Development of a High Throughput Screening Campaign for the Identification of New Pepsin Inhibitors
Mentor 1
Alexander Arnold
Location
Union Wisconsin Room
Start Date
24-4-2015 2:30 PM
End Date
24-4-2015 3:45 PM
Description
Laryngopharyngeal Reflux Disease (LPRD) is an extension of gastroesophageal reflux disease (GERD) where gastric contents are refluxed into extraesophageal tissues. Current acid suppression treatment with proton pump inhibitors (PPIs) has proven ineffective because unlike GERD, pepsin is the problem not stomach acid. When pepsin is refluxed into these tissues it digests healthy protein, damaging cells, leading to mutations and causing subsequent cancers. The objective of this research is to identify new pepsin inhibitors to completely and irreversibly inhibit pepsin’s activity when present in extraesophageal tissues. In order to find inhibitors, three different fluorescence assays were developed using labeled pepstatin, casein, and peptide probes to indicate varying levels of inhibition of the pepsin enzyme. Once optimized, these assays were used to screen the Library of Pharmacologically Active Compounds (LOPAC) for possible pepsin inhibitors. Compounds were identified as hits if they showed inhibition activity of ± 3 standard deviations away from the mean activity of all compounds. The assays proved to be sensitive and specific, turning up only a few hit compounds. The results of this research were encouraging and we look forward to moving these target compounds to cell based assays to test their viability as drug candidates.
Development of a High Throughput Screening Campaign for the Identification of New Pepsin Inhibitors
Union Wisconsin Room
Laryngopharyngeal Reflux Disease (LPRD) is an extension of gastroesophageal reflux disease (GERD) where gastric contents are refluxed into extraesophageal tissues. Current acid suppression treatment with proton pump inhibitors (PPIs) has proven ineffective because unlike GERD, pepsin is the problem not stomach acid. When pepsin is refluxed into these tissues it digests healthy protein, damaging cells, leading to mutations and causing subsequent cancers. The objective of this research is to identify new pepsin inhibitors to completely and irreversibly inhibit pepsin’s activity when present in extraesophageal tissues. In order to find inhibitors, three different fluorescence assays were developed using labeled pepstatin, casein, and peptide probes to indicate varying levels of inhibition of the pepsin enzyme. Once optimized, these assays were used to screen the Library of Pharmacologically Active Compounds (LOPAC) for possible pepsin inhibitors. Compounds were identified as hits if they showed inhibition activity of ± 3 standard deviations away from the mean activity of all compounds. The assays proved to be sensitive and specific, turning up only a few hit compounds. The results of this research were encouraging and we look forward to moving these target compounds to cell based assays to test their viability as drug candidates.
