Metabolism of the tryptamine‐derived new psychoactive substances 5‐MeO‐2‐Me‐DALT, 5‐MeO‐2‐Me‐ALCHT,and 5‐MeO‐2‐Me‐DIPT and their detectability in urine studied by GC–MS,LC–MSn,and LC‐HR‐MS/MS

Many N,N‐dialkylated tryptamines show psychoactive properties and were encountered as new psychoactive substances. The aims of the presented work were to study the phase I and II metabolism and the detectability in standard urine screening approaches (SUSA) of 5‐methoxy‐2‐methyl‐N,N‐diallyltryptamine (5‐MeO‐2‐Me‐DALT), 5‐methoxy‐2‐methyl‐N‐allyl‐N‐cyclohexyltryptamine (5‐MeO‐2‐Me‐ALCHT), and 5‐methoxy‐2‐methyl‐N,N‐diisopropyltryptamine (5‐MeO‐2‐Me‐DIPT) using gas chromatography–mass spectrometry (GC–MS), liquid chromatography coupled with multistage accurate mass spectrometry (LC–MSⁿ), and liquid chromatography‐high‐resolution tandem mass spectrometry (LC‐HR‐MS/MS). For metabolism studies, urine was collected over a 24 h period after administration of the compounds to male Wistar rats at 20 mg/kg body weight (BW). Phase I and II metabolites were identified after urine precipitation with acetonitrile by LC‐HR‐MS/MS. 5‐MeO‐2‐Me‐DALT (24 phase I and 12 phase II metabolites), 5‐MeO‐2‐Me‐ALCHT (24 phase I and 14 phase II metabolites), and 5‐MeO‐2‐Me‐DIPT (20 phase I and 11 phase II metabolites) were mainly metabolized by O‐demethylation, hydroxylation, N‐dealkylation, and combinations of them as well as by glucuronidation and sulfation of phase I metabolites. Incubations with mixtures of pooled human liver microsomes and cytosols (pHLM and pHLC) confirmed that the main metabolic reactions in humans and rats might be identical. Furthermore, initial CYP activity screenings revealed that CYP1A2, CYP2C19, CYP2D6, and CYP3A4 were involved in hydroxylation, CYP2C19 and CYP2D6 in O‐demethylation, and CYP2C19, CYP2D6, and CYP3A4 in N‐dealkylation. For SUSAs, GC–MS, LC‐MSⁿ, and LC‐HR‐MS/MS were applied to rat urine samples after 1 or 0.1 mg/kg BW doses, respectively. In contrast to the GC–MS SUSA, both LC–MS SUSAs were able to detect an intake of 5‐MeO‐2‐Me‐ALCHT and 5‐MeO‐2‐Me‐DIPT via their metabolites following 1 mg/kg BW administrations and 5‐MeO‐2‐Me‐DALT following 0.1 mg/kg BW dosage. Copyright © 2017 John Wiley & Sons, Ltd.     

Identification and analytical characterization of six synthetic cannabinoids NNL‐3, 5F–NPB‐22‐7N, 5F–AKB‐48‐7N, 5F–EDMB‐PINACA,EMB‐FUBINACA,and EG‐018

Clinical and forensic toxicology laboratories are continuously confronted by analytical challenges when dealing with the new psychoactive substances phenomenon. The number of synthetic cannabinoids, the chemical diversity, and the speed of emergence make this group of compounds particularly challenging in terms of detection, monitoring, and responding. Three indazole 7N positional isomer synthetic cannabinoids, two ethyl 2‐amino‐3‐methylbutanoate‐type synthetic cannabinoids, and one 9H –carbazole substituted synthetic cannabinoid were identified in seized materials. These six synthetic cannabinoid derivatives included: 1H –benzo[d ] [1,2,3]triazol‐1‐yl 1‐(5‐fluoropentyl)‐1H –pyrrolo[2,3‐b ]pyridine‐3‐carboxylate (NNL‐3, 1 ), quinolin‐8‐yl 1‐(5‐fluoropentyl)‐1H –pyrrolo[2,3‐b ]pyridine‐3‐carboxylate (5F–NPB‐22‐7N , 2 ), N ‐((1 s,3 s)‐adamantan‐1‐yl)‐1‐(5‐fluoropentyl)‐1H –pyrrolo[2,3‐b ]pyridine‐3‐carboxamide (5F–AKB‐48‐7N , 3 ), ethyl 2‐(1‐(5‐fluoropentyl)‐1H –indazole‐3‐carboxamido)‐3,3‐dimethylbutanoate (5F–EDMB‐PINACA, 4 ), ethyl 2‐(1‐(4‐fluorobenzyl)‐1H –indazole‐3‐carboxamido)‐3‐methylbutanoate (EMB‐FUBINACA, 5 ), and naphthalen‐1‐yl(9‐pentyl‐9H ‐carbazol‐3‐yl)methanone (EG‐018, 6 ). The identification was based on ultra‐high‐performance liquid chromatography‐quadrupole time‐of‐flight‐mass spectrometry (UHPLC‐QTOF‐MS), gas chromatography–mass spectrometry (GC–MS), and nuclear magnetic resonance spectroscopy (NMR). The analytical characterization of these six synthetic cannabinoids was described, so as to assist forensic laboratories in identifying these compounds or other substances with similar structure in their case work. To our knowledge, no analytical data about the compounds 1 – 5 have appeared until now, making this the first report on these compounds. The GC–MS data of 6 has been reported, but this study added the LC–MS, NMR, and Fourier transform infrared (FTIR), data to render the analytical data collection process more complete. Copyright © 2017 John Wiley & Sons, Ltd.     

