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CHM 114: Bioorganic Molecules Week 6 Laboratory
CHM 114: Bioorganic Molecules Week 6 Laboratory Azosulfonamides: Part 1. Synthesis of a Ligand for Protein Binding Studies1 Dyes and Serendipity: In the Persian fairy tale The Three Princes of Serendip, the title characters were forever discovering things they were not looking for at the time—thus, serendipity is the aptitude for for making happy discoveries by accident. The preparation of the first commercially important synthetic dye by William Henry Perkin (not of Perkin’s restaurant fame) in 1854 is a good example of a serendipitous discovery in science. During the nineteenth century quinine was the only drug known to be effective against malaria, and it could be obtained only from the bark of the cinchona tree, which grew in South America. French chemists had isolated pure quinine from cinchona bark in 1820, but the inaccessibility of the tree made natural quinine very expensive. Perkin, then an 18-year-old graduate student working for the eminent German chemist August Wilhelm von Hofmann, realized that anyone who could make synthetic quinine might well become rich and famous. Perkin knew nothing about the molecular structure of quinine—structural organic chemistry was in its infancy in the mid-1800’s—but he knew that quinine’s molecular formula was C20H24N2O2. Perkin prepared some allytoluidine (C10H13N), apparently thinking that two molecules of allyltoluidine plus three oxygen atoms minus a molecule of water would magically yield C20H24N2O2 –quinine! (Figure 1). H H N N +3[O] 2 OH H H C -H2O 3 H3CO Allyltoluidine N Quinine Figure 1. Perkin’s Idea for the Synthesis of Quinine Of course Perkin had attempted the impossible. -
Linezolid - Tigecycline
Linezolid - Tigecycline Paul M. Tulkens, MD, PhD Cellular and Molecular Pharmacology Louvain Drug Research Institute Catholic University of Louvain, Brussels, Belgium With the support of Wallonie-Bruxelles-International 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 1 Dong-A pharmaceuticals and tedizolid: step #1 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 2 Mode of action: • Protein synthesis inhibition: LZD binds to the 23S portion of the ribosomal 50S subunit (the centre of peptidyl transferase activity) → no initial complex 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 3 RNA interaction Karen L. Leach et al, Molecular Cell (2007) 26,393-402 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 4 Spectrum of activity No useful activity against other Gram-negative organisms because of constitutive efflux ! 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 5 Registered clinical indications Linezolid is often used off-label (endocarditis, osteomyelitis, ….) in pace of vancomycin 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 6 Linzezolid: mechanism of resistance 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 7 Can linzolid induce resistance ? 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 8 Linzolid can induce resistance… Locke et al. Antimicrob Agent Chemother 2009;53:5265-5274. 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 9 Linezolid pharmacokinetics 12-11-2015 WBI - HUP Cooperation - Bach Mai Hospital 10 Linezolid human pharmacokinetics Oral therapeutic doses (600mg linezolid q12h for 21 days) Linezolid Tedizolid MIC 90 MIC90 Muñoz et al. ECCMID 2010; P1594 Flanagan SD, et al. Pharmacotherapy 2014;34(3):240–250. -
Antimicrobial Resistance in Bacterial Pathogens and Detection of Carbapenemases in Klebsiella Pneumoniae Isolates from Hospital Wastewater
antibiotics Article Antimicrobial Resistance in Bacterial Pathogens and Detection of Carbapenemases in Klebsiella pneumoniae Isolates from Hospital Wastewater Hercules Sakkas * , Petros Bozidis , Afrodite Ilia, George Mpekoulis and Chrissanthy Papadopoulou Microbiology Department, Faculty of Medicine, School of Health Sciences, University of Ioannina, 45110 Ioannina, Greece * Correspondence: [email protected]; Tel.: +30-265-100-7769 Received: 21 May 2019; Accepted: 26 June 2019; Published: 27 June 2019 Abstract: During a six-month period (October 2017–March 2018), the prevalence and susceptibility of important pathogenic bacteria isolated from 12 hospital raw sewage samples in North Western Greece was investigated. The