T-butyl Nitrite Research Articles

Tunable Key [3 + 2] and [2 + 1] Cycloaddition of Enaminones and α-Diazo Compounds for the Synthesis of Isomeric Isoxazoles: Metal-Controlled Selectivity

The three-component reactions of enaminones, α-diazo esters/ketones, and t-butyl nitrite (TBN) for the switchable synthesis of isomeric isoxazoles have been realized. The catalysis with Cu(II) salt provides 3,4-disubsituted isoxazoles via [3 + 2] cycloaddition. On the other hand, the catalysis of Ag(I) with identical substrates leads to isomeric isoxazoles with reversed C3 and C4 substitution based on a key [2 + 1] cycloaddition.

Nitric Oxide Release and Antibacterial Efficacy Analyses of S-Nitroso-N-Acetyl-Penicillamine Conjugated to Titanium Dioxide Nanoparticles.

Therapeutic agents can be linked to nanoparticles to fortify their selectivity and targeted delivery while impeding systemic toxicity and efficacy loss. Titanium dioxide nanoparticles (TiNPs) owe their rise in biomedical sciences to their versatile applicability, although the lack of inherent antibacterial properties limits its application and necessitates the addition of bactericidal agents along with TiNPs. Structural modifications can improve TiNP's antibacterial impact. The antibacterial efficacy of nitric oxide (NO) against a broad spectrum of bacterial strains is well established. For the first time, S-nitroso-N-acetylpenicillamine (SNAP), an NO donor molecule, was covalently immobilized on TiNPs to form the NO-releasing TiNP-SNAP nanoparticles. The TiNPs were silanized with 3-aminopropyl triethoxysilane, and N-acetyl-d-penicillamine was grafted to them via an amide bond. The nitrosation was carried out by t-butyl nitrite to conjugate the NO-rich SNAP moiety to the surface. The total NO immobilization was measured to be 127.55 ± 4.68 nmol mg-1 using the gold standard chemiluminescence NO analyzer. The NO payload can be released from the TiNP-SNAP under physiological conditions for up to 20 h. The TiNP-SNAP exhibited a concentration-dependent antimicrobial efficiency. At 5 mg mL-1, more than 99.99 and 99.70% reduction in viable Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria, respectively, were observed. No significant cytotoxicity was observed against 3T3 mouse fibroblast cells at all the test concentrations determined by the CCK-8 assay. TiNP-SNAP is a promising and versatile nanoparticle that can significantly impact the usage of TiNPs in a wide variety of applications, such as biomaterial coatings, tissue engineering scaffolds, or wound dressings.

Ex vivo evaluation of blood coagulation on endothelial glycocalyx-inspired surfaces using thromboelastography

Present blood-contacting materials have not yet demonstrated to be effective in reducing blood coagulation without causing additional side effects clinically. We have developed an endothelial glycocalyx-inspired biomimetic surface that combines nanotopography, heparin presentation, and nitric oxide (NO)-releasing features. The resulting modified surfaces have already shown promise in reducing unfavorable blood-material interactions using platelet-rich plasma. In this study, the efficacy of modified surfaces for reducing coagulation of human whole blood was measured. In addition, the effects of leached polysaccharides and chemical modification of the modified surfaces were evaluated. Leached polysaccharides in the incubation solution were detected by a refractive index method to determine the potential influences of these modified surfaces on the blood coagulation observation. Chemical modifications by the nitrosation process on the polysaccharides in the modified surfaces were detected using attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). Clot formation parameters were measured using thromboelastography (TEG), a clinically relevant technique to evaluate whole blood coagulation. No polysaccharides were detected in the heparinized polyelectrolyte multilayer-coated titania nanotube array surface (TiO2NT + PEM) incubation solution; however, polysaccharides were detected from NO-releasing TiO2NT + PEM surface (TiO2NT + PEM + NO) incubation solution both after the nitrosation process and after all NO was released. The structures of thiolated chitosan and heparin were altered by t-butyl nitrite. All heparin-containing surfaces were shown to slow or inhibit clot formation. This study is the first to evaluate these endothelial glycocalyx-inspired surfaces using clinically relevant parameters, as well as proposing potential influences of these modified surfaces on the inhibition of clot formation.

