Synthesis, anti-microbial, toxicity and molecular docking studies of N-nitroso-N-phenylhydroxylamine (cupferron) and its derivatives.

Bacterial resistance to antimicrobial agents is increasing at an alarming rate globally and requires new lead compounds for antibiotics. In this study, N-phenyl-N-nitroso hydroxylamine (cupferron) and its derivatives have been synthesised using readily available starting materials. The compounds were obtained in high yield and purity. They show activity towards a range of Gram-positive and Gram-negative pathogenic bacteria, with minimum inhibitory concentration (MIC) values as low as 2 µg.mL-1 against the tested organisms, especially for Gram-positive species. Toxicity studies on the lead compound 3b indicate insignificant effects on healthy cell lines. Molecular docking studies on the lead compound identify possible binding modes of the compound, and the results obtained correlate with those of in vitro and MIC studies. The lead compound shows excellent drug-likeness properties.

Identification of precursors and mechanisms of tobacco-specific nitrosamine formation in water during chloramination.

We report here that tobacco-specific nitrosamines (TSNAs) are produced from specific tobacco alkaloids during water chloramination. To identify the specific precursors for the formation of specific TSNAs in drinking water, we have developed a solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) method for simultaneous determination of five TSNAs and three tobacco alkaloids. Using this method, we detected nicotine (NIC) at 15.1 ng/L in a source water. Chloramination of this source water resulted in the formation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) (0.05 ng/L) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) (0.2 ng/L) along with the reduction of NIC to 1.1 ng/L, suggesting that NNK and NNAL were formed from NIC. To confirm that tobacco alkaloids are the precursors of TSNAs, we chloraminated water-leaching samples of tobacco from three brands of cigarettes and found that the formation of TSNAs coincides with the reduction of the alkaloids. Chloramination of individual alkaloids confirms that NNK and NNAL are produced from NIC, N-nitrosonornicotine (NNN) from nornicotine (NOR), and N-nitrosoanabasine (NAB) from anabasine (ANA). Furthermore, we have identified specific intermediates of these reactions and proposed potential pathways of formation of TSNAs from specific alkaloids. These results confirm that NNK and NNAL are the disinfection byproducts (DBPs) resulting from NIC in raw water.

Transnitrosylation directs TRPA1 selectivity in N-nitrosamine activators.

S-Nitrosylation, the addition of a nitrosyl group to cysteine thiols, regulates various protein functions to mediate nitric oxide (NO) bioactivity. Recent studies have demonstrated that selectivity in protein S-nitrosylation signaling pathways is conferred through transnitrosylation, a transfer of the NO group, between proteins via interaction. We previously demonstrated that sensitivity to activation by synthetic NO-releasing agents via S-nitrosylation is a common feature of members of the transient receptor potential (TRP) family of Ca(2+)-permeable cation channels. However, strategies to confer subtype selectivity to nitrosylating agents targeted to TRP channels are yet to be developed. Here, we show selective activation of TRPA1 channels by novel NO donors derived from the ABBH (7-azabenzobicyclo[2.2.1]heptane) N-nitrosamines, which exhibit transnitrosylation reactivity to thiols without releasing NO. The NNO-ABBH1 (N-nitroso-2-exo,3-exo-ditrifluoromethyl-7-azabenzobicyclo[2.2.1]heptane) elicits S-nitrosylation of TRPA1 proteins, and dose-dependently induces robust Ca(2+) influx via both recombinant and native TRPA1 channels, but not via other NO-activated TRP channels. TRPA1 activation by NNO-ABBH1 is suppressed by specific cysteine mutations but not by NO scavenging, suggesting that cysteine transnitrosylation underlies the activation of TRPA1 by NNO-ABBH1. This is supported by the correlation of N-NO bond reactivity and TRPA1-activating potency in a congeneric series of ABBH N-nitrosamines. Interestingly, nonelectrophilic derivatives of ABBH also activate TRPA1 selectively, but less potently, compared with NNO-ABBH1. Thus, ABBH N-nitrosamines confer subtype selectivity on S-nitrosylation in TRP channels through synergetic effects of two chemical processes: cysteine transnitrosylation and molecular recognition of the nonelectrophilic moiety.

Nitrosamine formation in amine scrubbing at desorber temperatures.

