Isolation and Antibacterial Activity of Indole Alkaloids from Pseudomonas aeruginosa UWI-1.

In this study, we report the first isolation of three antibiotic indole alkaloid compounds from a Pseudomonad bacterium, Pseudomonas aeruginosa UWI-1. The bacterium was batch fermented in a modified Luria Broth medium and compounds were solvent extracted and isolated by bioassay-guided fractionation. The three compounds were identified as (1) tris(1H-indol-3-yl) methylium, (2) bis(indol-3-yl) phenylmethane, and (3) indolo (2, 1b) quinazoline-6, 12 dione. A combination of 1D and 2D NMR, high-resolution mass spectrometry data and comparison from related data from the literature was used to determine the chemical structures of the compounds. Compounds 1-3 were evaluated in vitro for their antimicrobial activities against a wide range of microorganisms using the broth microdilution technique. Compounds 1 and 2 displayed antibacterial activity against only Gram-positive pathogens, although 1 had significantly lower minimum inhibitory concentration (MIC) values than 2. Compound 3 displayed potent broad-spectrum antimicrobial activity against a range of Gram positive and negative bacteria. Several genes identified from the genome of P. aeruginosa UWI-1 were postulated to contribute to the biosynthesis of these compounds and we attempted to outline a possible route for bacterial synthesis. This study demonstrated the extended metabolic capability of Pseudomonas aeruginosa in synthesizing new chemotypes of bioactive compounds.

A preliminary assessment of heavy metals in sediments from the Cipero and South Oropouche Rivers in Trinidad, West Indies.

The increasing urbanization and industrial processes in Trinidad within recent years could pose a possible contamination threat to the aquatic environment. The southwestern part of the island houses numerous industrial activities, and the recent sightings of schools of dead fish and other marine organisms in that locality is cause for concern prompting research into this occurrence. Sediment and surface water samples from the Cipero and South Oropouche Rivers in South Trinidad were analyzed for their heavy metal content (Cd, Cr, Cu, Mn, Ni, Pb, and Zn). Another watercourse, the Moruga River, was selected as a control, based on its location away from significant anthropogenic sources, and the levels of heavy metals obtained for this location were considered as background concentrations for both surface waters and sediments. Cadmium, Ni, and Pb were not detected in surface water samples of both rivers. The corresponding order of metals in the Cipero River was Mn > Cr > Zn > Cu, while for the South Oropouche River, the order was Mn > Cr > Cu > Zn. The individual concentrations of metals in sediments found in the Cipero and South Oropouche Rivers varied according to the following orders: Mn > Zn > Cu > Pb > Ni > Cr > Cd and Mn > Zn > Pb > Cu > Ni > Cr > Cd, respectively. Assessments of the pollution status in sediments revealed that the Cipero River was considered polluted with a moderate degree of ecological pollution while the South Oropouche River was also deemed polluted; however, the degree of ecological pollution associated with that river was low. Principal component analysis (PCA) and cluster analysis (CA) confirmed that both anthropogenic and natural sources contributed to heavy metal contamination in sediments of both rivers.

Identification of broad-spectrum antiviral compounds and assessment of the druggability of their target for efficacy against respiratory syncytial virus (RSV).

The search for novel therapeutic interventions for viral disease is a challenging pursuit, hallmarked by the paucity of antiviral agents currently prescribed. Targeting of viral proteins has the inextricable challenge of rise of resistance. Safe and effective vaccines are not possible for many viral pathogens. New approaches are required to address the unmet medical need in this area. We undertook a cell-based high-throughput screen to identify leads for development of drugs to treat respiratory syncytial virus (RSV), a serious pediatric pathogen. We identified compounds that are potent (nanomolar) inhibitors of RSV in vitro in HEp-2 cells and in primary human bronchial epithelial cells and were shown to act postentry. Interestingly, two scaffolds exhibited broad-spectrum activity among multiple RNA viruses. Using the chemical matter as a probe, we identified the targets and identified a common cellular pathway: the de novo pyrimidine biosynthesis pathway. Both targets were validated in vitro and showed no significant cell cytotoxicity except for activity against proliferative B- and T-type lymphoid cells. Corollary to this finding was to understand the consequences of inhibition of the target to the host. An in vivo assessment for antiviral efficacy failed to demonstrate reduced viral load, but revealed microscopic changes and a trend toward reduced pyrimidine pools and findings in histopathology. We present here a discovery program that includes screen, target identification, validation, and druggability that can be broadly applied to identify and interrogate other host factors for antiviral effect starting from chemical matter of unknown target/mechanism of action.

Identification of a PCSK9-LDLR disruptor peptide with in vivo function.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) regulates plasma low-density lipoprotein cholesterol (LDL-C) levels by promoting hepatic LDL receptor (LDLR) degradation. Therapeutic antibodies that disrupt PCSK9-LDLR binding reduce LDL-C concentrations and cardiovascular disease risk. The epidermal growth factor precursor homology domain A (EGF-A) of the LDLR serves as a primary contact with PCSK9 via a flat interface, presenting a challenge for identifying small molecule PCSK9-LDLR disruptors. We employ an affinity-based screen of 1013in vitro-translated macrocyclic peptides to identify high-affinity PCSK9 ligands that utilize a unique, induced-fit pocket and partially disrupt the PCSK9-LDLR interaction. Structure-based design led to molecules with enhanced function and pharmacokinetic properties (e.g., 13PCSK9i). In mice, 13PCSK9i reduces plasma cholesterol levels and increases hepatic LDLR density in a dose-dependent manner. 13PCSK9i functions by a unique, allosteric mechanism and is the smallest molecule identified to date with in vivo PCSK9-LDLR disruptor function.

Inhibition of HIV-1 nuclear import via schiff base formation with arylene bis(methylketone) compounds.

Arylene bis(methylketone) compounds specifically block nuclear translocation of the HIV-1 pre-integration complex by forming Schiff-base adducts with contiguous lysines within nuclear localization signal.