Site-selective and metal-free C\u2013H nitration of biologically relevant N-heterocycles

The site-selective and metal-free C–H nitration reaction of quinoxalinones and pyrazinones as biologically important N-heterocycles with t-butyl nitrite is described. A wide range of quinoxalinones were efficiently applied in this transformation, providing C7-nitrated quinoxalinones without undergoing C3-nitration. From the view of mechanistic point, the radical addition reaction exclusively occurred at the electron-rich aromatic region beyond electron-deficient N-heterocycle ring. This is a first report on the C7–H functionalization of quinoxalinones under metal-free conditions. In contrast, the nitration reaction readily takes place at the C3-position of pyrazinones. This transformation is characterized by the scale-up compatibility, mild reaction conditions, and excellent functional group tolerance. The applicability of the developed method is showcased by the selective reduction of NO2 functionality on the C7-nitrated quinoxalinone product, providing aniline derivatives. Combined mechanistic investigations aided the elucidation of a plausible reaction mechanism.

Synthesis of α-sulfonyl ketoximes through a salicylic acid-catalyzed four-component reaction involving radical sulfonylation followed by arylsulfonylation and oximation

A four-component reaction of styrene derivatives, anilines, t-butyl nitrite, and DABSO was used to prepare α-sulfonyl ketoximes. The reaction was realized through a salicylic acid-catalyzed aryl radial formation and ensuing sulfonylation by DABSO, followed by arylsulfonylation and oximation of styrenes. Under the optimized conditions, the title compounds were obtained in good yields within 30 min.

Site-selective C–H nitration of N-aryl-7-azaindoles under palladium(II) catalysis

The site-selective C–H nitration reaction of 7-azaindoles with t-butyl nitrite under palladium catalysis is described. This protocol provides an efficient method for the construction of ortho-nitrated N-aryl-7-azaindoles with excellent site-selectivity and functional group compatibility. The formed 7-azaindole derivatives can be readily transformed into 7-azaindoles containing an aniline functional group under palladium-catalyzed hydrogenation conditions.

Saccharin: an efficient organocatalyst for the one-pot synthesis of 4-amidocinnolines under metal and halogen-free conditions

A mild and efficient protocol for the one-pot synthesis of 4-amidocinnolines was developed using saccharin as a cheap and eco-friendly organocatalyst and t-butyl nitrite as a metal-free diazotization reagent. The current method has advantages such as cost effectiveness, use of the biodegradable reagents, simple experimental procedure; afford the quantitative yield of the desired products, metal- and halogen-free as well as environmentally benign conditions.

Fabrication and Characterization of a Nitric Oxide-Releasing Nanofibrous Gelatin Matrix

Nitric oxide (NO) plays an important role in cardiovascular homeostasis, immune responses, and wound repair. The pro-angiogenic and antimicrobial properties of NO has stimulated the development of NO-releasing materials for wound dressings. Gelatin, an abundant natural biodegradable polymer derived from collagen, is able to promote wound repair. S-Nitroso-N-acetylpenicillamine (SNAP) can release NO under physiological conditions and when exposed to light. The objective of this project was to fabricate a NO-releasing gelatin-based nanofibrous matrix with precise light-controllable ability. Results showed that under controlled phase separation fabrication conditions, the gelatin formed a highly porous matrix with the nanofiber diameter ranging from 50 to 500 nm. Importantly, the removal of the trace amount of divalent metal ions within gelatin generated a more stable nanofibrous structure. N-acetyl-D-penicillamine (NAP) was functionalized onto the matrix and nitrosated with t-butyl nitrite, yielding a SNAP-gelatin matrix. Analysis of the photoinitiated NO-release showed that the SNAP-gelatin matrices released NO in a highly controllable manner. Application of increasing light intensities yielded increased NO flux from the matrices. In addition, the dried matrices stored in dark at 4 °C maintained stable NO storage capacity, and the purified (ion-removed) gelatin preserved higher NO-releasing capacity than nonpurified gelatin. The antibacterial effect from the SNAP-gelatin matrices was demonstrated by exposing Staphylococcus aureus ( S. aureus ) to a light-triggered NO flux. This controllable NO-releasing scaffold provides a potential antibacterial therapeutic approach to combat drug resistant bacteria.