samples were analyzed for methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), extended-spectrum beta-lactamase (ESBL) producing Escherichia coli, carbapenemase-producing Klebsiella pneumoniae (CKP), and multidrug-resistant Pseudomonas aeruginosa. Antimicrobial susceptibility testing was performed using the agar diffusion method according to the recommendations of the Clinical and Laboratory Standards Institute. The diversity of carbapenemases harboring K. pneumoniae was examined by two phenotyping screening methods (modified Hodge test and combined disk test), a new immunochromatographic rapid assay (RESIST-4 O.K.N.V.) and a polymerase chain reaction (PCR). The results demonstrated the prevalence of MRSA, vancomycin-resistant Staphylococcus aureus (VRSA), VRE, and CKP in the examined -
Tigecycline: a Igecycline
Molecules of the Millennium Tigecycline: A novel glycylcycline antibioticantibiotic Tetracycline antibiotics were first isolated at Lederle to occur.[5] Tigecycline is also active against organisms that Laboratories in 1945 and represented a significant display efflux-based resistance, which may be because of the advancement in the treatment of many infections. However, inability of the glycylcyclines to induce tetracycline efflux due to an increased incidence of resistance among various proteins, or because the efflux protein cannot export bacteria, the use of tetracyclines has been relegated to second tigecycline.[6] and third-line categories for most clinical indications. The two The binding site of tigecycline on the ribosome is common major mechanisms of resistance include tetracycline efflux to tetracyclines, but tigecycline binds 5-fold more strongly to and ribosomal protection, where tetracycline is prevented from the ribosome than tetracyclines and this enhanced binding is, binding to the ribosome. Research to find tetracycline probably, responsible for overcoming the ribosomal protection analogues, that circumvented these resistance mechanisms, mechanisms of tetracycline resistance.[5] The action of has led to the development of a novel group of drugs called tigecycline is bacteriostatic in nature, which is likely due to its glycylcyclines, the most promising compound being the 9-tert reversible interaction with the ribosome.[5] Its efficacy suggests butyl glycyclamido derivative of minocycline-tigecycline (GAR that traditional thinking about using bacteriostatic drugs in 936). treating serious infections needs to be revised.[7] Chemistry Antimicrobial activity The nucleus consists of four linear fused tetracyclic rings In vitro antibacterial activity of tigecycline has been and there is the addition of N, N-dimethylglycylamido (DMG) assessed against clinical isolates as a part of ongoing TEST group at C-9 position of minocycline.[1] initiative (Tigecycline Evaluation Surveillance Trial). -
From Prontosil Rubrum to Antipsychotics
IJRPC 2017, 7(3), 306-319 Edna S Suyenaga et al. ISSN: 22312781 INTERNATIONAL JOURNAL OF RESEARCH IN PHARMACY AND CHEMISTRY Available online at www.ijrpc.com Research Article GENESIS OF PHARMACEUTICALS: FROM PRONTOSIL RUBRUM TO ANTIPSYCHOTICS – A HISTORY OF SULFA DRUGS FROM THE PERSPECTIVE OF MEDICINAL CHEMISTRY Gabriela M Aver1, Vitória HM Mottin2, Olyr C Kreutz1 and Edna S Suyenaga1,2* 1Universidade Feevale, ERS-239, 2755, Vila Nova, 93525-000, Novo Hamburgo–RS, Brazil. 2Universidade Vale do Rio dos Sinos, Avenida Unisinos, 950, 93022-000, São Leopoldo–RS, Brazil. ABSTRACT Based on research conducted on dye Prontosil Rubrum, it was possible to obtain the first synthetic bacteriostatic, the sulfas, which present sulfanilamide as its pharmacophore. This prototype made possible to obtain pharmaceuticals of different pharmacological classes, such as diuretics, hypoglycemiants, anti-inflammatories, antipsychotics, anti-leprosy drugs and antiadrenergics. This paper intends to report, through a bibliographic review, the structural alterations that characterize the different therapeutic classes of sulfanilamide derivatives, through chemical structure versus pharmacological activity relationship analysis. As a result, it was observed that some chemical groups are essential to define the pharmacological action. Diuretic drugs derived from sulfanilamide present groups of benzothiazide, thiadiazole and associated to sulfamoylbenzoic acid in their chemical structure. As for hypoglycemiants, they present the group sulfonylurea in their chemical structure. In the chemical structure of oxicams, there is cyclization of the pharmacophore, 4- hydroxy-2-methyl-1,2-benzothiazine-3-carboxamide -1,1-dioxide. The coxibs present in their chemical structures two aromatic rings linked by a furan or trifluoromethyl pyrazole ring, the antipsychotics show the group N-(pyrrolidin-2-yl) -sulfonylbenzamide. -
