Rapid Detection of Broad-Spectrum Pathogenic Bacteria Based on Highly Sensitive Proton Response of the Nucleic Acid Amplification SPQC Platform.

Pathogenic bacteria significantly contribute to elevated morbidity and mortality rates, highlighting the urgent need for early and precise detection. Currently, there is a paucity of effective broad-spectrum methods for detecting pathogenic bacteria. We have developed an innovative proton-responsive series piezoelectric quartz crystal (PR-SPQC) platform for the broad-spectrum identification of pathogenic bacteria. This was achieved by retrieving and aligning sequences from the NCBI GenBank database to identify and validate 16S rRNA oligonucleotide sequences that are signatures of pathogenic bacteria but absent in humans or fungi. The hyperbranched rolling circle amplification, activated exclusively by the screened target, exponentially generates protons that are detected by SPQC through a 2D polyaniline (PANI) film. The PR-SPQC platform demonstrates broad-spectrum capabilities in detecting pathogenic bacteria, with a detection limit of 2 CFU/mL within 90 min. Clinical testing of blood samples yielded satisfactory results. With its advantages in miniaturization, cost efficiency, and suitability for point-of-care testing, PR-SPQC has the potential to be extensively used for the rapid identification of diverse pathogenic bacteria within clinical practice and public health sectors.

Synthesis of Selenium Nanoparticles and Their Effect on Pancreatic Functions and Acute Pancreatitis in Rats.

Acute pancreatitis (AP) have been documented to have severe impact on pancreatic function. Frequent incidence of AP can result in chronic pancreatitis and thereby it can increase the probability of pancreatic cancers. This study intended to examine the effect of selenium nanoparticles (Se-NPs) synthesized from Coleus forskohlii leaf extract on pancreatic function and AP in rat. Primarily, Se-NPs was fabricated using the C. forskohlii leaf extract. The synthesized nanomaterial was characterized through UV-visible, XRD, and FTIR spectroscopies. Notably, the zeta potential of Se-NPs was found to be -32.8 mV with a polydispersity index (PDI) of 0.18. Morphological analysis on SEM unveiled the spherical shape of Se-NP with an average particle size of 12.69 nm. Strikingly, cytotoxicity analysis on pancreatic cancer and normal cells unveiled the concentration-dependent toxicity profile. However, IC 50 value is lower in normal pancreatic cell lines in comparison to pancreatic cancer cells lines. Assessment of Se-NPs on AP rats revealed the positive impact of Se-NPs. It effectively decreased the amount of lipase, amylase, IL-1ß, MDA, NO, and Bcl-2 while increased the glucose, insulin, HOMA-ß and antioxidant potential in AP rats. In addition, an evaluation of Se-NPs in the pancreatic functions revealed the non-harmful effect of Se-NPs.

Bibliometric analysis of the 100 most-cited papers about the role of gut microbiota in irritable bowel syndrome from 2000 to 2021.

AIM: Over the last few decades, gut microbiota research has been the focus of intense research and this field has become particularly important. This research aimed to provide a quantitative evaluation of the 100 most-cited articles on gut microbiota and IBS and highlight the most important advances in this field. METHODS: The database Web of Science Core Collection was used to download the bibliometric information the top 100 most-cited papers. Microsoft Excel 2021, CiteSpace, VOSviewer, R software, and an online analytical platform ( https://bibliometric.com/ ) were was applied to perform bibliometric analysis of these papers. RESULTS: The total citation frequency in the top 100 article ranged from 274 to 2324, with an average citation of 556.57. A total of 24 countries/regions made contributions to the top 100 cited papers, and USA, Ireland, and China were the most top three productive countries. Cryan JF was the most frequently nominated author, and of the top 100 articles, 20 listed his name. Top-cited papers mainly came from the Gastroenterology (n = 13, citations = 6373) and Gut (n = 9, citations = 3903). There was a significant citation path, indicating publications in molecular/biology/immunology primarily cited journals in molecular/biology/genetics fields. Keywords analysis suggested that the main topics on gut microbiota and IBS were mechanisms of microbiome in brain-gut axis." Behavior" was the keyword with the strongest burst strength (2.36), followed by "anxiety like behavior" (2.24), "intestinal microbiota" (2.19), and "chain fatty acid" (1.99), and "maternal separation" (1.95). CONCLUSION: This study identified and provided the bibliometric information of the top 100 cited publications related to gut microbiota and IBS. The results provided a general overview of this topic and might help researchers to better understand the evolution, Influential findings and hotspots in researching gut microbiota and IBS, thus providing new perspectives and novel research ideas in this specific area.

