6-Amino-4-aryl-7-phenyl-3-(phenylimino)-4,7-dihydro-3H-[1,2]dithiolo[3,4-b]pyridine-5-carboxamides: Synthesis, Biological Activity, Quantum Chemical Studies and In Silico Docking Studies.

New [1,2]dithiolo[3,4-b]pyridine-5-carboxamides were synthesized through the reaction of dithiomalondianilide (N,N'-diphenyldithiomalondiamide) with 3-aryl-2-cyanoacrylamides or via a three-component reaction involving aromatic aldehydes, cyanoacetamide and dithiomalondianilide in the presence of morpholine. The structure of 6-amino-4-(2,4-dichloro- phenyl)-7-phenyl-3-(phenylimino)-4,7-dihydro-3H-[1,2]dithiolo[3,4-b]pyridine-5-carboxamide was confirmed using X-ray crystallography. To understand the reaction mechanism in detail, density functional theory (DFT) calculations were performed with a Grimme B97-3c composite computational scheme. The results revealed that the rate-limiting step is a cyclization process leading to the closure of the 1,4-dihydropyridine ring, with an activation barrier of 28.8 kcal/mol. Some of the dithiolo[3,4-b]pyridines exhibited moderate herbicide safening effects against 2,4-D. Additionally, ADMET (Absorption, Distribution, Metabolism, Excretion, Toxicity) parameters were calculated and molecular docking studies were performed to identify potential protein targets.

Cyanoacetohydrazide as a Novel Derivatization Agent for the Determination of UHPLC-HRMS Steroids in Urine.

The possibility of cyanoacetohydrazide usage as a novel derivatizing agent is demonstrated in the presented article, and a comparison with hydroxylamine as the most commonly used reagent is provided. Optimal conditions for steroid derivatization with cyanoacetohydrazide are provided. According to the collected data, the maximum yield of derivatives was observed at pH 2.8 within 70 min at 40 °C with 5 ng/mL limit of detection for all investigated analytes. It was shown that cyanoacetohydrazide derivatives produces both syn- and anti-forms as well as hydroxylamine, and their ratios were evaluated and shown in presented work. An efficiency enchantment from two to up to five times was achieved with a novel derivatization reagent. Its applicability for qualitative analysis of steroids in urine was presented at real samples. Additionally, the reproducible fragmentation of the derivatizing agent in collision-induced dissociation offers opportunities for simplified non-targeted steroidomic screening. Furthermore, cyanoacetohydrazide increases ionization efficiency in positive mode, which can eliminate the need for redundant high-resolution instrument runs required for both positive and negative mode analyses.

Selectivity of Exhaled Breath Biomarkers of Lung Cancer in Relation to Cancer of Other Localizations.

Lung cancer is a leading cause of death worldwide, mostly due to diagnostics in the advanced stage. Therefore, the development of a quick, simple, and non-invasive diagnostic tool to identify cancer is essential. However, the creation of a reliable diagnostic tool is possible only in case of selectivity to other diseases, particularly, cancer of other localizations. This paper is devoted to the study of the variability of exhaled breath samples among patients with lung cancer and cancer of other localizations, such as esophageal, breast, colorectal, kidney, stomach, prostate, cervix, and skin. For this, gas chromatography-mass spectrometry (GC-MS) was used. Two classification models were built. The first model separated patients with lung cancer and cancer of other localizations. The second model classified patients with lung, esophageal, breast, colorectal, and kidney cancer. Mann-Whitney U tests and Kruskal-Wallis H tests were applied to identify differences in investigated groups. Discriminant analysis (DA), gradient-boosted decision trees (GBDT), and artificial neural networks (ANN) were applied to create the models. In the case of classifying lung cancer and cancer of other localizations, average sensitivity and specificity were 68% and 69%, respectively. However, the accuracy of classifying groups of patients with lung, esophageal, breast, colorectal, and kidney cancer was poor.

Phthalylglycyl Chloride as a Derivatization Agent for UHPLC-MS/MS Determination of Adrenaline, Dopamine and Octopamine in Urine.

