Alleviating Cellular Oxidative Stress through Treatment with Superoxide-Triggered Persulfide Prodrugs

Overproduction of superoxide anion (O₂^.−), the primary cellular reactive oxygen species (ROS), is implicated in various human diseases. To reduce cellular oxidative stress caused by overproduction of superoxide, we developed a compound that reacts with O₂^.− to release a persulfide (RSSH), a type of reactive sulfur species related to the gasotransmitter hydrogen sulfide (H₂S). Termed SOPD-NAC , this persulfide donor reacts specifically with O₂^.−, decomposing to generate N-acetyl cysteine (NAC) persulfide. To enhance persulfide delivery to cells, we conjugated the SOPD motif to a short, self-assembling peptide (Bz-CFFE-NH₂) to make a superoxide-responsive, persulfide-donating peptide ( SOPD-Pep ). Both SOPD-NAC and SOPD-Pep delivered persulfides/H₂S to H9C2 cardiomyocytes and lowered ROS levels as confirmed by quantitative in vitro fluorescence imaging studies. Additional in vitro studies on RAW 264.7 macrophages showed that SOPD-Pep mitigated toxicity induced by phorbol 12-myristate 13-acetate (PMA) more effectively than SOPD-NAC and several control compounds, including common H₂S donors.     

Synergistic Anticancer Therapy by Ovalbumin Encapsulation-Enabled Tandem Reactive Oxygen Species Generation

The anticancer efficacy of photodynamic therapy (PDT) is limited due to the hypoxic features of solid tumors. We report synergistic PDT/chemotherapy with integrated tandem Fenton reactions mediated by ovalbumin encapsulation for improved in vivo anticancer therapy via an enhanced reactive oxygen species (ROS) generation mechanism. O₂^.− produced by the PDT is converted to H₂O₂ by superoxide dismutase, followed by the transformation of H₂O₂ to the highly toxic ^.OH via Fenton reactions by Fe²⁺ originating from the dissolution of co-loaded Fe₃O₄ nanoparticles. The PDT process further facilitates the endosomal/lysosomal escape of the active agents and enhances their intracellular delivery to the nucleus—even for drug-resistant cells. Cisplatin generates O₂^.− in the presence of nicotinamide adenine dinucleotide phosphate oxidase and thereby improves the treatment efficiency by serving as an additional O₂^.− source for production of ^.OH radicals. Improved anticancer efficiency is achieved under both hypoxic and normoxic conditions.     

Versatile Fluorescent Probes for Imaging the Superoxide Anion in Living Cells and In Vivo

The superoxide anion (O₂^.−) is widely engaged in the regulation of cell functions and is thereby intimately associated with the onset and progression of many diseases. To ascertain the pathological roles of O₂^.− in related diseases, developing effective methods for monitoring O₂^.− in biological systems is essential. Fluorescence imaging is a powerful tool for monitoring bioactive molecules in cells and in vivo owing to its high sensitivity and high temporal-spatial resolution. Therefore, increasing numbers of fluorescent imaging probes have been constructed to monitor O₂^.− inside live cells and small animals. In this minireview, we summarize the methods for design and application of O₂^.−-responsive fluorescent probes. Moreover, we present the challenges for detecting O₂^.− and suggestions for constructing new fluorescent probes that can indicate the production sites and concentration changes in O₂^.− as well as O₂^.−-associated active molecules in living cells and in vivo.     

Recent advances in fluorescent probes for the detection of reactive oxygen species

Reactive oxygen species (ROS) have captured the interest of many researchers in the chemical, biological, and medical fields since they are thought to be associated with various pathological conditions. Fluorescent probes for the detection of ROS are promising tools with which to enhance our understanding of the physiological roles of ROS, because they provide spatial and temporal information about target biomolecules in in vivo cellular systems. ROS probes, designed to detect specific ROS with a high selectivity, would be desirable, since it is now becoming clear that each ROS has its own unique physiological activity. However, dihydro-compounds such as 2′,7′-dichlorodihydrofluorescein (DCFH), which have traditionally been used for detecting ROS, tend to react with a wide variety of ROS and are not completely photostable. Some attractive fluorescent probes that exhibit a high degree of selectivity toward specific ROS have recently been reported, and these selective probes are expected to have great potential for elucidating unknown physiological mechanisms associated with their target ROS. This review focuses on the design, detection mechanism, and performance of fluorescent probes for the detection of singlet oxygen (¹O₂), hydrogen peroxide (H₂O₂), hydroxyl radicals (^.OH), or superoxide anion (O₂ ^−.), a field in which remarkable progress has been achieved in the last few years.     

