Insight into the mechanism of chlorinated nitroaromatic compounds anaerobic reduction with mackinawite (FeS) nanoparticles.

Anaerobic biotechnology for wastewaters treatment can nowadays be considered as state of the art methods. Nonetheless, this technology exhibits certain inherent limitations when employed for industrial wastewater treatment, encompassing elevated substrate consumption, diminished electron transfer efficiency, and compromised system stability. To address the above issues, increasing interest is being given to the potential of using conductive non-biological materials, e,g., iron sulfide (FeS), as a readily accessible electron donor and electron shuttle in the biological decontamination process. In this study, Mackinawite nanoparticles (FeS NPs) were studied for their ability to serve as electron donors for p-chloronitrobenzene (p-CNB) anaerobic reduction within a coupled system. This coupled system achieved an impressive p-CNB removal efficiency of 78.3 ± 2.9% at a FeS NPs dosage of 1 mg/L, surpassing the efficiencies of 62.1 ± 1.5% of abiotic and 30.6 ± 1.6% of biotic control systems, respectively. Notably, the coupled system exhibited exclusive formation of aniline (AN), indicating the partial dechlorination of p-CNB. The improvements observed in the coupled system were attributed to the increased activity in the electron transport system (ETS), which enhanced the sludge conductivity and nitroaromatic reductases activity. The analysis of equivalent electron donors confirmed that the S2- ions dominated the anaerobic reduction of p-CNB in the coupled system. However, the anaerobic reduction of p-CNB would be adversely inhibited when the FeS NPs dosage exceeded 5 g/L. In a continuous operation, the p-CNB concentration and HRT were optimized as 125 mg/L and 40 h, respectively, resulting in an outstanding p-CNB removal efficiency exceeding 94.0% after 160 days. During the anaerobic reduction process, as contributed by the predominant bacterium of Thiobacillus with a 6.6% relative abundance, a mass of p-chloroaniline (p-CAN) and AN were generated. Additionally, Desulfomonile was emerged with abundances ranging from 0.3 to 0.7%, which was also beneficial for the reduction of p-CNB to AN. The long-term stable performance of the coupled system highlighted that anaerobic technology mediated by FeS NPs has a promising potential for the treatment of wastewater containing chlorinated nitroaromatic compounds, especially without the aid of organic co-substrates.

Mechanism of quinone mediators modified polyurethane foam for enhanced nitrobenzene reduction and denitrification.

The nitrobenzene (NB) reduction and denitrification performance of the immobilized biofilm (I-BF) reactors based on 9,10-anthraquinone-2-sulfonyl chloride (ASC) modified polyurethane foam (PUF-ASC) carriers were investigated. Experiments demonstrated that the quinone mediators enhanced NB reduction and denitrification performance. The NB reduction rates increased by 1.46, while the NO3--N removal rates increased by 1.55 times in the PUF-0.1ASC system. The quinone mediators promote extracellular polymeric substances (EPS) secretion. Electrochemical tests indicated that quinone mediators enhanced the electron transfer of biofilm systems. NADH generation was accelerated and microbial electron transport system activity (ETSA) was promoted. The abundance of genera with electrochemical activity, NB degradation and denitrification ability (Pseudomonas sp., Diaphorobate sp., and Acinetobacter sp.) increased. Metabolic pathways relating to NO3--N and NB reduction were uploaded. In conclusion, electron acquisition by NO3--N and NB was facilitated, bacterial community structure and metabolic pathways were affected by the quinone mediators.

Photocatalytic performance of SiO2@TiO2 spheres in selective conversion of oxidation of benzyl alcohol to benzaldehyde and reduction of nitrobenzene to aniline.