Dopamine, adrenaline and octopamine are small polar molecules that play a vital role in regulatory systems. In this paper, phthalylglycyl chloride was proposed as a derivatization agent for octopamine, adrenaline and dopamine determination in urine for the first time. The derivatization procedure facilitated the use of reversed-phase liquid chromatography with positive electrospray ionization-high-resolution mass spectrometry. An LC-HRMS method was developed that provided quantification limits of 5 ng/mL and detection limits of 1.5 ng/mL for all analytes. The 95-97% yield of derivates was observed after a 10 min derivatization with phthalylglycyl chloride at pH 6.5 and 30 °C. The proposed method was successfully applied to the analysis of human urine samples. The obtained results were compared with those of conventional derivatization procedures with 9-fluorenyl-methoxycarbonyl chloride and dansyl chloride.

New 6'-Amino-5'-cyano-2-oxo-1,2-dihydro-1'H-spiro[indole-3,4'-pyridine]-3'-carboxamides: Synthesis, Reactions, Molecular Docking Studies and Biological Activity.

The purpose of this work was to prepare new isatin- and monothiomalondiamide-based indole derivatives, as well as to study the properties of the new compounds. The four-component reaction of 5-R-isatins (R = H, CH3), malononitrile, monothiomalonamide (3-amino-3-thioxo- propanamide) and triethylamine in hot EtOH yields a mixture of isomeric triethylammonium 6'-amino-3'-(aminocarbonyl)-5'-cyano-2-oxo-1,2-dihydro-1'H- and 6'-amino-3'-(aminocarbonyl)- 5'-cyano-2-oxo-1,2-dihydro-3'H-spiro[indole-3,4'-pyridine]-2'-thiolates. The reactivity and structure of the products was studied. We found that oxidation of spiro[indole-3,4'-pyridine]-2'-thiolates with DMSO-HCl system produced only acidification products, diastereomeric 6'-amino-5'-cyano-5-methyl-2-oxo-2'-thioxo-1,2,2',3'-tetrahydro-1'H-spiro-[indole-3,4'-pyridine]- 3'-carboxamides, instead of the expected isothiazolopyridines. The alkylation of the prepared spiro[indole-3,4'-pyridine]-2'-thiolates upon treatment with N-aryl α-chloroacetamides and α-bromoacetophenones proceeds in a regioselective way at the sulfur atom. In the case of α-bromoacetophenones, ring-chain tautomerism was observed for the S-alkylation products. According to NMR data, the compounds consist of a mixture of stereoisomers of 2'-amino-6'-[(2-aryl-2-oxoethyl)thio]-3'-cyano-2-oxo-1'H-spiro[indoline-3,4'-pyridine]-5'-carboxamides and 5'-amino-3'-aryl-6'-cyano-3'-hydroxy-2-oxo-2',3'-dihydrospiro[indoline-3,7'-thiazolo[3,2-a]pyridine]-8'-carboxamides in various ratios. The structure of the synthesized compounds was confirmed by IR spectroscopy, HRMS, 1H and 13C DEPTQ NMR studies and the results of 2D NMR experiments (1H-13C HSQC, 1H-13C HMBC). Molecular docking studies were performed to investigate suitable binding modes of some new compounds with respect to the transcriptional regulator protein PqsR of Pseudomonas aeruginosa. The docking studies revealed that the compounds have affinity for the bacterial regulator protein PqsR of Pseudomonas aeruginosa with a binding energy in the range of -5.8 to -8.2 kcal/mol. In addition, one of the new compounds, 2'-amino-3'-cyano-5-methyl-2-oxo-6'-{[2-oxo-2-(p-tolylamino)ethyl]thio}-1'H-spiro-[indoline-3,4'-pyridine]-5'-carboxamide, showed in vitro moderate antibacterial effect against Pseudomonas aeruginosa and good antioxidant properties in a test with 1,1-diphenyl-2-picrylhydrazyl radical. Finally, three of the new compounds were recognized as moderately active herbicide safeners with respect to herbicide 2,4-D in the laboratory experiments on sunflower seedlings.

Alkyl 4-Aryl-6-amino-7- phenyl-3-(phenylimino)-4,7-dihydro- 3H-[1,2]dithiolo[3,4-b]pyridine-5-carboxylates: Synthesis and Agrochemical Studies.