Alleviating Cellular Oxidative Stress through Treatment with Superoxide‐Triggered Persulfide Prodrugs

Overproduction of superoxide anion (O₂^.?), the primary cellular reactive oxygen species (ROS), is implicated in various human diseases. To reduce cellular oxidative stress caused by overproduction of superoxide, we developed a compound that reacts with O₂^.? to release a persulfide (RSSH), a type of reactive sulfur species related to the gasotransmitter hydrogen sulfide (H₂S). Termed SOPD‐NAC , this persulfide donor reacts specifically with O₂^.?, decomposing to generate N‐acetyl cysteine (NAC) persulfide. To enhance persulfide delivery to cells, we conjugated the SOPD motif to a short, self‐assembling peptide (Bz‐CFFE‐NH₂) to make a superoxide‐responsive, persulfide‐donating peptide ( SOPD‐Pep ). Both SOPD‐NAC and SOPD‐Pep delivered persulfides/H₂S to H9C2 cardiomyocytes and lowered ROS levels as confirmed by quantitative in vitro fluorescence imaging studies. Additional in vitro studies on RAW 264.7 macrophages showed that SOPD‐Pep mitigated toxicity induced by phorbol 12‐myristate 13‐acetate (PMA) more effectively than SOPD‐NAC and several control compounds, including common H₂S donors.     

Evaluation of chemiluminescence reagents for selective detection of reactive oxygen species

In order to evaluate the chemiluminescence (CL) reagents for selective detection of reactive oxygen species (ROS), we comprehensively measured the CL responses of 20 CL reagents (three luminol derivatives, two imidazopyrazinone derivatives, eight lophine derivatives, six acridinium ester derivatives and lucigenin) against six types of ROS (superoxide anion: O₂⁻, hydroxyl radical: OH, hydrogen peroxide: H₂O₂, hypochlorite anion: ClO⁻, singlet oxygen: ¹O₂, and nitric oxide: NO). As a result of the screening, it was found that nine CL reagents selectively detected O₂⁻ while one CL reagent selectively detected OH. However, no CL reagent had selectivity on the detection of H₂O₂, ClO⁻, ¹O₂ and NO. Our screening results could help to select the most suitable CL reagent for selective determination of different ROS.As an application study, 4-methoxyphenyl-10-methylacridinium-9-carboxylate (MMAC), one of the acridinium ester derivatives, showed high selectivity on the detection of O₂⁻, and thus was applied to the assay of superoxide dismutase (SOD) activity. The dynamic range and detection limit of the developed CL assay were 0.1-10 and 0.06 U mL⁻¹, respectively. Significant correlation (r = 0.997) was observed between the results by the CL assay using MMAC and the spectrophotometric assay using 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt.     

Indeno[2,1-a]fluorene-11,12-dione Radical Anions: Synthesis,Characterization, and Properties

The one-electron reduction of indeno[2,1-a]fluorene-11,12-dione ( IF ) with various alkali metals prepare the radical anion salts. The data about different structures, properties, and characterization was obtained by single-crystal X-ray diffraction, electron paramagnetic resonance (EPR) spectroscopy, superconducting quantum interference device (SQUID) measurements, and physical property measurement system (PPMS). Compound IF ^.−K⁺(18-c-6) is regarded as a one-dimensional magnetic chain through C−H⋅⋅⋅C interaction. Theoretical calculations and magnetic results showed that [ IF ^.−K⁺(15-c-5)]₂ is a dimer with an open-shell ground state. Compounds IF ^.−Na⁺(15-c-5) and IF ^.−K⁺(cryptand) are monoradical anion salts: IF ₂^.−Li⁺ possesses unique π-stack structure with an interplanar separation less than 3.46 Å, making it a semiconductor (δ_RT=1.9×10⁻⁴ S ⋅ cm⁻¹). This work gives insights into multifunctional radical anions, and describes the design and development of different functional radicals.     