Design,synthesis, and evaluation of the anticancer activity of 2‐amino‐aryl‐7‐aryl‐benzoxazole compounds

A series of 2‐amino‐aryl‐7‐aryl‐benzoxazole derivatives have been designed, synthesized, and evaluated as anticancer agents. Fourteen of the compounds exhibited cytotoxic effects toward human A549 lung cancer cells. We found 12l was the most potent with an EC₅₀ of 0.4 μm , equivalent to the anticancer drug doxorubicin, but had low selectivity following cross screening in monkey kidney Vero cells. Eight of the most potent or most selective compounds were further profiled in additional cell lines (MCF7, NCI‐H187, and KB) to better understand their cytotoxic activity. Only compound 12l had a measurable EC₅₀ in a single cell line (3.3 μm in the KB cell line). Taken together, this data suggest the series as a whole display specific cytotoxicity toward A549 cells. Cheminformatics searches pointed to JAK2 as a possible target. A subset of compounds assayed at this target showed IC₅₀s ranging from 10 to 0.08 μm ; however, no clear correlation between JAK2 potency and A549 cytotoxicity was observed.     

Identification of five substituted phenethylamine derivatives 5‐MAPDB, 5‐AEDB,MDMA methylene homolog, 6‐Br‐MDMA,and 5‐APB‐NBOMe

This paper reports analytical properties of five substituted phenethylamine derivatives seized from a clandestine laboratory. These five derivatives include 5‐(2‐methylaminopropyl)‐2,3‐dihydrobenzofuran (5‐MAPDB, 1 ), 5‐(2‐aminoethyl)‐2,3‐dihydrobenzofuran (5‐AEDB, 2 ), N ,2‐dimethyl‐3‐(3,4‐methylenedioxyphenyl)propan‐1‐amine (MDMA methylene homolog, 3 ), 6‐bromo‐3,4‐methylenedioxymethamphetamine (6‐Br‐MDMA, 4 ), and 1‐(benzofuran‐5‐yl)‐N ‐(2‐methoxybenzyl)propan‐2‐amine (5‐APB‐NBOMe, 5 ). These compounds were identified by liquid chromatography‐quadrupole time‐of‐flight mass spectrometry (LC‐QTOF‐MS), gas chromatography‐mass spectrometry (GC‐MS), and nuclear magnetic resonance spectroscopy (NMR). No analytical properties about compounds 1‐4 have appeared until now, making this the first report on these compounds. Copyright © 2016 John Wiley & Sons, Ltd.     

Design,Synthesis, and In‐Vivo Pharmacological Screening of N,3‐(Substituted Diphenyl)‐5‐phenyl‐1H‐pyrazoline‐1‐carbothioamide Derivatives

Various 3,5‐(substituted diphenyl)‐4,5‐dihydro‐pyrazole‐1‐carbothioic acid phenylamides were synthesized starting from substituted acetophenones. Structures of the compounds were confirmed on the basis of spectral data. The compounds were evaluated for their anticonvulsant and antidepressant activity. Interestingly, out of 26 compounds, four ( 3f , 3g , 3t , and 3u ) were found to protect 100% of the animals in the MES screen at a dose of 25 mg/kg. They were also found to have appreciable anticonvulsant activity in scPTZ screen. Two compounds, 3j and 3o , significantly reduced the duration of the immobility time at 25 mg/kg dose, when compared to control.     

