AID	Name	Type	Comment
2326	qHTS Assay for Inhibitors of Influenza NS1 Protein Function	confirmatory
623997	qHTS Assay for Inhibitors of Influenza NS1 Protein Function: Mammalian Cell Viability	other
602450	qHTS Assay for Inhibitors of Influenza NS1 Protein Function: Viral Replication TCID50 SAR for Probe	other
602454	qHTS Assay for Inhibitors of Influenza NS1 Protein Function: Viral Replication TCID50	other
602456	qHTS Assay for Inhibitors of Influenza NS1 Protein Function: RT-PCR	other
504647	Assay for Inhibitors of Influenza NS1 Protein Function: Influenza virus replication assay in MDCK cells infected with virus A/PR/8/34	other
492980	Assay for Inhibitors of Influenza NS1 Protein Function: Hit Validation	confirmatory
602452	qHTS Assay for Inhibitors of Influenza NS1 Protein Function: SAR Using NS1 and pYES strains	other

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: MH084878-01A1
Assay Submitter (PI): Daniel Engel

Influenza is a world-wide public health problem and emerging forms of the virus have the potential to cause a pandemic of equal or greater magnitude to the outbreaks recorded in 1918, 1957 or 1968. Vaccine development is proceeding and there also exist two classes of anti-influenza compounds. However these therapeutic modalities are neither fully effective nor widely enough available to fulfill global needs. In addition their potential usefulness against newly emergent strains is not known. Efforts are needed to develop novel agents against influenza virus, including broad-spectrum agents. Identification of small molecules that inhibit NS1 function either directly or by interfering with specific cellular pathways may be a key to increasing our defense against the virus.

We have miniaturized and optimized a cell based assay in which NS1 from influenza A is expressed in the budding yeast S. cerevisiae. NS1 is a multi-functional protein that counters the host innate immune response and facilitates viral versus cellular gene expression. Expression of NS1 causes a pronounced slow growth phenotype in yeast due to its intrinsic molecular activities. Small molecules that suppress the slow growth phenotype can be identified by a straightforward growth recovery assay using optical density (OD) as the measurement. The same yeast strain not expressing NS1 was used as positive control in the yeast growth recovery assay.

Please refer to AID 2326 for detailed assay protocols.

This summary is written for the purposes of summarizing the probe activities from the project. MLSCN probes are given a score of 100. Molecules in the literature are given a score of 80. Other, less active molecules in the same chemical series as the probe molecules are given a score of 50. Molecules pending validation are given a score of 10. Inactive analogues from these series are given a score of 0. No compounds from this project have yet been validated.

Grant Number: MH084878-01A1