Kinetics of S-Nitrosation Processes in Aqueous Polymer Solution for Controlled Nitric Oxide Loading: Toward Tunable Biomaterials

An understanding of the nitrosation processes that dictate S-nitrosothiol formation in the presence of a polymer is crucial toward the controlled synthesis of nitric oxide (NO)-releasing materials, an important class of biomaterials that mimic the natural function of cells. Herein, the kinetics of S-nitrosoglutathione (GSNO) formation in the presence of dextran under a variety of nitrosation conditions, including the nitrosating agent and the dextran concentration, are reported. When comparing nitrous acid and t-butyl nitrite as the nitrosating agent, the use of nitrous acid results in 100% nitrosation of the thiol sites within less than a minute and t-butyl nitrite requires more than 5 min to reach completion. This trend establishes nitrous acid as a highly efficient nitrosating agent. In the presence of increasing dextran concentration from 0 w/v% to 10 w/v%, the extent of nitrosation decreases by approximately 5% and 30% using nitrous acid and t-butyl nitrite, respectively. With sufficient reaction time, either reagent leads to 100% nitrosation. This indicates that t-butyl nitrite is the preferred reagent for fine-tuned NO loading of thiol sites as the extent of reaction is greatly impacted by the polymer concentration. Taken together, these studies provide valuable insights regarding the ability to tailor NO storage within biomaterials for use in a wide range of clinical applications.

S-Nitrosated biodegradable polymers for biomedical applications: synthesis, characterization and impact of thiol structure on the physicochemical properties

A new class of nitric oxide (NO)-releasing biodegradable polymers has been synthesized by derivatizing poly(lactic-co-glycolic-co-hydroxymethyl propionic acid) (PLGH) polymers with structurally unique thiol functionalities followed by nitrosation with t-butyl nitrite to yield pendant S-nitrosothiol moieties. The extent of thiolation was found to be dependent on the thiol moiety itself with the efficiency of incorporation as follows: cysteamine > cysteine > homocysteine. Glutathione and penicillamine were not incorporated to any significant extent. The structure and polymer environment associated with the pendant thiol has been related to the physicochemical properties of the resulting polymers. To quantify the extent of S-nitrosation, chemiluminescence and UV-visible spectroscopy techniques were employed in combination. The cysteamine and homocysteine derivatives were found to have the highest extent of nitrosation at 93 ± 3% and 96 ± 3%, respectively, followed by 43 ± 1% for cysteine. Thermal decomposition led to near-complete recovery of NO based upon the quantification of the RSNO formation for each nitrosated polymer. Our ability to exert control over the thiol structure, extent of incorporation and the subsequent nitrosation is crucial to the resulting range of NO release kinetics that were yielded. The functional utility of these materials is demonstrated in that these non-toxic polymers release NO under physiological conditions, have degradation profiles that are appropriate for tissue scaffolds and can be prepared as electrospun nanofibers, commonly used in tissue and bone regeneration applications.