Selective photocatalytic oxidation of benzyl alcohol to benzaldehyde and reduction of nitrobenzene to aniline reactions are investigated by using SiO2@TiO2 spheres produced in a simple route using chitosan as a template. The spheres are predominantly macroporous and, the XRD points out an amorphous crystallographic profile suggesting the uniform distribution of TiO2. Under low-power lighting for 4 hours, the conversions achieved was of the order of 49% and 99% for benzyl alcohol and nitrobenzene, respectively, with selectivity to benzaldehyde and aniline of 99% in both reactions. The study also follows the effects of the solvent and the presence of O2.

Research on the electrochemical treatment of nitrobenzene wastewater: The effects of process parameters and the mechanism of distinct degradation pathways.

Nitrobenzene is a typical organic pollutant of petroleum pollutant, which is a synthetic chemical not found naturally in the environment. Nitrobenzene in environment can cause toxic liver disease and respiratory failure in humans. Electrochemical technology provides an effective and efficient method for degrading nitrobenzene. This study, the effects of process parameter (e.g., electrolyte solution type, electrolyte concentration, current density and pH) and distinct reaction pathways for electrochemical treatment of nitrobenzene were investigated. As a result, available chlorine dominates the electrochemical oxidation process compared with hydroxyl radical, thus the electrolyte of NaCl is more suitable for the degradation of nitrobenzene than that of Na2SO4. The concentration and the existence form of available chlorine were mainly controlled by electrolyte concentration, current density and pH, which directly affect the removal of nitrobenzene. Cyclic voltammetry and mass spectrometric analyses suggested that electrochemical degradation of nitrobenzene included two important ways. Firstly, single oxidation: nitrobenzene → other forms of aromatic compounds→ NO-x + organic acids + mineralization products. Secondly, coordination of reduction and oxidation: nitrobenzene → aniline→ N2 + NO-x + organic acid + mineralization products. The results of this study will encourage us to further understand the electrochemical degradation mechanism of nitrobenzene and develop the efficient processes for nitrobenzene treatment.

Surfactant-enhanced reduction of soil-adsorbed nitrobenzene by carbon-coated nZVI: Enhanced desorption and mechanism.

The reduction process of pollutants by nano zero-valent iron (nZVI) is limited by mass transfer and its effective utilization, and previous studies have ignored the electron loss caused by its oxidative passivation. The carbon-coated structure can effectively inhibit the oxidation of nZVI, but the effectiveness of carbon-coated nZVI (Fe0@C) as a reducing agent in soil remediation is unclear. Therefore, in this study, the Fe0@C/surfactant system was used to remove soil-adsorbed nitrobenzene (NB) to simultaneously enhance the mass transfer process and effective utilization of nZVI. The results showed that the use of surfactants effectively promoted the desorption of NB adsorbed by the soil, and the desorption process was affected by factors such as the type and concentration of surfactants, water-soil ratio, and soil organic matter (SOM) content. The enhanced desorption of NB by the surfactant in the soil system promoted the effective contact between the composite and NB, thereby enhancing the reduction of NB by the composite. In addition, Fe0@C exhibited excellent performance for the reduction of soil-adsorbed NB compared with the conventional nZVI, and this advantage was more obvious in the potting soil system. However, the composite will be gradually passivated due to the alkaline environment during the reduction process, and this phenomenon was especially obvious in the campus soil system. When the pH value decreased from 9 to 3, the proportion of aniline (AN) generated in the campus soil system increased from 19.37 % to 69.29 %. In addition, in potting soil systems with high SOM content, the adsorption of soil particles to the composite and the high dissolved organic matter (DOM) content resulting from the high SOM content also negatively affected the reduction process. The conclusions of this study demonstrate the great potential of the Fe0@C/surfactant system for in-situ contaminated site remediation applications.

Electrochemical reduction of nitrobenzene via redox-mediated chronoamperometry.

Anilines are important feedstocks for pharmaceuticals, dyes, and other materials, but traditional approaches to their syntheses usually lack selectivity and environmental sustainability. Here, we describe the selective reduction of nitrobenzene to aniline under mild conditions, using water as the ultimate source of the required protons and electrons. We describe the electrochemical cell assembly, and detail steps for electrochemical reduction followed by organic extraction and analysis of the extracts using NMR. For complete details on the use and execution of this protocol, please refer to Stergiou and Symes (2022a).