Clinical value of the expression levels of protein tyrosine phosphatase non-receptor type 22.6 mRNA in peripheral blood mononuclear cells in Crohn's disease.

OBJECTIVE: To explore the relationship between the expression levels of protein tyrosine phosphatase non-receptor type (PTPN) 22.6 mRNA in peripheral blood mononuclear cells (PBMCs) and the disease activity as well as clinical characteristics in Crohn's disease (CD) patients. METHODS: A total of 480 subjects were enrolled. Data were collected including baseline information, expression levels of PTPN22.6 mRNA in PBMCs for all subjects, C-reactive protein (CRP) levels in serum, clinical characteristics, and disease activity for all patients. Expression levels of PTPN22.6 mRNA in PBMCs, CRP levels in serum, clinical characteristics according to Montreal Classification [8], and Crohn's disease activity index (CDAI) were the primary observation outcomes. RESULTS: The expression levels of PTPN22.6 mRNA (P = 0.032) in PBMCs and serum CRP levels (P < 0.001) were significantly higher in active CD patients than in inactive CD patients (P = 0.032). Correlation analysis showed that there was a positive correlation between expression levels of PTPN22.6 mRNA and CDAI value (r = 0.512, P = 0.003), as well as expression levels of PTPN22.6 mRNA and CRP levels in the CD group (r = 0.456, P = 0.006). There were significantly higher expression levels of PTPN22.6 mRNA in PBMCs in patients with structuring behavior than that in patients with non-stricturing and non-penetrating (NSNP) behaviors (P = 0.018) and penetrating behaviors (P = 0.024). CONCLUSIONS: The expression levels of PTPN22.6 mRNA can be used as an indicator to help predict CD diagnosis, disease activity, serum CRP level, and behavior type of CD disease.

Thirty minutes of moderate-intensity downhill or level running has no effect on postprandial lipemia: A randomized controlled trial.

Elevated postprandial triglyceride (TG) concentrations are linked to a relatively high risk of cardiovascular disease. Eccentric endurance exercise, such as downhill walking and running, can provide metabolic benefits similar to concentric exercise. However, whether eccentric exercise affects postprandial lipemia remains unknown. Nine healthy young men performed level running (trial) or downhill running (DR trial, -15% slope) at 60% [INSIDE:1]O2max or rest (CON trial) for 30 min in a randomized crossover design. The participants were fed a high-fat meal the next day. Blood and expired gas samples were collected before and 0.5, 1, 2, 3, 4, 5, and 6 h after the meal. Muscle soreness was measured using a visual analog scale. The DR trial induced mild muscle damage. During the 6-h postprandial period, serum TG concentrations and area under the curve (AUC) were similar across the three trials. The DR trial had a significantly higher AUC of nonesterified fatty acid concentrations and a significantly lower AUC of glucose concentrations than the CON trial. The results suggested that neither moderate-intensity DR nor running a level surface had a significant effect on lipemia after a high-fat meal. However, DR improved the postprandial glycemic response.

Molecular identification and relative abundance of microorganisms in douchi koji and salted egg white sufu during processing.

A novel animal protein-based douchi koji-inoculated steamed salted egg white sufu (SEWS) has been developed. This study determined the relative abundance of microorganisms in the douchi koji and semi-finished (5-day fermentation) and finished (5-day fermentation and 14-day ripening) SEWS by using 16S and 18S ribosomal DNA and gene-cloning methods. The results revealed that Bacillus spp. and Aspergillus oryzae were dominant in the douchi koji. In the semi-finished SEWS, the percentages of Bacillus amyloliquefaciens and Bacillus subtilis were considerably lower, whereas those of Enterococcus and Staphylococcus were substantially higher. In the finished SEWS, Bacillus spp. became dominant again and A. oryzae was the only fungus detected. In conclusion, by using molecular techniques, microbial population dynamics in SEWS can be evaluated. During processing, the relative abundance of microorganisms in SEWS changed and Bacillus spp. and A. oryzae remained dominant. This study provides crucial information for designing starter cultures for producing SEWS.

Phosphorylated­myosin light chain mediates the destruction of small intestinal epithelial tight junctions in mice with acute liver failure.

