Poster Presentation The 47th Lorne Conference on Protein Structure and Function 2022

Investigation of the structural and functional properties of the P2X1-purinoceptor for use in male contraception (#126)

Felix M. Bennetts 1 , Jesse I. Mobbs 1 , Alisa Glukhova 2 , Sab Ventura 1 , David M. Thal 1
  1. Drug Discovery Biology, Monash Institute Of Pharmaceutical Sciences, Melbourne, Victoria, Australia
  2. Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia

We aim to advance the finding that a dual genetic knockout of the P2X1-purinoceptor and α1A-adrenoceptor produces 100% infertility in male mice, by developing a pharmacological male contraceptive. A major barrier to this goal is the lack of drug-like P2X1-purinoceptor antagonists. Previous studies have identified P2X1-purinoceptor antagonists using high throughput screening or structure-activity relationship studies, but these compounds fall short due to poor potency, pharmacokinetics or selectivity. We hypothesise that a structure of the P2X1-purinoceptor will provide a wealth of biological information leading to numerous structure-based drug design opportunities at the P2X1-purinoceptor. To date, we have purified the full-length P2X1-purinoceptor and established pharmacological assays that validate the activity of the purified receptor. We have shown that the P2X1-purinoceptor is pure, homogeneous, and at suitable concentration for single-particle cryogenic electron microscopy, (cryo-EM) using SDS-PAGE, size exclusion chromatography, and 2D classifications from negative stain electron microscopy. Cryo-EM studies are underway to solve structures of the full-length P2X1-purinoceptor in apo and ligand-bound states. We are also integrating novel P2X1-purinoceptor antagonists and structural data to propel us towards a solid candidate for a male contraceptive pill.