The Susceptibility of Trypanosomatid Pathogens to PI3/mTOR Kinase Inhibitors Affords a New Opportunity for Drug Repurposing

In our study we describe the potency of established phosphoinositide-3-kinase (PI3K) and mammalian Target of Rapamycin (mTOR) kinase inhibitors against three trypanosomatid parasites: Trypanosoma brucei, T. cruzi, and Leishmania sp., which are the causative agents for African sleeping sickness, Chagas disease, and leishmaniases, respectively. We noted that these parasites and humans express similar kinase enzymes. Since these similar human targets have been pursued by the drug industry for many years in the discovery of cellular growth and proliferation inhibitors, compounds developed as human anti-cancer agents should also have effect on inhibiting growth and proliferation of the parasites. With that in mind, we selected eight established PI3K and mTOR inhibitors for profiling against these pathogens. Among these inhibitors is an advanced clinical candidate against cancer, NVP-BEZ235, which we demonstrate to be a highly potent trypanocide in parasite cultures, and in a mouse model of T. brucei infection. Additionally, we describe observations of these inhibitors' effects on parasite growth and other cellular characteristics

NK-CD11c+ Cell Crosstalk in Diabetes Enhances IL-6-Mediated Inflammation during Mycobacterium tuberculosis Infection

In this study, we developed a mouse model of type 2 diabetes mellitus (T2DM) using streptozotocin and nicotinamide and identified factors that increase susceptibility of T2DM mice to infection by Mycobacterium tuberculosis (Mtb). All Mtb-infected T2DM mice and 40% of uninfected T2DM mice died within 10 months, whereas all control mice survived. In Mtb-infected mice, T2DM increased the bacterial burden and pro- and anti-inflammatory cytokine and chemokine production in the lungs relative to those in uninfected T2DM mice and infected control mice. Levels of IL-6 also increased. Anti-IL-6 monoclonal antibody treatment of Mtb-infected acute- and chronic-T2DM mice increased survival (to 100%) and reduced pro- and anti-inflammatory cytokine expression. CD11c+ cells were the major source of IL-6 in Mtb-infected T2DM mice. Pulmonary natural killer (NK) cells in Mtb-infected T2DM mice further increased IL-6 production by autologous CD11c+ cells through their activating receptors. Anti-NK1.1 antibody treatment of Mtb-infected acute-T2DM mice increased survival and reduced pro- and anti-inflammatory cytokine expression. Furthermore, IL-6 increased inflammatory cytokine production by T lymphocytes in pulmonary tuberculosis patients with T2DM. Overall, the results suggest that NK-CD11c+ cell interactions increase IL-6 production, which in turn drives the pathological immune response and mortality associated with Mtb infection in diabetic mice

A New Synthetic Approach to C-2-Symmetric Octacyclic Cage Diol via Claisen Rearrangement and Ring-Closing Metathesis as Key Steps

A simple synthetic strategy to C-2-symmetric bisannulated octacyclic novel cage diol 20 is reported in eight linear steps starting with a readily available 1,4-hydroquinone 22. Here, an atom economic processes such as Claisen rearrangement, Diels-Alder reaction, intramolecular [2+2] photocycloaddition and ring-closing metathesis (RCM) were used as key steps. The synthetic approach demonstrated here opensup new opportunities to assembled highly functionalized complex cage molecules that are difficult to realize by conventional routes. The structure of the RCM product 19 was firmly established on the basis of single-crystal X-ray diffraction studies

Design and synthesis of polycyclic bisindoles via Fischer indolization and ring-closing metathesis as key steps

Several bisindoles have been synthesized starting with a readily available tetracyclic dione via Fischer indolization and ring-closing metathesis as key steps. The Fischer indolization was achieved under deep eutectic reaction conditions using L-tartaric acid and dimethyl urea (30:70). Various indole derivatives synthesized here are C-2-symmetrical in nature. Due to binding ability, most of the C-2-symmetrical indole moieties exhibit inhibitory activities against to HIV-1 protease, Gram-positive bacteria Bacillus subtilis and Micrococcus luteus. These C-2-symmetric new chemical entities play a special role in medicinal chemistry. (C) 2016 Elsevier Ltd. All rights reserved

Spiro[cyclopentane-1,11′-hexacyclo[7.6.0.01,6.06,13.08,12.010,14]pentadecane]-7′,15′-dione

An unusual rearrangement of spiro cage dione to a trishomocubane derivatives is reported by acid-catalysed rearrangement with the aid of BF3·OEt2 in benzene (solvent) reflux conditions. Here, the molecular structure of cage molecule C19H22O2 (major product) consists of five-membered rings, which adopt an envelope conformation and six-membered rings adopt a chair or boat conformation. The Cremer & Pople puckering parameters of all four six-membered rings are calculated

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Figure S2. Type 2 diabetes enhances pro- and anti-inflammatory responses during Mtb infection. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv. One and six months p.i., lung homogenates were collected from uninfected control and diabetic mice and from Mtb-infected control and diabetic mice and subjected to multiplex ELISA to determine the various cytokines and chemokines. Mean values, p-values and SEs are shown. *P < 0.05, **P < 0.01, ***P < 0.001

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Figure S4. CD11c+ dendritic cells are the major source of IL-6 in Mtb-infected type 2 diabetic mice. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv. A. Six months p.i., pooled CD4+, CD8+, DX5+ and CD11c+ cells from the lung, spleen and lymph nodes were isolated by magnetic selection. IL-6 mRNA expression was determined by real-time PCR. The results of three independent experiments are shown. B & C. Splenic DX5+ and CD11C+ cells were isolated by magnetic selection and cultured (1 NK cell and 4 dendritic cells) with or without g-irradiated Mtb (10 µg/ml). Some of the g-irradiated Mtb H37Rv cultured cells were cultured in the presence of neutralizing antibodies (10 µg/ml) against DNAM-1 or rat IgG2a, κ (the isotype control antibody for the anti-DNAM-1 antibody) or against NKG2D or rat IgG1, κ (the isotype control antibody for the anti-NKG2D antibody). After 18 hours, cell-free culture supernatants were collected and IL-6 levels were measured by ELISA. Mean values, p-values and SEs are shown. *P ≤ 0.05, **P ≤0.01, ***P ≤ 0.001

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Figure S5 Natural killer and dendritic cell interaction enhanced IL-6 production in Mtb-infected type 2 diabetic mice. Control and T2DM mice were infected with 50-100 CFU of aerosolized Mtb H37Rv. Six months p.i., lungs from Mtb-infected control and T2DM mice were isolated and formalin-fixed. Paraffin-embedded tissue sections were prepared and confocal microscopy analysis was performed to determine NK (pink), IL-6+ (green) and dendritic (red) cell co-localization. Scale bar: 20 µm (yellow bar) and 5 µm (white bar). Representative images of staining pattern of three independent experiments, each with three per group were shown