NMR Studies Research Articles

Chiral Diselenophosphoric Acids for Ion Pair Catalysis: A Novel Approach to Enhance Both Proton Donating and Proton Accepting Properties.

The activation of poorly reactive substrates via strong chiral acids is a central topic in asymmetric ion pair catalysis these days. Despite highly successful scaffolds such as N-triflylphosphoramides, these catalysts either lack C2-symmetry or provide multiple H-bond acceptor sites, leading to lower ee values for certain reactions. We present BINOL-based diselenophosphoric acids (DSA) as an extremely promising alternative. Using an intertwined approach of synthesis and NMR studies, we developed a synthetic approach to DSA with up to 98% NMR yield. The obtained acids provide both very high proton donor and proton acceptor properties, a bifunctionality, which is key to catalytic applications. Indeed, first reactivity test proved the much higher acidity of DSA and its ability to initiate Mukaiyama-Mannich reaction and protodesilylation of silyl ethers. Together with their C2-symmetry, the single donor and single acceptor situation, the decreased tendency of self-association, and the straightforward synthesis with potential 3,3'-substitution, the DSA provide all features ideal for the further development of ion pair catalysis.

Facile Synthesis of Rigid Binuclear Manganese Complexes for Magnetic Resonance Angiography and SLC39A14-Mediated Hepatic Imaging.

Manganese(II)-based contrast agents (MBCAs) are potential candidates for gadolinium-free enhanced magnetic resonance imaging (MRI). In this work, a rigid binuclear MBCA (Mn2-PhDTA2) with a zero-length linker was developed via facile synthetic routes, while the other dimer (Mn2-TPA-PhDTA2) with a longer rigid linker was also synthesized via more complex steps. Although the molecular weight of Mn2-PhDTA2 is lower than that of Mn2-TPA-PhDTA2, their T1 relaxivities are similar, being increased by over 71% compared to the mononuclear Mn-PhDTA. In the presence of serum albumin, the relaxivity of Mn2-PhDTA2 was slightly lower than that of Mn2-TPA-PhDTA2, possibly due to the lower affinity constant. The transmetalation reaction with copper(II) ions confirmed that Mn2-PhDTA2 has an ideal kinetic inertness with a dissociation half-life of approximately 10.4 h under physiological conditions. In the variable-temperature 17O NMR study, both Mn-PhDTA and Mn2-PhDTA2 demonstrated a similar estimated q close to 1, indicating the formation of monohydrated complexes with each manganese(II) ion. In addition, Mn2-PhDTA2 demonstrated a superior contrast enhancement to Mn-PhDTA in in vivo vascular and hepatic MRI and can be rapidly cleared through a dual hepatic and renal excretion pattern. The hepatic uptake mechanism of Mn2-PhDTA2 mediated by SLC39A14 was validated in cellular uptake studies.

Fluorinated tags to study protein conformation and interactions using19F NMR.

The incorporation of fluorine atoms into a biomacromolecule provides a background-free and environmentally sensitive reporter of structure, conformation and interactions using19F NMR. There are several methods to introduce the19F reporter - either by synthetic incorporationviasolid phase peptide synthesis; by suppressing the incorporation or biosynthesis of a natural amino acid and supplementing the growth media with a fluorinated counterpart during protein expression; and by genetic code expansion to add new amino acids to the amino acid alphabet. This review aims to discuss progress in the field of introducing fluorinated handles into biomolecules for NMR studies by post-translational bioconjugation or 'fluorine-tagging'. We will discuss the range of chemical tagging 'warheads' that have been used, explore the applications of fluorine tags, discuss ways to enhance reporter sensitivity and how the signal to noiseratioscan be boosted. Finally, we consider some key challenges of the field and offer some ideas for future directions.

Structural basis for the regulation of plant transcription factor WRKY33 by the VQ protein SIB1

The WRKY transcription factors play essential roles in a variety of plant signaling pathways associated with biotic and abiotic stress response. The transcriptional activity of many WRKY members are regulated by a class of intrinsically disordered VQ proteins. While it is known that VQ proteins interact with the WRKY DNA-binding domains (DBDs), also termed as the WRKY domains, structural information regarding VQ-WRKY interaction is lacking and the regulation mechanism remains unknown. Herein we report a solution NMR study of the interaction between Arabidopsis WRKY33 and its regulatory VQ protein partner SIB1. We uncover a SIB1 minimal sequence neccessary for forming a stable complex with WRKY33 DBD, which comprises not only the consensus “FxxhVQxhTG” VQ motif but also its preceding region. We demonstrate that the βN-strand and the extended βN-β1 loop of WRKY33 DBD form the SIB1 docking site, and build a structural model of the complex based on the NMR paramagnetic relaxation enhancement and mutagenesis data. Based on this model, we further identify a cluster of positively-charged residues in the N-terminal region of SIB1 to be essential for the formation of a SIB1-WRKY33-DNA ternary complex. These results provide a framework for the mechanism of SIB1-enhanced WRKY33 transcriptional activity.

