Unveiling the Collaborative Effect at the Cucurbit[8]uril-MoS2 Hybrid Interface for Electrochemical Melatonin Determination.

Host-guest interactions are of paramount importance in supramolecular chemistry and in a wide range of applications. Particularly well known is the ability of cucurbit[n]urils (CB[n]) to selectively host small molecules. We show that the charge transfer and complexation capabilities of CB[n] are retained on the surface of 2D transition metal dichalcogenides (TMDs), allowing the development of efficient electrochemical sensing platforms. We unveil the mechanisms of host-guest recognition between the MoS2 -CB[8] hybrid interface and melatonin (MLT), an important molecular regulator of vital constants in vertebrates. We find that CB[8] on MoS2 organizes the receptor portals perpendicularly to the surface, facilitating MLT complexation. This advantageous adsorption geometry is specific to TMDs and favours MLT electro-oxidation, as opposed to other 2D platforms like graphene, where one receptor portal is closed. This study rationalises the cooperative interaction in 2D hybrid systems to improve the efficiency and selectivity of electrochemical sensing platforms.

Unveiling the Collaborative Effect at the Cucurbit [8] uril-MoS2 Hybrid Interface for Electrochemical Melatonin Determination.

Invited for the cover of this issue are two collaborating groups: one at the Universidad Autónoma de Madrid and the other at the Instituto de Ciencia de Materiales de Madrid. The image depicts Cucurbit[8]uril adsorbed on a transition metal dichalcogenide surface letting the cavity open for complex formation with melatonin and allowing efficient electrochemical sensing. Read the full text of the article at 10.1002/chem.202203244.

Differential pulse voltammetric determination of the carcinogenic diamine 4,4'-oxydianiline by electrochemical preconcentration on a MoS2 based sensor.

An electrochemical sensor for the carcinogen 4,4'-oxydianiline (Oxy) is described. The method is based on the ability of MoS2 nanosheets to preconcentrate Oxy. A glassy carbon electrode (GCE) was covered, by drop-casting, with MoS2 nanosheets that were obtained by exfoliation. X-Ray photoemission spectroscopy indicates that Oxy accumulates on the MoS2 nanosheets through an electropolymerization process similar to that reported for aniline. Both electrochemical impedance spectroscopy and atomic force microscopy were used to characterize the electrode surface at the different stages of device fabrication. Employing the current measured at +0.27 V vs. Ag/AgCl after Oxy adsorption, the modified GCE enables the voltammetric detection of Oxy at 80 nM levels with relative errors and relative standard deviations of <8.3 and <5.6%, respectively, at all the concentrations studied. The method was applied to the selective determination of Oxy in spiked river water samples. Very good selectivity and recoveries of around 95% in average are found. Graphical abstractSchematic representation of 4,4-oxydianiline electrochemical polymerization and preconcentration onto molybdenum disulfide nanosheets for the diamine determination in river waters.

Synergistic effect of MoS2 and diamond nanoparticles in electrochemical sensors: determination of the anticonvulsant drug valproic acid.

The authors describe an electrochemical sensor based on the use of diamond nanoparticles (DNPs) and molybdenum disulfide (MoS2) platelets. The sensor was applied to the voltammetric determination of the anticonvulsant valproic acid which was previously derivatized with ferrocene. The MoS2 platelets were obtained by an exfoliation method, and the DNPs were directly dispersed in water and subsequently deposited on a glassy carbon electrode (GCE). The sensor response was optimized in terms of the solvent employed for dispersing the MoS2 nanomaterial and the method for modifying the GCE. Sensors consisting of a first layer of MoS2 dispersed in ethanol/water and a second layer of DNPs give better response. The single steps of sensor construction were characterized by atomic force microscopy and electrochemical impedance spectroscopy. The differential pulse voltammetric response of the GCE (measured at +0.18 V vs. Ag/AgCl) was compared to that of sensors incorporating only one of the nanomateriales (DNPs or MoS2). The formation of a hybrid MoS2-DNP structure clearly improves performance. The GCE containing both nanomaterials exhibits high sensitivity (740 µA ⋅ mM-1 ⋅ cm-2), a 0.27 µM detection limit, and an 8% reproducibility (RSD). The sensor retained 99% of its initial response after 45 days of storage. Graphical abstract Electrochemical sensor by co-immobilization of MoS2 and diamond nanoparticles (DNP). The formation of a hybrid MoS2-DNP structure enhances the performance of the sensor towards valproic acid derivatized with a ferrocene group, when compared with sensors incorporating only DNP or MoS2.