Mono-reduced Corannulene: To Couple and Not to Couple in One Crystal

One-electron reduction of corannulene, C₂₀H₁₀, with Li metal in diglyme resulted in crystallization of [{Li⁺(diglyme)₂}₄(C₂₀H₁₀^.−)₂(C₂₀H₁₀-C₂₀H₁₀)²⁻] ( 1 ), as revealed by single-crystal X-ray diffraction. This hybrid product contains two corannulene monoanion-radicals along with a dianionic dimer, crystallized with four Li⁺ ions wrapped by diglyme molecules. The dimeric (C₂₀H₁₀-C₂₀H₁₀)²⁻ anion provides the first crystallographically confirmed example of spontaneous radical dimerization for C₂₀H₁₀^.−. The C−C bond length between the two C₂₀H₁₀^.− bowls of 1.588(5) Å is consistent with the single σ-bond character of the linker. The trans-disposition of two bowls in the centrosymmetric (C₂₀H₁₀-C₂₀H₁₀)²⁻ dimer is observed with the torsion angle around the central C−C bond of 180°. Comprehensive theoretical analysis of formation/decomposition processes of the dimeric dianion has been carried out in order to evaluate the nature of bonding and energetics of the C₂₀H₁₀^.− coupling. It is found that such σ-bonded dimers are thermodynamically unstable due to large preparation energy and repulsive Pauli component of the bonding, but kinetically persistent due to a high energy barrier provided by the existing spin-crossing point.     

Mediated bioelectrocatalytic O2 reduction to water at highly positive electrode potentials near neutral pH

Combinations of bilirubin oxidase and metal complexes: [W(CN)₈]^3−/4−, [Os(CN)₆]^3−/4− and [Mo(CN)₈]^3−/4− (the formal potentials, E^0′(M), being 0.320, 0.448, and 0.584 V vs. Ag|AgCl, respectively, at pH 7.0), allowed bioelectrocatalytic reduction of O₂ to water at their formal potentials near neutral pH. The O₂ reduction current appeared even at the standard potential of the O₂/H₂O redox couple, E^0′(O₂/H₂O), when [Mo(CN)₈]^3−/4− was used at pH 7.4, though the magnitude was small. The magnitude of the bioelectrocatalytic current systematically decreased with the decrease in the potential difference between E^0′(O₂/H₂O) and E^0′(M). A limiting current as large as 17 mA/cm² of a projected electrode surface area was obtained at 0.25 V (−0.37 V vs. E^0′(O₂/H₂O)) for the O₂ reduction at pH 7.0 with a carbon felt electrode modified with electrostatically entrapped bilirubin oxidase and [W(CN)₈]^3−/4− at the electrode rotation rate of 4000 rpm.     

On the nature of the peroxoborate ion in solution

The Raman spectra of alkaline lithium metaborate solutions containing H₂O₂ reveal lines attributable to a peroxoborate anion, B(OH)₃OOH^?, in equilibrium with B(OH)₄^? and H₂O₂. There is no evidence for the formation of peroxoboric acids at low pH.     

Hydrogen Sulfide Donors Activated by Reactive Oxygen Species

Hydrogen sulfide (H₂S) exhibits promising protective effects in many (patho)physiological processes, as evidenced by recent reports using synthetic H₂S donors in different biological models. Herein, we report the design and evaluation of compounds denoted PeroxyTCM, which are the first class of reactive oxygen species (ROS)‐triggered H₂S donors. These donors are engineered to release carbonyl sulfide (COS) upon activation, which is quickly hydrolyzed to H₂S by the ubiquitous enzyme carbonic anhydrase (CA). The donors are stable in aqueous solution and do not release H₂S until triggered by ROS, such as hydrogen peroxide (H₂O₂), superoxide (O₂^?), and peroxynitrite (ONOO^?). We demonstrate ROS‐triggered H₂S donation in live cells and also demonstrate that PeroxyTCM‐1 provides protection against H₂O₂‐induced oxidative damage, suggesting potential future applications of PeroxyTCM and similar scaffolds in H₂S‐related therapies.     

