Simultaneous Ligand-Directed Cytotoxic Toxin and Endosome Disruptor Delivery to Ablate Prostate Cancer
Abstract
In this grant we proposed to develop toxins that targeted GnRH-R and endosome disruptors that targeted bombesin 2 receptors to treat prostate cancer. In this first year, we have made GnRH-Gelonin and GnRH-Saporin conjugates. We have also established that GnRH-Gelonin and GnRH-Saporin conjugates induce apoptosis in a metastatic human prostate cancer cell lines (PC-3). We have further shown that targeting the endosome disruptor, listeriolysin O (LLO), to PC-3 cells using the kisspeptin receptor agonist (metastin) induces apoptosis. Metastin targeted LLO increases apoptosis induced by GnRH-Gelonin and/or GnRH-Saporin conjugates. We have developed a 2-step FPLC based purification for a his-tagged LLO that includes hydrophobic interaction chromatography followed by nickel affinity chromatography. We have also developed a 2-step FPLC based purification of his-tagged Gelonin purification, which includes nickel affinity chromatography followed by size exclusion chromatography. We have synthesized D-Lys6 GnRH and the Bombesin 2 receptor agonist, D-Phe6-Nle14 Bombesin7-14. These will be conjugated to LLO for additional in vitro tests. As proposed in the initial application, in vivo model development and testing will begin this year and will take full advantage of current and future in vitro results.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2018
- Accession Number
- AD1063939
Entities
People
- Benjamin J. Renquist
- Josef Vagner
- Ramesh Selvaraj
Organizations
- University of Georgia Research Foundation