Complex Formation Research Articles

Optimized Second-Generation IL-4Rα Inhibition: Structural and Molecular Dynamics Properties of Rademikibart Fab-IL-4Rα Complex

Introduction: Global Burden of Disease studies showed atopic dermatitis (AD) and asthma are the top two immune-mediated inflammatory diseases at younger age [1]. Dupilumab was the initial first-generation interleukin-4 receptor alpha (IL-4Rα) inhibitor for treating both type I and type II IL-4Rα-dependent inflammatory disorders. Following recent clinical trials, rademikibart (previously, CBP-201) emerges as an optimized next-generation human monoclonal antibody with higher binding affinity to IL-4Rα compared to dupilumab [2]. It demonstrated better effect in inhibiting STAT6 intracellular signaling in vitro and provided similar potency inhibiting both IL-4 induced TARC release and IL-4 induced B cell activation [2]. Materials & Methods: X-ray crystallography was used to determine the atomic resolution 3D structure of rademikibart fragment antigen binding (Fab) bound to IL-4Rα. This structure was analyzed and compared computationally with the 2.82 Å resolution crystal structure of dupilumab Fab bound to IL-4Rα (Protein Data Bank Code 6WGL). Molecular dynamics studies on rademikibart and dupilumab bound to IL-4Rα examined the stability of the complexes and effects of amino acid mutations on complex formation. Results: The x-ray crystal structure of rademikibart Fab bound to IL-4Rα was determined at 2.71Å and compared to the complex of dupilumab Fab and IL-4Rα. The rotation angle between dupilumab and rademikibart bound to IL- 4Rα is 59.17°. This rotation enables the epitope of rademikibart, but not dupilumab, on IL-4Rα to overlap more closely with the conserved binding interface utilized by IL-4 and IL-13 cytokines. Molecular dynamics simulations of rademikibart Fab and dupilumab Fab complexed with IL-4Rα showed the third interface loop (residues 148 to 152 in domain 2) of IL-4Rα interacts directly with rademikibart, which is absent in dupilumab/IL- 4Rα complex. This finding is confirmed by analysis of the hydrogen bond interactions at the interface between the antibodies and IL-4Rα, demonstrating superior binding energy for rademikibart. Through single amino acid mutation analysis on rademikibart, we identified residue Y50 on rademikibart as the key residue interacting with IL- 4Rα’s third interface loop. Conclusion: Our data provide a molecular and structural rationale for the enhanced IL-4Rα inhibition by rademikibart over dupilumab, confirming rademikibart as an optimized second-generation IL-4Rα inhibitor.

Institutional Complexity and Social Innovation: The Case of Chinese Social Enterprises

Abstract Social enterprises (SEs) have emerged throughout the world to address societal challenges through market-based activities and innovation. Research has focused on how SEs manage the tensions arising from the combination of social-welfare and market logics, neglecting the institutional complexity that arises in authoritarian regimes where the state plays a dominant role. Based on the institutional logics perspective and interview data of 42 SE leaders in China, we find that key tensions arise from the state and social-welfare logics and that state logic (re)configures the social–market relationships to be compatible. Chinese SEs employed depoliticization and localization to adapt to this unique form of institutional complexity in their social innovation efforts. This study advances research on SEs, institutional complexity, and hybrid organizing.

Concept of architectural formation of construction production complexes in the cities of Donbass

The scientific research is a continuation of the author’s publications and for the first time provides a complete conceptual set of principles and techniques, systematically covering all levels of the architectural and planning organization of construction production complexes. A concept for the architectural formation of construction industrial complexes in the cities of Donbass is proposed, which includes recommendations for the formation of these complexes. The urban planning approach to the selection of territories of non-operating coal mines is generalized. An analysis of existing buildings and structures of the coal mine and the features of construction production complexes was also carried out.

