacturing process. Commercial carob syrup samples from Cyprus were purchased and analyzed employing Ultra-High Performance Liquid Chromatography-Quadrupole Time-of-Flight Electrospray Ionization Mass Spectrometry (UHPLC-QTOF & IEcy;SI-MS) and Nuclear Magnetic Resonance (NMR) spectroscopy. LC-MS based suspect screening resulted in the identification of 39 metabolites, i.e. amino acids, fatty acyls/fatty acids and conjugates, sugars, flavonols and flavonoids, organic acids, nucleotides and their derivatives, and alkaloids. Moreover, the quantification of 16 compounds was achieved through the utilization of analytical standards. Additionally, NMR revealed the distinctiveness of aromatic and aliphatic regions, and the abundance of sugars (pinitol, sucrose, glucose, and fructose as majors and mannose and lactose as minors). The results demonstrate that the analyzed syrup shows a consistent chemical profile, which justifies its utilization as nutraceutical. The combination of UHPLC-QTOF-ESI/ MS and NMR analyses contributed to the identification of pinitol in carob, which has significant antidiabetic activity.

Donepezil (DH), a selective acetylcholinesterase inhibitor, is widely used to manage symptoms of mild to moderate Alzheimer’s disease by enhancing cholinergic neurotransmission and preventing acetylcholine breakdown. Despite the effectiveness of oral formulations, extensive hepatic metabolism and low systemic bioavailability have driven the search for alternative delivery systems. This study focuses on nasal delivery as a non-parenteral substitute, utilizing hydroxypropyl methylcellulose (HPMC) for its mucoadhesive properties and methyl-β-cyclodextrin (Me-β-CD) for its ability to enhance permeability and form inclusion complexes with drugs. Prior studies demonstrated the potential of HPMC-based nasal films for nose-to-brain delivery of donepezil and highlighted Me-β-CD’s role in improving drug solubility. Building on this, transparent gel formulations containing DH, HPMC, and 2,6 Me-β-CD were developed to investigate molecular interactions within two- and three-component systems. This study utilized a combination of nuclear magnetic resonance (NMR) spectroscopy and density functional theory (DFT) to provide detailed insights into the interactions between DH, 2,6-Me-β-CD, and HPMC. The findings provide critical insights into drug–excipient interactions, aiding the optimization of stability, solubility, and controlled release. This advances the rational design of nanotechnology-based drug delivery systems for enhanced therapeutic efficacy.

Myasthenia Gravis, a chronic autoimmune disease, is primarily treated with acetylcholinesterase inhibitors. However, these drugs are not specific, and their mechanism of action against the disease has not been elucidated. They have a propensity to act on different targets, and their therapeutic action is symptomatic. For this reason, we have studied the interactions of commercially available drugs against Myasthenia Gravis to various enzyme targets to examine if there is any selectivity in their action and possibly to reveal any potential use for other diseases. In particular, the Computational Chemistry programs, AutoDock and Maestro, were used to assess the binding of azathioprine, prednisone, and pyridostigmine to different classes of enzymes, such as: cyclooxygenases (COX-1, COX-2), monoamine oxidases (MAO-A, MAO-B), angiotensin receptors (AT1, AT2), and lipoxygenases (LOX-1, 5-LOX). Molecular Dynamics simulations were employed to further analyze the stability and interactions of the most effective compounds. Using in silico platforms it was found that these drugs are not toxic, they do not produce unwanted adverse eaffects, and that pyridostigmine seems to be the best compound according to the ADME results. Additionally, Saturation Transfer Difference NMR experiments were carried out and confirmed the binding of azathioprine to LOX-5 at both the molecular and atomic levels. The in vitro evaluation of azathioprine and prednisone also revealed important inhibition of human 15-LOX-1 over general lipoxygenase activity. Finally, it was found that these drugs have potential for use in various biological and pharmacological applications such as CNS drugs.

Background: In this study, two chalcone analogs were synthesized through in silico and experimental methods, and their potential to inhibit the lipoxygenase enzyme, which plays a role in the inflammation pathway, was assessed. Specifically, this study is a continuation of previous research in which chalcone derivatives were synthesized and characterized. Objectives/Methods: In the current work, we present the re-synthesis of two chalcones, with a focus on their docking studies, NMR analysis, and dynamic simulations. The structure of each chalcone was elucidated through a combination of Nuclear Magnetic Resonance (NMR) and Density Functional Theory (DFT). The substituent effect on the absorption spectrum of the two chalcone derivatives was studied. Results: A “LOX–chalcone” complex, predicted by docking studies, was further examined using molecular dynamics (MD) simulations to evaluate the stability of the complex. After fully characterizing the “LOX–chalcone” complexes in silico, the atomic details of each chalcone’s interaction with LOX-1 and 5-LOX were revealed through Saturation Transfer Difference (STD) NMR (Nuclear Magnetic Resonance). Finally, their selectivity profile was investigated against human 15-LOX-1 and general Lipoxidase activity. Conclusions: The in silico methods suggest that chalcones could be promising lead compounds for drug designs targeting the LOX enzyme.