Development of High-Efficiency, Magnetically Separable Palladium-Decorated Manganese-Ferrite Catalyst for Nitrobenzene Hydrogenation.

Aniline (AN) is one of the most important compounds in the chemical industry and is prepared by the catalytic hydrogenation of nitrobenzene (NB). The development of novel, multifunctional catalysts which are easily recoverable from the reaction mixture is, therefore, of paramount importance. Compared to conventional filtration, magnetic separation is favored because it is cheaper and more facile. For satisfying these requirements, we developed manganese ferrite (MnFe2O4)-supported, magnetically separable palladium catalysts with high catalytic activity in the hydrogenation of nitrobenzene to aniline. In addition to high NB conversion and AN yield, remarkable aniline selectivity (above 96 n/n%) was achieved. Surprisingly, the magnetic support alone also shows moderate catalytic activity even without noble metals, and thus, up to 94 n/n% nitrobenzene conversion, along with 47 n/n% aniline yield, are attainable. After adding palladium nanoparticles to the support, the combined catalytic activity of the two nanomaterials yielded a fast, efficient, and highly selective catalyst. During the test of the Pd/MnFe2O4 catalyst in NB hydrogenation, no by-products were detected, and consequently, above 96 n/n% aniline yield and 96 n/n% selectivity were achieved. The activity of the Pd/MnFe2O4 catalyst was not particularly sensitive to the hydrogenation temperature, and reuse tests indicate its applicability in at least four cycles without regeneration. The remarkable catalytic activity and other favorable properties can make our catalyst potentially applicable to both NB hydrogenation and other similar or slightly different reactions.

Effects of different modifiers on the sorption and structural properties of biochar derived from wheat stalk.

Nitrobenzene is a widespread contaminant in water. Biochar (BC) is a promising material for removing organic pollutants, but the adsorption capacity of pristine BC is low. Chemical modification is often used to improve the adsorption performance, but information on the sorption of nitrobenzene by modified BC is rare. In this study, BCs pyrolyzed at 300, 500, and 700 °C were modified by hydrochloric acid (HCl), sulfuric acid (H2SO4), sodium hydroxide (NaOH), hydrogen peroxide (H2O2), and nitric acid (HNO3), respectively. The properties, nitrobenzene sorption behaviors, and sorption mechanisms of different BCs were analyzed. The results showed that chemical modification decreased the sorption of nitrobenzene on BCs pyrolyzed at 300 °C, possibly due to the loss of the partition phase and the increase in polarity after modification. Regarding BCs pyrolyzed at 500 and 700 °C, the NaOH and HCl modifications significantly increased the sorption capacity by 19% and 60%, 18%, and 41%, respectively, possibly due to the increase in surface area, available pores, and aromaticity, while HNO3 modification decreased the sorption capacity by 41% and 31%. Two reasons were probably responsible for the decrease: one was the decrease in surface area after HNO3 modification due to the destruction of pore walls and the continuity of holes; the other was the strong repulsion between the nitro groups formed on the surface of BC and the nitro groups of nitrobenzene that drove nitrobenzene molecules away from the surface. A principal component-based comprehensive evaluation of the BC properties, which were significantly correlated with the sorption isotherm parameters, was used to evaluate the nitrobenzene sorption performance of the modified BC. Overall, BC pyrolyzed at 700 °C modified with NaOH or HCl were proposed as effective sorption materials for the removal of nitrobenzene in environment, which also provided a chemical modified method of biochar derived from agricultural waste.

Enhanced nitrobenzene removal in soil by biochar supported sulfidated nano zerovalent iron: Solubilization effect and mechanism.

