Affinity selection of double-click triazole libraries for rapid discovery of allosteric modulators for GLP-1 receptor.

The recently developed double-click reaction sequence [G. Meng et al., Nature 574, 86-89 (2019)] is expected to vastly expand the number and diversity of synthetically accessible 1,2,3-triazole derivatives. However, it remains elusive how to rapidly navigate the extensive chemical space created by double-click chemistry for bioactive compound discovery. In this study, we selected a particularly challenging drug target, the glucagon-like-peptide-1 receptor (GLP-1R), to benchmark our new platform for the design, synthesis, and screening of double-click triazole libraries. First, we achieved a streamlined synthesis of customized triazole libraries on an unprecedented scale (composed of 38,400 new compounds). By interfacing affinity-selection mass spectrometry and functional assays, we identified a series of positive allosteric modulators (PAMs) with unreported scaffolds that can selectively and robustly enhance the signaling activity of the endogenous GLP-1(9-36) peptide. Intriguingly, we further revealed an unexpected binding mode of new PAMs which likely act as a molecular glue between the receptor and the peptide agonist. We anticipate the merger of double-click library synthesis with the hybrid screening platform allows for efficient and economic discovery of drug candidates or chemical probes for various therapeutic targets.

Modular click chemistry libraries for functional screens using a diazotizing reagent.

Click chemistry is a concept in which modular synthesis is used to rapidly find new molecules with desirable properties1. Copper(I)-catalysed azide-alkyne cycloaddition (CuAAC) triazole annulation and sulfur(VI) fluoride exchange (SuFEx) catalysis are widely regarded as click reactions2-4, providing rapid access to their products in yields approaching 100% while being largely orthogonal to other reactions. However, in the case of CuAAC reactions, the availability of azide reagents is limited owing to their potential toxicity and the risk of explosion involved in their preparation. Here we report another reaction to add to the click reaction family: the formation of azides from primary amines, one of the most abundant functional groups5. The reaction uses just one equivalent of a simple diazotizing species, fluorosulfuryl azide6-11 (FSO2N3), and enables the preparation of over 1,200 azides on 96-well plates in a safe and practical manner. This reliable transformation is a powerful tool for the CuAAC triazole annulation, the most widely used click reaction at present. This method greatly expands the number of accessible azides and 1,2,3-triazoles and, given the ubiquity of the CuAAC reaction, it should find application in organic synthesis, medicinal chemistry, chemical biology and materials science.

Identification of simple arylfluorosulfates as potent agents against resistant bacteria.

Sulfur fluoride exchange (SuFEx), a next generation of click chemistry, opens an avenue for drug discovery. We report here the discovery and structure-activity relationship studies of a series of arylfluorosulfates, synthesized via SuFEx, as antibacterial agents. Arylfluorosulfates 3, 81, and 101 showed potency to overcome multidrug resistance and were not susceptible to the generation of resistance. They exhibited rapid bactericidal potency and selectively killed gram-positive bacterial strains. These compounds also exhibited the ability to disrupt established bacterial biofilm and kill persisters derived from biofilm. Furthermore, arylfluorosulfate 3 had a synergistic effect with streptomycin and gentamicin. In addition, their anti-MRSA potency was evaluated and determined by the Caenorhabditis elegans model.

Genetic and cultural adaptations underlie the establishment of dairy pastoralism in the Tibetan Plateau.

BACKGROUND: Domestication and introduction of dairy animals facilitated the permanent human occupation of the Tibetan Plateau. Yet the history of dairy pastoralism in the Tibetan Plateau remains poorly understood. Little is known how Tibetans adapted to milk and dairy products. RESULTS: We integrated archeological evidence and genetic analysis to show the picture that the dairy ruminants, together with dogs, were introduced from West Eurasia into the Tibetan Plateau since ~ 3600 years ago. The genetic admixture between the exotic and indigenous dogs enriched the candidate lactase persistence (LP) allele 10974A > G of West Eurasian origin in Tibetan dogs. In vitro experiments demonstrate that - 13838G > A functions as a LP allele in Tibetans. Unlike multiple LP alleles presenting selective signatures in West Eurasians and South Asians, the de novo origin of Tibetan-specific LP allele - 13838G > A with low frequency (~ 6-7%) and absence of selection corresponds - 13910C > T in pastoralists across eastern Eurasia steppe. CONCLUSIONS: Results depict a novel scenario of genetic and cultural adaptations to diet and expand current understanding of the establishment of dairy pastoralism in the Tibetan Plateau.

