Suzanne S. Stokes
AstraZeneca
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Featured researches published by Suzanne S. Stokes.
Antimicrobial Agents and Chemotherapy | 2011
Scott D. Mills; Ann E. Eakin; Ed T. Buurman; Joseph V. Newman; Ning Gao; Hoan Huynh; Kenneth D. Johnson; Sushmita D. Lahiri; Adam B. Shapiro; Grant K. Walkup; Wei Yang; Suzanne S. Stokes
ABSTRACT DNA ligases are indispensable enzymes playing a critical role in DNA replication, recombination, and repair in all living organisms. Bacterial NAD+-dependent DNA ligase (LigA) was evaluated for its potential as a broad-spectrum antibacterial target. A novel class of substituted adenosine analogs was discovered by target-based high-throughput screening (HTS), and these compounds were optimized to render them more effective and selective inhibitors of LigA. The adenosine analogs inhibited the LigA activities of Escherichia coli, Haemophilus influenzae, Mycoplasma pneumoniae, Streptococcus pneumoniae, and Staphylococcus aureus, with inhibitory activities in the nanomolar range. They were selective for bacterial NAD+-dependent DNA ligases, showing no inhibitory activity against ATP-dependent human DNA ligase 1 or bacteriophage T4 ligase. Enzyme kinetic measurements demonstrated that the compounds bind competitively with NAD+. X-ray crystallography demonstrated that the adenosine analogs bind in the AMP-binding pocket of the LigA adenylation domain. Antibacterial activity was observed against pathogenic Gram-positive and atypical bacteria, such as S. aureus, S. pneumoniae, Streptococcus pyogenes, and M. pneumoniae, as well as against Gram-negative pathogens, such as H. influenzae and Moraxella catarrhalis. The mode of action was verified using recombinant strains with altered LigA expression, an Okazaki fragment accumulation assay, and the isolation of resistant strains with ligA mutations. In vivo efficacy was demonstrated in a murine S. aureus thigh infection model and a murine S. pneumoniae lung infection model. Treatment with the adenosine analogs reduced the bacterial burden (expressed in CFU) in the corresponding infected organ tissue as much as 1,000-fold, thus validating LigA as a target for antibacterial therapy.
Bioorganic & Medicinal Chemistry Letters | 2012
Suzanne S. Stokes; Robert Albert; Ed T. Buurman; Beth Andrews; Adam B. Shapiro; Oluyinka Green; Andrew R. McKenzie; Ludovic R. Otterbein
A previously described aryl sulfonamide series, originally found through HTS, targets GlmU, a bifunctional essential enzyme involved in bacterial cell wall synthesis. Using structure-guided design, the potency of enzyme inhibition was increased in multiple isozymes from different bacterial species. Unsuitable physical properties (low LogD and high molecular weight) of those compounds prevented them from entering the cytoplasm of bacteria and inhibiting cell growth. Further modifications described herein led to compounds that possessed antibacterial activity, which was shown to occur through inhibition of GlmU. The left-hand side amide and the right-hand side sulfonamides were modified such that enzyme inhibitory activity was maintained (IC(50) <0.1 μM against GlmU isozymes from Gram-negative organisms), and the lipophilicity was increased giving compounds with LogD -1 to 3. Antibacterial activity in an efflux-pump deficient mutant of Haemophilus influenzae resulted for compounds such as 13.
Journal of Biological Chemistry | 2011
Ed T. Buurman; Beth Andrews; Ning Gao; Jun Hu; Thomas A. Keating; Sushmita D. Lahiri; Ludovic R. Otterbein; Arthur Patten; Suzanne S. Stokes; Adam B. Shapiro
Background: A search was initiated to identify inhibitors of the acetyltransferase domain of GlmU that could be exploited as starting points for new antimicrobials. Results: Sulfonamide inhibitors were identified that upon chemical modification displayed antimicrobial activity mediated via GlmU. Conclusion: Enzymatic inhibition of GlmU can lead to antimicrobial activity. Significance: For the first time, GlmU was validated as an antimicrobial target in vitro. GlmU is a bifunctional enzyme that is essential for bacterial growth, converting d-glucosamine 1-phosphate into UDP-GlcNAc via acetylation and subsequent uridyl transfer. A biochemical screen of AstraZenecas compound library using GlmU of Escherichia coli identified novel sulfonamide inhibitors of the acetyltransferase reaction. Steady-state kinetics, ligand-observe NMR, isothermal titration calorimetry, and x-ray crystallography showed that the inhibitors were competitive with acetyl-CoA substrate. Iterative chemistry efforts improved biochemical potency against Gram-negative isozymes 300-fold and afforded antimicrobial activity against a strain of Haemophilus influenzae lacking its major efflux pump. Inhibition of precursor incorporation into bacterial macromolecules was consistent with the antimicrobial activity being caused by disruption of peptidoglycan and fatty acid biosyntheses. Isolation and characterization of two different resistant mutant strains identified the GlmU acetyltransferase domain as the molecular target. These data, along with x-ray co-crystal structures, confirmed the binding mode of the inhibitors and explained their relative lack of potency against Gram-positive GlmU isozymes. This is the first example of antimicrobial compounds mediating their growth inhibitory effects specifically via GlmU.