Sulfidated nano zerovalent iron (S-nZVI) is reported to be effective in removal of aqueous organic contaminants. However, little is known about its potential use in reductive degradation of soil-sorbed contaminants. In this study, biochar (BC) supported S-nZVI (S-nZVI@BC) was successfully synthesized through sulfidation and carbon loading modification, which effectively combined the solubilization characteristics of BC and high reduction characteristics of S-nZVI. Transmission electron microscopy (TEM) with an energy-dispersive X-ray spectroscopy (EDS) analysis suggested that sulfur and iron were evenly distributed throughout BC matrix. The degradation of nitrobenzene (NB) in soil was achieved more efficiently with the as-synthesized S-nZVI@BC composites. Results indicated that S-nZVI@BC with S-nZVI/BC mass ratio of 3:1, dosage of 10 mg/g exhibited superior NB removal (98%) and aniline (AN) formation (90%) efficiency within 24 h without formation of other intermediates, higher than those of S-nZVI. Meanwhile, the surface FeSX layer enhanced the antioxidant capacity of S-nZVI@BC and participated in the reduction of NB. The soil-sorbed NB decreased from 14% to 1.4%, indicating that the addition of BC played an important role in solubilization of NB from soil. Solubilization-reduction was the dominant mechanism for NB removal. This research indicated that S-nZVI@BC held the potential to enhance in-situ remediation of NB-contaminated soil.

Synthesis, biological evaluation, and molecular docking of new series of antitumor and apoptosis inducers designed as VEGFR-2 inhibitors.

Based on quinazoline, quinoxaline, and nitrobenzene scaffolds and on pharmacophoric features of VEGFR-2 inhibitors, 17 novel compounds were designed and synthesised. VEGFR-2 IC50 values ranged from 60.00 to 123.85 nM for the new derivatives compared to 54.00 nM for sorafenib. Compounds 15a, 15b, and 15d showed IC50 from 17.39 to 47.10 µM against human cancer cell lines; hepatocellular carcinoma (HepG2), prostate cancer (PC3), and breast cancer (MCF-7). Meanwhile, the first in terms of VEGFR-2 inhibition was compound 15d which came second with regard to antitumor assay with IC50 = 24.10, 40.90, and 33.40 µM against aforementioned cell lines, respectively. Furthermore, Compound 15d increased apoptosis rate of HepG2 from 1.20 to 12.46% as it significantly increased levels of Caspase-3, BAX, and P53 from 49.6274, 40.62, and 42.84 to 561.427, 395.04, and 415.027 pg/mL, respectively. Moreover, 15d showed IC50 of 253 and 381 nM against HER2 and FGFR, respectively.

The discovery of a novel series of potential ERRα inverse agonists based on p-nitrobenzenesulfonamide template for triple-negative breast cancer in vivo.

Oestrogen related receptor α participated in the regulation of oxidative metabolism and mitochondrial biogenesis, and was overexpressed in many cancers including triple-negative breast cancer. A set of new ERRα inverse agonists based on p-nitrobenzenesulfonamide template were discovered and compound 11 with high potent activity (IC50 = 0.80 µM) could significantly inhibit the transcription of ERRα-regulated target genes. By regulating the downstream signalling pathway, compound 11 could suppress the migration and invasion of the ER-negative MDA-MB-231 cell line. Furthermore, compound 11 demonstrated a significant growth suppression of breast cancer xenograft tumours in vivo (inhibition rate 23.58%). The docking results showed that compound 11 could form hydrogen bonds with Glu331 and Arg372 in addition to its hydrophobic interaction with ligand-binding domain. Our data implied that compound 11 represented a novel and effective ERRα inverse agonist, which had broad application prospects in the treatment of triple-negative breast cancer.

4,6-Dichloro-5-Nitrobenzofuroxan: Different Polymorphisms and DFT Investigation of Its Reactivity with Nucleophiles.

