ELISA screen of novel compounds that inhibit S100P and RAGE binding to identify new candidate drugs as a new therapy for pancreatic cancer

Deborah Ogbeni, Rama Camara, David Chau, Stuart Kirton, Sharon Rossiter, Louise MacKenzie

Research output: Contribution to conferenceAbstract

Abstract

Introduction: Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal human cancers with a 5-year survival rate of less than 5%. There is a high prevalence of S100P in PDAC; the calcium-binding protein S100P is highly metastasis-promoting through its extracellular interaction with the receptor for advanced glycation end-products (RAGE). Once bound by S100P, RAGE stimulates pancreatic tumour proliferation, invasion and metastasis progression in vivo.
Methods: Using computational chemistry methods, our laboratory has designed inhibitors that prevent RAGE-S100P binding. A total of 86 compounds were synthesized or purchased from a commercial supplier for screening. Development of an ELISA was based on a published study by (Padilla et al, 2014); 2µM S100P (made using recombinant human S100P plasmid in E.coli) was coated onto 96 well plates, and increasing concentrations of RAGE added to the wells. Cromolyn (10µM), a known inhibitor of S100P-RAGE binding was used as a positive control; after incubation with goat anti human antibody HRP conjugate and activation with tetramethylbenzidine (TMB), RAGE binding was ascertained at 450nm using a multiskan spectrophotometer. All 86 compounds were screened in triplicate, with a further two tests conducted with compounds that indicated inhibitory properties.
Results: This ELISA screen identified 18 lead compounds (Figure 1); 5 of which have shown anti-proliferative effects in pancreatic cell lines.
Conclusions: Results from this project will ultimately lead on to further screens for anti-migratory effects of the novel compounds on pancreatic cancer, and will be used as scaffolds for optimisation and further development of an S100P-RAGE inhibitor.
Padilla, L. et al (2014). S100 to receptor for advanced glycation end-products binding assay: Looking for inhibitors. Biochemical and Biophysical Research Communications, 446(1), 404-409.
Funded by Worldwide Cancer Research
Original languageEnglish
Publication statusPublished - 2015
EventPharmacology 2015 - London, United Kingdom
Duration: 16 Dec 2015 → …

Conference

ConferencePharmacology 2015
CountryUnited Kingdom
CityLondon
Period16/12/15 → …

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    Ogbeni, D., Camara, R., Chau, D., Kirton, S., Rossiter, S., & MacKenzie, L. (2015). ELISA screen of novel compounds that inhibit S100P and RAGE binding to identify new candidate drugs as a new therapy for pancreatic cancer. Abstract from Pharmacology 2015, London, United Kingdom.