scholarly journals High-Throughput Hit Screening Cascade to Identify Respiratory Syncytial Virus (RSV) Inhibitors

2015 ◽  
Vol 20 (5) ◽  
pp. 597-605 ◽  
Author(s):  
Helen Plant ◽  
Clare Stacey ◽  
Choi-Lai Tiong-Yip ◽  
Jarrod Walsh ◽  
Qin Yu ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
High Throughput ◽  
High Throughput Screening ◽  
Compound Library ◽  
Viral Inhibition ◽  
The United Kingdom ◽  
Physiological Relevance ◽  
Syncytial Virus ◽  
Infant Hospitalization ◽  
Screening Cascade

Respiratory syncytial virus (RSV) infects 99% of children by age 2 years and is a leading cause of serious lower respiratory tract infection (LRTI) and infant hospitalization in the United Kingdom. Identification of efficacious RSV therapeutics has been hindered by the lack of a robust and appropriate primary assay for high-throughput screening (HTS). Here we report an HTS cascade that identified inhibitors of RSV replication using a robust RSV replicon luminescence-reporter assay for the primary campaign. The performance of the assay was consistent and reliable at scale, with Z′ of 0.55 ± 0.08 across 150 assay plates and signal-to-background ratios >40. The HTS assay was used to screen the AstraZeneca compound library of 1 million compounds at a single concentration of 10 µM. Hits specifically targeting the RSV replicon were determined using a series of hit generation assays. Compounds nonspecifically causing cell toxicity were removed, and hits were confirmed in live viral inhibition assays exhibiting greater physiological relevance than the primary assay. In summary, we developed a robust screening cascade that identified hit molecules that specifically targeted RSV replication.

scholarly journals High-throughput screening of active compounds against human respiratory syncytial virus

Virology ◽  
2019 ◽  
Vol 535 ◽  
pp. 171-178
Author(s):  
Yuan-Hui Fu ◽  
Zhu-Xin Xu ◽  
Nan Jiang ◽  
Yan-Peng Zheng ◽  
Marie-Anne Rameix-Welti ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
High Throughput ◽  
High Throughput Screening ◽  
Human Respiratory Syncytial Virus ◽  
Active Compounds ◽  
Syncytial Virus

scholarly journals A cell based high-throughput screening approach for the discovery of new inhibitors of respiratory syncytial virus

2013 ◽  
Vol 10 (1) ◽  
pp. 19 ◽  
Author(s):  
Dong-Hoon Chung ◽  
Blake P Moore ◽  
Daljit S Matharu ◽  
Jennifer E Golden ◽  
Clinton Maddox ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
High Throughput ◽  
High Throughput Screening ◽  
Screening Approach ◽  
A Cell ◽  
Syncytial Virus

Characterization of novel respiratory syncytial virus inhibitors identified by high throughput screen

2015 ◽  
Vol 115 ◽  
pp. 71-74 ◽  
Author(s):  
Valerie A. Laganas ◽  
Ewan F. Dunn ◽  
Robert E. McLaughlin ◽  
Choi Lai Tiong-Yip ◽  
Olga Yuzhakov ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
High Throughput ◽  
High Throughput Screen ◽  
Syncytial Virus

scholarly journals Respiratory Syncytial Virus Prophylaxis in Special Populations: Is it Something Worth Considering in Cystic Fibrosis and Immunosuppression?

2011 ◽  
Vol 16 (2) ◽  
pp. 77-86
Author(s):  
William A. Prescott ◽  
David J. Hutchinson
Keyword(s):  
Respiratory Syncytial Virus ◽  
Cost Benefit ◽  
The United States ◽  
Randomized Controlled ◽  
Increased Risk ◽  
Rsv Infection ◽  
Syncytial Virus ◽  
The Cost ◽  
Infant Hospitalization ◽  
Infants And Young Children

ABSTRACT Respiratory syncytial virus (RSV) bronchiolitis is the leading cause of infant hospitalization in the United States. Prophylaxis with palivizumab is effective in reducing RSV hospitalizations in premature infants and in infants or children with chronic lung disease or congenital heart disease. Patients with CF or those who are immunocompromised may be at increased risk for RSV infection–related complications; hence, prophylaxis may prove beneficial to these populations. The extent of palivizumab use in the CF and immunocompromised populations is variable. Palivizumab appears to be safe and may be effective in infants and young children with CF and immunocompromise. However, well-designed, randomized, controlled trials published in peer-reviewed journals are lacking, and its routine use can therefore not be recommended at this time. If used in patients with CF or those who are immunocompromised, RSV prophylaxis should be restricted to peak outbreak months in order to optimize the cost benefit of palivizumab.

scholarly journals Development of a Simple Assay Protocol for High-Throughput Screening of Mycobacterium tuberculosis Glutamine Synthetase for the Identification of Novel Inhibitors

2005 ◽  
Vol 10 (7) ◽  
pp. 725-729 ◽  
Author(s):  
Upasana Singh ◽  
Vinita Panchanadikar ◽  
Dhiman Sarkar
Keyword(s):  
Escherichia Coli ◽  
Mycobacterium Tuberculosis ◽  
Glutamine Synthetase ◽  
Small Molecules ◽  
High Throughput ◽  
High Throughput Screening ◽  
Coli Strain ◽  
Compound Library ◽  
Essential Enzyme ◽  
Assay Protocol