Amine scrubbing is a thermodynamically efficient and industrially proven method for carbon capture, but amine solvents can nitrosate in the desorber, forming potentially carcinogenic nitrosamines. The kinetics of reactions involving nitrite and monoethanolamine (MEA), diethanolamine (DEA), methylethanolamine (MMEA), and methyldiethanolamine (MDEA) were determined under desorber conditions. The nitrosations of MEA, DEA, and MMEA are first order in nitrite, carbamate species, and hydronium ion. Nitrosation of MDEA, a tertiary amine, is not catalyzed by the addition of CO2 since it cannot form a stable carbamate. Concentrated and CO2 loaded MEA was blended with low concentrations of N-(2-hydroxyethyl) glycine (HeGly), hydroxyethyl-ethylenediamine (HEEDA), and DEA, secondary amines common in MEA degradation. Nitrosamine yield was proportional to the concentration of secondary amine and was a function of CO2 loading and temperature. Blends of tertiary amines with piperazine (PZ) showed n-nitrosopiperazine (MNPZ) yields close to unity, validating the slow nitrosation rates hypothesized for tertiary amines. These results provide a useful tool for estimating nitrosamine accumulation over a range of amine solvents.

Effects of flue gas compositions on nitrosamine and nitramine formation in postcombustion CO2 capture systems.

Amine-based technologies are emerging as the prime contender for postcombustion CO2 capture. However, concerns have arisen over the health impacts of amine-based CO2 capture associated with the release of nitrosamines and nitramines, which are byproducts from the reactions between flue gas NOx and solvent amines. In this study, flue gas compositions were systematically varied to evaluate their effects on the formation of nitrosamines and nitramines in a lab-scale CO2 capture reactor with morpholine as a model solvent amine. The accumulation of N-nitrosomorpholine in both the absorber and washwater increased linearly with both NO and NO2 for concentrations up to ∼20 ppmv. These correlations could be extrapolated to estimate N-nitrosomorpholine accumulation at extremely low NOx levels (0.3 ppmv NO2 and 1.5 ppmv NO). NO played a particularly important role in driving N-nitrosomorpholine formation in the washwater, likely following partial oxidation to NO2 by O2. The accumulation of N-nitromorpholine in both the absorber and washwater positively correlated with flue gas NO2 concentration, but not with NO concentration. Both N-nitrosomorpholine and N-nitromorpholine accumulated fastest in the absence of CO2. Flue gas humidity did not affect nitrosamine accumulation in either the absorber or the washwater unit. These results provide a basis for estimating the effects of flue gas composition on nitrosamine and nitramine accumulation in postcombustion CO2 capture systems.

Efficient, scalable and economical preparation of tris(deuterium)- and 13C-labelled N-methyl-N-nitroso-p-toluenesulfonamide (Diazald®) and their conversion to labelled diazomethane.

A method for the preparation of multi-gramme quantities of N-methyl-d3-N-nitroso-p-toluenesulfonamide (Diazald-d3) and N-methyl-(13)C-N-nitroso-p-toluenesulfonamide (Diazald-(13)C) and their conversion to diazomethane-d2 and diazomethane-(13) C, respectively, is presented. This approach uses robust and reliable chemistry, and critically, employs readily commercially available and inexpensive methanol as the label source. Several reactions of labelled diazomethane are also reported, including alkene cyclopropanation, phenol methylation and α-diazoketone formation, as well as deuterium scrambling in the preparation of diazomethane-d2 and subsequent methyl esterification of benzoic acid.

Occurrence and fate of N-nitrosamines and their formation potential in three wastewater treatment plants in Japan.

N-nitrosamines are well known as carcinogens present in the environment. However, studies of the occurrence and fate of N-nitrosamines and their N-nitrosamine formation potential (FP) in wastewater treatment plants (WWTPs) are lacking. Therefore, the objectives of this study were to determine the occurrence of N-nitrosamines in WWTPs, the FP of N-nitrosamines on chloramination during wastewater treatment, and the efficiency of FPNH2Cl reduction by biological treatment. Also, the residual FPNH2Cl in the final discharge was investigated. The efficiencies of removal of N-nitrosamines ranged from 35 to 94% (WWTP O; residential area), from 58 to 98% (WWTP E; industrial area), and from 58% to >99% (WWTP N; industrial area). In WWTP O, the rates of production of N-nitrosodimethylamine (NDMA) (42%) and NDBA (58%) were the highest. In WWTP E, only NDBA (34%) was produced. In WWTP N, NPYR and NPIP (both >99%) were produced. NDMA FPNH2Cl values were very high in influent in all WWTPs. The efficiencies of removal of N-nitrosamine FPNH2Cl during biological treatment ranged from 20% to >99%, but there is no obvious explanation for this variability. Residual N-nitrosamine FPNH2Cl ranged from 2 to 22 ng L(-1). Thus, N-nitrosamines could be produced in water purification or reclamation plants using discharge from WWTPs.