Design,Synthesis, and Antitumor Activity of (E,Z)‐1‐(dihydrobenzofuran‐5‐yl)‐3‐phenyl‐2‐(1,2,4‐triazol‐1‐yl)‐2‐propen‐1‐ones

A series of (E,Z)‐1‐(dihydrobenzofuran‐5‐yl)‐3‐phenyl‐2‐(1,2,4‐triazol‐1‐yl)‐2‐propen‐1‐ones ( C1 – C35 ) were designed and synthesized, and the structures of compounds (Z)‐ C27 and (Z)‐ C29 were confirmed by single‐crystal X‐ray diffraction. The antitumor activities of these novel compounds against cervical cancer (HeLa), lung cancer (A549), and breast cancer (MCF‐7) cell lines were evaluated in vitro. Majority of the title compounds exhibited strong antitumor activities and were much more promising than the positive control Taxol, which were also accompanied by lower cytotoxicity to normal cells. In particular, compounds (E,Z)‐ C24 exhibited the most consistent potent activities against three neoplastic cells with IC₅₀ values ranging from 3.2 to 7.1 μm . Further researches demonstrated that compounds (E,Z)‐ C24 could induce cell apoptosis and arrest cell cycle at the G2/M and S phases. Meanwhile, the structure–activity relationship between the configurations and cytotoxicity of the compounds was also investigated.     

Synthesis,anticonvulsant, and antinociceptive activity of new 3‐(3‐methyl‐2,5‐dioxo‐3‐phenylpyrrolidin‐1‐yl)propanamides and 3‐phenyl‐butanamides

A focused library of new 3‐(3‐methyl‐2,5‐dioxo‐3‐phenylpyrrolidin‐1‐yl)propanamides and their nonimide analogs were synthesized and tested for anticonvulsant activity. These compounds were obtained through the coupling reaction of the starting carboxylic acids with appropriate amines. The initial anticonvulsant screening was performed in mice (intraperitoneal administration) using the maximal electroshock seizure (MES) and the subcutaneous pentylenetetrazole (scPTZ) seizure models. The most promising compound 6 showed more potent protection in the MES and scPTZ tests than valproic acid, which is still recognized as one of the most relevant first‐line anticonvulsants. The structure–activity relationship analysis revealed that the presence of the pyrrolidine‐2,5‐dione ring is important but not indispensable to retain anticonvulsant activity. Additionally, compound 6 showed potent antinociceptive properties in the oxaliplatin‐induced neuropathic pain model in mice. The most plausible mechanism of action for compound 6 may result from its influence on the neuronal sodium channel (Site 2) and the high‐voltage‐activated L‐type calcium channel.     

Design,Synthesis, and Potential Antidepressant‐like Activity of 7‐prenyloxy‐2,3‐dihydroflavanone Derivatives

A series of 7‐prenyloxy‐2, 3‐dihydroflavanone derivatives were synthesized and screened for their antidepressant‐like activity. Among them, it was observed that compounds 5j and 5k were found to be the most antidepressant‐like activity. In addition, it was found that compounds 5j and 5k significantly increased the concentrations of the main neurotransmitters 5‐HT and NE in the hippocampus, hypothalamus, and cortex. Compounds 5j and 5k also significantly increased the contents of 5‐HIAA in the hippocampus and cortex, shut down 5‐HT metabolism compared with mice treated with stress vehicle. These results suggested that compounds 5j and 5k displayed potent antidepressant‐like properties that were mediated via neurochemical systems.     

Attenuation of the LPS‐induced,ERK‐mediated upregulation of fibrosis‐related factors FGF‐2, uPA,MMP‐2, and MMP‐9 by Carthamus tinctorius L in cardiomyoblasts

Severe and potentially fatal hypotension and cardiac contractile dysfunction are common symptoms in patients with sepsis. LPS was previously found to dramatically upregulate expression of fibrosis‐related factors FGF‐2, uPA, MMP‐2, and MMP‐9 in primary cardiac fibroblasts. MMPs are capable of denaturing and degrading fibrillar collagens and other components of the extracellular matrix (ECM). Studies have shown that dysregulation of expression of MMPs is associated with development of myocardial extracellular matrix remodeling and cardiac fibrosis, which contribute to progression of heart failure. In this study, H9c2 cells and cardiac fibroblasts were divided into five treatment groups: control, LPS (1 μg/mL) and three concentrations of FC_EtOH (Carthami Flos ethanolic extract) (31.25, 62.5, and 125 μg/mL). Phosphorylation of ERK‐1/2 was observed to be rapidly induced upon treatment with LPS. In contrast, it was significantly suppressed by the administration of FC_EtOH (125 μg/mL). Effects of FC_EtOH on LPS‐induced MMP‐2 and MMP‐9 expression in H9c2 cells occurred directly through ERK1/2 were determined. H9c2 cells were therefore pretreated with EGF‐R to activate ERK pathway. Both protein levels of MMP‐2 and MMP‐9 and immunefluorescent signals of MMP‐9 were significantly enhanced by EGFR. In contrast, MMP‐2 and MMP‐9 were significantly reduced after FC_EtOH administration. Based on these findings, the authors concluded that FC_EtOH elicits a protective effect against LPS‐induced cardio‐fibrosis through the ERK1/2 pathway. Carthamus tinctorius L may potentially serve as a cardio‐protective agent against LPS‐ induced cardiac fibrosis. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 754–763, 2017.     