This research focuses on the X-ray structure of 4,6-dichloro-5-nitrobenzofuroxan 1 and of some of its amino derivatives (4a, 4e, 4g, and 4l) and on DFT calculations concerning the nucleophilic reactivity of 1. We have found that by changing the solvent used for crystallization, it is possible to obtain 4,6-dichloro-5-nitrobenzofuroxan (1) in different polymorphic structures. Moreover, the different torsional angles observed for the nitro group in 1 and in its amino derivatives (4a, 4e, 4g, and 4l) are strictly dependent on the steric hindrance of the substituent at C-4. DFT calculations on the course of the nucleophilic substitution confirm the role of the condensed furoxan ring in altering the aromaticity of the carbocyclic frame, while chlorine atoms strongly influence the dihedral angle and the rotational barrier of the nitro group. These results corroborate previous observations based on experimental kinetic data and give a deep picture of the reaction with amines, which proceeds via a "non-aromatic" nucleophilic substitution.

Development of hypoxia-activated PROTAC exerting a more potent effect in tumor hypoxia than in normoxia.

Hypoxia is a hallmark of many solid tumors, and it causes the overexpression of a variety of proteins including the epidermal growth factor receptor (EGFR). Many antitumor prodrugs have been designed to target hypoxia. Here we report the identification of a kind of hypoxia-activated proteolysis targeting chimera (ha-PROTAC) by introducing the hypoxia-activated leaving group (1-methyl-2-nitro-1H-imidazol-5-yl)methyl or 4-nitrobenzyl into the structure of an EGFRDel19-based PROTAC. Among the obtained molecules, ha-PROTAC 13 exhibits a more potent degradation activity for EGFRDel19 in hypoxia than in normoxia in HCC4006 cells. This is the first example of identifying a PROTAC to selectively act on tumors utilizing the characteristic of tumor hypoxia and provides a new approach for PROTAC development.

Investigation of NPB Analogs That Target Phosphorylation of BAD-Ser99 in Human Mammary Carcinoma Cells.

The design and development of a small molecule named NPB [3-{(4(2,3-dichlorophenyl)piperazin-1-yl}{2-hydroxyphenyl)methyl}-N-cyclopentylbenzamide], which specifically inhibited the phosphorylation of BAD at Ser99 in human carcinoma cells has been previously reported. Herein, the synthesis, characterization, and effect on cancer cell viability of NPB analogs, and the single-crystal X-ray crystallographic studies of an example compound (4r), which was grown via slow-solvent evaporation technique is reported. Screening for loss of viability in mammary carcinoma cells revealed that compounds such as 2[(4(2,3-dichlorophenyl)piperazin-1-yl][naphthalen-1-yl]methyl)phenol (4e), 5[(4(2,3-dichlorophenyl)piperazin-1-yl][2-hydroxyphenyl)methyl)uran-2-carbaldehyde (4f), 3[(2-hydroxyphenyl][4(p-tolyl)piperazin-1-yl)methyl)benzaldehyde (4i), and NPB inhibited the viability of MCF-7 cells with IC50 values of 5.90, 3.11, 7.68, and 6.5 µM, respectively. The loss of cell viability was enhanced by the NPB analogs synthesized by adding newer rings such as naphthalene and furan-2-carbaldehyde in place of N-cyclopentyl-benzamide of NPB. Furthermore, these compounds decreased Ser99 phosphorylation of hBAD. Additional in silico density functional theory calculations suggested possibilities for other analogs of NPB that may be more suitable for further development.

PIII/PV═O-Catalyzed Intermolecular N-N Bond Formation: Cross-Selective Reductive Coupling of Nitroarenes and Anilines.

An organophosphorus-catalyzed method for the synthesis of unsymmetrical hydrazines by cross-selective intermolecular N-N reductive coupling is reported. This method employs a small ring phosphacycle (phosphetane) catalyst together with hydrosilane as the terminal reductant to drive reductive coupling of nitroarenes and anilines with good chemoselectivity and functional group tolerance. Mechanistic investigations support an autotandem catalytic reaction cascade in which the organophosphorus catalyst drives two sequential and mechanistically distinct reduction events via PIII/PV═O cycling in order to furnish the target N-N bond.