Nitromethane with IBX/TBAF as a nitrosating agent: synthesis of nitrosamines from secondary or tertiary amines under mild conditions.

Aliphatic or aromatic N,N-disubstituted nitrosamine was generated in fair to excellent yield from the reaction of a secondary or tertiary amine with o-iodoxybenzoic acid (IBX) or o-iodosylbenzoic acid (IBA)/R(4)NX (X = halide) and nitromethane. The product yield was strongly influenced by both the halide of R(4)NX and iodanes. IBX gave a higher yield than IBA, while the halides follow F(-) > Cl(-) > Br(-) ∼ I(-). Nitrous acid formed in situ from nitromethane and IBX (or IBA)/halides is likely responsible for the observed reaction.

N-nitrosamines formation from secondary amines by nitrogen fixation on the surface of activated carbon.

Our previous study demonstrated that many commercial activated carbon (AC) particles may catalyze transformation of secondary amines to yield trace levels of N-nitrosamines under ambient aerobic conditions. Because of the widespread usage of AC materials in numerous analytical and environmental applications, it is imperative to understand the reaction mechanism responsible for formation of nitrosamine on the surface of ACs to minimize their occurrence in water treatment systems and during analytical methods employing ACs. The study results show that the AC-catalyzed nitrosamine formation requires both atmospheric oxygen and nitrogen. AC's surface reactive sites react with molecular oxygen to form reactive oxygen species (ROS), which facilitate fixation of molecular nitrogen on the carbon surfaces to generate reactive nitrogen species (RNS) likely nitrous oxide and hydroxylamine that can react with adsorbed amines to form nitrosamines. AC's properties play a crucial role as more nitrosamine formation is associated with carbon surfaces with higher surface area, more surface defects, reduced surface properties, higher O(2) uptake capacity, and higher carbonyl group content. This study is a first of its kind on the nitrosamine formation mechanism involving nitrogen fixation on AC surfaces, and the information will be useful for minimization of nitrosamines in AC-based processes.

Identifying urotropine derivatives as co-donors of formaldehyde and nitric oxide for improving antitumor therapy.

A pharmacophore integration strategy was utilized to develop the first co-donor of formaldehyde and nitric oxide (FANO), composed of urotropine derived nitramine/nitrosamine. FANO simultaneously generated formaldehyde and nitric oxide on-demand, resulting in synergistic anticancer effects. Importantly, liposomal formulation of FANO effectively inhibited tumor growth with minimal side-effects, providing a potent combined nitric oxide therapy for malignancy.

Celecoxib prodrugs possessing a diazen-1-ium-1,2-diolate nitric oxide donor moiety: synthesis, biological evaluation and nitric oxide release studies.

A new class of anti-inflammatory (AI) cupferron prodrugs was synthesized wherein a diazen-1-ium-1,2-diolato ammonium salt, and its O(2)-methyl and O(2)-acetoxyethyl derivatives, nitric oxide (NO) donor moieties were attached directly to an aryl carbon on a celecoxib template. The percentage of NO released from the O(2)-methyl and O(2)-acetoxyethyl compounds was higher (18.0-37.8% of the theoretical maximal release of one molecule of NO/molecule of the parent compound) upon incubation in the presence of rat serum, relative to incubation with phosphate buffer saline (PBS) at pH 7.4 (3.8-11.6% range). All compounds exhibited weak inhibition of the COX-1 isozyme (IC(50)=5.8-17.0 microM range) in conjunction with weak or modest inhibition of the COX-2 isozyme (IC(50)=1.6-14.4 microM range). The most potent AI agent 5-[4-(O(2)-ammonium diazen-1-ium-1,2-diolato)phenyl]-1-(4-sulfamoylphenyl)-3-trifluoromethyl-1H-pyrazole exhibited a potency that was about fourfold and twofold greater than that observed for the respective reference drugs aspirin and ibuprofen. These studies indicate that use of a cupferron template constitutes a plausible drug design approach targeted toward the development of AI drugs that do not cause gastric irritation, or elevate blood pressure and induce platelet aggregation that have been associated with the use of some selective COX-2 inhibitors.