Development of Photoinitiated Nitric Oxide Releasing Polymer Films for Controlled Drug Delivery

Nitric Oxide (NO) is small, free radical gas that has been shown to have a wide variety of physiological functions, including the ability to hinder tumor angiogenesis at high, but non lethal, concentrations [1]. Previous work suggests that if NO could be effectively delivered in vivo to tumors of patients currently undergoing chemotherapy treatments at the appropriate levels, less damaging chemotherapy treatments could be used against cancer [2]. This could increase the overall survivability of cancer patients, especially in those prone to the harmful effects of chemotherapy: children, elderly, and those of weak immune systems. If NO is especially successful at preventing and eliminating tumor growth, angiogenesis, and carcinogenesis the need for stressful chemotherapy treatments could be eliminated altogether. This project is focused on developing novel photosensitive NO donors that can be incorporated into polymeric systems and used in a fiber optic drug delivery system. Development of these NO-releasing polymers will allow continued investigation of NO's role in tumor death by precisely controlling the surface flux of NO that cells are exposed to. Generating specific surface fluxes of NO from polymer films has been demonstrated by using polymer films that contain photoinitiated NO donors [3], prepared by synthesizing S-nitrosothiol (RSNO) derivitized polymer fillers that are blended into hydrophobic polymers and cast into a film. These films generate and sustain a surface flux of NO based on the wavelength and intensity of light used [3]. Polymers releasing NO are more promising as an NO donor than simply injecting NO into samples because they allow for spatial and temporal control of NO delivery. The specific concentration of NO needed to produce desirable effects on tumor cells (i.e., apoptosis) is not known. Data will be presented that show the synthesis and NO-release properties of novel RSNOs based on the nitrosation of benzyl mercaptan thiols. Specifically, UV-Vis spectrum of benzyl mercaptan in toluene and S-nitrosobenzyl mercaptan after the addition of t-butyl nitrite will be presented. We are currently investigating the effects of varying NO-surface fluxes generated from photolytic NO donating polymer films on aortic smooth muscle cell cultures obtained from mice. Once we have established that we can quantitatively determine the effects of different levels of NO on the proliferation of smooth muscle cell cultures, work will begin to apply this methodology and these novel NO-releasing polymeric systems to begin investigating what durations and surface fluxes of NO are necessary to have tumorcidal effects on specific cancer cells.

Development of Photoinitiated Nitric Oxide Releasing Polymer Films for Controlled Drug Delivery

Nitric Oxide (NO) is small, free radical gas that has been shown to have a wide variety of physiological functions, including the ability to hinder tumor angiogenesis at high, but non lethal, concentrations. Previous work suggests that if NO could be effectively delivered in vivo to tumors of patients currently undergoing chemotherapy treatments at the appropriate levels, less damaging chemotherapy treatments could be used against cancer. This could increase the overall survivability of cancer patients, especially in those prone to the harmful effects of chemotherapy: children, elderly, and those of weak immune systems. If NO is especially successful at preventing and eliminating tumor growth, angiogenesis, and carcinogenesis the need for stressful chemotherapy treatments could be eliminated altogether. This project is focused on developing novel photosensitive NO donors that can be incorporated into polymeric systems and used in a fiber optic drug delivery system. Development of these NO-releasing polymers will allow continued investigation of NO's role in tumor death by precisely controlling the surface flux of NO that cells are exposed to. Generating specific surface fluxes of NO from polymer films has been demonstrated by using polymer films that contain photoinitiated NO donors, prepared by synthesizing S-nitrosothiol (RSNO) derivitized polymer fillers that are blended into hydrophobic polymers and cast into a film. These films generate and sustain a surface flux of NO based on the wavelength and intensity of light used. Polymers releasing NO are more promising as an NO donor than simply injecting NO into samples because they allow for spatial and temporal control of NO delivery. The specific concentration of NO needed to produce desirable effects on tumor cells (i.e. apoptosis) is not known. Data will be presented that show the synthesis and NO-release properties of novel RSNOs based on the nitrosation of benzyl mercaptan thiols. Specifically, UV-Vis spectrum of benzyl mercaptan in toluene and S-nitrosobenzyl mercaptan after the addition of t-butyl nitrite will be presented. We are currently investigating the effects of varying NO-surface fluxes generated from photolytic NO donating polymer films on aortic smooth muscle cell cultures obtained from mice. Once we have established that we can quantitatively determine the effects of different levels of NO on the proliferation of smooth muscle cell cultures, work will begin to apply this methodology and these novel NO-releasing polymeric systems to begin investigating what durations and surface fluxes of NO are necessary to have tumorcidal effects on specific cancer cells.