A 12‐week,randomized, parallel‐group,proof‐of‐concept study of tulobuterol patch and salmeterol inhaler as add‐on therapy in adult‐onset mild‐to‐moderate asthma

Patch formulation of tulobuterol has been used in asthma treatment as a long‐acting β₂‐agonist (LABA) through sustained skin absorption. Its treatment efficacy, especially in small airways, remains poorly understood. The study aim was to investigate LABA add‐on effects of tulobuterol patch (TP) and salmeterol inhaler (SA) on pulmonary function, asthma control and health status. Patients who had adult‐onset under‐control asthma, despite taking inhaled corticosteroids, were enrolled in a randomized, open‐label, parallel‐group, proof‐of‐concept study of 12‐week add‐on treatment with TP (n=16) or SA (n=17). Spirometry, impulse oscillometry (IOS), exhaled nitric oxide levels, and clinical questionnaires of asthma control, health status (St. George's Respiratory Questionnaire: SGRQ), and symptoms were evaluated every 4 weeks. Add‐on treatment of SA significantly improved the spirometric indices of small airway obstruction (forced expiratory flow between 25% and 75% of FVC: FEF_25‐75, and maximum expiratory flow at 25% of FVC: MEF₂₅) and IOS indices of whole respiratory resistance (resistance at 5 Hz) as compared to TP. In intra‐group comparisons, add‐on treatment of TP improved the scores of the asthma control test and the total SGRQ, as well as the symptom and impact components of the SGRQ. SA add‐on treatment improved FEV₁ and IOS parameters of resistance at 20 Hz and reactance at 5 Hz. Neither of the treatments improved exhaled nitric oxide levels. In conclusion, add‐on treatment of TP improved asthma control and health status, whereas SA improved pulmonary function measures associated with large and small airway involvement among patients with adult‐onset mild‐to‐moderate asthma.     

Synthesis of empagliflozin,a novel and selective sodium‐glucose co‐transporter‐2 inhibitor,labeled with carbon‐14 and carbon‐13

Empagliflozin, (2S,3R,4R,5S,6R)‐2‐[4‐chloro‐3‐[[4‐[(3S)‐oxolan‐3‐yl]oxyphenyl]methyl]phenyl]‐6‐(hydroxymethyl)oxane‐3,4,5‐triol was recently approved by the FDA for the treatment of chronic type 2 diabetes mellitus. Herein, we report the synthesis of carbon‐13 and carbon‐14 labeled empagliflozin. Carbon‐13 labeled empagliflozin was prepared in five steps and in 34% overall chemical yield starting from the commercially available α‐D‐glucose‐[¹³C₆]. For the radiosynthesis, the carbon‐14 atom was introduced in three different positions of the molecule. In the first synthesis, Carbon‐14 D‐(+)‐gluconic acid δ‐lactone was used to prepare specifically labeled empagliflozin in carbon‐1 of the sugar moiety in four steps and in 19% overall radiochemical yield. Carbon‐14 labeled empagliflozin with the radioactive atom in the benzylic position was obtained in eight steps and in 7% overall radiochemical yield. In the last synthesis carbon‐14 uniformly labeled phenol was used to give [¹⁴C]empagliflozin in eight steps and in 18% overall radiochemical yield. In all these radiosyntheses, the specific activities of the final compounds were higher than 53 mCi/mmol, and the radiochemical purities were above 98.5%.     