[Investigation on relationship between flavonoid content and inhibitory ability to nitrosamine synthesis for eleven Chinese herbal medicines].

The eleven Chinese herbal medicine containing flavonoids are applied as raw materials to explore the relationship between the inhibitory ratio of nitrosamine synthesis, the scavenging ratio of nitrite and the flavonoid content in the samples. The inhibitory ratio of nitrosamine synthesis and the scavenging ratio of nitrite of the 11 herbal medicines, Vit C and rutin were determined in intro compare with Vit C and the standard ample of rutin. The results indicate that each sample exhibits certain ability to inhibitiory nitrosamine synthesis. Among these samples, Honeysuckle flower is found to be of best effects, its inhibitory ratio and scavenging ratio reaches 78.5% and 60.5%, respectively. Except kudzuvine root, the other samples with higher content of flavonoid result in a higher inhibitory or scavenging ratio, and the relative coefficient reaches a value of 0.9338 and 0.9272, respectively, displaying notable positive correlation. The concentrations of IC50 (g x L(-1)) of flavonoid extracted from honeysuckle, rutin and VC were 0.013, 0.022 and 0.187, respectively. While the inhibitory ratio of synthesis of nitrosamines reached 50%, and those were 0.042, 0.024 and 0.041, respectively. While scavenging ratio of nitrite reaches 50%. The inhibitory ratio of synthesis of nitrosamine of flavonoids extracted from honeysuckle flower is higher than that of Vit C and rutin, and the scavenging ratio of nitrite is similar to that of Vit C.

An improved synthesis of (+/-)-N'-nitrosonornicotine 5'-acetate.

A concise and efficient route to (+/-)-N'-nitrosonornicotine 5'-acetate (NNN-5'-OAc), a stable precursor of the active metabolite 5'-hydroxy(+/-)-N'-nitrosonornicotine NNN-5'-OH is reported. The synthesis utilizes sulfinimine chemistry to give (+/-)-NNN-5'-OAc in 26% yield over 4 steps.

Efficient system for the preparation of [13N]labeled nitrosamines.

In the present Letter, a fast and reproducible method for the synthesis of N-[(13)N]nitrosamines is reported. The labeling strategy is based on trapping [(13)N]NO2- in an anion exchange resin. The reaction with secondary amines in the presence of Ph(3)P and Br(2) led to the formation of the desired nitrosamines in short reaction times (2 min) with excellent radiochemical conversion (>45%). Final radiotracers were obtained after purification in good radiochemical yields (>30%, decay corrected). Radiochemical purity was above 99% in all cases.

Removal Characteristics of N-Nitrosamines and Their Precursors by Pilot-Scale Integrated Membrane Systems for Water Reuse.

This study investigated the removal characteristics of N-Nitrosamines and their precursors at three pilot-scale water reclamation plants. These plants applies different integrated membrane systems: (1) microfiltration (MF)/nanofiltration (NF)/reverse osmosis (RO) membrane; (2) sand filtration/three-stage RO; and (3) ultrafiltration (UF)/NF and UF/RO. Variable removal of N-Nitrosodimethylamine (NDMA) by the RO processes could be attributed to membrane fouling and the feed water temperature. The effect of membrane fouling on N-Nitrosamine removal was extensively evaluated at one of the plants by conducting one month of operation and chemical cleaning of the RO element. Membrane fouling enhanced N-Nitrosamine removal by the pilot-scale RO process. This finding contributes to better understanding of the variable removal of NDMA by RO processes. This study also investigated the removal characteristics of N-Nitrosamine precursors. The NF and RO processes greatly reduced NDMA formation potential (FP), but the UF process had little effect. The contributions of MF, NF, and RO processes for reducing FPs of NDMA, N-Nitrosopyrrolidine and N-Nitrosodiethylamine were different, suggesting different size distributions of their precursors.

Formation of N-nitrosamines by micelle-catalysed nitrosation of aliphatic secondary amines.