Uranium-Mediated Thiourea/Urea Conversion on Chelating Ligands

2,6-Dipicolinoylbis(N,N-dialkylthioureas) and H2LR2 react with uranyl salts and a supporting base (e.g., NEt3) under formation of monomeric or oligomeric complexes of the compositions [UO2(LR2)(solv)] (solv = donor solvents) or [{UO2(LR2)(µ2-OMe)}2]2–. In such complexes, the uranyl ions are commonly coordinated by the “hard” O,N,O or N,N,N donor atom sets of the central ligand unit and the lateral sulfur donor atoms remain uncoordinated. Their individual structures, however, depend on the reaction conditions, particularly on the equivalents of NEt3 used. An unprecedented, selective hydrolysis of the uranium-coordinating bis(thioureato) ligands results in an S/O donor atom exchange at exclusively one thiourea side-arm, when an excess of NEt3 is used. The resulting trimeric uranyl complexes are isolated in fair yields and have a composition of [(UO2)3(L2Et2)2(µ2–OR)(µ3-O)]–. H2L2Et2 represents the newly formed 2,6-dipicolinoyl(N,N-diethylthiourea)(N,N-diethylurea) and R = H, Me, or Et. {L2Et2}2– binds to the uranyl units via the pyridine ring, the dialkylurea arm, and the central carbonyl groups, while the thiourea unit remains uncoordinated. The central cores of the products consist of oxido-centered triangular {(UO2)3O}4+ units. The observed reactivity is metal-driven and corresponds mechanistically most probably to a classical metal-catalyzed hydrodesulfurization. The hydrolytic thiourea/urea conversion is only observed in the presence of uranyl ions. The products were isolated in crystalline form and studied spectroscopically and by X-ray diffraction. The experimental findings are accompanied by DFT calculations, which help to understand the energetic implications in such systems.

A New Geological Map of the Marginal Basins of Eastern Papua New Guinea: Implications for Crustal Accretion and Mineral Endowment at Arc–Continent Collisions

Abstract Accretion of island arc terranes is a fundamental process of crustal growth and the formation of new continents. Convergent margin tectonics, both compressional and extensional, in accretionary orogens also control the origin and distribution of their contained mineral resources, including many of the world’s important Cu and Au deposits. However, the details of crustal growth and accretion are often lost because of deformation and selective preservation during subduction. The Melanesian Borderland, which includes the offshore regions of eastern Papua New Guinea and the Solomon Islands, contains several active and relict arc and backarc systems that have formed in response to more than 50 Ma of subduction and complex plate tectonic adjustments. The composite terrane is a region of some of the fastest growing crust on Earth and also spectacular mineral endowment, including three of the top ten porphyry Cu and epithermal Au deposits in the world. However, more than 80% of the belt is submerged, and so little is known about its geological evolution and makeup. Here, we present the first detailed geological map of the region in one map sheet, including the marginal deep ocean basins. The map identifies and groups the key lithostratigraphic formations and correlates associated tectonic events across the belt. The final compilation is presented at 1:1,000,000 scale, which is sufficient to allow quantitative analysis of crustal growth and accretion during ocean–continent collision throughout the region. The map shows the diversity of assemblages in accreting terranes that may eventually become part of a growing continent and highlights their complex formation and structural relationships. Because so much of that history has occurred offshore, the new map presents the first complete picture of the geology of the region in the critical period leading up to its eventual incorporation in the Australian continent.

Evaluation of structurally related acyclic ligands OBETA, EHDTA, and EGTA for stable Mn2+ complex formation.

In recent years, significant research efforts have been dedicated to finding efficient and safe alternatives to the currently used gadolinium (Gd)-based MRI contrast agents. Among the most explored alternatives are paramagnetic chelates of the Earth-abundant Mn2+, which form a prominent class of metal complexes. The design of Mn2+ complexes with enhanced relaxation properties and improved safety profiles hinges on a delicate balance between thermodynamic and kinetic stability, as well as the presence of coordinated water molecules. In this study, we present a comprehensive investigation into the coordination chemistry of three structurally related polyetheraminocarboxylic chelating agents. Our aim is to elucidate the structural features, paramagnetic properties, and thermodynamic and kinetic inertness of the corresponding Mn2+ complexes. The most significant finding is the considerable difference in the dissociation rates of the complexes, with the octadentate EGTA complex being the most labile. The observed dissociation rates correlate well with the nitrogen inversion dynamics, as assessed through NMR spectral analysis of the analogous Zn2+ complexes.

Control of Starch Molecular Weight by Enzyme Treatment Facilitates the Formation of V-Type Starch-Resveratrol Complexes in a High-Pressure Homogenization Environment and Their Modulation Effects on the Gut Microbiota.

