Rationally designed novel Phenyloxazoline Synthase Inhibitors: chemical synthesis and biological evaluation to accelerate the discovery of new antimycobacterial antibiotics

The uncontrolled spread of drug-resistant tuberculosis (DR-TB) clinical cases necessitates the urgent discovery of newer chemotypes with novel mechanisms of action. Here, we report the chemical synthesis of rationally designed novel transition-state analogues (TSAs) by targeting the cyclization (Cy) domain of phenyloxazoline synthase (MbtB), a key enzyme of the conditionally essential siderophore biosynthesis pathway. Following bio-assay-guided evaluation of TSA analogues preferentially in iron-deprived and iron-rich media to understand target preferentiality against a panel of pathogenic and non-pathogenic mycobacteria strains, we identified a hit, i.e., TSA-5. Molecular docking, dynamics, and MMPBSA calculations enabled us to comprehend TSA-5’s stable binding at the active site pocket of MbtB_Cy and the results imply that the MbtB_Cy binding pocket has a strong affinity for electron-withdrawing functional groups and contributes to stable polar interactions between enzyme and ligand. Furthermore, enhanced intracellular killing efficacy (8 μg/mL) of TSA-5 against Mycobacterium aurum in infected macrophages is noted in comparison to moderate in vitro antimycobacterial efficacy (64 μg/mL) against M. aurum. TSA-5 also demonstrates whole-cell efflux pump inhibitory activity against Mycobacterium smegmatis. Identification of TSA-5 by focusing on the modular MbtB_Cy domain paves the way for accelerating novel anti-TB antibiotic discoveries

Synthetic Biology Open Language Visual (SBOL Visual) Version 2.1

People who are engineering biological organisms often find it useful to communicate in diagrams, both about the structure of the nucleic acid sequences that they are engineering and about the functional relationships between sequence features and other molecular species . Some typical practices and conventions have begun to emerge for such diagrams. The Synthetic Biology Open Language Visual (SBOL Visual) has been developed as a standard for organizing and systematizing such conventions in order to produce a coherent language for expressing the structure and function of genetic designs. This document details version 2.1 of SBOL Visual, which builds on the prior SBOL Visual 2.0 standard by expanding diagram syntax to include methods for showing modular structure and mappings between elements of a system, interactions arrows that can split or join (with the glyph at the split or join indicating either superposition or a chemical process), and adding new glyphs for indicating genomic context (e.g., integration into a plasmid or genome) and for stop codons

Mycobactin analogue interacting with siderophore efflux-pump protein: insights from molecular dynamics simulations and whole-cell assays

IntroductionIn response to continued public health emergency of antimicrobial resistance (AMR), a significant key strategy is the discovery of novel mycobacterial efflux-pump inhibitors (EPIs) as potential adjuvants in combination drug therapy. Interest in identifying new chemotypes which could potentially synergize with the existing antibiotics and can be deployed as part of a combination therapy. This strategy could delay the emergence of resistance to existing antibiotics and increase their efficacy against resistant strains of mycobacterial species. In recent decades, notable approaches have been accounted for EPI development and have resulted in the discovery of several EPIs including SQ109 and AU1235. In context, to accelerate newer EPIs with novel mode of action here we have discussed mycobactin analogues and highlighted in silico binding orientation with siderophore efflux-pump proteins MmpL4/5.Methods3-(2-hydroxyphenyl)-5-(aryl)-pyrazoline series was investigated for whole-cell efflux-pump inhibitory activity against Mycobacterium smegmatis and Mycobacterium abscessus. Machine learning and molecular dynamics were performed to construct a MmpL4/5 complex embedded in a lipid bilayer to identify the putative binding site and to predict ligand-protein binding energetics. Furthermore, the identified HIT compound was investigated in synergistic assay with bedaquiline.ResultsCompound Il, 2-(5-(4-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl)phenol, was identified as the most potent efflux pump inhibitor against M. smegmatis in whole-cell efflux-pump investigation. Followed HIT Il employed against M. abscessus for efflux-pump inhibition investigations and notable whole-cell efflux-pump inhibitory profile has been observed. The theoretical investigations predicted compound Il to be selective towards MmpL4, with significant hydrogen bonding and π-π stacking interactions effectively blocking a critical Asp-Tyr dyad interaction network necessary for proton translocation. Compound Il with bedaquiline highlighted an additive profile against the M. abscessus pathogen.ConclusionsMD simulations and whole-cell assays are indicating potential development of compound Il as an adjunct to the existing therapeutic regimen against mycobacterial infections