Chapter 12 Antimicrobial Therapy Antibiotics
3/31/2010 Chapter 12 Antimicrobial Therapy Topics: • Ehrlich (1900’s) – compound 606 to - Antimicrobial Therapy treat syphilis. Coined the term - Selective Toxicity “selective toxicity” - Survey of Antimicrobial Drug • Fleming (1928) – discovered - Microbial Drug Resistance penicillin - Drug and Host Interaction • Domagk (1930s) – “prontosil” was modified in the body to an active compound (first sulfa drug!) An ideal antimicrobic: Chemotherapy is the use of any chemical - soluble in body fluids, agent in the treatment of disease. - selectively toxic , - nonallergenic, An antibiotic agent is usually considered to - reasonable half life (maintained at a be a chemical substance made by a constant therapeutic concentration) microorganism that can inhibit the growth or - unlikely to elicit resistance, kill another microorganisms. - has a long shelf life, An antimicrobic or antimicrobial agent is - reasonably priced. a chemical substance similar to an There is no ideal antimicrobic!!! antibiotic, but may be synthetic. Selective Toxicity - Drugs that specifically target microbial processes, and not the human host’s. Antibiotics Spectrum of antibiotics and targets • Naturally occurring antimicrobials – Metabolic products of bacteria and fungi – Reduce competition for nutrients and space • Bacteria – Streptomyces, Bacillus, • Molds – Penicillium, Cephalosporium * * 1 3/31/2010 The mechanism of action for different 5 General Mechanisms of Action for antimicrobial drug targets in bacterial cells Antibiotics - Inhibition of Cell Wall Synthesis - Disruption of Cell Membrane Function - Inhibition of Protein Synthesis - Inhibition of Nucleic Acid Synthesis - Anti-metabolic activity Antibiotics weaken the cell wall, and cause the cell to lyse . Cell wall synthesis • Bactericidal • Vancomycin – hinders peptidoglycan elongation • Penicillin and cephalosporins – binds and blocks peptidases involved in cross-linking the glycan molecules Fig. -
Progress in Pharmacokinetics and Pharmacodynamics - I
274 Abstracts Progress in pharmacokinetics and pharmacodynamics - I P1022 Pharmacokinetics of telithromycin in plasma and was higher in young women than in young men (21% difference), soft tissue after single-dose administration in healthy volunteers with only a 4% difference between elderly women and men. At the target clinical dose of 100 mg load infused over 30–60 min fol- R. Gattringer, F. Traunmueller, E. Urbauer, M. Zeitlinger, lowed by 50 mg q12h, Cmax and AUCss (mean Æ SD) were M. Mueller, C. Joukhadar 621 Æ 93 ng/mL and 3069 Æ 381 ng h/mL, respectively. Vienna, A Objectives: Telithromycin was described to reach high concentra- Dose (mg), with MD given q 12h tions levels in inflammatory fluid, in bronchopulmonary tissues and in tonsillar tissue. Because of these data telithromycin is spe- Pk parameter 12.5 25 50 75 100 200 300 culated to be a new option in the therapy of skin and soft tissue infections. To determine the concentration of telithromycin in the SD CLt 0.29 Æ 0.20 0.20 Æ 0.10 0.28 Æ 0.04 0.29 Æ 0.04 0.30 Æ 0.08 0.23 Æ 0.04 0.25 Æ 0.03 interstitial space fluid, the pharmacokinetics of this new antibiotic (L/hr/kg) (n ¼ 6) (n ¼ 6) (n ¼ 6) (n ¼ 6) (n ¼ 57) (n ¼ 24) (n ¼ 12) were assessed after single dose administration in young healthy MD CLt ÁÁÁ 0.20 Æ 0.04 0.20 Æ 0.02 ÁÁÁ 0.24 Æ 0.045 ÁÁÁ ÁÁÁ (L/hr/kg) (n ¼ 5) (n ¼ 5) (n ¼ 3) volunteers by the use of microdialysis. -
Supplementary Information Table of Contents Table S1