Harnessing two-dimensional nanomaterials for diagnosis and therapy in neurodegenerative diseases: Advances, challenges and prospects.

Neurodegenerative diseases (NDDs) are specific brain disorders characterized by the progressive deterioration of different motor activities as well as several cognitive functions. Current conventional therapeutic options for NDDs are limited in addressing underlying causes, delivering drugs to specific neuronal targets, and promoting tissue repair following brain injury. Due to the paucity of plausible theranostic options for NDDs, nanobiotechnology has emerged as a promising field, offering an interdisciplinary approach to create nanomaterials with high diagnostic and therapeutic efficacy for these diseases. Recently, two-dimensional nanomaterials (2D-NMs) have gained significant attention in biomedical and pharmaceutical applications due to their precise drug-loading capabilities, controlled release mechanisms, enhanced stability, improved biodegradability, and reduced cell toxicity. Although various studies have explored the diagnostic and therapeutic potential of different nanomaterials in NDDs, there is a lack of comprehensive review addressing the theranostic applications of 2D-NMs in these neuronal disorders. Therefore, this concise review aims to provide a state-of-the-art understanding of the need for these ultrathin 2D-NMs and their potential applications in biosensing and bioimaging, targeted drug delivery, tissue engineering, and regenerative medicine for NDDs.

Enhanced Electrocatalysis on Copper Nanostructures: Role of the Oxidation State in Sulfite Oxidation.

The influence of surface morphology and the oxidation state on the electrocatalytic activity of nanostructured electrodes is well recognized, yet disentangling their individual roles in specific reactions remains challenging. Here, we investigated the electrooxidation of sulfite ions in an alkaline environment using cyclic voltammetry on copper oxide nanostructured electrodes with different oxidation states and morphologies but with similar active areas. To this aim, we synthesized nanostructured Cu films made of nanoparticles or nanorods on top of glassy carbon electrodes. Our findings showed an enhanced sensitivity and a lower detection threshold when utilizing Cu(I) over Cu(II). Density functional theory-based thermochemical analysis revealed the underlying oxidation mechanism, indicating that while the energy gain associated with the process is comparable for both oxide surfaces, the desorption energy barrier for the resulting sulfate molecules is three times higher on Cu(II). This becomes the limiting step of the reaction kinetics and diminishes the overall electrooxidation efficiency. Our proposed mechanism relies on the tautomerization of hydroxyl groups confined on the surface of Cu-based electrodes. This mechanism might be applicable to electrochemical reactions involving other sulfur compounds that hold technological significance.

2D-ReS2 & diamond nanoparticles-based sensor for the simultaneous determination of sunset yellow and tartrazine in a multiple-pulse amperometry FIA system.

We present a flow injection system with a multiple pulse amperometric detection (FIA-MPA)-based methodology for the simultaneous analysis of sunset yellow and tartrazine. As transducer, we have developed a novel electrochemical sensor based on the synergistic effect of ReS2 nanosheets and diamond nanoparticles (DNPs). Among several transition dichalcogenides for the sensor development, we have selected ReS2 nanosheets since it yields a better response towards both colourants. Scanning probe microscopy characterization shows that the surface sensor is composed by scattered and stacked ReS2 flakes and large aggregates of DNPs. With this system, the gap between the oxidation potential values of sunset yellow and tartrazine is wide enough to allow the simultaneous determination of both dyes. Under the optimum potential pulse conditions (0.8 and 1.2 V) during 250 ms, a flow rate of 3 mL/min and a volume injection of 250 µL, detection limits of 3.51 × 10-7 M and 2.39 × 10-7 M for sunset yellow and tartrazine, respectively, were obtained. This method exhibits good accuracy and precision with Er minor than 13% and RSD lower than 8% with a sampling frequency of 66 samples per hour. Pineapple jelly samples were analyzed by the standard addition method, obtaining 53.7 mg/kg and 29.0 mg/kg of sunset yellow and tartrazine, respectively. From the analysis of fortified samples, recoveries of 94% and 105% were obtained.