Improving the Stability and Kinetic Inertness of Mn(II) Complexes by Increasing the Bridge Length in Bicyclic CDTA-Like Ligands.

Kinetic inertness of Mn(II)-based MRI contrast agents can be improved by increasing the rigidity of the polydentate ligand that tightly coordinate the metal ion. Taking inspiration from the remarkable increase in kinetic inertness of [Mn(CDTA)]2- compared to [Mn(EDTA)]2- due to the cyclohexyl backbone rigidity, we devised that bicyclic ligands would further improve the kinetic inertness of the Mn(II) complexes. The length of the alkyl bridge on the cyclohexane ring was varied from methylene (BCH-DTA), ethylene (BCO-DTA) to propylene (BCN-DTA) to evaluate the influence of the different trans-diaminotetraacetate ligands on relaxometric, thermodynamic and kinetic properties of the Mn(II) complexes. 1H and 17O NMR relaxometric studies showed a slight increase in relaxivity and a faster water exchange rate in these Mn(II)-complexes with respect to [Mn(CDTA)]2-. Solution studies revealed that the conditional stability (pMn) and dissociation half-life (t1/2) at pH 7.4 follow the order [Mn(BCH-DTA)]2-<[Mn(BCO-DTA)]2-<[Mn(BCN-DTA)]2- highlighting the effect of the bridge length on the overall stability of the Mn(II) complexes. Remarkably, [Mn(BCN-DTA)]2- shows an improved pMn value and a 7-times higher kinetic inertness than [Mn(CDTA)]2-. NMR studies on the Zn(II) analogues confirm the rigidity of the bicyclic complexes with an isomerization process at >313 K for the smaller bridged complex [Zn(BCH-DTA)]2-.

A discussion of the paper “Low-field [formula omitted]H NMR study on geopolymers: The effect of paramagnetic Fe(III)” by Z. Yu, R. de Oliveira-Silva, Y. Pontikes, and D. Sakellariou

The recent paper “Low-field 1H NMR study on geopolymers: The effect of paramagnetic Fe(III)” is discussed. After examining the NMR test method and results, it is deduced that, the relaxation signals of the geopolymers with various Fe(III) contents were inconsistently obtained and interpreted without attention to total intensity. The calibration of surface relaxivity was also questionable. These inappropriate interpretations not only make the experimental results about the effect of Fe(III) loading on spin relaxation unreliable, but also make the discussion about its effect on pore structure of the geopolymers too ambitious, which is further incompatible with current understandings of geopolymerization.

Computational and Spectroscopic Studies on the Formation of Halogen-Bonded Complexes Between Tertiary Amines and CBr4 and Application in the Light-mediated Amino Acid Coupling.

In recent years, halogen-bonded complexes (XBCs), in solution, have played a pivotal role in inducing photochemical organic reactions. In this work, we explore the ability of various tertiary amines to act as XB acceptors in the presence of the XB donor CBr4 by computational and spectroscopic studies. DFT studies clearly showcase the formation of XBCs between the studied tertiary amines and CBr4. Simultaneously, computational and experimental UV-Vis studies display intense red shifts that are consistent with charge transfer observed from tertiary amines to CBr4. A detailed NMR study revealed a clear chemical shift of the carbon carrying the bromine atoms upon mixing the XB acceptor with the donor, suggesting that this spectroscopic technique is indeed an experimental tool to identify the generation of XBCs. An application of the ability of such XBCs to activate a carboxylic acid under UVA irradiation or sunlight is presented for amino acid coupling. Among the various tertiary amines studied, the pair DABCO-CBr4 was found to work well for the photochemical amide bond formation. Direct infusion-HRMS studies allowed us to propose a general mechanism for the photochemical amino acid coupling in the presence of a tertiary amine and CBr4, initiated by the photoactivation of an XBC.