The reaction between dithiomalondianilide (N,N'-diphenyldithiomalondiamide) and alkyl 3-aryl-2-cyanoacrylates in the presence of morpholine in the air atmosphere leads to the formation of alkyl 6-amino-4-aryl-7-phenyl-3-(phenylimino)-4,7-dihydro-3H-[1,2]dithiolo[3,4-b]- pyridine-5-carboxylates in 37-72% yields. The same compounds were prepared in 23-65% yields by ternary condensation of aromatic aldehydes, ethyl(methyl) cyanoacetate and dithiomalondianilide. The reaction mechanism is discussed. The structure of ethyl 6-amino-4-(4-methoxyphenyl)-7-phenyl-3-(phenylimino)-4,7-dihydro-3H-[1,2]dithiolo[3,4-b]pyridine-5-carboxylate was confirmed by X-ray crystallography. Two of the prepared compounds showed a moderate growth-stimulating effect on sunflower seedlings. Three of the new compounds were recognized as strong herbicide safeners with respect to herbicide 2,4-D in the laboratory and field experiments on sunflower.

GC-MS/MS Determination of Steroid Hormones in Urine Using Solid-Phase Derivatization as an Alternative to Conventional Methods.

Solid-phase analytical derivatization (SPAD) is a promising hybrid sample preparation technique combining the clean-up and preconcentration of the sample in a single step. In this work, a novel SPAD method based on the preparation of trimethylsilyl (TMS) derivatives of steroid hormones (testosterone, estrone, DHT, estriol, estradiol, and progesterone) in Phenomenex Strata C18-E (100 mg, 1 mL) cartridges has been developed and applied for their GC-MS/MS determination in human urine samples. The proposed procedure allows the detection and quantification of steroids with limits of 1.0-2.5 and 2.5-5 ng/mL, respectively. These characteristics are comparable with those obtained with a conventional liquid-liquid extraction, while the recovery of analytes in the proposed SPAD procedure is higher. The major advantages of SPAD are a short derivatization time, high efficiency, and the possibility to automatize the procedure. However, its cost-effectiveness in routine practice is still questionable.

Accessing Perfluoroaryl Sulfonimidamides and Sulfoximines via Photogenerated Perfluoroaryl Nitrenes: Synthesis and Application as a Chiral Auxiliary.

Sulfonimidamides (SIAs) and sulfoximines (SOIs) have attracted attention due to their potential in agriculture and in medicinal chemistry as bioisosteres of biologically active compounds, and new synthetic methods are needed to access and explore these compounds. Herein, we present a light-promoted generation of perfluorinated aromatic nitrenes, from perfluorinated azides, that subsequently are allowed to react with sulfinamides and sulfoxides, generating achiral and chiral SIAs and SOIs. One of the enantiopure SIAs was evaluated as a novel chiral auxiliary in Grignard additions to the imines yielding the product in up to 96:4 diastereomeric ratio.

A novel approach to the quantification of urinary aryl-propionamide-derived SARMs by UHPLC-MS/MS.

A simple and sensitive procedure for the quantification of two commonly abused aryl-propionamide-derived selective androgen receptor modulators (SARMs), namely S-4 (GTx-007, andarine) and S-22 (GTx-024, MK-2866, ostarine, enobosarm), has been described. Urine samples were prepared for analysis by means of a dispersive liquid-liquid microextraction using methanol and chloroform as dispersive and extracting solvents, respectively. Factors that might influence the extraction process as well as their optimum conditions were evaluated by Box-Benken and central composite designs. After extraction, the analytes were quantified by UHPLC-MS/MS. The proposed procedure was validated on human urine samples. As a result, for both SARMs the detection limits were observed at 0.05 ng/mL and calibration curves were linear in the concentration range of 0.25-50 ng/mL with the coefficient of determination of 0.998.

Diagnosis of Lung Cancer Through Exhaled Breath: A Comprehensive Study.