Enabling Modular Click Chemistry Library through Sequential Ligations of Carboxylic Acids and Amines.

High-throughput synthesis and screening of chemical libraries play pivotal roles in drug discovery. Click chemistry has emerged as a powerful strategy for constructing highly modular chemical libraries. However, the development of new click reactions and unlocking new clickable building blocks remain exceedingly challenging. Herein, we describe a double-click strategy that enables the sequential ligations of widely available carboxylic acids and amines with fluorosulfuryl isocyanate (FSO2NCO) via a modular amidation/SuFEx (sulfur-fluoride exchange) process. This method provides facile access to chemical libraries of N-fluorosulfonyl amides (RCONHSO2F) and N-acylsulfamides (RCONHSO2NR'R'') in near-quantitative yields under simple and practical conditions. The robustness and efficiency of this double click strategy is showcased by the facile construction of chemical libraries in 96-well microtiter plates from a large number of carboxylic acids and amines. Preliminary biological activity screening reveals that some compounds exhibit high antimicrobial activities against Gram-positive bacterium S. aureus and drug-resistant MRSA (MIC up to 6.25 µg ⋅ mL-1). These results provide compelling evidence for the potential application of modular click chemistry library as an enabling technology in high-throughput medicinal chemistry.

Changes in liver function, fluctuations in blood glucose, insulin secretion and gender differences in patients with hyperthyroidism after treatment with propranolol hydrochloride tablets coalition with methimazole tablets.

To investigate liver function changes, blood glucose fluctuation, insulin secretion and gender differences in hyperthyroidism patients before and after propranolol with methimazole. Clinical data 110 hyperthyroidism patients admitted to Zhangzhou Affiliated Hospital of Fujian Medical University from February 2023 to February 2024 were retrospectively analyzed. They were categorized into the methimazole group (methimazole, n = 55) and the coalition medication group (Methimazole with propranolol, n = 55). The therapeutic effects of both groups were observed. Pre- and post-treatment liver function dynamic blood glucose parameters and insulin secretion characteristics were analyzed between the two groups. Gender differences prior to treatment were also examined. Overall efficacy was significantly higher in the oalition group (96.35%) than in the methimazole group (83.64%) (P<0.05); Post-treatment, the TBiL, AST, ALT, FT3, FT4, FBG, P1BG, HOMA-IR, HOMA-ß, postprandial blood glucose peak value, LAGE, MAGE, MODD and SDBG levels in the coalition group were lower compared to the methimazole group, while TSH was higher (P<0.05). Female patients exhibited significantly lower LAGE, MAGE, MODD and SDBG levels compared to the male group (P<0.05). The combination of methimazole and propranolol enhances thyroid and liver functions for hyperthyroid patients while improving insulin resistance along with reducing postprandial blood glucose variability.

Antileukemic effect of venetoclax and hypomethylating agents via caspase-3/GSDME-mediated pyroptosis.

BACKGROUND: The identifying of B-cell lymphoma 2 (Bcl-2) as a therapeutic target has led to a paradigm shift in acute myeloid leukemia (AML) treatment. Pyroptosis is a novel antitumor therapeutic mechanism due to its cytotoxic and immunogenic effects. The combination of venetoclax and hypomethylating agents (HMAs) has been shown to lead to durable responses and significantly improve prognosis in patients with AML. However, our understanding of the mechanisms underlying this combinatorial activity is evolving. METHODS: We investigated whether the Bcl-2 inhibitor venetoclax induces AML cell pyroptosis and identified pyroptosis effector proteins. Via using western blotting, immunoprecipitation, RNA interference, CCK8 assays, and LDH assays, we explored the mechanism underlying the pyroptotic effect. The relationship between the expression of the pyroptosis effector protein GSDME and AML prognosis was investigated. The effect of GSDME demethylation combined with venetoclax treatment on pyroptosis was investigated and confirmed in mouse models and clinical samples. RESULTS: Venetoclax induces pyroptosis that is mediated by caspase-3-dependent GSDME cleavage. Mechanistically, venetoclax upregulates caspase-3 and GSDME cleavage by activating the intrinsic apoptotic pathway. GSDME is downregulated in AML by promoter methylation, and low GSDME expression is significantly associated with poor prognosis, based on public databases and patient sample analysis. In vivo and in vitro experiments showed that GSDME overexpression or HMAs-mediated restoration of GSDME expression significantly increased venetoclax-induced pyroptosis in AML. CONCLUSION: GSDME-mediated pyroptosis may be a novel aspect of the antileukemic effect of Bcl-2 inhibitors. This finding offers new insights into potential biomarkers and therapeutic strategies, identifying an important mechanism explaining the clinical activity of venetoclax and HMAs in AML.