Chiral Pyclen-Based Heptadentate Chelates as Highly Stable MRI Contrast Agents.

In recent years, pyclen-based complexes have attracted a great deal of interest as magnetic resonance imaging (MRI) contrast agents (CAs) and luminescent materials, as well as radiopharmaceuticals. Remarkably, gadopiclenol, a Gd(III) bishydrated complex featuring a pyclen-based heptadentate ligand, received approval as a novel contrast agent for clinical MRI application in 2022. To maximize stability and efficiency, two novel chiral pyclen-based chelators and their complexes were developed in this study. Gd-X-PCTA-2 showed significant enhancements in both thermodynamic and kinetic stabilities compared to those of the achiral parent derivative Gd-PCTA. 1H NMRD profiles reveal that both chiral gadolinium complexes (Gd-X-PCTA-1 and Gd-X-PCTA-2) have a higher relaxivity than Gd-PCTA, while variable-temperature 17O NMR studies show that the two inner-sphere water molecules have distinct residence times τMa and τMb. Furthermore, in vivo imaging demonstrates that Gd-X-PCTA-2 enhances the signal in the heart and kidneys of the mice, and the chiral Gd complexes exhibit the ability to distinguish between tumors and normal tissues in a 4T1 mouse model more efficiently than that of the clinical agent gadobutrol. Biodistribution studies show that Gd-PCTA and Gd-X-PCTA-2 are primarily cleared by a renal pathway, with 24 h residues of Gd-X-PCTA-2 in the liver and kidney being lower than those of Gd-PCTA.

Design of Triazolium-Grafted Peptidomimetic Macrocycles with Facial Amphipathicity to Target Pathogenic Bacteria.

Access to 1,2,3-triazolium-grafted peptoid macrocycles was developed by macrocyclization and multivalent postmodification of linear peptoid oligomers carrying an alternance of benzylic and propargyl groups as side chains. X-ray analysis and NMR studies revealed a conformational preference for constrained hairpin-shaped structures leading to the facial amphipathic character of these macrocycles. A preliminary evaluation showed the antimicrobial activities of these new cationic amphipathic architectures.

Stability and dynamics of extradenticle modulates its function

Extradenticle (EXD) is a partner protein of the HOX transcription factors and plays an important role in the development of Drosophila. It confers increased affinity and specificity of DNA-binding to the HOX proteins. However, the DNA-binding homeodomain of EXD has a significantly weaker affinity to DNA compared to the HOX homeodomains. Here, we show that a glycine residue (G290) in the middle of the EXD DNA-binding helix primarily results in this weaker binding. Glycine destabilizes helices. To probe its role in the stability and function of the protein, G290 was mutated to alanine. The intrinsic stability of the DNA-binding helix increased in the G290A mutant as observed by NMR studies and molecular dynamics (MD) simulation. Also, NMR dynamics and MD simulation show that dynamic motions present in the wild-type protein are quenched in the mutant. This in turn resulted in increased stability of the entire homeodomain (ΔΔGG→A of −2.6 kcal/mol). Increased protein stability resulted in three-fold better DNA-binding affinity of the mutant as compared to the wild-type protein. Molecular mechanics with generalized Born and surface area solvation (MMGBSA) analysis of our MD simulation on DNA-bound models of both wild-type and mutant proteins shows that the contribution to binding is enhanced for most of the interface residues in the mutant compared to the wild-type. Interestingly, the flexible N-terminal arm makes more stable contact with the DNA minor groove in the mutant. We found that the two interaction sites i.e. the DNA-binding helix and the unstructured N-terminal arm influence each other via the bound DNA. These results provide an interesting conundrum: alanine at position 290 enhances both the stability and the DNA-binding affinity of the protein, however, evolution prefers glycine at this position. We have provided several plausible explanations for this apparent conundrum. The function of the EXD as a HOX co-factor requires its ability to discriminate similar DNA sequences, which is most likely comprom

Investigation of the Dyeing Ability of Some Reactive Triazine Azo Dyes Containing Pyrazole Fragment.

The IR, 1H NMR, and DFT studies indicated that the prepared reactive disperse dyes predominately exist as hydrazone tautomers. The electronic absorption spectra in methanol were observed and compared to those computed using B3LYP/6-311G(d,p). The dyeing efficiency of the produced dispersed reactive dyes was examined on polyester, cotton, and polyester/cotton blended fabrics. The degree of exhaustion and the quick properties of the dyed samples in terms of perspiration, washing, scorch and light fastness were assessed. It was found that reactive disperse dyes under investigation have a higher affinity for dyeing polyester textiles than cotton textiles. Moreover, the reflectance and color strength of the synthesized dyes were measured and discussed.