OBJECTIVES: Exhaled breath analysis is an attractive lung cancer diagnostic tool. However, various factors that are not related to the disease status, comorbidities, and other diseases must be considered to obtain a reliable diagnostic model. METHODS: Exhaled breath samples from 646 individuals including 273 patients with lung cancer (LC), 90 patients with cancer of other localizations (OC), 150 patients with noncancer lung diseases (NLD), and 133 healthy controls (HC) were analyzed using gas chromatography-mass spectrometry (GC-MS). The samples were collected in Tedlar bags. Volatile organic compounds (VOCs) were preconcentrated on Tenax TA sorbent tubes with subsequent two-stage thermal desorption followed by GC-MS analysis. The influence of age, gender, smoking status, time since last food consumption, and comorbidities on exhaled breath were evaluated. Also, the effect of histology, TNM, tumor localization, treatment status, and the presence of a tumor on VOC profile of patients with lung cancer were assessed. Intergroup statistics were estimated, diagnostic models were created using artificial neural networks (ANNs) and gradient boosted decision trees (GBDTs). RESULTS: Smoking status and food consumption affect exhaled breath VOC profile: benzene, ethylbenzene, toluene, 1,3-pentadiene 1,4-pentadiene acetonitrile, and some ratios are significantly different in exhaled breath of smokers and nonsmokers; the ratios 2,3-butandione/2-pentanone, 2,3-butandione/dimethylsulfide, and 2-butanone/2-pentanone are affected by time since last food consumption. Exhaled breath of LC is affected by the form of the disease and comorbidities. One-pentanol and 2-butanone were different in exhaled breath of patients with various tumor localization; 2-butanone was different in exhaled breath of patients before and during treatment. Diabetes as a comorbidity affects the pentanal level in exhaled breath; obesity affects the ratios of 2,3-butanedione/dimethylsulfide and 2-butanone/isoprene. Sensitivity and specificity of diagnostic models aimed to discriminate LC and HC, OC, and NLD were 78.7% and 51.0%, 62.2% and 53.4%, and 60.4% and 58.0%, respectively. HC and patients, regardless of the disease, can be classified with sensitivity of 76.6% and specificity of 68.2%. CONCLUSIONS: The models created to diagnose lung cancer can also classify OC and NLD as patients with lung cancer. Additionally, the influence of comorbidities and factors not related to the disease status must be considered before the creation of diagnostic models to avoid false results.

Quantification of cancer biomarkers in urine using volatilomic approach.

Urine analysis is an attractive approach for non-invasive cancer diagnostics. In this study, a procedure for the determination of volatile organic compounds (VOCs) in human urine (acetone, acetonitrile, dimethylsulfide, dimethyl disulfide, dimethyl trisulfide, hexane, benzene, toluene, 2-butanone, 2-pentanone, pentanal) has been described including sample preparation using preconcentration of analytes in sorbent tubes followed by gas chromatography with mass spectrometry (GC-MS). Fractional factorial design and constrained surfaces design were used to optimize preconcentration of VOCs in sorbent tubes. The procedure was validated by analysis of synthetic urine containing VOC standards in the concentration range of 1-5000 ng/mL. Optimized procedure was applied to analyze urine samples of 89 healthy volunteers and 85 patients with cancer of various localizations: 42 patients with lung cancer, 25 - colon, 3 - stomach, 2 - prostate, 2 - esophageal, 2 - pancreas, 2 - kidney, 1 - ovarian, 1 - cervical, 1 - skin, 1 - liver. Concentrations of 2-butanone, 2-pentanone, acetonitrile, and benzene were found different in urine of patients with cancer and healthy individuals. Influence of cancer localization and tumor, nodule, metastasis stage on urine VOC profile was considered. The approach of using ratios of VOCs to the main ones instead of concentrations was considered. A diagnostic model based on significantly different VOC ratios was created to classify healthy individuals and patients with cancer using artificial neural network (ANN). The model was validated during construction by means of 3-fold cross-validation. Average area under receiver operating characteristic (ROC) curve on test dataset was 0.886. Average sensitivity and specificity of the created model were 91 % and 82 %.

Correction to "Non-invasive Exhaled Breath and Skin Analysis to Diagnose Lung Cancer: Study of Age Effect on Diagnostic Accuracy".

[This corrects the article DOI: 10.1021/acsomega.2c06132.].

Non-Invasive Lung Cancer Diagnostics through Metabolites in Exhaled Breath: Influence of the Disease Variability and Comorbidities.