NPLC0393 from Gynostemma pentaphyllum ameliorates Alzheimer's disease-like pathology in mice by targeting protein phosphatase magnesium-dependent 1A phosphatase.

Alzheimer's disease (AD) is a neurodegenerative disease with clinical hallmarks of progressive cognitive impairment and memory loss. Gynostemma pentaphyllum ameliorates cognitive impairment, but the mechanisms remain obscure. Here, we determine the effect of triterpene saponin NPLC0393 from G. pentaphyllum on AD-like pathology in 3×Tg-AD mice and elucidate the underlying mechanisms. NPLC0393 was administered daily in vivo by intraperitoneal injection for 3 months and its amelioration on the cognitive impairment in 3×Tg-AD mice was assessed by new object recognition (NOR), Y-maze, Morris water maze (MWM), and elevated plus-maze (EPM) tests. The mechanisms were investigated by RT-PCR, western blot, and immunohistochemistry techniques, while verified by the 3×Tg-AD mice with protein phosphatase magnesium-dependent 1A (PPM1A) knockdown (KD) through brain-specific injection of adeno-associated virus (AAV)-ePHP-KD-PPM1A. NPLC0393 ameliorated AD-like pathology targeting PPM1A. It repressed microglial NLRP3 inflammasome activation by reducing NLRP3 transcription during priming and promoting PPM1A binding to NLRP3 to disrupt NLRP3 assembly with apoptosis-associated speck-like protein containing a CARD and pro-caspase-1. Moreover, NPLC0393 suppressed tauopathy by inhibiting tau hyperphosphorylation through PPM1A/NLRP3/tau axis and promoting microglial phagocytosis of tau oligomers through PPM1A/nuclear factor-κB/CX3CR1 pathway. PPM1A mediates microglia/neurons crosstalk in AD pathology, whose activation by NPLC0393 represents a promising therapeutic strategy for AD.

The effect of head rotation on OSA is associated with disease severity: a cross-sectional study.

PURPOSE: Head rotation is an effective positional therapy for obstructive sleep apnea (OSA). However, not all OSA patients benefit from head rotation. This study aimed to explore the clinical phenotype of OSA patients who can benefit from head rotation. METHODS: We performed a retrospective review of 184 consecutive OSA patients who underwent polysomnography. Head rotation in supine position was determined by high-quality video recording. According to the changes in apnea-hypopnea index (AHI) after head rotation, OSA patients were divided into two groups: patients with response to head rotation(HR) and patients without response to head rotation(NHR). Demographic factors and overnight polysomnography were analyzed. RESULTS: Compared with NHR group, HR group showed significantly lower AHI (51.8 vs 31.5, p < 0.01), time spent with oxygen saturation below 90%(5.3% vs 0.51%, p < 0.01), and higher lowest oxygen saturation(80% vs 86%, p < 0.05). Logistic regression showed that AHI was an independent factor to predict the decrease of AHI in head rotation (OR 0.985, 95% CI 0.970-0.979, p < 0.05). Among mild to moderate group (AHI < 30/h), severe group (30/h ≤ AHI < 60/h), and extremely severe group (AHI ≥ 60/h), the percentage decrease of AHI in head rotation was 18.5%, 9.5%, and 2.6%, respectively. Surprisingly, the percentage decrease of AHI of 6 responders in mild to moderate group was more than 50%. CONCLUSION: OSA patients who respond well to head rotation have less severe disease, and patients with mild to moderate OSA are more likely to improve and benefit from this position. Our research provides potential strategies and insights into the individual treatment of OSA patients.