Valorization and kinetic modeling of biosurfactant produced by Kocuria marina DAGII: substitute for chemical surfactant

Microbial biosurfactant has wide applications in industrial sectors owing to their eco-friendly and biocompatible nature giving, an alternative to chemically synthesized surfactants. In our study, Kocuria marina DAG II, an isolated bacterium was observed to produce a biosurfactant. The production of biosurfactant was established by oil-spreading (3.72 ±0.15 cm), drop collapse and CTAB (Cetyltrimethyl ammonium bromide) assay. Biosurfactant activity was re-confirmed by the reduction in surface tension of uninoculated broth from 68.52 mN/m to 47.40±0.57 mN/m.Chemicalcharacterization study of the biosurfactant was done by TLC (thin-layer chromatography) indicating the presence of proteins (Rf= 0.78±0.08), lipids (Rf=0.79±0.09), and polysaccharides (Rf=0.81±0.06).FTIR and NMR (1HNMR and 13CNMR) studies confirmed the presence of different functional groups and revalidating the presence of amino acids, lipids and polysaccharides in the biosurfactant.From the optimization study, the highest production of biosurfactant having a yield of 3.05g/L ± 0.05was obtained attemperature of 31°C and pH 8, when kept for an incubation period of 88 hrs. Addition of rancidand waste frying mustard oil and used engine oil could enhance the production of biosurfactant by Kocuria marinaDAGII hence substantiating the concept of value added biosurfactant production from waste.Kinetic modeling showed good fit between experimental and predicted values using Gompertz model with R2 values of 0.99 and 0.99. The biosurfactant exhibited radical scavenging activity through DPPHand ABTS assays with IC50 values of 1.29 mg/ml and 0.56 mg/ml respectively.The biosurfactant produced by Kocuria marina DAG II could be utilized for various applications, bypassing the need of a synthetic surfactant.

Thermodynamic and spectroscopic investigation of the molecular interactions among vitamin B5, calcium salt, and cyclodextrins in aqueous media

The pharmaceutical industry aims to enhance drug development processes by gaining a deeper understanding of the physiochemical properties of drug molecules. This study provides a comprehensive analysis of the volumetric and compressibility properties of the drug, calcium-D-pantothenate (CDP) in aqueous solutions of cyclodextrins (β-CD/HP-β-CD). By measuring density and speed of sound, we have evaluated various parameters, including apparent molar volume, compressibility, and their corresponding infinite dilution parameters. by measuring density and speed of sound. In addition to this, positive transfer parameters such as ΔtrVs,φo and ΔtrKs,φo of solute provide insights into solute-co-solute and solute–solvent interactions. Also, Hepler’s constant offers valuable information on the structure-breaking characteristics of the studied drug in cyclodextrin medium. The decrease in molar conductance of binary system on addition of cyclodextrin supports our theoretical observation from volumetric and compressibility studies. Positive values of activation energy favours spontaneity and solubilization process. Furthermore, the 1H NMR study demonstrates that the β-CD molecule interacts more strongly with CDP molecules as compared to HP-β-CD.

Inhibition of Human Colorectal Cancer by a Natural Product 7-Acetylhorminone and Interactions with BSA/HSA: Multispectral Analysis and In Silico and In Vitro Studies.

We have semi-synthesized a natural product 7-acetylhorminone from crude extract of Premna obtusifolia (Indian headache tree), which is active against colorectal cancer after probation through computational screening methods as it passed through the set parameters of pharmacokinetics (most important nonblood-brain barrier permeant) and drug likeliness (e.g., Lipinski's, Ghose's, Veber's rule) which most other phytoconstituents failed to pass combined with docking with EGFR protein which is highly upregulated in the colorectal carcinoma cell. The structure of 7-acetylhorminone was confirmed by single crystal X-ray diffraction studies and 1H NMR, 13C NMR, and COSY studies. To validate the theoretical studies, first, in vitro experiments were carried out against human colorectal carcinoma cell lines (HCT116) which revealed the potent cytotoxic efficacy of 7-acetylhorminone and verified preliminary investigation. Second, the drugability of 7-acetylhorminone interaction with serum albumin proteins (HSA and BSA) is evaluated both theoretically and experimentally via steady-state fluorescence spectroscopic studies, circular dichroism, isothermal titration calorimetry, and molecular docking. In summary, this study reveals the applicability of 7-acetylhorminone as a potent drug candidate or as a combinatorial drug against colorectal cancer.