As the concept of precision nutrition has been gradually popularized in recent years, the relationship between the structure of starch-polyphenol complexes with significant health effects and their nutritional functions has been progressively investigated. In this study, G50 high-amylose maize starch with different molecular weights was first prepared by pullulanase and α-amylase, and their effects on the structural formation, digestion properties, and release behaviors of the starch-resveratrol (RA) complex were discussed. The results confirmed that enzyme-treated starch could enhance intermolecular hydrogen bonding and hydrophobic interactions between starch and RA in a high-pressure homogeneous (HPH) environment, forming stable single-helix and V-type crystalline structures while reducing the B-type crystalline structures. Meanwhile, the in vitro experiment showed that when the RA addition was 3%, the resistant starch content of the starch-RA complex could reach 60.3%, and its RA colonic transport rate could reach more than 97%. Interestingly, the starch-RA complex with a relatively higher V-type crystalline structure content contributed to the production of short-chain fatty acids (SCFAs), especially butyrate, and it might be effective in carbohydrate metabolism and immunometabolism by promoting the functions of Phascolarctobacteriu and Alistipes. These findings provide new ideas for the design of the nutritional functions of RS.

Cation-lone pair interaction in Alkali/alkaline earth metal ion-heavier borazine analogue complexes.

The present study is the first report on the formation of alkali/alkaline earth metal ion-heavier borazine analogue complexes via cation-lone pair interaction. Density functional calculations are performed in scrutinizing the complex formation between alkali (Li+, Na+, K+)/alkaline earth (Be2+, Mg2+, Ca2+) metal ions and heavier borazine analogues (HBA) viz. B3P3H6, Al3N3H6, Al3P3H6, Al3As3H6, and Ga3P3-H6. The complexes are found to be stable in gas phase with stabilization energies within the range 26.40 to 324.74 kcalmol-1. The stability can be attributed to the polarizing power of the involved metal ions. Presence of solvent phase exerted notable impact on the stability of the complexes; stability is reduced significantly with the increase in solvent polarity. The process of complexation is exothermic and spontaneous. QTAIM analysis indicated the presence of both ionic and covalent interaction between HBAs and metal ions. HOMO energy, Wiberg bond index, NCI-isosurface and RDG plot analysis revealed the major role of cation-lone pair interaction in the complexation process.

Platelet activation through CD62P and the formation of platelet-monocyte complexes are associated with the exacerbation of mucosal inflammation in patients with ulcerative colitis.

Ulcerative colitis (UC) is a refractory, chronic inflammatory bowel disease of unknown etiology. Although platelets are activated in UC, their relevance in pathophysiology remains unclear. We analyzed the correlation of platelet activation and platelet-monocyte complexes (PMCs) with severity of mucosal inflammation using the Mayo endoscopic subscore (MES). Platelet activation marker, CD62P was upregulated in patients with UC compared with that in healthy controls (P < 0.05). CD62P expression was significantly higher in patients with MES3 (severe inflammation) than in those with MES ≤ 2 (endoscopic remission to moderate inflammation) (P < 0.001). The concentration of sCD62P in patients with MES0 (endoscopic remission) was significantly higher than in those with MES ≥ 1 (P < 0.01). The expression of CD40L, CD63, PAC-1, annexin V, and CD36, and the concentrations of sCD40L, PF4, and RANTES did not correlate with MES. The proportion of PMCs in patients with MES3 was higher than in those with MES ≤ 2 (P < 0.05). CD16 expression on monocytes with platelets was significantly higher than in monocytes without platelets (P < 0.001). Patients with complete remission after treatment showed significant reduction in PMCs 3 months after treatment (P < 0.05) but had no change in CD62P and sCD62P. Our data suggest that platelet activation via the CD62P-PMC axis is involved in UC pathophysiology.

Noncanonical Amino Acid Incorporation in Animals and Animal Cells.

Noncanonical amino acids (ncAAs) are synthetic building blocks that, when incorporated into proteins, confer novel functions and enable precise control over biological processes. These small yet powerful tools offer unprecedented opportunities to investigate and manipulate various complex life forms. In particular, ncAA incorporation technology has garnered significant attention in the study of animals and their constituent cells, which serve as invaluable model organisms for gaining insights into human physiology, genetics, and diseases. This review will provide a comprehensive discussion on the applications of ncAA incorporation technology in animals and animal cells, covering past achievements, current developments, and future perspectives.