Tight junction dysregulation and epithelial damage contribute to intestinal barrier loss in patients with acute liver failure (ALF); however, the regulatory mechanisms of these processes remain poorly understood. The aim of the present study was to investigate the changes of intestinal tight junction and intestinal mucosa in mice with ALF and their mechanisms. In the present study, ALF was induced in mice through an intraperitoneal injection of D­galactosamine and lipopolysaccharide (D­GalN/LPS), and the morphological changes of the liver or small intestine were analyzed using hematoxylin and eosin staining, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The intestinal tissues and isolated serum were analyzed using western blotting, immunofluorescence staining and ELISA. D­GalN/LPS­induced mice exhibited signs of hepatocyte necrosis, alongside inflammatory cell infiltration into the liver tissue and partial microvilli detachment in the small intestinal mucosa. TEM demonstrated that the intestinal epithelial tight junctions were impaired, whereas SEM micrographs revealed the presence of abnormal microvilli in D­GalN/LPS­induced mice. In addition, the expression levels of phosphorylated (p)­myosin light chain (MLC), MLC kinase (MLCK) and Rho­associated kinase (ROCK) were significantly increased in the D­GalN/LPS­induced mice compared with those in the control mice, whereas the subsequent inhibition of MLCK or ROCK significantly reduced p­MLC expression levels. Conversely, the expression levels of occludin and zonula occludens­1 (ZO­1) were significantly decreased in the D­GalN/LPS­induced mice, and the inhibition of MLCK or ROCK significantly increased occludin and ZO­1 protein expression levels compared with those in the control group. Changes in the serum levels of tumor necrosis factor­α (TNF­α) and interleukin (IL)­6 were similar to the trend observed in p­MLC expression levels. In conclusion, the findings of the present study suggested that in a D­GalN/LPS­induced ALF model, TNF­α and IL­6 signaling may increase MLCK and ROCK expression levels, further mediate phosphorylation of MLC, which may result in tight junction dysregulation and intestinal barrier dysfunction.

Deep learning for detection and segmentation of artefact and disease instances in gastrointestinal endoscopy.

The Endoscopy Computer Vision Challenge (EndoCV) is a crowd-sourcing initiative to address eminent problems in developing reliable computer aided detection and diagnosis endoscopy systems and suggest a pathway for clinical translation of technologies. Whilst endoscopy is a widely used diagnostic and treatment tool for hollow-organs, there are several core challenges often faced by endoscopists, mainly: 1) presence of multi-class artefacts that hinder their visual interpretation, and 2) difficulty in identifying subtle precancerous precursors and cancer abnormalities. Artefacts often affect the robustness of deep learning methods applied to the gastrointestinal tract organs as they can be confused with tissue of interest. EndoCV2020 challenges are designed to address research questions in these remits. In this paper, we present a summary of methods developed by the top 17 teams and provide an objective comparison of state-of-the-art methods and methods designed by the participants for two sub-challenges: i) artefact detection and segmentation (EAD2020), and ii) disease detection and segmentation (EDD2020). Multi-center, multi-organ, multi-class, and multi-modal clinical endoscopy datasets were compiled for both EAD2020 and EDD2020 sub-challenges. The out-of-sample generalization ability of detection algorithms was also evaluated. Whilst most teams focused on accuracy improvements, only a few methods hold credibility for clinical usability. The best performing teams provided solutions to tackle class imbalance, and variabilities in size, origin, modality and occurrences by exploring data augmentation, data fusion, and optimal class thresholding techniques.

Clinical characteristics and risk factors for liver injury in COVID-19 patients in Wuhan.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has become a worldwide pandemic. We investigated the clinical characteristics and risk factors for liver injury in COVID-19 patients in Wuhan by retrospectively analyzing the epidemiological, clinical, and laboratory data for 218 COVID-19 patients and identifying the risk factors for liver injury by multivariate analysis. AIM: To investigate the clinical characteristics and risk factors for liver injury in COVID-19 patients in Wuhan. METHODS: The 218 patients included 94 males (43.1%), aged 22 to 94 (50.1 ± 18.4) years. Elevated aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were present in 42 (53.2%) and 36 (45.6%) cases, respectively, and 79 (36.2%) patients had abnormally elevated transaminase levels at admission. Patients with liver injury were older than those with normal liver function by a median of 12 years, with a significantly higher frequency of males (68.4% vs 28.8%, P < 0.001) and more coexisting illnesses (48.1% vs 27.3%, P = 0.002). Significantly more patients had fever and shortness of breath (87.3% vs 69.8% and 29.1% vs 14.4%, respectively) in the liver injury group. Only 12 (15.2%) patients had elevated total bilirubin. ALT and AST levels were mildly elevated [1-3 × upper limit of normal (ULN)] in 86.1% and 92.9% of cases, respectively. Only two (2.5%) patients had an ALT or AST level > 5 × ULN. Elevated γ-glutamyl transpeptidase was present in 45 (57.0%) patients, and 86.7% of these had a γ-glutamyl-transpeptidase level < 135 U/L (3 × ULN). Serum alkaline phosphatase levels were almost normal in all patients. Patients with severe liver injury had a significantly higher frequency of abnormal transaminases than non-severe patients, but only one case had very high levels of aminotransferases. RESULTS: Multivariate analysis revealed that male sex, high D-dimer level, and high neutrophil percentage were linked to a higher risk of liver injury. The early stage of COVID-19 may be associated with mildly elevated aminotransferase levels in patients in Wuhan. Male sex and high D-dimer level and neutrophil percentage may be important predictors of liver injury in patients with COVID-19. CONCLUSION: Male sex and high D-dimer level and neutrophil percentage may be important predictors of liver injury in patients with COVID-19.

