Chemical library screening and structural and biochemical characterization of ligands of novel molecular targets for the treatment of inflammatory diseases.

• Grant number: 19/10473-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: July 01, 2019
• End date: March 31, 2023
• Field of knowledge: Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
• Principal Investigator: Thiago Mattar Cunha
• Grantee: Amanda Araujo Souza
• 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

Inflammatory diseases are a complex and heterogeneous group of diseases that causes morbidity and mortality and represent an important socioeconomic problem. The mission of the Center of Research in Inflammatory Diseases (CRID) is to perform translational research, basic and clinical research in the area of inflammatory diseases. In the last years several studies in CRID labs have characterized the role of some molecules in the pathophysiology of inflammatory diseases. Based on these targets, one aim of CRID is to discover novel chemical modulators (small molecule ligands) of hKMO and hPAD4. The kynurenine 3-monooxygenase (KMO) catalyses L-kynurenine (L-Kyn) hydroxylation to form 3-hydroxy-L- kynurenine (L-3OHKyn). As an enzyme of the kynurenine (Kyn) pathway KMO is an attractive target for neurodegenerative and neuroinflammatory diseases3,4. The protein-arginine deiminase type-4 (PAD4) catalyzes the citrullination and deamination of arginin residues in histones, regulating transcription acting on chromatin. PAD4 is considered a target for rheumatoid arthritis5.We have already established the protocols for preparing hKMO and PAD4 for structural studies (protein X-ray crystallography) and have developed an in vitro assay for accessing commercial KMO activity from Pseudomonas fluorescens. In this context, from the previous results, now the focus will be on establishing the methods for screening assays of the hKMO enzyme using the perform high throughput screening assay (HTS), followed by the screening of chemical libraries and biochemical and structural characterization of the inhibitors. Once performed will be given to the establishment of the PAD4 assay, screening and characterization of small chemical modulators.