Abstract: FR-PO566
Artificial Intelligence (AI)-Based Prospective Repurposing Studies for the Identification of New Promising Vasopressin V2 Receptor Ligands
Session Information
- Fluid, Electrolyte, and Acid-Base Disorders: Basic
October 25, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Fluid, Electrolytes, and Acid-Base Disorders
- 1101 Fluid, Electrolyte, and Acid-Base Disorders: Basic
Authors
- Angelini, Ines, Department of Biosciences, Biotechnologies and Environment, University of Bari “Aldo Moro”, Bari, Italy
- Trisciuzzi, Daniela, Department of Pharmacy - Pharmaceutical Science, University of Bari Aldo Moro, Bari, Italy
- Centrone, Mariangela, Department of Biosciences, Biotechnologies and Environment, University of Bari “Aldo Moro”, Bari, Italy
- Di Mise, Annarita, Department of Biosciences, Biotechnologies and Environment, University of Bari “Aldo Moro”, Bari, Italy
- Ranieri, Marianna, Department of Biosciences, Biotechnologies and Environment, University of Bari “Aldo Moro”, Bari, Italy
- Valenti, Giovanna, Department of Biosciences, Biotechnologies and Environment, University of Bari “Aldo Moro”, Bari, Italy
- Altomare, Cosimo Damiano, Department of Pharmacy - Pharmaceutical Science, University of Bari Aldo Moro, Bari, Italy
- Cotecchia, Susanna, Department of Biosciences, Biotechnologies and Environment, University of Bari “Aldo Moro”, Bari, Italy
- Nicolotti, Orazio, Department of Pharmacy - Pharmaceutical Science, University of Bari Aldo Moro, Bari, Italy
- Tamma, Grazia, Department of Biosciences, Biotechnologies and Environment, University of Bari “Aldo Moro”, Bari, Italy
Background
The vasopressin V2 receptor (V2R) controls renal water balance. As a G protein-coupled receptor (GPCR), the V2R transduces the vasopressin signals activating the cAMP/PKA pathway. Besides water reabsorption, the activation of V2R is involved in abnormal cell proliferation, cancer, and cyst enlargement in polycystic kidney disease. In this perspective, we performed an inverse screening of a large collection of known drugs to identify novel V2R ligands that might modulate different receptor-mediated effects.
Methods
The inverse screening campaign was run by using PLATO, our homemade target fishing platform. Structure-based studies were also carried out. Renal collecting duct MCD4 cells, stably expressing human V2R and aquaporin-2 (AQP2), were used as an experimental model to test the effects of drugs. Fluorescence Resonance Energy Transfer (FRET), and calcein fluorescence quenching (CFQ) were applied to evaluate changes in intracellular cAMP and DDAVP-induced water flux.
Results
Our prospective repurposing studies identified five promising drugs as potential V2R ligands (perphenazine, cloxacillin, clopidogrel, cabergoline, and F2544). FRET studies were conducted to test whether these compounds affect the DDAVP-induced cAMP responses. Interestingly, only one of them (F2544) at 1nM concentration significantly reduced the DDAVP-dependent cAMP production. Functional CFQ studies revealed that this modulator reduced the DDAVP-induced water reabsorption, with effects comparable to tolvaptan, a well-known V2R antagonist. In this respect, an in-depth computational investigation showed a nice overlap of the molecular interaction fields generated from the binding sites of V2R and F2544. Finally, molecular docking simulations returned a promising posing and scoring of F2544 in the V2R binding site.
Conclusion
The present findings identify for the first time new V2R ligands by applying an AI-based approach. Moreover, by combining in-depth computational investigations and functional studies F2544 was prioritized for being repurposed for treating diseases associated with abnormal V2R signaling.
Funding
- Government Support – Non-U.S.