Structural studies and molecular screening for the development of PKM2 and related proteins inhibitors

• Grant number: 20/13742-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: November 01, 2020
• End date: January 31, 2025
• Field of knowledge: Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
• Principal Investigator: José Carlos Farias Alves Filho
• Grantee: Andrew Albert de Oliveira
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 13/08216-2 - CRID - Center for Research in Inflammatory Diseases, AP.CEPID

Abstract

Inflammation is classically described as a response to infection or injury. Although occasional changes in inflammation are vital for survival, recent research suggests that certain social, environmental, and lifestyle factors can facilitate the development of chronic systemic inflammation, contributing collectively to various diseases of disability and mortality worldwide, such as cardiovascular dysfunction, cancer, diabetes mellitus, chronic liver diseases, among others. Therefore, inflammatory diseases represent a complex and heterogeneous group of disorders that cause morbidity and mortality, thus portraying a public health problem to be combated. In this context, the center of excellence in research on inflammatory diseases (CRID/USP) conducts studies in a translational way, involving basal and clinical research in the various areas of inflammatory diseases. In recent results, the center has identified multiple molecular targets that are involved in promoting these diseases. Therefore, the group aims to discover and plan new molecules capable of satisfactorily modulating these target proteins' activity. This work plan focuses on the identification and characterization of modulators of two selected enzymes as targets in the CRID: i. the enzyme Pyruvate kinase isoform two human (hPKM2), which catalyzes the conversion of phosphoenolpyruvate to pyruvate and, in addition to its metabolic role, is involved in the regulation of gene expression in the nucleus in the context of inflammatory diseases mediated by autoimmunity and cancer; and ii. human Peptidyl arginine deaminase type 4 (hPAD4), which catalyzes the citrullination and deamination of arginine residues in different proteins that act on chromatin regulating transcription production of NETS in the inflammation perspective, including those derived from viral infections. Preliminary results achieved at CRID involve the expression, purification, and preparation conditions of the hPAD4 enzyme for structural studies by macromolecule crystallography. In this proposal, it is intended to establish such routines for the hPKM2 target and proceed with the activities of discovery and development of new molecules with modulatory potential of the indicated targets (hPAD4 and hPKM2) through structural biology and biochemical and biophysical characterization of protein-ligand interaction. (AU)