Supplementary Information Table of Contents Table S1. Matched high-throughput sequencing reads of antibiotic resistance genes (ARGs) in sludge fed with 0 mg/L tetracycline. S1 Table S2. Matched high-throughput sequencing reads of antibiotic resistance genes (ARGs) in sludge fed with 20 mg/L tetracycline. S12 Table S3. Matched high-throughput sequencing reads of plasmids in sludge cultured with 0 mg/L tetracycline. S21 Table S4. Matched high-throughput sequencing reads of plasmids in sludge cultured with 20 mg/L tetracycline. S32 Table S5. PCR primers of the 15 tetracycline resistance genes detected in this study. S40 Figure S1. Occurrence patterns of 11 tet genes in activated sludge analyzed by electrophoresis of PCR products. S41 Table S1. Matched high-throughput sequencing reads of antibiotic resistance genes (ARGs) in sludge fed with 0 mg/L tetracycline of Day 6 against antibiotic resistance database (ARDB). ARDB Accession Identity Hit Length Number ARG No. E Value t Function Related Antibiotics Number (%) n (AA) u of Reads Name 1 CAE53425 90.62 25 7.0 × 10−8 1628 sul2 Sulfonamide-resistant dihydropteroate synthase sulfonamide 2 YP_001969930 90 28 6.0 × 10−9 182 sul2 Sulfonamide-resistant dihydropteroate synthase sulfonamide 3 YP_002112964 100 27 1.0 × 10−11 29 aph33ib Aminoglycoside O-phosphotransferase streptomycin 4 YP_001571041 90 28 5.0 × 10−9 21 macB Macrolide-specific efflux system macrolide 5 YP_002029849 90.32 27 2.0 × 10−8 21 smeE Multidrug resistance efflux pump fluoroquinolone 6 YP_002890644 90.32 31 3.0 × 10−10 21 bacA -
COMPLETE GENOME SEQUENCING of Pseudomonas Syringae Pv. Actinidiae BIOVAR 3, P155, KIWIFRUIT PATHOGEN ORIGINATING from CHINA SEQU
2220 Bioscience Journal Original Article COMPLETE GENOME SEQUENCING OF Pseudomonas syringae pv. actinidiae BIOVAR 3, P155, KIWIFRUIT PATHOGEN ORIGINATING FROM CHINA SEQUENCIAMENTO COMPLETO DO GENOMA Pseudomonas syringae pv. actinidiae BIOVAR 3, P155, AGENTE PATOGÊNICO DO KIWI, ORIGINÁRIO DA CHINA Xin PAN1,2; Siyue ZHAO3; Yongzhi WANG2; Mingzhang LI2; Liqin HE2*; Qiguo ZHUANG2* 1. College of Tourism and Town and Country Planning, Chengdu University of Technology, Chengdu, Sichuan, China; 2. Kiwifruit Breeding and Utilization Key Laboratory, Sichuan Provincial Academy of Natural Resource Sciences, Chengdu, China; 3. Department of Applied Microbiology, College of Resources, Sichuan Agricultural University, Chengdu, Sichuan, China.*Corresponding author: Qiguo Zhuang, [email protected]; Liqin HE, [email protected] ABSTRACT: Pseudomonas syringae pv. actinidiae is a bacterial pathogen of kiwifruit. Based on the results of the pathogenicity assay, we sequenced the strain Pseudomonas syringae (Psa3) P155 which possesses a series of virulence and resistance genes, CRISPR candidate elements, prophage related sequences, methylation modifications, genomic islands as well as one plasmid. Most importantly, the copper resistance genes copA, copB, copC, copD, and copZ as well as aminoglycoside resistance gene ksgA were identified in strain P155, which would pose a threat to kiwifruit production. The complete sequence we reported here will provide valuable information for a better understanding of the genetic structure and pathogenic characteristics of the genome of P155. KEYWORDS: Actinidia sp. Bacterial canker. Complete sequence. Pathogenicity. Pac-Bio platform. INTRODUCTION the development of whole genome sequencing, virulence and antimicrobial resistance profile of Kiwifruit (Actinidia sp.), an economically bacteria can be predicted. Additionally, the sequence important fruit, is cultivated worldwide for its taste data also provide a level of strain discrimination and and nutritional value (Fujikawa and Sawada 2016). -
Protein Synthesis Inhibitors Lecture Outline
Protein Synthesis Inhibitors • Macrolides - Lincosamides • Aminoglycosides • Tetracyclines • Chloramphenicol • Oxazolidinones • Streptogramins Lecture Outline • Description of protein synthesis - translation • Antibiotics – Structure - function - classification – Mechanism(s) of action – Mechanism(s) of resistance – Spectrum of activity/Indications for use – Pharmacology – Toxicity • Clinical examples 1 Overview of Translation (1) Initiation: • 30S binds RBS of mRNA • AA binds tRNA using aminoacyl-tRNA synthetase • IF2 and fmet-tRNA binds 30S at P site • 50S binds complex 70S resulilting in th e f ormati on of the initiation complex Overview of Translation (2) Initiation – tRNA + AA binds translation elongation factor – Enters ribosome and attaches at the A site 2 Overview of Translation (3) Amino Acid Transfer – Petidyltransferase on 50S ribosome attaches the next AA to the polypeptide – Met added to Leu at A site Overview of Translation (4) Elongation tRNA moved to P site by EF-G creating room at A site for next tRNA Translation termination Occurs at nonsense codon sites e.g. UAA Release factors Ribosome dissociates 3 Mechanisms of Action - Protein Synthesis Inhibitors Macrolides • Broad spectrum antibiotics • Original agent: erythromycin • Azalides: azithromycin and clarithromycin – seltdtiilected antimicrobi bildhal and pharmacoki kitinetic advantages 4 Large 14 member macrolactone ring with one or more deoxy sugars attached. Inhibits formation of 50S ribosome blocking trans- peptidation or translocation. Large 14 member lactone ring -