A Metabolic-Tag-Based Method for Assessing the Permeation of Small Molecules Across the Mycomembrane in Live Mycobacteria.

The general lack of permeability of small molecules observed for Mycobacterium tuberculosis (Mtb) is most ascribed to its unique cell envelope. More specifically, the outer mycomembrane is hypothesized to be the principal determinant for access of antibiotics to their molecular targets. We describe a novel assay that combines metabolic tagging of the peptidoglycan, which sits directly beneath the mycomembrane, click chemistry of test molecules, and a fluorescent labeling chase step, to measure the permeation of small molecules. We showed that the assay workflow was robust and compatible with high-throughput analysis in mycobacteria by testing a small panel of azide-tagged molecules. The general trend is similar across the two types of mycobacteria with some notable exceptions. We anticipate that this assay platform will lay the foundation for medicinal chemistry efforts to understand and improve uptake of both existing drugs and newly-discovered compounds into mycobacteria.

Sestrin2 attenuates renal damage by regulating Hippo pathway in diabetic nephropathy.

Glomerular mesangial cell proliferation and extracellular matrix accumulation contribute to the progression of diabetic nephropathy (DN). As a conserved stress-inducible protein, sestrin2 (Sesn2) plays critical role in the regulation of oxidative stress, inflammation, autophagy, metabolism, and endoplasmic reticulum stress. In this study, we investigated the role of Sesn2 on renal damage in diabetic kidney using transgenic mice overexpressing Sesn2 and the effect of Sesn2 on mesangial cell proliferation and extracellular matrix accumulation in diabetic conditions and the possible molecular mechanisms involved. Sesn2 overexpression improved renal function and decreased glomerular hypertrophy, albuminuria, mesangial expansion, extracellular matrix accumulation, and TGF-ß1 expression, as well as oxidative stress in diabetic mice. In vitro experiments, using human mesangial cells (HMCs), revealed that Sesn2 overexpression inhibited high glucose (HG)-induced proliferation, fibronectin and collagen IV production, and ROS generation. Meanwhile, Sesn2 overexpression restored phosphorylation levels of Lats1 and YAP and inhibited TEAD1 expression. Inhibition of Lats1 accelerated HG-induced proliferation and expression of fibronectin and collagen IV. Verteporfin, an inhibitor of YAP, suppressed HG-induced proliferation and expression of fibronectin and collagen IV. However, Sesn2 overexpression reversed Lats1 deficiency-induced Lats1 and YAP phosphorylation, nuclear expression levels of YAP and TEAD1, and proliferation and fibronectin and collagen IV expressions in HMCs exposed to HG. In addition, antioxidant NAC or tempol treatment promoted phosphorylation of Lats1 and YAP and inhibited TEAD1 expression, proliferation, and fibronectin and collagen IV accumulation in HG-treated HMCs. Taken together, Sesn2 overexpression inhibited mesangial cell proliferation and fibrosis via regulating Hippo pathway in diabetic nephropathy. Induction of Sesn2 may be a potential therapeutic target in diabetic nephropathy.

Olfactory dysfunction is associated with cognitive impairment in patients with obstructive sleep apnea: a cross-sectional study.

PURPOSE: Obstructive sleep apnea (OSA) increases the risk for olfactory dysfunction. However, the relationship between olfactory function and cognition in OSA patients is unclear. The present study aimed to investigate the relationship between cognition and olfactory dysfunction (OD) in patients with OSA. METHOD: This was a cross-sectional study in which 74 patients with OSA and 22 controls were recruited. All subjects completed polysomnography, Sniffin' Sticks, and -neurocognitive assessments. According to results of Sniffin' Sticks, OSA patients were divided into two groups: OSA with OD (53 cases) and OSA without OD (21 cases). Neurocognitive function was assessed by Montreal Cognitive Assessment (MoCA), Memory and Executive Screening (MES), and Shape Trail Test (STT). Cognition was compared between OSA with and without OD. Correlation between olfactory parameters and respiratory sleep parameters and neurocognitive assessments was analyzed. RESULTS: Compared with OSA without OD, OSA with OD showed significantly decreased neurocognitive scores of MoCA (29-27 vs 27-23, p < 0.01), MES-5R (45-40.1 vs 43-33.5, p < 0.01) and increased consuming time of STT-B (91.66 vs 121.63, p < 0.01). A positive correlation was found between the scores of MoCA and MES-5R and all olfactory parameters. In addition, a negative correlation was present between the time consumed for STT-B and odor thresholds (r = - 0.344, p < 0.01), odor identification (r = - 0.335, p < 0.01), and threshold-discrimination-identification scores (r = - 0.448, p < 0.01). CONCLUSION: Olfactory function is associated cognitive function in patients with OSA and may provide a new direction for early treatment interventions in OSA patients at risk for cognitive impairment.