Enhancing Cannabinoid Bioavailability in Pain Management: The Role of Cyclodextrins

Chronic pain (CP), including pain related to cancer, affects approximately 2 billion people worldwide, significantly diminishing quality of life and imposing socio-economic burdens. Current treatments often provide limited relief and may cause adverse effects, demanding more effective alternatives. Natural compounds from Cannabis sativa L., particularly cannabinoids like THC and CBD, exhibit analgesic and anti-inflammatory properties, but their therapeutic use is restricted by poor solubility and low bioavailability. Cyclodextrins (CDs) and cyclic oligosaccharides may encapsulate hydrophobic drugs in order to enhance their solubility and stability, offering a promising solution to these challenges. This study explores the formation of CD inclusion complexes with cannabinoids and specific terpenes, such as D-limonene (LIM), beta-caryophyllene (BCP), and gamma-terpinene (γ-TPN), aiming to improve pharmacokinetic profiles and therapeutic efficacy. We discuss analytical techniques for characterizing these complexes and their mechanisms of action, highlighting the potential of CDs to optimize drug formulations. The integration of CDs in cannabinoid therapies may enhance patient compliance and treatment outcomes in CP management. Future research should focus on innovative formulations and delivery systems to maximize the clinical applications of those compounds.

Improving Subnational Opinion Estimation from Cluster-Sampled Polls

Abstract The development of multilevel regression and poststratification (MRP) has allowed scholars to more accurately estimate subnational public opinion using national polls. However, MRP generally recovers less accurate estimates from polls whose respondents are selected using cluster sampling – also called area-probability sampling. This is in part because cluster-sampled polls rely on a complex form of random sampling focused on national representativeness that may result in small or unrepresentative subsamples in subnational geographies. This has limited MRP’s usefulness in subnational opinion estimation in several contexts, including historical polls in the US, where cluster-sampling was common into the 1980s, and large academic studies in many countries today. In this paper, I propose two approaches to improve estimation from MRP with cluster-sampled polls. The first is pooling data from multiple surveys to produce a larger sample of clusters. The second is clustered MRP (CMRP), which extends MRP by modeling opinion using the geographic information included in a survey’s cluster-sampling procedure. Using simulations, I show that both methods improve upon traditional MRP, and I validate them using historical polls in the US

Analysis of Macromolecular Size Distributions in Concentrated Solutions

AbstractThe solution state of macromolecules in concentrated solutions impacts fields ranging from cell biology, to colloid chemistry and engineering of protein pharmaceuticals. Dependent on the interplay between repulsive and weakly attractive forces, proteins may exhibit oligomerization, aggregation, crystallization, liquid‐liquid phase separation, or the formation of multiprotein complexes. The particle size‐distribution is a key characteristic, but difficult to determine when interparticle distances are on the order of their size and macromolecular motion is coupled through hydrodynamic interactions. Here we extend sedimentation velocity analytical ultracentrifugation to measure macromolecular size distributions under these conditions: We apply results from statistical fluid mechanics for the concentration‐dependence of hindered settling and diffusion, embedded in a mean‐field approximation that can resolve coupled sedimentation and diffusion processes of different sized species given experimental sedimentation data. This is combined with a description of transient optical aberrations from lensing in the refractive index gradients associated with sedimentation boundaries (Wiener skewing). We demonstrate this approach in the application to protein solutions with macromolecular volume fractions up to ≈10 %, for example, resolving monomers and dimers of albumin at 140 mg/ml. This enables size‐distribution analysis of proteins at concentrations of therapeutic antibody formations and close to physiological concentration in serum and cells.

DOENÇA RENAL CRÔNICA CONCOMITANTE À ERLIQUIOSE: RELATO DE CASO

Kidney disease can range from acute kidney injury to chronic kidney disease, often caused by inflammation that impairs filtration. When kidney function is compromised, toxins accumulate, which can lead to kidney failure. In advanced cases, dialysis treatment becomes necessary. Other associated pathologies, such as ehrlichiosis, can make the condition irreversible. Ehrlichiosis triggers an inflammatory response affecting the kidneys, with the formation of immune complexes during infection. These complexes deposit in the renal capillaries, causing glomerulonephritis, an inflammation that impairs the function of the glomeruli, responsible for blood filtration. The purpose of this report is to present a case where kidney disease developed concomitantly with ehrlichiosis. In this case, an animal was referred to a veterinary hospital for dialysis treatment due to kidney disease. Laboratory tests were conducted, including a complete blood count, biochemical analysis, rapid ehrlichiosis test, and PCR, which confirmed the presence of active ehrlichiosis infection. Additionally, hemodialysis and dialysis sessions were performed, among other treatments. This case highlights the complexity and challenges of treating chronic kidney disease aggravated by ehrlichiosis, emphasizing the importance of advanced therapies and the need for ethical planning to balance available resources and the animal's quality of life. Raising awareness among pet owners about the risks, the importance of early diagnosis, correct treatment, and continuous monitoring are fundamental measures that can help in the early detection of kidney impairment and increase the chances of successful treatment.