Cartilage Oligomeric Matrix Protein promotes epithelial-mesenchymal transition by interacting with Transgelin in Colorectal Cancer.

Background and Purpose: The role of the cartilage oligomeric matrix protein (COMP) in epithelial-mesenchymal transition (EMT) in tumor progression has been studied, but its exact regulatory mechanism remains unknown. Methods: The interaction between COMP and the actin-binding protein transgelin (TAGLN) was identified by interaction protein prediction and co-immunoprecipitation and verified through the stochastic optical reconstruction microscopy (STORM) and duolink experiments. Western blot and immunofluorescence analyses were conducted to detect the changes in EMT-related markers after COMP overexpression and knockdown. Molecular docking and Biacore of the interaction interface of COMP/TAGLN revealed that Chrysin directly targeted COMP. The promotion of COMP and the Chrysin inhibition of EMT were detected through the cell migration, invasion, apoptosis, and xenotransplantation of nude mice. Results: COMP interacts with TAGLN in EMT in colorectal cancer to regulate cytoskeletal remodeling and promote malignant progression. COMP is highly expressed in highly malignant colorectal cancer and positively correlated with TAGLN expression. COMP knockdown can inhibit colorectal cancer metastasis and invasion, whereas COMP overexpression promotes EMT in colorectal cancer. Through virtual screening of the protein interaction interface, Chrysin, a flavonoid compound extracted from Oroxylum indicum, was found to have the highest docking score to the COMP/TAGLN complex. Chrysin inhibited COMP, thereby preventing EMT and the malignant progression of colorectal cancer. Conclusions: This study illustrated the role of COMP in EMT and suggested that COMP/TAGLN may be a potential tumor therapeutic target. Chrysin exhibits obvious antitumor effects. This work provides a preliminary antitumor therapy to target COMP or its interaction protein to inhibit EMT.

Clinical characteristics of coronavirus disease (COVID-19) patients with gastrointestinal symptoms: A report of 164 cases.

Objective: To explore the clinical characteristics of Coronavirus Disease (COVID-19) patients with gastrointestinal symptoms. Methods: The clinical data of 164 COVID-19 patients with gastrointestinal symptoms were extracted and analysed retrospectively. Results: In total, 505 COVID-19 patients were divided into two groups: those with gastrointestinal symptoms (G group) and those without gastrointestinal symptoms (NG group). Common gastrointestinal symptoms included inappetence, diarrhoea, nausea, abdominal pain, and vomiting. Significantly higher proportions of patients with fever, dizziness, myalgia, and fatigue were noted in group G than in group NG. Compared with patients without fever, there was a significant difference between G group and NG group in moderate fever or above, while there was no significant difference between the two groups in low fever. The laboratory results showed that patients in the G group had significantly higher C-reactive protein, lactate dehydrogenase, and α-hydroxybutyrate dehydrogenase levels than those in the NG group. Moreover, the proportion of patients with severe pneumonia was significantly higher in the G group than in the NG group. Conclusion: In Wuhan, the proportion of COVID-19 patients who experience gastrointestinal symptoms is relatively high. Patients who experience gastrointestinal symptoms are more likely to suffer from severe pneumonia, which may help clinicians identify patients at high risk of COVID-19 and thus reduce the incidence of this condition.

Effects of miR-135a-5p and miR-141 on proliferation, invasion and apoptosis of colorectal cancer SW620 cells.