A Sterically Hindered Derivative of 2,1,3-Benzotelluradiazole: A Way to the First Structurally Characterised Monomeric Tellurium–Nitrogen Radical Anion

Interaction of the tetradentate redox-active 6,6′-[1,2-phenylenebis(azanediyl)]bis(2,4-di-tert-butylphenol) (H₄L) with TeCl₄ leads to neutral diamagnetic compound TeL ( 1 ) in high yield. The molecule of 1 has a nearly planar TeN₂O₂ fragment, which suggests the formulation of 1 as Te^IIL²⁻, in agreement with the results of DFT calculations and QTAIM and NBO analyses. Reduction of 1 with one equivalent of [CoCp₂] leads to quantitative formation of the paramagnetic salt [CoCp₂]⁺[ 1 ]^.−, which was characterised by single-crystal XRD. The solution EPR spectrum of [CoCp₂]⁺[ 1 ]^.− at room temperature features a quintet due to splitting on two equivalent ¹⁴N nuclei. Below 150 K it turns into a broad singlet line with two weak satellites due to the splitting on the ¹²⁵Te nucleus. Two-component relativistic DFT calculations perfectly reproduce the a(¹⁴N) HFI constants and A_∥(¹²⁵Te) value responsible for the low-temperature satellite splitting. Calculations predict that the additional electron in 1 ^.− is localised mainly on L, while the spin density is delocalised over the whole molecule with significant localisation on the Te atom (≥30 %). All these data suggest that 1 ^.− can be regarded as the first example of a structurally characterised monomeric tellurium–nitrogen radical anion.     

Synthesis and Structure of an o-Carboranyl-Substituted Three-Coordinate Borane Radical Anion

Bis(1-(4-tolyl)-carboran-2-yl)-(4-tolyl)-borane [(1-(4-MeC₆H₄)-closo-1,2-C₂B₁₀H₁₀-2-)₂(4-MeC₆H₄)B] ( 1 ), a new bis(o-carboranyl)-(R)-borane was synthesised by lithiation of the o-carboranyl precursor and subsequent salt metathesis reaction with (4-tolyl)BBr₂. Cyclic voltammetry experiments on 1 show multiple distinct reduction events with a one-electron first reduction. In a selective reduction experiment the corresponding paramagnetic radical anion 1^.− was isolated and characterized. Single-crystal structure analyses allow an in-depth comparison of 1 , 1^.− , their calculated geometries, and the S₁ excited state of 1 . Photophysical studies of 1 show a charge transfer (CT) emission with low quantum yield in solution but a strong increase in the solid state. TD-DFT calculations were used to identify transition-relevant orbitals.     

Synthesis of sydnonimine derivatives as potential trypanocidal agents

N‐(p‐Nitrophenoxy)carbonyl‐3‐morpholino‐sydnonimine (NCMS) has been prepared from 3‐morpholinosydnonimine hydrochloride. Using the Griess assay and the superoxide‐mediated reduction of ferricytochrome c, the nitric oxide (NO?) and superoxide anion (O₂?^‐) ‐ releasing properties in phosphate buffer pH 7.4 of this novel peroxynitrite donor was studied and compared with the known 3‐morpholino‐sydnonimine (SIN‐1). From compound NCMS, a series of N‐substituted sydnonimine derivatives were easily prepared that contain purine or melaminophenyl groups which specify a recognition by a trypanosomal purine transporter. The ability of these new sydnonimines to inhibit the uptake of [2³H]adenosine on Trypanosoma equiperdum was studied.     

Tetrabutylammonium Salts of Aluminum(III) and Gallium(III) Phthalocyanine Radical Anions Bonded with Fluoren‐9‐olato− Anions and Indium(III) Phthalocyanine Bromide Radical Anions