MoS2 quantum dots-based optical sensing platform for the analysis of synthetic colorants. Application to quinoline yellow determination.

In this work, a novel fluorescence sensor has been designed to solve the actual need of new fast and inexpensive sensing platforms for the analysis of synthetic colorants. It is based on MoS2 quantum dots obtained by a hydrothermal method and incorporated as fluorophore into the matrix of PVC membranes, which are deposited on quartz substrates by spin-coating. It was proven, as in these conditions, MoS2 quantum dots maintain the fluorescent properties that they present in solution. Experiments carried out in solution displayed a maximum emission when they were excited under 310 nm. This initial fluorescence decreases linearly in presence of increasing concentrations of various synthetic colorants namely quinoline yellow, tartrazine, sunset yellow, allura red, ponceau 4R and carmoisine. The two possible mechanisms that can explain this quenching effect, colorants absorbing photons emitted by quantum dots and/or competing with the nanomaterial for photons coming from the excitation source, were evaluated. The most pronounced effect was observed with quinoline yellow, as a result of a mixed mechanism. The optimized methodology developed for the determination of quinoline yellow showed a linear concentration range between 5.4 and 55.0 µg with a limit of detection of 1.6 µg. The sensor was applied to the determination of quinoline yellow in a food colour paste obtaining results in good agreement with those obtained by HPLC-UV-vis measurements.

Synergistic effect of manganese (II) phosphate & diamond nanoparticles in electrochemical sensors for reactive oxygen species determination in seminal plasma.

In this work, we explore the ability of manganese (II) phosphate (MnP) as a catalytic element for the determination of reactive oxygen species (ROS) in seminal plasma, when MnP is employed as modifier of a glassy carbon electrode. The electrochemical response of the manganese (II) phosphate-modified electrode shows a wave at around +0.65 V due to the oxidation of Mn2+ to MnO2+, which is clearly enhanced after addition of superoxide, the molecule considered as the mother of ROS. Once proved the suitability of manganese (II) phosphate as catalyst, we evaluate the effect of including a 0D (diamond nanoparticles) or a 2D (ReS2) nanomaterial in the sensor design. The system consisting of manganese (II) phosphate and diamond nanoparticles yielded the largest improvement of the response. The morphological characterization of the sensor surface was performed by scanning electron microscopy and atomic force microscopy, while cyclic and differential pulse voltammetry were employed for the electrochemical characterization of the sensor. After optimizing the sensor construction, calibration procedures by chronoamperometry were performed, leading to a linear relation between peak intensity and the superoxide concentration in the range of 1.1 10-4 M - 1.0 10-3 M with a limit of detection of 3.2 10-5 M. Seminal plasma samples were analysed by the standard addition method. Moreover, the analysis of samples fortified with superoxide at the µM level leads to recoveries of 95%.

Electrochemical sensor based on the synergy between Cucurbit[8]uril and 2D-MoS2 for enhanced melatonin quantification.

We present the development of an electrochemical sensor towards melatonin determination based on the synergistic effect between MoS2 nanosheets and cucurbit[8]uril. For the sensor construction cucurbit[8]uril suspensions were prepared in water, and MoS2 nanosheets were obtained by liquid exfoliation in ethanol:water. The sensing platform was topographically characterized by Atomic Force Microscopy. Electrochemical Impedance Spectroscopy experiments allowed us to study the charge transfer process during melatonin oxidation. Moreover, stoichiometry of the resulting complex has also been determined. After the optimization of the sensor construction and the experimental variables involved in the Differential Pulse Voltammetric response of melatonin, detection limit of 3.80 × 10-7 M, relative errors minor than 3.8% and relative standard deviation lower than 4.4% were obtained. The proposed sensor has been successfully applied to melatonin determination in pharmaceutical and biological samples as human urine and serum, with very good recoveries ranging from 90 to 102%.