Simultaneous CK2/TNIK/DYRK1 inhibition by 108600 suppresses triple negative breast cancer stem cells and chemotherapy-resistant disease.

Triple negative breast cancer (TNBC) remains challenging because of heterogeneous responses to chemotherapy. Incomplete response is associated with a greater risk of metastatic progression. Therefore, treatments that target chemotherapy-resistant TNBC and enhance chemosensitivity would improve outcomes for these high-risk patients. Breast cancer stem cell-like cells (BCSCs) have been proposed to represent a chemotherapy-resistant subpopulation responsible for tumor initiation, progression and metastases. Targeting this population could lead to improved TNBC disease control. Here, we describe a novel multi-kinase inhibitor, 108600, that targets the TNBC BCSC population. 108600 treatment suppresses growth, colony and mammosphere forming capacity of BCSCs and induces G2M arrest and apoptosis of TNBC cells. In vivo, 108600 treatment of mice bearing triple negative tumors results in the induction of apoptosis and overcomes chemotherapy resistance. Finally, treatment with 108600 and chemotherapy suppresses growth of pre-established TNBC metastases, providing additional support for the clinical translation of this agent to clinical trials.

Wavelength-Selective Softening of Hydrogel Networks.

Photoresponsive hydrogels hold key potential in advanced biomedical applications including tissue engineering, regenerative medicine, and drug delivery, as well as intricately engineered functions such as biosensing, soft robotics, and bioelectronics. Herein, the wavelength-dependent degradation of bio-orthogonal poly(ethylene glycol) hydrogels is reported, using three selective activation levels. Specifically, three chromophores are exploited, that is, ortho-nitrobenzene, dimethyl aminobenzene, and bimane, each absorbing light at different wavelengths. By examining their photochemical action plots, the wavelength-dependent reactivity of the photocleavable moieties is determined. The wavelength-selective addressability of individual photoreactive units is subsequently translated into hydrogel design, enabling wavelength-dependent cleavage of the hydrogel networks on-demand. Critically, this platform technology allows for the fabrication of various hydrogels, whose mechanical properties can be fine-tuned using different colors of light to reach a predefined value, according to the chromophore ratios used. The softening is shown to influence the spreading of pre-osteoblastic cells adhering to the gels as a demonstration of their potential utility. Furthermore, the materials and photodegradation processes are non-toxic to cells, making this platform attractive for biomaterials engineering.

A Recently Assembled Degradation Pathway for 2,3-Dichloronitrobenzene in Diaphorobacter sp. Strain JS3051.

Diaphorobacter sp. strain JS3051 utilizes 2,3-dichloronitrobenzene (23DCNB), a toxic anthropogenic compound, as the sole carbon, nitrogen, and energy source for growth, but the metabolic pathway and its origins are unknown. Here, we establish that a gene cluster (dcb), encoding a Nag-like dioxygenase, is responsible for the initial oxidation of the 23DCNB molecule. The 2,3-dichloronitrobenzene dioxygenase system (DcbAaAbAcAd) catalyzes conversion of 23DCNB to 3,4-dichlorocatechol (34DCC). Site-directed mutagenesis studies indicated that residue 204 of DcbAc is crucial for the substrate specificity of 23DCNB dioxygenase. The presence of glutamic acid at position 204 of 23DCNB dioxygenase is unique among Nag-like dioxygenases. Genetic, biochemical, and structural evidence indicate that the 23DCNB dioxygenase is more closely related to 2-nitrotoluene dioxygenase from Acidovorax sp. strain JS42 than to the 34DCNB dioxygenase from Diaphorobacter sp. strain JS3050, which was isolated from the same site as strain JS3051. A gene cluster (dcc) encoding the enzymes for 34DCC catabolism, homologous to a clc operon in Pseudomonas knackmussii strain B13, is also on the chromosome at a distance of 2.5 Mb from the dcb genes. Heterologously expressed DccA catalyzed ring cleavage of 34DCC with high affinity and catalytic efficiency. This work not only establishes the molecular mechanism for 23DCNB mineralization, but also enhances the understanding of the recent evolution of the catabolic pathways for nitroarenes. IMPORTANCE Because anthropogenic nitroaromatic compounds have entered the biosphere relatively recently, exploration of the recently evolved catabolic pathways can provide clues for adaptive evolutionary mechanisms in bacteria. The concept that nitroarene dioxygenases shared a common ancestor with naphthalene dioxygenase is well established. But their phylogeny and how they evolved in response to novel nitroaromatic compounds are largely unknown. Elucidation of the molecular basis for 23DCNB degradation revealed that the catabolic pathways of two DCNB isomers in different isolates from the same site were derived from different recent origins. Integrating structural models of catalytic subunits and enzymatic activities data provided new insight about how recently modified enzymes were selected depending on the structure of new substrates. This study enhances understanding and prediction of adaptive evolution of catabolic pathways in bacteria in response to new chemicals.