Reduction of aluminum(III), gallium(III), and indium(III) phthalocyanine chlorides by sodium fluorenone ketyl in the presence of tetrabutylammonium cations yielded crystalline salts of the type (Bu₄N⁺)₂[M^III(HFl−O⁻)(Pc^.3−)]^.−(Br⁻) ⋅ 1.5 C₆H₄Cl₂ [M=Al ( 1 ), Ga ( 2 ); HFl−O⁻=fluoren‐9‐olato⁻ anion; Pc=phthalocyanine] and (Bu₄N⁺) [In^IIIBr(Pc^.3−)]^.− ⋅ 0.875 C₆H₄Cl₂ ⋅ 0.125 C₆H₁₄ ( 3 ). The salts were found to contain Pc^.3− radical anions with negatively charged phthalocyanine macrocycles, as evidenced by the presence of intense bands of Pc^.3− in the near‐IR region and a noticeable blueshift in both the Q and Soret bands of phthalocyanine. The metal(III) atoms coordinate HFl−O⁻ anions in 1 and 2 with short Al−O and Ga−O bond lengths of 1.749(2) and 1.836(6) Å, respectively. The C−O bonds [1.402(3) and 1.391(11) Å in 1 and 2 , respectively] in the HFl−O⁻ anions are longer than the same bond in the fluorenone ketyl (1.27–1.31 Å). Salts 1 – 3 show effective magnetic moments of 1.72, 1.66, and 1.79 μ_B at 300 K, respectively, owing to the presence of unpaired S= 1/2 spins on Pc^.3−. These spins are coupled antiferromagnetically with Weiss temperatures of −22, −14, and −30 K for 1 – 3 , respectively. Coupling can occur in the corrugated two‐dimensional phthalocyanine layers of 1 and 2 with an exchange interaction of J /k _B=−0.9 and −1.1 K, respectively, and in the π‐stacking {[In^IIIBr(Pc^.3−)]^.−}₂ dimers of 3 with an exchange interaction of J /k _B=−10.8 K. The salts show intense electron paramagnetic resonance (EPR) signals attributed to Pc^.3−. It was found that increasing the size of the central metal atom strongly broadened these EPR signals.     

Solid-State Properties of Hexaazatriphenylenehexacarbonitrile HAT(CN)6.− Radical Anions in Crystalline Salts Containing Cryptand(M+) and Crystal Violet Cations

Crystalline {Cryptand[2.2.2](Na⁺)}{HAT(CN)₆^.−}⋅0.5C₆H₄Cl₂ ( 1 ), {Cryptand[2.2.2](K⁺)}{HAT(CN)₆^.−} ( 2 ), (CV⁺){HAT(CN)₆^.−} ( 3 ), and (CV⁺){HAT(CN)₆^.−}⋅2C₆H₄Cl₂ ( 4 ) salts (where CV⁺ is the crystal violet cation) containing hexaazatriphenylenehexacarbonitrile radical anions have been obtained. The solid-state molecular structure as well as the optical and magnetic properties of HAT(CN)₆^.− are studied. The formation of HAT(CN)₆^.− in 1 – 4 leads to the appearance of new bands in the visible range, at 694 and 740 nm. The HAT(CN)₆^.− radical anions have spin state S=1/2 and are packed in one-dimensional stacks containing the {HAT(CN)₆^.−}₂ dimers alternated with weaker interacting pairs of HAT(CN)₆^.− in 1 and nearly isolated {HAT(CN)₆^.−}₂ dimers in 2 . The {HAT(CN)₆^.−}₂ dimers are diamagnetic in 1 but they effectively mediate one-dimensional antiferromagnetic coupling of spins within the stacks with moderate exchange interaction of J/k_B = −80 K. The behaviour of salt 2 is described by a singlet–triplet model for the {HAT(CN)₆^.−}₂ dimers with an energy gap of 434(±7) K. Magnetic behaviour of both salts agree well with the data of extended Hückel calculations. Salts 3 and 4 contain isolated stacks of alternated HAT(CN)₆^.− and CV⁺ ions, and in this case, nearly paramagnetic behaviour is observed with Weiss temperatures of −1 and −7 K, respectively. Narrow Lorentzian EPR signals with g = 2.0033–2.0039 were found for the HAT(CN)₆^.− radical anions in 1 and 4 but in solution g-factor shifts to 1.9964. The electronic structure of HAT(CN)₆^.− is analysed based on X-ray diffraction data for 2 , showing a Jahn–Teller distortion of the radical anion that reduces the symmetry from D_3h to C_s and splits the initially degenerated LUMOs.     