Rapid identification of ETP-46992, orally bioavailable PI3K inhibitor, selective versus mTOR.

Phosphoinositide-3-kinases (PI3K) are a family of lipid kinases mediating numerous cell processes such as proliferation, migration and differentiation. PI3K is an important target for cancer therapeutics due to the deregulation of this signaling pathway in a wide variety of human cancers. Herein, we describe the rapid identification of ETP-46992, within 2-aminocarbonyl imidazo [1,2-a] pyrazine series, with suitable pharmacokinetic (PK) properties that allows the establishment of mechanism of action and efficacy in vivo studies. ETP-46992 showed tumor growth inhibition in a GEMM mouse tumor model driven by a K-Ras(G12V) oncogenic mutation and in tumor xenograft models with PI3K pathway deregulated (BT474).

Identification of ETP-46321, a potent and orally bioavailable PI3K α, δ inhibitor.

Phosphoinositide-3-kinase (PI3K) is an important target for cancer therapeutics due to the deregulation of this signaling pathway in a wide variety of human cancers. Herein, we describe the optimization of imidazo [1,2-a] pyrazines, which allow us to identify compound 14 (ETP-46321), with potent biochemical and cellular activity and good pharmacokinetic properties (PK) after oral dosing. ETP-46321 PK/PD studies showed time dependent downregulation of AKT(Ser473) phosphorylation, which correlates with compound levels in tumor tissue and demonstrating to be efficacious in a GEMM mouse tumor model driven by a K-Ras(G12V) oncogenic mutation. Treatment with ETP-46321 resulted in significant tumor growth inhibition.

Lactate biosensing based on covalent immobilization of lactate oxidase onto chevron-like graphene nanoribbons via diazotization-coupling reaction.

We have designed and prepared an electrochemical biosensor for lactate determination. Through a diazotation process, the enzyme lactate oxidase (LOx) is anchored onto chevron-like graphene nanoribbons (GNR), previously synthesized by a solution-based chemical route, and used as modifiers of glassy carbon electrodes. In a first step, we have performed the grafting of a 4-carboxyphenyl film, by electrochemical reduction of the corresponding 4-carboxyphenyl diazonium salt, on the GNR-modified electrode surface. In this way, the carboxylic groups are exposed to the solution, enabling the covalent immobilization of the enzyme through the formation of an amide bond between these carboxylic groups and the amine groups of the enzyme. The biosensor design was optimized through the morphological and electrochemical characterization of each construction step by atomic force microscopy, scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy.The cyclic voltammetric response of the biosensor in a solution of hydroxymethylferrocene in presence of l-lactate evidenced a clear electrocatalytic effect powered by the specific design of the biosensing platform with LOx covalently attached to the GNR layer. From the calibration procedures employed for l-lactate determination, a linear concentration range of 3.4 · 10-5- 2.8 · 10-4 M and a detection limit of 11 µM were obtained, with relative errors and relative standard deviations less than 6.0% and 8.4%, respectively. The applicability of the biosensor was tested by determining lactate in apple juices, leading to results that are in good agreement with those obtained with a well-established enzymatic spectrophotometric assay kit.

A supramolecular hybrid sensor based on cucurbit[8]uril, 2D-molibdenum disulphide and diamond nanoparticles towards methyl viologen analysis.