Antimicrobial Effects of Minocycline, Tigecycline, Ciprofloxacin
antibiotics Article Antimicrobial Effects of Minocycline, Tigecycline, Ciprofloxacin, and Levofloxacin against Elizabethkingia anophelis Using In Vitro Time-Kill Assays and In Vivo Zebrafish Animal Models Jiun-Nong Lin 1,2,3,* , Chung-Hsu Lai 2,3, Yi-Han Huang 3 and Chih-Hui Yang 4 1 Department of Critical Care Medicine, E-Da Hospital, I-Shou University, Kaohsiung 824, Taiwan 2 Division of Infectious Diseases, Department of Internal Medicine, E-Da Hospital, I-Shou University, Kaohsiung 824, Taiwan; [email protected] 3 School of Medicine, College of Medicine, I-Shou University, Kaohsiung 824, Taiwan; [email protected] 4 Department of Biological Science and Technology, Meiho University, Pingtung 912, Taiwan; [email protected] * Correspondence: [email protected] Abstract: Elizabethkingia anophelis is a multidrug-resistant pathogen. This study evaluated the antimicro- bial activity of minocycline, tigecycline, ciprofloxacin, and levofloxacin using in vitro time-kill assays and in vivo zebrafish animal models. The E. anophelis strain ED853-49 was arbitrarily selected from a bacterial collection which was concomitantly susceptible to minocycline, tigecycline, ciprofloxacin, and levofloxacin. The antibacterial activities of single agents at 0.5–4 × minimum inhibitory concentration Citation: Lin, J.-N.; Lai, C.-H.; (MIC) and dual-agent combinations at 2 × MIC using time-kill assays were investigated. The therapeutic Huang, Y.-H.; Yang, C.-H. Antimicrobial Effects of Minocycline, effects of antibiotics in E. anophelis-infected zebrafish were examined. Both minocycline and tigecycline Tigecycline, Ciprofloxacin, and demonstrated bacteriostatic effects but no bactericidal effect. Minocycline at concentrations ≥2 × Levofloxacin against Elizabethkingia MIC and tigecycline at concentrations ≥3 × MIC exhibited a long-standing inhibitory effect for 48 h. -
Antibiotic Prescribing – Quality Indicators
Clinical Microbiology and Infection, Volume 12, Supplement 4, 2006 Antibiotic prescribing – quality indicators P1460 Methods: Medical records from all patients with positive blood Is self-medication with antibiotics in Europe cultures in 2001 were analysed retrospectively. Factors driven by prescribed use? predisposing to infections, results of blood cultures, antibiotic use, and outcome were recorded. L. Grigoryan, F.M. Haaijer-Ruskamp, J.G.M. Burgerhof, Results: The antibiotic use in 226 episodes of true bacteraemia J.E. Degener, R. Deschepper, D. Monnet, A. Di Matteo, were analysed. According to guidelines empirical antibiotic ˚ and the SAR E.A. Scicluna, A. Bara, C. Stalsby Lundborg, J. Birkin treatment should be adjusted in 166 episodes. Antibiotic use was group adjusted in 146 (88%) of these 166 episodes, which led to a Objectives: The occurrence of self-medication with antibiotics narrowing of therapy in 118 (80%) episodes. Compared to has been described in the US and Europe, a possible empirical therapy there was a 22% reduction in the number of contributing factor to increased antibiotic resistance. An antibiotics. Adjustment of therapy was more often performed in important reason for using self-medication can be past Gram-negative bacteraemia and polymicrobial cultures than in experience with antibiotics prescribed by health professionals. Gram-positive bacteraemia. In bacteraemia caused by We investigated whether self-medication in Europe follows the ampicillin-resistant E. coli, ampicillin was mostly replaced by same pattern as prescribed use. ciprofloxacin. The cost for 7 days adjusted therapy was 19800 Methods: A population survey was conducted in: North and EUR (23%) less than for 7 days of empirical therapy.