N-Nitrosamines are an important class of potent human carcinogens and mutagens that can be present in water and wastewater. For instance, N-nitrosamines can be formed by reaction of nitrosating agents such as NO+ or N2O3 formed from nitrite under acidic conditions with secondary amine precursors by an acid-catalysed nitrosation pathway. This study investigates the catalytic effect of cationic and anionic micelles on the nitrosation of secondary aliphatic amines in the presence of nitrite at different pH values. The results of this study demonstrate that the nitrosation of hydrophobic secondary amines (e.g., dipropylamine and dibutylamine) by nitrite was significantly enhanced in the presence of micelles of the cationic surfactant cetyltrimethylammonium chloride whereas anionic micelles formed by sodium dodecylsulfate did not significantly enhance the formation of N-nitrosamines. Rate enhancements of up to 100-fold were observed for the formation of N-nitrosodibutylamine in the presence of cetyltrimethylammonium chloride. The magnitude of the catalytic effect of cationic micelles on the nitrosation reaction depended mainly of the hydrophobicity of the amine precursors (i.e., alkyl chain length), the stability and the charge of the micelles and pH. One important enhancement factor is the lowering of the pKa of the precursor alkylammonium ion due to the electrical potential at the micelle-water interface by up to ∼2.5 pH units. These results suggest that cationic micelle-forming surfactants might play a role in the formation of N-nitrosamines in wastewater, consumer products and in industrial processes using high concentrations of cationic surfactants.

Computational investigation of the nitrosation mechanism of piperazine in CO2 capture.

Quantum chemistry calculations and kinetic modeling were performed to investigate the nitrosation mechanism and kinetics of diamine piperazine (PZ), an alternative solvent for widely used monoethanolamine in postcombustion CO2 capture (PCCC), by two typical nitrosating agents, NO2- and N2O3, in the presence of CO2. Various PZ species and nitrosating agents formed by the reactions of PZ, NO2-, and N2O3 with CO2 were considered. The results indicated that the reactions of PZ species having NH group with N2O3 contribute the most to the formation of nitrosamines in the absorber unit of PCCC and follow a novel three-step nitrosation mechanism, which is initiated by the formation of a charge-transfer complex. The reactions of all PZ species with NO2- proceed more slowly than the reactions of PZ species with ONOCO2-, formed by the reaction of NO2- with CO2. Therefore, the reactions of PZ species with ONOCO2- contribute more to the formation of nitrosamines in the desorber unit of PCCC. In view of CO2 effect on the nitrosation reaction of PZ, the effect through the reaction of PZ with CO2 shows a completely different tendency for different nitrosating agents. More importantly, CO2 can greatly accelerate the nitrosation reactions of PZ by NO2- through the formation of ONOCO2- in the reaction of CO2 with NO2-. This work can help to better understand the nitrosation mechanism of diamines and in the search for efficient methods to prevent the formation of carcinogenic nitrosamines in CO2 capture unit.

Inhibitory effects of Orostachys japonicus extracts on the formation of N-nitrosodimethylamine.

In Korea, Orostachys japonicus has been used traditionally as a drug and health food. The aim of this study was to investigate possible inhibitory effects of O. japonicus extracts on the formation of N-nitrosodimethylamines (NDMA). Chloroform extraction was the most effective method for recovering the highest number of phenolic compounds and flavonoids; in these extracts the greatest nitrite-scavenging activity and inhibition of NDMA formation occurred at pH 2.5. The chloroform extract was separated into 10 fractions (J1-J10); fraction J4 inhibited NDMA formation by 90.1 +/- 0.4%. This fraction was then separated into five subfractions (J4-1-J4-5) using a silica gel column. Subfractions J4-2 [(+)-catechin] and J4-4 (3,4-dihdroxybenzoic acid) inhibited NDMA formation by 89.5 +/- 0.9 and 77.6 +/- 0.8%, respectively.

Origin in tobacco smoke of N'-nitrosonornicotine, a tobacco-specific carcinogen.

To evaluate risk factors and to approach methods of reduction of the carcinogenic potential of cigarette smoke, the transfer rate of N-nitrosonornicotine in a popular U.S. blended cigarette into mainstream smoke was quantitatively determined. The mean transfer rate was 11.3%; thus approximately 46% of the tobacco-specific carcinogen in the smoke came from the tobacco, and the remainder was synthesized during smoking.

Kinetics of nitrosamine formation in the presence of micelle-forming surfactants.

The nitrosation of dihexylamine by nitrite was catalyzed in the presence of micelles of the cationic surfactant decyltrimethylammonium bromide. Rate enhancements up to 800-fold were observed. The magnitude of the rate enhancements was dependent on the amine structure, particularly the alkyl chain length. Rate enhancements for nitrosation were also observed in the presence of micelles of phosphatidylcholine and Triton X-100. We present a mechanistic interpretation of these catalytic effects.