We develop an electrochemical sensor by using 2D-transition metal dichalcogenides (TMD), specifically MoS2, and nanoparticles stabilized with cucurbit[8]uril (CB[8]) incorporated together with them. Two different nanoparticles are assayed: diamond nanoparticles (DNPs) and gold nanoparticles (AuNp). 0D materials, together with TMD, provide increased conductivity and active surface while the macrocycle CB[8] affords selectivity towards the guest methyl viologen (MV2+), also named paraquat. Glassy Carbon (GC) electrodes are modified by drop-casting of suspensions of MoS2, followed by either a CB[8]-DNPs hybrid dispersion or a CB[8]-AuNp suspension. Atomic force microscopy is employed for the morphological characterization of the electrochemical sensor surface while cyclic voltammetry and electrochemical impedance spectroscopy techniques allow the electrochemical characterization of the sensor. The well-stablished signals of CB[8]-encapsulated MV2+ arise in voltammetric measurements when the macrocycle modifies the 0D-materials. Once the sensor construction and differential pulse voltammetry parameters have been optimized for quantification purposes, calibration procedures are performed with the platform GC/MoS2/CB[8]-DNPs. This sensing platform shows linear relations between peak intensity and the MV2+ concentration in the linear concentration range of (0.73-8.0) · 10-6 M with a limit of detection of 2.2 · 10-7 M. Analyses of river water samples fortified with MV2+ at the µM level shows recoveries of 100% with RSD values of 6.4% (n = 3).

MoS2 quantum dots for on-line fluorescence determination of the food additive allura red.

This work presents an on-line fluorescence method for the allura red (AR) determination. The method is based on the fluorescence quenching of dots of MoS2 because of their interaction with the non-fluorescence dye. MoS2-dots were synthetized and characterized by spectroscopic techniques and High Resolution Transmission Electronic Microscopy (HR-TEM). The simultaneous injection of the nanomaterial and the dye in a flow injection analysis system allows the determination of allura red at 1.7 × 10-6 M concentration level with a very good accuracy and precision (Er minor than 10% and RSD lower than 8%) and a sampling frequency of 180 samples per hour. Moreover, the interaction fluorophore-quencher results a dynamic inhibition mechanism. The proposed methodology was applied to the AR analysis in soft beverages and powders for gelatine preparation. Colourant concentrations of 63 ± 2 mg/L (n = 3) and 0.30 ± 0.01 mg/g (n = 3) were found, respectively. These results were validated by high performance liquid chromatography technique.

Macrocyclization as a Source of Desired Polypharmacology. Discovery of Triple PI3K/mTOR/PIM Inhibitors.

The PI3K/AKT/mTOR and PIM kinase pathways contribute to the development of several hallmarks of cancer. Cotargeting of these pathways has exhibited promising synergistic therapeutic effects in liquid and solid tumor types. To identify molecules with combined activities, we cross-screened our collection of PI3K/(±mTOR) macrocycles (MCXs) and identified the MCX thieno[3,2-d]pyrimidine derivative 2 as a moderate dual PI3K/PIM-1 inhibitor. We report the medicinal chemistry exploration and biological characterization of a series of thieno[3,2-d]pyrimidine MCXs, which led to the discovery of IBL-302 (31), a potent, selective, and orally bioavailable triple PI3K/mTOR/PIM inhibitor. IBL-302, currently in late preclinical development (AUM302), has recently demonstrated efficacy in neuroblastoma and breast cancer xenografts. Additionally, during the course of our experiments, we observed that macrocyclization was essential to obtain the desired multitarget profile. As a matter of example, the open precursors 35-37 were inactive against PIM whereas MCX 28 displayed low nanomolar activity.

AFM, SECM and QCM as useful analytical tools in the characterization of enzyme-based bioanalytical platforms.

One of the key issues to develop biosensing platforms concerns the processes involved in enzyme immobilization on surfaces. The understanding of their fundamentals is crucial to obtain stable and catalytically active protein layers for developing successful biosensing devices. In this respect, the advent and development of new characterization techniques, in particular at the submicron level, has allowed the study of these processes with high resolution, which has opened new routes to improve, and eventually control, enzyme immobilization on electrode surfaces. This review focuses on the application of Atomic Force Microscopy (AFM), Scanning Electrochemical Microscopy (SECM) and Quartz Crystal Microbalance (QCM) techniques in the characterization of the successive immobilization steps involved in the development of bioanalytical platforms. A common advantage of these techniques is their ability to provide important information without damaging the immobilized biological sample due to the possibility of performing measurements under physiological conditions close to the native environment of the specimens. A particular emphasis is placed on the application of these techniques to the characterization of the immobilization of enzymes on different modified and unmodified surfaces as well as on the study of protein interactions, which is a more recent and less current application.

