Identification of compounds that interact with the ClpC1 enzyme of the ClpP1P2C1 protein complex of Mycobacterium tuberculosis by fragment screening

• Grant number: 24/15711-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2025
• End date: December 31, 2025
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Marcio Vinicius Bertacine Dias
• Grantee: Pedro Henrique Campos Ribeiro
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 21/10577-0 - Biology of Bacteria and Bacteriophages Research Center, AP.CEPID

Abstract

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis and affects around 10 million people a year worldwide, making it one of the main causes of death from infections. The therapy used to treat tuberculosis is complex and requires at least three different relatively toxic drugs over a period of approximately 6 months, which makes it easier for patients to give up and for resistant strains to emerge. Only recently, after a period of around 40 years, have two new drugs been introduced onto the market for the treatment of multidrug-resistant tuberculosis. Thus, new therapy strategies against tuberculosis are needed and the AAA+ degradation systems present interesting targets for the development of new antimicrobials, since this is an essential process for the survival of the mycobacterium, and should cause damage to the bacterial cell if this process is deregulated either positively or negatively. Thus, the ClpC1 protein of Mycobacterium tuberculosis (MtbClpC), which is a hexameric AAA+ unfoldase and participates in the degradation of porteins by the ClpC1/P1P2 system, is an essential and viable target for the development of new drugs. This system has been little explored as a target for drug development using the fragment approach, especially from a structural point of view. Therefore, in this project, we intend to identify compounds that interact with the N-terminal domain of ClpC1 using biophysical techniques. This will be done by overexpressing this domain of ClpPC1 in a heterologous system. The protein will be purified using affinity and gel filtration columns. ClpC1 will then be screened against a library of low molecular weight molecules. The interaction will be validated using alternative orthogonal biophysical techniques and attempts will be made to obtain the structure of the N-terminal domain of ClpC1 in complexes with the molecules identified. The results obtained in this project will be the starting point for the synthesis of compounds that show activity that deregulates the activity of ClpC1 and consequently candidates for the development of new molecules with anti-tuberculosis activity.