International Journal of Antimicrobial Agents | 2013
Maria Uria-Nickelsen; Georg Neckermann; Shubha Sriram; Beth Andrews; John Irvin Manchester; Dan Carcanague; Suzanne S. Stokes; Kenneth Gregory Hull
Pyrimidine compounds were identified as inhibitors of DNA topoisomerase IV through high-throughput screening. This study was designed to exemplify the in vitro activity of the pyrimidines against Gram-positive and Gram-negative microorganisms, to reveal the mode of action of these compounds and to demonstrate their in vivo efficacy. Frequencies of resistance to pyrimidines among Staphylococcus aureus and Streptococcus pneumoniae were <10(-10) at four times their minimum inhibitory concentrations (MICs). These compounds exhibited a dual mode of action through inhibition of the ParE subunit of DNA topoisomerase IV as well as the GyrB subunit of DNA gyrase, a homologue of DNA topoisomerase IV. Pyrimidines were shown to have MIC(90) values (MIC that inhibited 90% of the strains tested) of ≤2 mg/L against Gram-positive pathogens, including meticillin-resistant S. aureus, quinolone- and meticillin-resistant S. aureus, vancomycin-resistant enterococci, penicillin-non-susceptible S. pneumoniae and Streptococcus pyogenes, and MIC(90) values of 2- to >16 mg/L and ≤0.5 mg/L against the Gram-negative pathogens Haemophilus influenzae and Moraxella catarrhalis, respectively. The pyrimidines were bactericidal and exhibited a ca. 1000-fold reduction of the bacterial counts at 300 mg/kg in a S. pneumoniae lung infection model. The microbiological properties and in vivo efficacy of pyrimidines underscore their potential as candidates for the treatment of soft-tissue infections and hospital-acquired pneumonia.
Bioorganic & Medicinal Chemistry Letters | 2012
Suzanne S. Stokes; Madhusudhan Gowravaram; Hoan Huynh; Min Lu; George Mullen; Brendan Chen; Robert Albert; Thomas J. O’Shea; Michael T. Rooney; Haiqing Hu; Joseph V. Newman; Scott D. Mills
Optimization of clearance of adenosine inhibitors of bacterial NAD(+)-dependent DNA ligase is discussed. To reduce Cytochrome P-450-mediated metabolic clearance, many strategies were explored; however, most modifications resulted in compounds with reduced antibacterial activity and/or unchanged total clearance. The alkyl side chains of the 2-cycloalkoxyadenosines were fluorinated, and compounds with moderate antibacterial activity and favorable pharmacokinetic properties in rat and dog were identified.
Bioorganic & Medicinal Chemistry Letters | 2012
Oluyinka Green; Andrew R. McKenzie; Adam B. Shapiro; Ludovic R. Otterbein; Haihong Ni; Arthur Patten; Suzanne S. Stokes; Robert Albert; Sameer Kawatkar; Jason Breed
A novel arylsulfonamide-containing series of compounds represented by 1, discovered by highthroughput screening, inhibit the acetyltransferase domain of N-acetylglucosamine-1-phosphate-uridyltransferase/glucosamine-1-phosphate-acetyltransferase (GlmU). X-ray structure determination confirmed that inhibitor binds at the site occupied by acetyl-CoA, indicating that series is competitive with this substrate. This letter documents our early hit-to-lead evaluation of the chemical series and some of the findings that led to improvement in in-vitro potency against Gram-negative and Gram-positive bacterial isozymes, exemplified by compound 40.
Bioorganic & Medicinal Chemistry Letters | 2014
Kerry E. Murphy-Benenato; Hongming Wang; Helen M. McGuire; Hajnalka E. Davis; Ning Gao; D. Bryan Prince; Haris Jahić; Suzanne S. Stokes; P. Ann Boriack-Sjodin
In an attempt to identify novel inhibitors of NAD(+)-dependent DNA ligase (LigA) that are not affected by a known resistance mutation in the adenosine binding pocket, a detailed analysis of the binding sites of a variety of bacterial ligases was performed. This analysis revealed several similarities to the adenine binding region of kinases, which enabled a virtual screen of known kinase inhibitors. From this screen, a thienopyridine scaffold was identified that was shown to inhibit bacterial ligase. Further characterization through structure and enzymology revealed the compound was not affected by a previously disclosed resistance mutation in Streptococcus pneumoniae LigA, Leu75Phe. A subsequent medicinal chemistry program identified substitutions that resulted in an inhibitor with moderate activity across various Gram-positive bacterial LigA enzymes.
Bioorganic & Medicinal Chemistry Letters | 2011
Suzanne S. Stokes; Hoan Huynh; Madhusudhan Gowravaram; Robert Albert; Marta Cavero-Tomas; Brendan Chen; Jenna Harang; James T. Loch; Min Lu; George Mullen; Shannon Zhao; Ce-Feng Liu; Scott D. Mills
Archive | 2009
Ann Boriack-Sjodin; Daniel Robert Carcanague; Daemian Dussault; Holia Hatoum-Mokdad; Kenneth G. Hull; Georgine Ioannidis; John Irvin Manchester; Helen M. McGuire; David Charles Mckinney; Suzanne S. Stokes
Archive | 2009
Ann Boriack-Sjodin; Daniel Robert Carcanague; Daemian Dussault; Holia Hatoum-Mokdad; Kenneth G. Hull; Georgine Ioannidis; John Irvin Manchester; Helen M. McGuire; David Charles Mckinney; Suzanne S. Stokes