Evaluation of oxidative stress: Nanoparticle-based electrochemical sensors for hydrogen peroxide determination in human semen samples.

We have developed electrochemical sensors for the determination of H2O2 in a complex matrix such as human semen as a method to evaluate oxidative stress related to male infertility. Our sensors are based on the modification of conventional electrode surfaces with nanoparticles. We used diamond nanoparticles (DNp) either on glassy carbon or gold surfaces (GC/DNp and Au/DNp sensors, respectively), and copper nanoparticles electrochemically generated directly on glassy carbon surfaces (GC/CuNp). The morphology of the modified electrode surfaces was characterized by Atomic Force Microscopy (AFM), and the H2O2 determination performance evaluated by chronoamperometric measurements at different applied potentials. The best results are obtained for GC/DNp at +1.0 V, Au/DNp at -0.6 V and GC/CuNp at +0.2 V with detection limits (LD) of 1.1 µM, 2.4 µM and 2.6 µM, respectively. The analysis of H2O2 in doped synthetic semen using the GC/CuNp sensor shows the best recoveries, reaching a mean value of 103%. The GC/CuNp sensor was successfully applied to H2O2 analysis in real human semen. In this case, a H2O2 concentration of 1.42 ± 0.05 mM is found and recoveries of 102% on average are obtained.

Chemically synthesized chevron-like graphene nanoribbons for electrochemical sensors development: determination of epinephrine.

We employ chevron-like graphene nanoribbons (GNRs) synthesized by a solution-based chemical route to develop a novel electrochemical sensor for determination of the neurotransmitter epinephrine (EPI). The sensor surface, a glassy carbon electrode modified with GNRs, is characterized by atomic force microscopy, scanning electron microscopy and Raman spectroscopy, which show that the electrode surface modification comprises of bi-dimensional multilayer-stacked GNRs that retain their molecular structure. The charge transfer process occurring at the electrode interface is evaluated by electrochemical impedance spectroscopy. The sensor is applied to the determination of EPI, employing as an analytical signal the reduction peak corresponding to the epinephrinechrome-leucoepinephrinechrome transition (E = - 0.25 V) instead of the oxidation peak usually employed in the literature (E = + 0.6 V) in order to minimize interferences. The results obtained demonstrate that chevron-like nanoribbons synthesized by solution methods exhibit reliable electrocatalytic activity for EPI determination. Using differential pulse voltammetry, we obtain a linear concentration range from 6.4 × 10-6 to 1.0 × 10-4 M and a detection limit of 2.1 × 10-6 M. The applicability of the sensor was evaluated by determining EPI in pharmaceutical samples with satisfactory results.

Screening protocol for the identification of modulators by immunofluorescent cell-based assay.

High-throughput assays are a common strategy for the identification of compounds able to modulate a certain cellular activity. Here, we show an automatized analysis platform for the quantification of nuclear foci as inhibitory effect of compounds on a target protein labeled by fluorescent antibodies. Our experience led us to a fast analysis platform that combines cell-based assays, high-content screening, and confocal microscopy, with an automatic and user-friendly statistical analysis of plate-based assays including positive and negative controls, able to identify inhibitory effect of compounds tested together with the Z-prime and Window of individual plate-based assays to assess the reliability of the results. The platform integrates a web-based tool implemented in Pipeline Pilot and R, and allows computing the inhibition values of different parameters obtained from the high-content screening and confocal microscopy analysis. This facilitates the exploration of the results using the different parameters, providing information at different levels as the number of foci observed, the sum of intensity of foci, area of foci, etc, the detection and filtering of outliers over the assay plate, and finally providing a set of statistics of the parameters studied together with a set of plots that we believe significantly helps to the interpretation of the assay results.