[Clinical features and prognosis in 20 children with rhabdomyosarcoma]. / 20例横纹肌肉瘤患儿的临床特点及预后分析.

OBJECTIVES: To study the clinical features of children with rhabdomyosarcoma (RMS) and the influencing factors for prognosis. METHODS: A retrospective analysis was performed on the clinical and follow-up data of 20 children with RMS who were admitted to the Department of Pediatric Hematology, Xiangya Hospital of Central South University, from June 2014 to September 2020. RESULTS: The most common clinical symptoms of the 20 children with RMS at the first visit were painless mass (13/20, 65%), exophthalmos (4/20, 20%), and abdominal pain (3/20, 15%). According to the staging criteria of Intergroup Rhabdomyosarcoma Study Group (IRSG), there was 1 child (5%) with stage I RMS, 4 (20%) with stage II RMS, 9 (45%) with stage III RMS, and 6 (30%) with stage IV RMS. The median follow-up time was 19 months for the 20 children (range: 3-93 months), with a 2-year overall survival (OS) rate of 79.5% (95%CI: 20.1-24.3) and a 2-year event-free survival (EFS) rate of 72.0% (95%CI: 19.5-23.9). Pleomorphic RMS was associated with the reduced 2-year OS rate (P<0.05), and distant metastasis, IRSG stage IV RMS, and high-risk RMS were associated with the reduced 2-year EFS rate (P<0.05). CONCLUSIONS: RMS has no specific clinical symptoms at the first visit, with painless mass as the most common symptom. Distant metastasis, IRSG stage, and risk degree may be associated with the prognosis of children with RMS.

Crazy-paving patterns as rare radiological manifestations of pulmonary cryptococcosis: a case report.

BACKGROUND: Crazy-paving patterns are rarely reported as radiological manifestations of pulmonary cryptococcosis. CASE PRESENTATION: Herein, we presented a very rare case of a crazy-paving pattern as a radiological manifestation of pulmonary cryptococcosis in a patient with primary ciliary dyskinesia. The diagnosis of pulmonary cryptococcosis and primary ciliary dyskinesia was ultimately confirmed by bronchoscopic biopsy, fungus culture, whole exome sequencing of blood, etc. The patient received flucytosine (PO, 5 g per day) and amphotericin B (IV, 70 mg per day) during hospitalization and sequential therapy with voriconazole (PO, 200 mg twice a day) after discharge. He recovered during follow-up. CONCLUSIONS: We concluded that pulmonary cryptococcosis should be considered a possible cause of crazy-paving patterns in chest CT scans.

Comparative pharmacokinetic and bioavailability study of lobetyolin in rats after administration of lobetyolin and Codonopsis pilosula extract by ultra-performance LC-tandem mass spectrometry.

Codonopsis pilosula (CP) is a traditional Chinese medicine used to invigorate spleen, replenish lung, nourish blood and engender fluid. A rapid, selective and sensitive ultra-performance LC-tandem mass spectrometry method was developed and validated to determine lobetyolin in rat plasma. The calibration curve showed good linearity over a concentration range of 0.46-1000 ng/mL for lobetyolin. The extraction recovery ranged from 72.5% to 89.1% with matrix effects of 81.6%-107.8%. The intra- and inter-batch precision and accuracy were 0.02-14.4% and -13.9% to -1.36%, respectively. The method was successfully applied for the bioavailability study of lobetyolin in rats after oral administration of pure lobetyolin and CP extract. Results showed that the elimination half-time (t1/2 ) and the area under the concentration-time curve from zero to infinity of lobetyolin in CP extract were statistically different from those of the pure monomer (P < 0.05). However, the time to reach the maximum plasma concentration (Tmax ) and the maximum concentration (Cmax ) showed no significant differences between the two treatments. Furthermore, the bioavailability of lobetyolin in the experimental group was only 3.90%, significantly lower than that of the CP extract group (6.97%). The low bioavailability indicated that this component may be absorbed poorly or metabolized extensively in rats. Our results will provide useful information for further preclinical studies and formulation preparation of lobetyolin.