Effects of miR-135a-5p and miR-141 on the biological function of colorectal cancer SW620 cells were investigated. Fifty-four specimens of cancer tissues and 54 specimens of corresponding adjacent tissues in colon cancer patients who were treated in The Central Hospital of Wuhan from March 2014 to March 2015 were collected. RT-PCR was used to detect the expression levels of miR-135a-5p and miR-141 in cancer tissues and adjacent tissues. The miR-135a-5p inhibitor and miR-141 mimic carriers were established. The cell proliferation was detected by CCK8, the invasion ability of cells in vitro was evaluated by Transwell chamber, and cell apoptosis of each group was detected by flow cytometry. The results of RT-qPCR showed that expression levels of miR-135a-5p in colorectal cancer tissues were significantly higher than those in adjacent tissues, the expression levels of miR-141 in colorectal cancer tissues were significantly lower than those in adjacent tissues, and the difference was statistically significant (P<0.001). The cell survival rates of the miR-135a-5p inhibitor group and the miR-141 mimic group were significantly lower than those of the NC group and the blank group 48 and 72 h after transfection (P<0.001). The number of invasive cells in the miR-135a-5p inhibitor group and the miR-141 mimic group was significantly lower than that in the blank group and the NC group (P<0.001). Apoptosis rate was significantly higher than that of the NC group and the blank group (P<0.001). In conclusion, low expression levels of miR-135a-5p and miR-141 in colorectal adenomas suggested that miR-135a-5p and miR-141 could act as tumor suppressors in the development of colorectal adenomas; miR-135a-5p and miR-141 inhibited the proliferation and invasion of colon cancer SW620 cells and promoted apoptosis of colon cancer cells.

Cytotoxin-associated gene A increases carcinogenicity of helicobacter pylori in colorectal adenoma.

OBJECTIVE: This study aimed to investigate the correlation of Helicobacter pylori (Hp) infection with disease risk and severity of colorectal adenoma, also to explore the association of cytotoxin-associated gene A (CagA) positive (CagA+)-Hp infection with gastrin and ki-67 expressions in colorectal adenoma patients. METHODS: There were 1000 colorectal adenoma patients and 1500 controls consecutively enrolled, then Hp infection status was determined by 14C urea breath test and rapid urease test. Also, serum CagA expression and gastrin expression of colorectal adenoma patients were determined by enzyme-linked immunosorbent assay. Ki-67 expression in adenoma tissue of colorectal adenoma patients was assessed using immunohistochemistry. RESULTS: Hp+ rate in colorectal adenoma patients (623 (62.3%)) was more elevated than that in controls (814 (54.3%)). Multivariate logistic regression model analysis disclosed that Hp+ was an independent risk factor for colorectal adenoma. Additionally, Hp+ was positively associated with tumor size and high-grade intraepithelial neoplasia in colorectal adenoma patients. Also, serum gastrin expression and intratumoral ki-67 expression were higher in Hp+ CagA+ patients and Hp+ CagA- patients compared to Hp- patients, and they were also higher in Hp+ CagA+ patients compared to Hp+ CagA- patients. CONCLUSION: Hp infection positively associates with higher disease risk and worse disease conditions of colorectal adenoma, and CagA enhances the carcinogenicity of Hp in colorectal adenoma.

Small bowel gastrointestinal stromal tumor: a retrospective study of 32 cases at a single center and review of the literature.

BACKGROUND: Gastrointestinal stromal tumor (GIST) is a rare tumor of the small bowel, which can be difficult to diagnose and has a varied clinical outcome. PURPOSE: This is a retrospective review of the diagnosis, management, and clinical outcome of 32 patients diagnosed with primary small bowel GIST from a single center and a comparison of the findings with previously published cases. PATIENTS AND METHODS: Retrospective review of data from patient clinical records, endoscopic and imaging findings, surgical procedures, tumor histology and immunohistochemistry, and clinical outcome was conducted. RESULTS: Data of 32 patients with a median age of 56 years including 50% men and women were reviewed. The majority (29/32) were symptomatic at presentation, with the main symptom being gastrointestinal bleeding (15/32). Imaging detection rates included ultrasound (0%), magnetic resonance imaging (0%), computed tomography (54.8%), computed tomography angiography (71.4%), and double-balloon enteroscopy (88.9%). The mean tumor diameter was 5.3 cm; 4 tumors were located in the duodenum, 21 in the jejunum, and 7 in the ileum. Based on the tumor size and mitotic index, 5 (15.6%), 15 (46.9%), 0 (0%), and 12 (37.5%) patients were classified into very low-risk, low-risk, intermediate-risk, and high-risk groups. Immunohistochemistry showed positive expression for CD117 (100%), CD34 (81.2%), DOG1 (93.8%), smooth muscle actin (37.5%), S100 (9.4%), and desmin (6.2%). Twenty-five patients (78.1%) were treated with open surgical tumor resection; seven patients (21.9%) underwent laparoscopic surgery. Postoperative complications that occurred in seven patients (21.9%) were resolved with conservative management. Four patients were treated with postoperative imatinib. At median follow-up of 30 months, two patients were died. CONCLUSION: The findings from this case series, combined with the findings from previously published cases, provide an update on the current status of the diagnosis and the therapeutic approaches that might lead to improvement in prognosis for patients who present with primary small bowel GIST.