Environment-Sensitive Fluorescence of 7-Nitrobenz-2-oxa-1,3-diazol-4-yl (NBD)-Labeled Ligands for Serotonin Receptors.

Serotonin is a neurotransmitter that plays a crucial role in the regulation of several behavioral and cognitive functions by binding to a number of different serotonin receptors present on the cell surface. We report here the synthesis and characterization of several novel fluorescent analogs of serotonin in which the fluorescent NBD (7-nitrobenz-2-oxa-1,3-diazol-4-yl) group is covalently attached to serotonin. The fluorescent ligands compete with the serotonin1A receptor specific radiolabeled agonist for binding to the receptor. Interestingly, these fluorescent ligands display a high environmental sensitivity of their fluorescence. Importantly, the human serotonin1A receptor stably expressed in CHO-K1 cells could be specifically labeled with one of the fluorescent ligands with minimal nonspecific labeling. Interestingly, we show by spectral imaging that the NBD-labeled ligand exhibits a red edge excitation shift (REES) of 29 nm when bound to the receptor, implying that it is localized in a restricted microenvironment. Taken together, our results show that NBD-labeled serotonin analogs offer an attractive fluorescent approach for elucidating the molecular environment of the serotonin binding site in serotonin receptors. In view of the multiple roles played by the serotonergic systems in the central and peripheral nervous systems, these fluorescent ligands would be useful in future studies involving serotonin receptors.

Design, Synthesis, and Evaluation of o-(Biphenyl-3-ylmethoxy)nitrophenyl Derivatives as PD-1/PD-L1 Inhibitors with Potent Anticancer Efficacy In Vivo.

Two series of novel o-(biphenyl-3-ylmethoxy)nitrophenyl compounds (A1-31 and B1-17) were designed as programmed cell death protein 1 (PD-1)/PD-ligand 1 (PD-L1) inhibitors. All compounds showed significant inhibitory activity with IC50 values ranging from 2.7 to 87.4 nM except compound A17, and compound B2 displayed the best activity. Further experiments showed that B2 bound to the PD-L1 protein without obvious toxicity in Lewis lung carcinoma (LLC) cells. Furthermore, B2 significantly promoted interferon-gamma secretion in a dose-dependent manner in vitro and in vivo. Especially, B2 exhibited potent in vivo anticancer efficacy in an LLC-bearing allograft mouse model at a low dose of 5 mg/kg, which was more active than BMS-1018 (tumor growth inhibition rate: 48.5% vs 17.8%). A panel of immunohistochemistry and flow cytometry assays demonstrated that B2 effectively counteracted PD-1-induced immunosuppression in the tumor microenvironment, thereby triggering antitumor immunity. These results indicate that B2 is a promising PD-1/PD-L1 inhibitor worthy of further development.