FTIR and computational studies of gas-phase hydrogen atom abstraction kinetics by t-butoxy radical

By using Fourier-Transform Infrared (FTIR) absorption spectroscopy, rate coefficients in the range of 10 −16 to 10 −14 cm 3 molecule −1 s −1 have been determined for the hydrogen atom abstraction reactions of several substrates including halogenated organic compounds and amines by t-butoxy radical generated from the uv photolysis of t-butyl nitrite in the gas phase. Arrhenius parameters for selected reactions have been measured in the temperature range 299–318 K. Transition states and activation barriers for such reactions have been computed with the help of G aussian 03 software and found to match very well with the experimental values.

Competing rearrangement reactions in small gas-phase ionic complexes: The internal S N2 and nitro-nitrite rearrangements in nitroalkane proton-bound pairs

The dissociation of metastable proton-bound pairs, (R 1NO 2)(R 2NO 2)H + (R 1 and R 2 = CH 3, CH 3CH 2, (CH 3) 2CH and (CH 3) 3C) have been investigated by mass spectrometry and density functional theory calculations. The proton-bound pairs can dissociate via hydrogen-bond cleavage into protonated and neutral nitroalkanes. Methyl substitution of the nitroalkanes (R 1 and R 2 = (CH 3) 2CH, (CH 3) 3C) permits a rearrangement process to compete with the H-bond cleavage on the microsecond timescale. The rearrangement reaction results in an isomer that then loses nitrous acid and involves an internal S N2-type mechanism in which (R 1NO 2)(R 2NO 2)H + isomerizes to R 1NO 2⋯R 2NO 2H + via TS1 and then subsequently to R 1NO 2R 2 +⋯HONO via TS2 prior to dissociation. The process is favoured by stabilization of the charge in TS2 by methyl substitution. The stability of the t-butyl ion changes the mechanism in ((CH 3) 3CNO 2) 2H + to one that involves a two-step alkyl cation transfer. An investigation of nitro-nitrite rearrangement in protonated nitroalkanes at the B3-LYP/6-31 + G(d) level of theory found that the rearrangement barrier is lowered to the point that (CH 3) 3CNO 2H + can easily interconvert into (CH 3) 3CO(H)NO + in the gas phase and leads to the conclusion that the proton-bound pairs involving (CH 3) 3CNO 2 are a mixture of nitro-nitro and nitro-nitrite proton-bound pairs. The nitrite isomer can dissociate into protonated t-butyl nitrite and neutral nitroalkane via a simple hydrogen-bond cleavage. A more favourable competing dissociation process leads to the loss of t-butanol to form the ((CH 3) 3CNO 2)(NO) + complex.

A Specific Gas-Phase Substitution Reaction between Enol Radical Cations and t-Butyl Nitrite

Ion–molecule reactions between ionized carbonyl compounds or ionized enols with t-butyl nitrite allow a clear-cut distinction to be made between both these isomeric structures. The most relevant difference between the observed reactions is the formal substitution of a hydrogen atom by nitric oxide in the enol ions. Collisional activation experiments on the product ions are interpreted in terms of α-nitroso carbonyl structures. However, a quantum chemical investigation at the B3LYP/6-311++G(d,p) level of theory shows that ionized α-nitroso carbonyl and vinyl nitrite structures may isomerize provided they contain ca 60 kJ mol−1 of internal energy. It is, therefore, possible that both structures are generated in the substitution reaction.