Design and Synthesis of a Novel Series of Orally Bioavailable, CNS-Penetrant, Isoform Selective Phosphoinositide 3-Kinase γ (PI3Kγ) Inhibitors with Potential for the Treatment of Multiple Sclerosis (MS).

The lipid kinase phosphoinositide 3-kinase γ (PI3Kγ) has attracted attention as a potential target to treat a variety of autoimmune disorders, including multiple sclerosis, due to its role in immune modulation and microglial activation. By minimizing the number of hydrogen bond donors while targeting a previously uncovered selectivity pocket adjacent to the ATP binding site of PI3Kγ, we discovered a series of azaisoindolinones as selective, brain penetrant inhibitors of PI3Kγ. This ultimately led to the discovery of 16, an orally bioavailable compound that showed efficacy in murine experimental autoimmune encephalomyelitis (EAE), a preclinical model of multiple sclerosis.

Clinical, endoscopic and pathological characteristics of colorectal polyps in elderly patients: Single-center experience.

Increasing age is a risk factor for the development of colorectal adenomas and advanced adenomas. However, few studies have been published on the features of colorectal polyps in the elderly. The present study aimed to investigate the clinical, enteroscopic and pathological characteristics of colorectal polyps in Chinese elderly patients in a single center (The Central Hospital of Wuhan, Hubei, China). The endoscopic and pathological reports of colonoscopies performed in our center were retrospectively analyzed. A total of 7,795 consecutive patients referred for colonoscopy were evaluated between January 2013 and December 2014. Of the 297 who met the inclusion criteria, 279 polyps were observed in men and 230 in women. Of all the polyps, 263 were non-adenomatous polyps, 104 were non-advanced adenomas and 142 were advanced adenomas. 336 polyps were left-sided and 173 were right-sided. Polyps ≥10 mm were more likely to exhibit an adenomatous component and advanced features, and these findings continued to hold true when the size cut-off was set at 5 mm. The data shown in the present study have revealed that a significant number of polyps lie proximal to the splenic flexure. Thus, evaluation of the whole bowel is particularly important in elderly patients who are undergoing colonoscopy. In addition, the polyp size was associated with the presence of adenoma, and advanced component, diminutive and small polyps should not be ignored in elderly patients.

Discovery of Highly Isoform Selective Thiazolopiperidine Inhibitors of Phosphoinositide 3-Kinase γ.

A series of high affinity second-generation thiazolopiperidine inhibitors of PI3Kγ were designed based on some general observations around lipid kinase structure. Optimization of the alkylimidazole group led to inhibitors with higher levels of PI3Kγ selectivity. Additional insights into PI3K isoform selectivity related to sequence differences in a known distal hydrophobic pocket are also described.

Second-generation antibacterial benzimidazole ureas: discovery of a preclinical candidate with reduced metabolic liability.

Compound 3 is a potent aminobenzimidazole urea with broad-spectrum Gram-positive antibacterial activity resulting from dual inhibition of bacterial gyrase (GyrB) and topoisomerase IV (ParE), and it demonstrates efficacy in rodent models of bacterial infection. Preclinical in vitro and in vivo studies showed that compound 3 covalently labels liver proteins, presumably via formation of a reactive metabolite, and hence presented a potential safety liability. The urea moiety in compound 3 was identified as being potentially responsible for reactive metabolite formation, but its replacement resulted in loss of antibacterial activity and/or oral exposure due to poor physicochemical parameters. To identify second-generation aminobenzimidazole ureas devoid of reactive metabolite formation potential, we implemented a metabolic shift strategy, which focused on shifting metabolism away from the urea moiety by introducing metabolic soft spots elsewhere in the molecule. Aminobenzimidazole urea 34, identified through this strategy, exhibits similar antibacterial activity as that of 3 and did not label liver proteins in vivo, indicating reduced/no potential for reactive metabolite formation.