The Effect of Kappa Carrageenan Composition on the Characteristics of Chitosan-Kappa Carrageenan Hydrogel Film

Chitosan-Kappa Carrageenan (Chit-KC) hydrogel films have been made using a casting method with different compositions. This research aimed to investigate the effects of different compositions on the chemical, physical, and mechanical characteristics of the Chit-KC hydrogel films. The compositions of the kappa carrageenan in the Chit-KC films were 0%, 16.67%, 33.33%, 50.0%, and 100.0%. The films were characterized using FTIR, SEM, DTA-TGA, and XRD instruments to analyze their functional groups, morphology, thermal properties, and crystallinity. They were also tested for their solubility, swelling, water vapor permeability (WVP), and mechanical characteristics. The results revealed that the addition of kappa-carrageenan induced the formation of a polyelectrolyte complex. The polyelectrolyte complex caused an increase in swelling, crystallinity, and WVP values and a decrease in solubility, tensile strength, elongation, and elasticity. It also caused the formation of film layers and granules on the surface of the hydrogel film. The IR spectra exhibited that the Chit-KC hydrogel films contained O-H, C-H, C=O, S=O, C-O, and C-O-C groups. The DTA-TGA test results revealed that the film began decomposing at around 200°C and re-decomposing at 300°C.

Single-cell Trajectories and Pseudo-time Analysis Reveal Additional Insights into Rheumatoid Arthritis Disease Progression

As the chicken embryo grows, its increasing metabolic demand drives the development of the cardiovascular system, enabling efficient nutrient delivery through complex molecular and structural formation. Key extracellular matrix (ECM) components, such as hyaluronic acid (HA) and collagen type I alpha 1 (COL1A1), play a crucial role in shaping the biomechanical environment of the heart, particularly the sinoatrial node (SAN). These ECM elements regulate cell adhesion, migration, and the maturation of cardiac progenitor cells (CPCs), essential for the heart's electrical conductivity and rhythmicity. Based on the hypothesis that ECM mechanical properties influence the spatial patterning of conduction-related genes, we analyzed spatial transcriptomics data from chicken hearts at different developmental stages. Collagen-related genes associated with a sturdy ECM showed an increasing trend over time, while soft ECM-related genes responsible for producing proteoglycans and breaking down collagen decreased. Although conduction-related genes did not fully align with ECM trends, intriguing patterns were observed, pointing to potential interactions between ECM remodeling and the construction of the cardiac conduction system. Our findings suggest significant ECM r7emodeling between days 4 and 7, which may influence the heart's structural and functional development.

Structures of complete extracellular assemblies of type I and type II Oncostatin M receptor complexes.

Oncostatin M (OSM) is a unique Interleukin 6 (IL-6) family cytokine that plays pivotal roles in numerous biological events by signaling via two types of receptor complexes. While type I OSM receptor complex is formed by glycoprotein 130 (gp130) heterodimerization with Leukemia Inhibitory Factor receptor (LIFR), type II OSM receptor complex is composed of gp130 and OSM receptor (OSMR). OSM is an important contributor to multiple inflammatory diseases and cancers while OSM inhibition has been shown to be effective at reducing symptoms, making OSM an attractive therapeutic target. Using cryogenic electron microscopy (cryo-EM), we characterize full extracellular assemblies of human type I OSM receptor complex and mouse type II OSM receptor complex. The juxtamembrane domains of both complexes are situated in close proximity due to acute bends of the receptors. The rigid N-terminal extension of OSM contributes to gp130 binding and OSM signaling. Neither glycosylation nor pro-domain cleavage of OSM affects its activity. Mutagenesis identifies multiple OSM and OSMR residues crucial for complex formation and signaling. Our data reveal the structural basis for the assemblies of both type I and type II OSM receptor complexes and provide insights for modulation of OSM signaling in therapeutics.

The mutual interaction of TRPC5 channel with polycystin proteins.