Thermal Properties oft-Butyl Nitrite (TBN) on Cu(111)

Reactions of coadsorbed t-butoxy and NO, generated by thermal dissociation of t-butyl nitrite dosed onto Cu(111), were investigated using time-of-flight mass spectrometry, temperature-programmed desorption, and Auger electron spectroscopy. In the high-temperature regime above 500 K, t-butyl alcohol, isobutene, carbon oxides, and molecular nitrogen desorb. In the more complex regime below 500 K, nitrous oxide, molecular nitrogen, hydrazine, water, carbon oxides, and acetylene desorb. The proposed low-temperature reaction pathways involve rearrangement of adsorbed nitric oxide to form N2O and N2, reaction between adsorbed (CH3)3CO(a) and O(a) or N(a) to form a metallocyclic intermediate (1) and either adsorbed O(a)H or N(a)H. The intermediate (1) undergoes multiple C−C bond cleavages in a series of steps to form carbon oxides and adsorbed methylenes. The latter couple to form di-σ-bonded ethylene, which can either dehydrogenate to acetylene or be partially oxidized to acetate that dissociates in the high-te...

Mechanistic Studies on Nitrosation—Deaminocyclization of Mono‐Carbamoylated Vicinal Amino Alcohols and Diols: A New Preparative in situ Formation of Ethanediazo Hydroxide for the Ethylation of Carboxylates under Mild Conditions.

AbstractFor Abstract see ChemInform Abstract in Full Text.

Mechanistic Studies on Nitrosation–Deaminocyclization of Mono-Carbamoylated Vicinal Amino Alcohols and Diols: A New Preparative In Situ Formation of Ethanediazo Hydroxide for the Ethylation of Carboxylates under Mild Conditions

Abstract While the cyclization of N-carbamoylamino alcohols into oxazolidinones via the activation with NO+ underwent smoothly, we found that similar reactions of vicinal diol monocarbamates were very slow. Mechanistic studies by means of time-resolved IR measurements of the former reaction suggested that the initial O-nitrosation was the rate-determining step. Indeed, the introduction of an ethyl group on the nitrogen terminus of diol monocarbamate promoted the desired cyclic carbonate formation. The concomitantly formed ethanediazo hydroxide, the precursor of the protonated form of diazoethane, was evidenced by trapping with p-nitrobenzoic acid as an ethyl ester. The formation of ethyl ester accelerates the reaction in an irreversible manner. Based on an elaboration of the substrates and reaction conditions, 2,3-dimethyl-2,3-butanediol mono-N-ethyl-N-nitrosocarbamate, which is easily prepared in situ from the corresponding ethylcarbamate and t-butyl nitrite, was developed as a new ethylation reagent of various carboxylic acids under mild conditions.

Gas phase nitrosation of substituted benzenes

Using a combination of tandem mass spectrometric experiments (ion–molecule reactions, collisional activation, neutralization–reionization, MS/MS/MS) and theoretical calculations, protonated substituted benzenes are demonstrated to readily react with neutral t-butyl nitrite by the formation of stable complexes linking ionized nitric oxide to the benzene derivatives. The overall process is proposed to involve the concomitant elimination of neutral 2-methyl-2-propanol. Proton-bound dimers are proposed to intervene as the key-intermediates in these reactions, which also competitively produce protonated t-butyl nitrite. All the experiments were performed in a single hybrid tandem mass spectrometer of sector-quadrupole-sector configuration.

T-Butyl nitrite adsorbed on Ag([formula omitted]): thermally activated structure changes

Dosed on Ag(111) at 80 K, t-butyl nitrite ((CH3)3CONO, TBN) adsorbs and desorbs without dissociation but during subsequent thermal activation, reflection absorption infrared spectra, reveal significant structural changes within both the multilayer and monolayer. Vibrational peaks of TBN dosed at 80 K and of matrix-isolated TBN lie within 15 cm−1 of each other. Reflecting local ordering within the multilayer upon annealing to 110 K, the vibrational peaks sharpen and shift slightly. As the annealing temperature increases further, the NO stretch to C–H stretch intensity ratio drops dramatically revealing alignment changes with respect to the silver surface in both multilayer and monolayer. The relative intensities of other modes also change and, using symmetry assignments, we conclude that the ensemble average angle between the CONO plane of TBN and the Ag(111) surface decreases as the annealing temperature increases. The potential impact of such alignment changes on ejection of NO during photodissociation of TBN is discussed.