Comparative pharmacokinetic analysis of raw and steamed Panax notoginseng roots in rats by UPLC-MS/MS for simultaneously quantifying seven saponins.

CONTEXT: After being steamed, the restorative effects of Panax notoginseng (Burk.) F. H. Chen (Araliaceae) will be strengthened. However, the underlying mechanism remains elusive. OBJECTIVE: To compare the pharmacokinetics of ginsenosides Rg1, Rb1, Rd, Re, Rg5, Rk1, notoginsenoside R1 (GRg1, GRb1, GRd, GRe, GRg5, GRk1 and NGR1) in the raw and steam-processed P. notoginseng (RPN and SPN). MATERIALS AND METHODS: The pharmacokinetics of seven components after oral administration of SPN and RPN extracts (1.0 g/kg) were investigated, respectively, in SD rats (two groups, n = 6) using UPLC-MS/MS. RESULTS: The approach elicited good linear regression (r2 > 0.991). The accuracy, precision and stability were all within ± 15%. The extraction recoveries and matrix effects were 75.0-100.8% and 85.1-110.3%, respectively. Compared with the RPN group, AUC0-t of GRg1 (176.63 ± 42.49 ng/h/mL), GRb1 (5094.06 ± 1453.14 ng/h/mL), GRd (1396.89 ± 595.14 ng/h/mL), and NGR1 (135.95 ± 54.32 ng/h/mL), along with Cmax of GRg1 (17.41 ± 5.43 ng/mL), GRb1 (361.48 ± 165.57 ng/mL), GRd (62.47 ± 33.65 ng/mL) and NGR1 (23.97 ± 16.77 ng/mL) decreased remarkably with oral administration of the SPN extracts, while GRe showed no significantly difference. Of note, GRg5 and GRk1 could not be detected in the plasma. CONCLUSIONS: Influence of the processing reduced the systemic exposure levels to GRg1, GRb1, GRd and NGR1. It is the first report of comparative pharmacokinetic study of multiple saponins analysis after oral administration of RPN and SPN extract, which might be helpful for further studies on its steam-processing mechanism.

Fluorosulfuryl Isocyanate Enabled SuFEx Ligation of Alcohols and Amines.

Fluorosulfuryl isocyanate (FSI, FSO2 NCO) is established as a reliable bis-electrophilic linker for stepwise attachment of an alcohol bearing module to an amine bearing module and thence a new module RO-C(=O)-NH-SO2 -NR'R'' is created. FSI's isocyanate motif fuses directly and quickly with alcohols and phenols, affording fluorosulfuryl carbamates in nearly quantitative yield. A new reagent and process to deliver the FSI-derived fluorosulfuryl carbamate fragment to amines are also developed. The resulting SVI -F motifs from step-1 are remarkably stable, given the great structural complexities in diverse products. In the step-2 reaction with amines, the best yield of the S-N linked products arise with water alone. This "on water" interfacial reactivity phenomenon is crucial, revealing the latent reactivity of SVI -F probe for potential covalent capture of proteins in vivo which is important in today's drug discovery. The scope of the SuFEx chemistry is largely expanded thereby and the facile entry to these phosphate-like connections should prove useful to click chemistry across diverse fields.

Metal-Free Synthesis of Functional 1-Substituted-1,2,3-Triazoles from Ethenesulfonyl Fluoride and Organic Azides.