Mycobacterium tuberculosis glutamine synthetase (GS) is an essential enzyme involved in the pathogenicity of the organism. The screening of a compound library using a robust high-throughput screening (HTS) assay is currently thought to be the most efficient way of getting lead molecules, which are potent inhibitors for this enzyme. The authors have purified the enzyme to a >90% level from the recombinant Escherichia coli strain YMC21E, and it was used for partial characterization as well as standardization experiments. The results indicated that the Kmof the enzyme for L-glutamine and hydroxylamine were 60 mM and 8.3 mM, respectively. The Km for ADP, arsenate, and Mn2+ were 2 [.proportional]M, 5 [.proportional]M, and 25 [.proportional]M, respectively. When the components were adjusted according to their Km values, the activity remained constant for at least 3 h at both 25° C and 37° C. The Z′ factor determined in microplate format indicated robustness of the assay. When the signal/noise ratios were determined for different assay volumes, it was observed that the 200-[.proportional]l volume was found to be optimum. The DMSO tolerance of the enzyme was checked up to 10%, with minimal inhibition. The IC50 value determined for L-methionine S-sulfoximine on the enzyme activity was 3 mM. Approximately 18,000 small molecules could be screened per day using this protocol by a Beckman Coulter HTS setup.

Recommendations for the use of palivizumab as prophylaxis against respiratory syncytial virus in infants with congenital cardiac disease

2003 ◽  
Vol 13 (5) ◽  
pp. 420-423 ◽  
Author(s):  
Robert Tulloh ◽  
Michael Marsh ◽  
Michael Blackburn ◽  
Frank Casey ◽  
Warren Lenney ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Cardiac Disease ◽  
Pulmonary Blood Flow ◽  
Surgical Intervention ◽  
Randomised Trial ◽  
Pulmonary Complications ◽  
The United Kingdom ◽  
Congenital Cardiac Disease ◽  
Syncytial Virus

New data are emerging on the use of palivizumab as prophylaxis against infection with the respiratory syncytial virus in infants with congenital cardiac disease. Following a 4-year multicentre randomised trial, it was shown that prophylactic injections with palivizumab were effective and safe for such children. Prophylaxis consists of 5, monthly, intramuscular injections of palivizumab, at a dose of 15 mg/kg, given during the season for infection with the respiratory syncytial virus. Timing is at the discretion of the physician, depending on the onset of the season locally. It is suggested that, in the United Kingdom, this should be commenced in mid-September. To help clinicians to identify appropriate candidates for palivizumab, a working group of the British Paediatric Cardiac Association has developed recommendations.Infants, namely those under 1 year old, with congenital cardiac disease likely to benefit from prophylaxis include those with haemodynamically significant lesions, particularly increased pulmonary blood flow with or without cyanosis; pulmonary venous congestion, pulmonary hypertension or long-term pulmonary complications, residual haemodynamic abnormalities following medical or surgical intervention (patients who have undergone cardiopulmonary bypass should receive an injection as soon as they are medically stable), cardiomyopathy requiring treatment, and congenital cardiac disease likely to need hospital admission for medical or surgical intervention during the season of infection with the virus. Prophylaxis with palivizumab may also be indicated, at the discretion of the physician, in some children with complex cardiac disease over the age of 1 year. Children less likely to benefit from prophylaxis are those with haemodynamically insignificant disease, or those with lesions adequately corrected by medical or surgical intervention.

scholarly journals High-Throughput Screening of a 100,000-Compound Library for Inhibitors of Influenza A Virus (H3N2)

2008 ◽  
Vol 13 (9) ◽  
pp. 879-887 ◽  
Author(s):  
William E. Severson ◽  
Michael McDowell ◽  
Subramaniam Ananthan ◽  
Dong-Hoon Chung ◽  
Lynn Rasmussen ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Influenza A ◽  
Moderate Activity ◽  
Compound Library ◽  
Highly Active ◽  
Virus Life Cycle ◽  
Influenza Activity ◽  
Compound Concentration ◽  
Viral Cytopathic Effect

Using a highly reproducible and robust cell-based high-throughput screening (HTS) assay, the authors screened a 100,000-compound library at 14- and 114-µM compound concentration against influenza strain A/Udorn/72 (H3N2). The “hit” rates (>50% inhibition of the viral cytopathic effect) from the 14- and 114-µM screens were 0.022% and 0.38%, respectively. The hits were evaluated for their antiviral activity, cell toxicity, and selectivity in dose-response experiments. The screen at the lower concentration yielded 3 compounds, which displayed moderate activity (SI50 = 10-49). Intriguingly, the screen at the higher concentration revealed several additional hits. Two of these hits were highly active with an SI50 > 50. Time of addition experiments revealed 1 compound that inhibited early and 4 other compounds that inhibited late in the virus life cycle, suggesting they affect entry and replication, respectively. The active compounds represent several different classes of molecules such as carboxanilides, 1-benzoyl-3-arylthioureas, sulfonamides, and benzothiazinones, which have not been previously identified as having antiviral/anti-influenza activity. ( Journal of Biomolecular Screening 2008:879-887)

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