PKD1 regulates a number of cellular processes through the formation of complexes with the PKD2 ion channel or transient receptor potential classical (TRPC) 4 in the endothelial cells. Although Ca2+ modulation by polycystins has been reported between PKD1 and TRPC4 channel or TRPC1 and PKD2, the function with TRPC subfamily regulated by PKD2 has remained elusive. We confirmed TRPC4 or TRPC5 channel activation via PKD1 by modulating G-protein signaling without change in TRPC4/C5 translocation. The activation of TRPC4/C5 channels by intracellular 0.2 mM GTPγS was not significantly different regardless of the presence or absence of PKD1. Furthermore, the C-terminal fragment (CTF) of PKD1 did not affect TRPC4/C5 activity, likely due to the loss of the N-terminus that contains the G-protein coupled receptor proteolytic site (GPS). We also investigated whether TRPC1/C4/C5 can form a heterodimeric channel with PKD2, despite PKD2 being primarily retained in the endoplasmic reticulum (ER). Our findings show that PKD2 is targeted to the plasma membrane, particularly by TRPC5, but not by TRPC1. However, PKD2 did not coimmunoprecipitate with TRPC5 as well as with TRPC1. PKD2 decreased both basal and La3+-induced TRPC5 currents but increased M3R-mediated TRPC5 currents. Interestingly, PKD2 increased STAT3 phosphorylation with TRPC5 and decreased STAT1 phosphorylation with TRPC1. To be specific, PKD2 and TRPC1 compete to bind with TRPC5 to modulate intracellular Ca2+ signaling and reach the plasma membrane. This interaction suggests a new therapeutic target in TRPC5 channels for improving vascular endothelial function in polycystic kidney disease.

Mixed Adsorption Mono- and Multilayers of ß-Lactoglobulin Fibrils and Sodium Polystyrene Sulfonate

The formation of beta-lactoglobulin (BLG)/sodium polystyrene sulfonate (PSS) complexes decelerates the change in the surface properties of the mixed solutions with the surface age and increases the steady-state dilational surface elasticity in a narrow PSS concentration range. At the same time, the changes in the surface properties are accelerated in the dispersions of BLG fibrils with and without PSS due to the influence of small peptides coexisting with fibrils. A decrease in the peptide concentration as a result of the dispersion purification leads to slower changes in the surface properties at low PSS concentrations. The increase in the polyelectrolyte concentration results in an increase in the steady-state surface elasticity due to the fibril/PSS complex formation and in very slow changes in the surface properties if the polyelectrolyte exceeds a certain critical value. The latter effect is a consequence of the formation of large aggregates and of an increase in the electrostatic adsorption barrier. The consecutive adsorption of BLG fibrils and PSS leads to the formation of regular multilayers at the liquid–gas interface. The multilayer properties change noticeably with an increase in the number of layers from four to six in agreement with previous results on the multilayers of PSS with an oppositely charged synthetic polyelectrolyte, presumably due to the heterogeneity of the first PSS layer. The dynamic elasticity of the multilayers approaches 250 mN/m, indicating that they can effectively stabilize foams and emulsions.

Dendritic Molecular Baskets for Selective Binding of Toxic Methotrexate.

We describe the preparation, assembly, recognition characteristics, and bioactivity of dendritic basket 612-. This novel cavitand has a deep aromatic pocket with three (S)-glutamic acid dendrons at the rim to amplify water solubility and prevent self-association. 1H NMR spectroscopy, calorimetry (ITC), and mass spectrometry (ESI-MS) measurements validate the formation of an inclusion complex between 612- and anticancer drug methotrexate (MTX2-) in water (Kd = 9.2 μM). To identify the docking pose, a comparison of computed (DFT and MM) and experimental 1H NMR chemical shifts suggests that MTX2- folds inside 612- (π···π), forming HBs with the peptidic dendrons while anchoring (C-H···π) to the aromatic pocket through its N-methyl group. In consequence, 612- selectively binds MTX2- in competition with structurally similar folic acid and leucovorin (reversal poisoning agent). While the host is biocompatible (HEK293; IC50 > 150 μM) and produces inclusion complex [MTX⊂6]14- in cell media, it experiences limitation in pharmacokinetic sequestration of MTX2- as dihydrofolate reductase's affinity to the drug is suggested to prevail over that of 612-. Nonetheless, considering the basket's biocompatibility, tunability, and chemoselectivity, it stands as the leading candidate in the pursuit of an effective abiotic antidote for methotrexate poisoning.