The boom in growth of 1,4-disubstituted triazole products, in particular, since the early 2000's, can be largely attributed to the birth of click chemistry and the discovery of the CuI -catalyzed azide-alkyne cycloaddition (CuAAC). Yet the synthesis of relatively simple, albeit important, 1-substituted-1,2,3-triazoles has been surprisingly more challenging. Reported here is a straightforward and scalable click-inspired protocol for the synthesis of 1-substituted-1,2,3-triazoles from organic azides and the bench stable acetylene surrogate ethenesulfonyl fluoride (ESF). The new transformation tolerates a wide selection of substrates and proceeds smoothly under metal-free conditions to give the products in excellent yield. Under controlled acidic conditions, the 1-substituted-1,2,3-triazole products undergo a Michael addition reaction with a second equivalent of ESF to give the unprecedented 1-substituted triazolium sulfonyl fluoride salts.

Expression of the Wnt ligands gene family and its relationship to prognosis in hepatocellular carcinoma.

BACKGROUND: The Wnt gene family members are known to participate regulating various normal and pathological processes including tumorigenesis. However, the association between Wnt ligands gene family and prognosis in hepatocellular carcinoma has not been systematically studied. Therefore, we evaluated the role of Wnt ligands gene family in hepatocellular carcinoma using publicly available data from The Cancer Genome Atlas (TCGA). METHODS: Clinical information and RNA-Seq mRNA expression data were derived from TCGA hepatocellular carcinoma cohort. Differences in overall survival (OS) and disease-free survival (DFS) between increased and decreased expression groups (defined by X-tile analyses) of Wnt ligands gene family were compared using Kaplan-Meier method and Cox regression model, with p-values calculated via log-rank test. Gene Set Enrichment Analysis (GSEA) was performed. RESULTS: Multivariate analysis adjusted for patient age, sex, BMI, tumor grade, and TMN stage revealed that Wnt1, Wnt3 and Wnt5B expressions were independent prognostic factors for OS and DFS (OS: HR = 0.58, P = 0.006; HR = 0.65, P = 0.03; HR = 0.56, P = 0.023, respectively; DFS: HR = 0.52, P < 0.001; HR = 1.93, P = 0.003; HR = 0.59, P = 0.011, respectively). Furthermore, expression of Wnt1 and Wnt5B was significantly associated with TMN stage (P = 0.02 and P = 0.03 for OS; P = 0.02 and P = 0.02 for DFS). GSEA showed that nucleotide excision repair was differentially enriched in Wnt1 low expression phenotype and aminoacyl trna biosynthesis and basal transcription factors were differentially enriched in Wnt5B low expression phenotype. CONCLUSIONS: Our results identified associations of several Wnt ligands with prognosis of HCC patients, indicating that these genes could serve as prognostic biomarkers of HCC.

Mechanism and chemoselectivity origins of bioconjugation of cysteine with Au(iii)-aryl reagents.

Organometallic reagents, in particular Pd(ii)- and Au(iii)-aryl reagents, have recently emerged as an efficient tool for bioconjugation. However, the detailed mechanism and origins of chemoselectivity are not well established, but are highly desirable from both synthetic and theoretical viewpoints. In this paper, we report that a computational study dealing with the reaction mechanism of Au(iii)-aryl reagents enabled selective cysteine S-arylation of peptides and proteins developed by Maynard and Spokoyny et al. (J. Am. Chem. Soc., 2018, 140, 7065). Our calculation results suggest that the reaction proceeds by a cationic Au(iii)/Au(i) pathway involving elementary steps of (a) binding of the SH residue to the Au(iii) center, (b) deprotonation of the SH residue, and (c) reductive elimination from a key four-coordinate square planar (L)Au(iii)(thiolate)(Ar) (L is a P,N-bidentate ligand) intermediate. Furthermore, the chemoselectivity of S-arylation against arylation of other nucleophilic residues can be rationalized in terms of energy demand of the three elementary steps. For instance, amine N-arylation is more difficult than S-arylation due majorly to the much higher energy required for deprotonation of much more basic N-H bonds than for deprotonation of weakly acidic S-H bonds. Carboxylate O-arylation is challenging due to the high activation energy of reductive elimination from LAu(iii)(carboxylate)(aryl), because carboxylate is much less nucleophilic than thiolate. These results thus identify acidity and nucleophilicity of the residue as two inherent factors for bioconjugation. This study provides a useful and convenient approach for predicting and rationalizing the feasibility and chemoselectivity of related bioconjugation reactions.