Synthesis,In Vitro Protoporphyrinogen Oxidase Inhibition,and Herbicidal Activity of N‐(Benzothiazol‐5‐yl)hexahydro‐1H‐isoindole‐1,3‐diones and N‐(Benzothiazol‐5‐yl)hexahydro‐1H‐isoindol‐1‐ones

Protoporphyrinogen oxidase ( EC 1.3.3.4 ) is one of the most significant targets for a large family of herbicides. As part of our continuous efforts to search for novel protoporphyrinogen oxidase‐inhibiting herbicides, N‐(benzothiazol‐5‐yl)tetrahydroisoindole‐1,3‐dione was selected as a lead compound for structural optimization, leading to the syntheses of a series of novel N‐(benzothiazol‐5‐yl)hexahydro‐1H‐isoindole‐1,3‐diones ( 1a – o ) and N‐(benzothiazol‐5‐yl)hexahydro‐1H‐isoindol‐1‐ones ( 2a – i ). These newly prepared compounds were characterized by elemental analyses, ¹H NMR, and ESI‐MS, and the structures of 1h and 2h were further confirmed by X‐ray diffraction analyses. The bioassays indicated that some compounds displayed comparable or higher protoporphyrinogen oxidase inhibition activities in comparison with the commercial control. Very promising, compound 2a , ethyl 2‐((6‐fluoro‐5‐(4,5,6,7‐tetrahydro‐1‐oxo‐1H‐isoindol‐2(3H)‐yl)benzo[d]thiazol‐2‐yl)‐sulfanyl)acetate, was recognized as the most potent candidate with K_i value of 0.0091 μm . Further greenhouse screening results demonstrated that some compounds exhibited good herbicidal activity against Chenopodium album at the dosage of 150 g/ha.     

Synthesis,characterization, and antimicrobial activity of some new N‐aryl‐N’‐(2‐oxoindolin‐3‐ylidene)‐benzohydrazonamides

A green approach was developed for synthesizing a series of (isatin‐3‐ylidene)‐hydrazonamides 3a–j  from the reaction between isatin, (isatin‐3‐ylidene)malononitrile, or 2‐cyano‐2‐(2‐isatin‐3‐ylidene)acetate and benzohydrazonamide in ethyl acetate solutions at ambient temperature. The structures of the new compounds were confirmed on the basis of spectral data. In this eco‐friendly medium, a variety of (isatin‐3‐ylidene)hydrazonamides were obtained free of catalyst in good to excellent yields. All the synthesized products were evaluated for their antimicrobial activity. Among the compounds tested, 3b and 3d exhibited good antibacterial activity against Staphylococcus aureus, whereas others responded moderately with reference to the standard drug ciprofloxacin.     

Design,Synthesis, and Biological Evaluation of Novel 4‐Aminopiperidinyl‐linked 3,5‐Disubstituted‐1,2,6‐thiadiazine‐1,1‐dione Derivatives as HIV‐1 NNRTIs

Based on the hybridization of the privileged fragments in DABO and DAPY‐typed HIV‐1 NNRTIs, a novel series of 4‐aminopiperidinyl‐linked 3,5‐disubstituted‐1,2,6‐thiadiazine‐1,1‐dione derivatives were designed, synthesized, and evaluated for their in vitro anti‐HIV activities in MT‐4 cells. Most of the target compounds showed weak inhibitory activity against WT HIV‐1. In order to confirm the mode of action of the target compounds, representative compounds Ba8 and Bb8 were selected to perform the HIV‐1 RT inhibitory assay. In this assay, Ba8 and Bb8 displayed good activity with IC₅₀ values of 3.15 and 1.52 μm , respectively. Additionally, preliminary structure–activity relationships (SARs) analysis and molecular docking studies of newly synthesized compounds are also discussed.     

Synthesis,molecular modeling,and biological evaluation of 4‐[5‐aryl‐3‐(thiophen‐2‐yl)‐4,5‐dihydro‐1H‐pyrazol‐1‐yl] benzenesulfonamides toward acetylcholinesterase,carbonic anhydrase I and II enzymes

In this study, 4‐[5‐aryl‐3‐(thiophen‐2‐yl)‐4,5‐dihydro‐1H‐pyrazol‐1‐yl] benzenesulfonamides were synthesized, and inhibition effects on AChE, hCA I, and hCA II were evaluated. K_i values of the compounds toward hCA I were in the range of 24.2 ± 4.6‐49.8 ± 12.8 nm , while they were in the range of 37.3 ± 9.0‐65.3 ± 16.7 nm toward hCA II. K_i values of the acetazolamide were 282.1 ± 19.7 nm and 103.60 ± 27.6 nm toward both isoenzymes, respectively. The compounds inhibited AChE with K_i in the range of 22.7 ± 10.3‐109.1 ± 27.0 nm , whereas the tacrine had K_i value of 66.5 ± 13.8 nm . Electronic structure calculations at M06‐L/6‐31 + G(d,p)//AM1 level and molecular docking studies were also performed to enlighten inhibition mechanism and to support experimental findings. Results obtained from calculations of molecular properties showed that the compounds obey drug‐likeness properties. The experimental and computational findings obtained in this study might be useful in the design of novel inhibitors against hCA I, hCA II, and AChE.