Ethnic differentials in early childhood mortality in Nepal

For more about the East-West Center, see http://www.eastwestcenter.org/</a

The Mycobactin Biosynthesis Pathway: A Prospective Therapeutic Target In The Battle Against Tuberculosis

The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link.The alarming rise in drug-resistant clinical cases of tuberculosis (TB) has necessitated the rapid development of newer chemotherapeutic agents with novel mechanisms of action. The mycobactin biosynthesis pathway, conserved only among the mycolata family of actinobacteria, a group of intracellularly surviving bacterial pathogens that includes Mycobacterium tuberculosis, generates a salicyl-capped peptide mycobactin under ironstress conditions in host macrophages to support the iron demands of the pathogen. This in vivo essentiality makes this less explored mycobactin biosynthesis pathway a promising endogenous target for novel lead-compounds discovery. In this Perspective, we have provided an up-to-date account of drug discovery efforts targeting selected enzymes (MbtI, MbtA, MbtM, and PPTase) from the mbt gene cluster (mbtA-mbtN). Furthermore, a succinct discussion on non-specific mycobactin biosynthesis inhibitors and the Trojan horse approach adopted to impair iron metabolism in mycobacteria has also been included in this Perspectiv

On-line multi-slice computed tomography interactive overlay with conventional X-ray : a new and advanced imaging fusion concept

Abstract Background Computed tomography (CT) has revolutionized noninvasive cardiovascular evaluations. Complicated percutaneous procedures require precise imaging guidance that conventional X-ray is often unable to provide. By combining X-ray imaging with real-time, interactive, CT-based landmarks, interventional procedures could be facilitated. We describe two cases using the first CT/Live X-ray overlay in which this technology shows its potential. Case reports A 31-year-old male with an anatomically complicated atrial septal defect (ASD) was referred for percutaneous closure. Transesophageal echocardiography (TEE) revealed an inferior location of the ASD complicated by it's proximity to a prominent Eustachian ridge. The CT was used to create a patient-specific physical model in preparation for the procedure and an in-lab real-time CT overlay allowing successful closure. A second case of a 41-year-old male with coronary artery disease status-post coronary artery bypass, aortic valve replacement (AVR), and aortic root replacement with an abnormal coronary computed tomography angiogram (CTA). In a prior procedure years ago the saphenous vein graft (SVG) to the left anterior descending artery (LAD) could not be cannulated during invasive angiography, given the patient's complicated and unusual anatomy. Using CT overlay, the superiorly and anteriorly located SVG was cannulated successfully. Discussion CT/Live X-ray overlay provided an adequate anatomical intra-procedural ASD evaluation, defect sizing, and guidance in one case and localization of an anatomically challenging graft ostium in the other case. Adding the CT landmarks as an overlay to traditional X-ray techniques provides a revolutionary and advanced imaging fusion concept that should improve procedural success. Keywords: Computed tomography; Angiography; Structural heart disease; Image fusion; OverlayGarcia J.A., Bhakta S., Kay J., Chan K.-C., Wink O., Ruijters D., Carroll J.D., ''On-line multi-slice computed tomography interactive overlay with conventional X-ray : a new and advanced imaging fusion concept'', International journal of cardiology, vol. 133, no. 3, pp. e101-e105, 2009.status: publishe