Non-invasive, simple, and fast tests for lung cancer diagnostics are one of the urgent needs for clinical practice. The work describes the results of exhaled breath analysis of 112 lung cancer patients and 120 healthy individuals using gas chromatography-mass spectrometry (GC-MS). Volatile organic compound (VOC) peak areas and their ratios were considered for data analysis. VOC profiles of patients with various histological types, tumor localization, TNM stage, and treatment status were considered. The effect of non-pulmonary comorbidities (chronic heart failure, hypertension, anemia, acute cerebrovascular accident, obesity, diabetes) on exhaled breath composition of lung cancer patients was studied for the first time. Significant correlations between some VOC peak areas and their ratios and these factors were found. Diagnostic models were created using gradient boosted decision trees (GBDT) and artificial neural network (ANN). The performance of developed models was compared. ANN model was the most accurate: 82-88% sensitivity and 80-86% specificity on the test data.

Non-invasive Exhaled Breath and Skin Analysis to Diagnose Lung Cancer: Study of Age Effect on Diagnostic Accuracy.

Development of simple, fast, and non-invasive tests for lung cancer diagnostics is essential for clinical practice. In this paper, exhaled breath and skin were studied as potential objects to diagnose lung cancer. The influence of age on the performance of diagnostic models was studied. Gas chromatography in combination with mass spectrometry (MS) was used to analyze the exhaled breath of 110 lung cancer patients and 212 healthy individuals of various ages. Peak area ratios of volatile organic compounds (VOCs) were used for data analysis instead of VOC peak areas. Various machine learning algorithms were applied to create diagnostic models, and their performance was compared. The best results on the test data set were achieved using artificial neural networks (ANNs): classification of patients with lung cancer and young healthy volunteers: 88 ± 4% sensitivity and 83 ± 3% specificity; classification of patients with lung cancer and old healthy individuals: 81 ± 3% sensitivity and 85 ± 1% specificity. The difference between performance of models based on young and old healthy groups was minor. The results obtained have shown that metabolic dysregulation driven by the disease biology is too high, which significantly overlaps the age effect. The influence of tumor localization and histological type on exhaled breath samples of lung cancer patients was studied. Statistically significant differences between some parameters in these samples were observed. A possibility of assessing the disease status by skin analysis in the Zakharyin-Ged zones using an electronic nose based on the quartz crystal microbalance sensor system was evaluated. Diagnostic models created using ANNs allow us to classify the skin composition of patients with lung cancer and healthy subjects of different ages with a sensitivity of 69 ± 2% and a specificity of 68 ± 8% for the young healthy group and a sensitivity of 74 ± 7% and a specificity of 66 ± 6% for the old healthy group. Primary results of skin analysis in the Zakharyin-Ged zones for the lung cancer diagnosis have shown its utility, but further investigation is required to confirm the results obtained.

Highly congested spiro-compounds via photoredox-mediated dearomative annulation cascade.

Photo-mediated radical dearomatization involving 5-exo-trig cyclizations has proven to be an important route to accessing spirocyclic compounds, whereas 6-exo-trig spirocyclization has been much less explored. In this work, a dearomative annulation cascade is realized through photoredox-mediated C-O bond activation of aromatic carboxylic acids to produce two kinds of spirocyclic frameworks. Mechanistically, the acyl radical is formed through oxidation of triphenylphosphine and subsequent C-O bond cleavage, followed by a 6-exo-trig cyclization/SET/protonation sequence to generate the spiro-chromanone products in an intramolecular manner. Furthermore, the protocol was extended to more challenging intermolecular tandem sequences consisting of C-O bond cleavage, radical addition to an alkene substrate, and 5-exo-trig cyclization to yield complex spirocyclic lactams.

Study of confounding factors influence on lung cancer diagnostics effectiveness using gas chromatography-mass spectrometry analysis of exhaled breath.

Aim: The purpose of this study was to estimate volatile organic compounds (VOCs) ability to distinguish exhaled breath samples of lung cancer patients and healthy volunteers and to assess the effect of smoking status and gender on parameters. Patients & methods: Exhaled breath samples of 40 lung cancer patients and 40 healthy individuals were analyzed using gas chromatography-mass spectrometry. Influence of other factors on the exhaled breath VOCs profile was investigated. Results: Some parameters correlating with the disease status were affected by other factors. Excluding these parameters allows creating a logistic regression diagnostic model with 83% sensitivity and 81% specificity. Conclusion: Influence of other factors on the exhaled breath VOCs profile has to be taken into account to avoid misleading results.