Aqueous Reactive Oxygen Species Induced by He + O<Subscript>2</Subscript> Plasmas: Chemistry Pathways and Dosage Control Approaches

Plasma–liquid interaction has already been a hotspot in the research field of plasma medicine. Aqueous reactive oxygen species (ROS) generated in this process are widely accepted playing a crucial role in plasma biomedical effects. In this paper, chemistry pathways among various aqueous ROS induced by He + O₂ plasmas are investigated by a numerical model. Simulation results show that these aqueous ROS can be classified into two groups according to their production ways: the group of species including O, ¹O₂ and e directly produced in plasma, and the other group of species including O₂ ^?, H₂O₂, O₃, etc. produced by liquid reactions. A key reaction chain of e → O₂ ^? → HO₂(→ HO₂ ^?) → H₂O₂ is found to be important in the plasma-induced liquid chemistry. Furthermore, impacts of changes in plasma and solution conditions on aqueous ROS concentrations are studied as well. It is found that changes in plasma conditions (O₂ ratio in the discharge gas/power density) can globally influence the concentrations of almost every aqueous ROS, while conditions changes of the treated liquid (pH/dissolved oxygen) only partially influence the concentrations of some specific species including O₂ ^?/HO₂, O₃ ^?/HO₃ and H₂O₂. The revelations of the liquid chemistry pathways and the dependence of ROS dosage on the treatment conditions offer a better understanding on the plasma–liquid interactions, as well as provide optimized dosage control approaches for biomedical applications.     

SYNTHESIS AND CHARACTERISATION OF A NEW NIOBATE COMPLEX WITH CHROMIUM(III)

Abstract

A new hexaniobate complex with chromium(III) was prepared and characterised by analytical and spectroscopic techniques. The compound is a violet, non-crystalline solid, formulated as [Cr₂(μ-OH)(H₃Nb₆O₁₉)(en)₃(H₂O)]. It was synthesised in aqueous solution by reaction of [Cr(en)₃]³⁺, en = ethylenediammine, with the hexaniobate anion [Nb₆O₁₉]^8-, at pH 8. Thermal decomposition yielded a mixture of CrNbO₄ and Nb₂O₅. The synthesis of this compound shows that the hexaniobate anion may form complexes at lower pH values than previously reported.     

Electrochemical characterization of glassy carbon electrodes modified with hybrid inorganic-organic single-layer of α-Keggin type polyoxotungstates

It has been demonstrated, for the first time, that an adsorbed single-layer of the hybrid salts (TBA)₄H₃PW₁₁O₃₉, (TBA)₄PW₁₁Fe(H₂O)O₃₉, (TBA)₄PW₁₁Mn(H₂O)O₃₉ and (TBA)₄HPW₁₁Co(H₂O)O₃₉ can be fabricated on the surface of a glassy carbon electrode by the droplet evaporation methodology. These chemically modified electrodes were stable and their preparation was reproducible and easy to perform. The electrochemical features of the immobilized polyanions were different from those of the corresponding soluble species, namely in what concerns the peak potential values. The effect of the scan rate and of pH on the voltammetric features led to the conclusion that the first W reduction process for all immobilized polyanions was diffusion-controlled. For TBA-PW₁₁, TBA-PW₁₁Fe and TBA-PW₁₁Co the two-electron reductions at the first W waves are accompanied by the uptake of protons (2 H⁺ for the PW₁₁ anion and 4 H⁺ for the Fe-substituted and Co-substituted species). For the PW₁₁Mn-modified electrode, the reduction at the first W wave was a 1 e/2 H⁺ process. Additionally, the results obtained in the presence of Na₂SO₄ in the solution highlighted the role of the ions in the supporting electrolyte in the redox features of the immobilized hybrid phosphotungstates.     

Co Single-Atom Catalysts Boost Chemiluminescence

Co single-atom catalysts (SACs) with good aqueous solubility and abundant labelling functional groups were prepared in Co/Fe bimetallic metal-organic frameworks by a facile solvothermal method without high-temperature calcination. In contrast to traditional chemiluminescence (CL) catalysts, Co SACs accelerated decomposition of H₂O₂ to produce a large amount of singlet oxygen (¹O₂) rather than superoxide (O₂^.−) and hydroxyl radical (OH^.). They were found to dramatically enhance the CL emission of the luminol-H₂O₂ reaction by 1349 times, and, therefore, were employed as very sensitive signal probes for conducting CL immunoassay of cardiac troponin I. The detection limit of the target analyte was as low as 3.3 pg mL⁻¹. It is the first time that employment of SACs for boosting CL reactions has been validated. The Co SACs can also be employed to trace other biorecognition events with high sensitivity.