Dehydrated mycelia (Cordyceps militaris, Grifola frondosa, Hericium erinaceus and Laricifomes officinalis) as Novel Foods: A comprehensive NMR study

Several medicinal mushrooms have been approved as European Novel Foods or have the potential to become Novel Foods. However, their chemical characterization is limited, being mainly focused on specific compounds.Here, the dehydrated mycelia (isolated from sporophores collected in Italy) of Grifola frondosa, Hericium erinaceus, recently defined as Novel Foods, and Cordyceps militaris and Laricifomes officinalis, with potential Novel Food use, were analyzed by NMR methodology to define their metabolite and nutritional profile. Amino acids and derivatives, sugars and polyols, organic acids, fatty acids, lipid polar moieties, nitrogen-containing compounds were identified and quantified in hydroalcoholic and organic extracts.C. militaris showed a high amount of glutamate, citrate and mannitol, also involved in mushroom taste, being the only species presenting tri-unsaturated fatty acids. L. officinalis was characterized by the highest amount of TAA and EAA, and the lowest sugars and polyols content. G. frondosa and H. erinaceus were characterized by high amounts of trehalose (13–14 % of dried weight).Each dehydrated mycelia potentially represents an innovative food matrix alone, for culinary, nutritional and nutraceutical needs, or mixed together to obtain new products. Moreover, these mycelia can be of interest as sources of isolated bioactive compounds for supplements with particular nutritional functions.

Differential impact on oral cancer cell viability for three carboxylate-stabilized rhenium(I) tricarbonyl centers supported by 2,2′-bipyridine: One-pot synthesis, a structure, and proton-catalyzed carboxylate ligand substitution

A set of six carboxylate-stabilized rhenium(I) tricarbonyl complexes supported by a 2,2′-bipyridine (bpy) ligand, Re(O2CR)(CO)3(bpy) (R = H, CH3, CHF2, R- or S-CHBrCH(CH3)2, and C5H11), were prepared by acidolysis of the complex Re(OCO2C5H11)(CO)3(bpy) with the appropriate carboxylic acid and characterized by 1H and 13C-{1H} NMR and IR spectroscopy. The crystal structure of the complex, Re[R-O2CCHBrCH(CH3)2](CO)3(bpy), was determined by X-ray crystallography. An alternate one-pot route to the carboxylate-stabilized rhenium(I) complexes Re(O2CR’)(CO)3(bpy) (R’ = CH3 or C6H5) and Re(O2CCH3)(CO)3(1,10-pheanthroline), which starts with Re2(CO)10 and in which an ester solvent serves as the source of the carboxylate ligand, was also developed. Cell viability tests on three carboxylate-stabilized rhenium(I) complexes (R = H, CH3, or CHF2), using the HSC-2 oral cancer cell line, found different levels of cytotoxicity for each complex. NMR studies of the carboxylate ligand substitution reaction found that the reaction is catalyzed by protons. In a chloride-rich NMR solution, substitution of the carboxylate ligand leads to either a chloride-stabilized neutral complex (major product) or to a water-stabilized cation (minor product). Cytotoxicity results correlate positively with the Kb value of the carboxylate ligand. Apparently, the more substitutionally inert the carboxylate-stabilized complex is in a chloride-rich environment (similar to extracellular fluid) the greater the amount of cytotoxic [Re(CO)3(bpy)(H2O)]+ that forms in the cytosol.

Synthesis and characterization of a unique fused spiro-dihydropyran based chemodosimeter for selective visual and fluorescent detection of Cu2+ ion in aqueous medium by “turn-on” mode

A realistically newly designed Cu2+ ion specific fluorescent chemodosimeter has been synthesized from α,α'-(E,E)-bis(benzylidene)cyclohexanone having a fused spiro-dihydropyran moiety characterized by 2D NMR and X-Ray Crystallographic study. It also demonstrates an irreversible, instant direct visualisation by unaided eye chemosensing property in aqueous buffer (pH 7.4) with a colour change from yellow to orange upto 0.3 × 10−6 M.