Exhaled breath analysis using GC-MS and an electronic nose for lung cancer diagnostics.

Exhaled breath analysis is an interesting and promising approach for the diagnostics of various diseases. Being non-invasive, convenient and simple, this approach has tremendous potential utility for further translation into clinical practice. In this study, gas chromatography-mass spectrometry (GC-MS) and quartz microbalance sensor-based "electronic nose" were applied for analysis of the exhaled breath of 40 lung cancer patients and 40 healthy individuals. It was found that the electronic nose was unable to distinguish the samples of different groups. However, the application of GC-MS allowed identifying statistically significant differences in compound peak areas and their ratios for investigated groups. Diagnostic models were created using random forest classifier based on peak areas and their ratios with the sensitivity and specificity of peak areas (ratios) of 85.7-96.5% (75.0-93.1%) and 73.3-85.1% (90.0-92.5%) on training data and 63.6-75.0% (72.7-100.0%) and 50.0-69.2% (76.9-84.6%) on test data, respectively. The exhaled breath samples of lung cancer patients and healthy volunteers could be distinguished by GC-MS with the use of individual compounds, but application of their ratios could help to determine specific differences between investigated groups and the level the influence of individual metabolism features alternating from one person to another as well as daily instrument reproducibility deviations. The electronic nose has to be significantly improved to apply it to lung cancer diagnostics of exhaled breath analysis and the influence of water vapour has to be lowered to increase the sensitivity of the sensors to detect lung cancer biomarkers.

Stereoselective synthesis of unnatural α-amino acid derivatives through photoredox catalysis.

A protocol for stereoselective C-radical addition to a chiral glyoxylate-derived N-sulfinyl imine was developed through visible light-promoted photoredox catalysis, providing a convenient method for the synthesis of unnatural α-amino acids. The developed protocol allows the use of ubiquitous carboxylic acids as radical precursors without prior derivatization. The protocol utilizes near-stoichiometric amounts of the imine and the acid radical precursor in combination with a catalytic amount of an organic acridinium-based photocatalyst. Alternative mechanisms for the developed transformation are discussed and corroborated by experimental and computational studies.

Quantification of steroid hormones in human urine by DLLME and UHPLC-HRMS detection.

A procedure for the quantification of steroid hormones of various classes in human urine (androgens, estrogens, progestins, corticosteroids) has been described consisting of sample preparation by means of dispersive liquid-liquid extraction after enzymatic hydrolysis with ß-glucuronidase from E. Coli followed by ultra-high performance liquid chromatography-high resolution mass spectrometry (quadrupole time-of-flight) detection. Both one-variable-at-a-time and multivariate approaches (full factorial and Box-Behnken designs) were applied to optimize sample preparation conditions. The procedure was validated using synthetic urine in the concentration range of 0.25-500 ng/mL. Then, it was applied to the analysis of real urine samples and the results were compared with those of a common liquid-liquid extraction procedure. The results obtained proved its applicability to the quantification of steroid hormones in human urine with high sensitivity and accuracy.

Investigation of different approaches for exhaled breath and tumor tissue analyses to identify lung cancer biomarkers.

Development of early noninvasive methods for lung cancer diagnosis is among the most promising technologies, especially using exhaled breath as an object of analysis. Simple sample collection combined with easy and quick sample preparation, as well as the long-term stability of the samples, make it an ideal choice for routine analysis. The conditions of exhaled breath analysis by preconcentrating volatile organic compounds (VOCs) in sorbent tubes, two-stage thermal desorption and gas-chromatographic determination with flame-ionization detection have been optimized. These conditions were applied to estimate differences in exhaled breath VOC profiles of lung cancer patients and healthy volunteers. The combination of statistical methods was used to evaluate the ability of VOCs and their ratios to classify lung cancer patients and healthy volunteers. The performance of diagnostic models on the test data set was greater than 90 % for both VOC peak areas and their ratios. Some of the exhaled breath samples were analyzed using gas chromatography coupled with mass spectrometry (GC-MS) to identify VOCs present in exhaled breath at lower concentration levels. To confirm the endogenous origin of VOCs found in exhaled breath, GC-MS analysis of tumor tissues was conducted. Some of the VOCs identified in exhaled breath were found in tumor tissues, but their frequency of occurrence was significantly lower than in the case of exhaled breath.