Novel antifungal compound from Prunella vulgaris: An in-silico approach towards the discovery of potent inhibitor to combat phytopathogenic fungi

Phytopathogenic fungi wreak havoc on crops, causing substantial losses globally. Numerous chemical fungicides are employed to manage the pathogen, yet effectiveness is limited and resistance concerns are rising. The current study aimed to identify the antifungal compound from the spikes of a significant medicinal herb Prunella vulgaris subsp. hispida (Benth.) Hultén as an alternative to combat the fungi. In vitro, antifungal assay revealed that the ethyl acetate extract of P. vulgaris emerged as a potent inhibitor against selected fungal phytopathogens Fusarium oxysporum, Alternaria solani and Penicillium digitatum. Through chromatographic analysis, 32 compounds were identified from the active fraction of ethyl acetate extract. Notably, five compounds were reported for the first time from P. vulgaris. They include dichloroacetic acid, tridec-2-ynyl ester (1), phenol, 3,5-bis(1,1-dimethylethyl)- (2), 2-thiopheneacetic acid, 3-tetradecyl ester (3), 1H-1,2,4-triazol-3-amine, 1-ethyl- (4) and sulfurous acid, octadecyl 2-propyl ester (5). Compound 1 was identified as the most abundant. NMR studies confirmed the presence of compound 1, prompting its selection for in silico antifungal analysis. Various computational models showcased that Compound 1 achieved a high bioactive score and met the criteria for medicinal chemistry friendliness. Additionally, a molecular docking study revealed that compound 1 displayed a good binding affinity with the multiple target fungal enzymes such as scytalone dehydratase and cytochrome P450 which is a prime target for antifungal drugs. Remarkably, this study marks the first detection of compound 1’s effectiveness against phytopathogenic fungi. Therefore, compound 1 holds the potential to pave the way for the development of a potent novel fungicide.

Anion Binding to Ammonium and Guanidinium Hosts: Implications for the Reverse Hofmeister Effects Induced by Lysine and Arginine Residues.

Anions have a profound effect on the properties of soluble proteins. Such Hofmeister effects have implications in biologics stability, protein aggregation, amyloidogenesis, and crystallization. However, the interplay between the important noncovalent interactions (NCIs) responsible for Hofmeister effects is poorly understood. To contribute to improving this state of affairs, we report on the NCIs between anions and ammonium and guanidinium hosts 1 and 2, and the consequences of these. Specifically, we investigate the properties of cavitands designed to mimic two prime residues for anion-protein NCIs─lysines and arginines─and the solubility consequences of complex formation. Thus, we report NMR and ITC affinity studies, X-ray analysis, MD simulations, and anion-induced critical precipitation concentrations. Our findings emphasize the multitude of NCIs that guanidiniums can form and how this repertoire qualitatively surpasses that of ammoniums. Additionally, our studies demonstrate the ease by which anions can dispense with a fraction of their hydration-shell waters, rearrange those that remain, and form direct NCIs with the hosts. This raises many questions concerning how solvent shell plasticity varies as a function of anion, how the energetics of this impact the different NCIs between anions and ammoniums/guanidiniums, and how this affects the aggregation of solutes at high anion concentrations.

The Bifunctional Effects of Lactoferrin (LFcinB11) in Inhibiting Neural Cell Adhesive Molecule (NCAM) Polysialylation and the Release of Neutrophil Extracellular Traps (NETs).

The expression of polysialic acid (polySia) on the neuronal cell adhesion molecule (NCAM) is called NCAM-polysialylation, which is strongly related to the migration and invasion of tumor cells and aggressive clinical status. Thus, it is important to select a proper drug to block tumor cell migration during clinical treatment. In this study, we proposed that lactoferrin (LFcinB11) may be a better candidate for inhibiting NCAM polysialylation when compared with CMP and low-molecular-weight heparin (LMWH), which were determined based on our NMR studies. Furthermore, neutrophil extracellular traps (NETs) represent the most dramatic stage in the cell death process, and the release of NETs is related to the pathogenesis of autoimmune and inflammatory disorders, with proposed involvement in glomerulonephritis, chronic lung disease, sepsis, and vascular disorders. In this study, the molecular mechanisms involved in the inhibition of NET release using LFcinB11 as an inhibitor were also determined. Based on these results, LFcinB11 is proposed as being a bifunctional inhibitor for inhibiting both NCAM polysialylation and the release of NETs.