Investigating Enzymatic Kinetics and Compound Screening for Inhibitor Candidates of Pyruvate Kinase II (PKII) from Plasmodium falciparum

• Grant number: 23/10023-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2023
• End date: July 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Rafael Victorio Carvalho Guido
• Grantee: João Pedro Salazar Martins
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated research grant: 13/07600-3 - CIBFar - Center for Innovation in Biodiversity and Drug Discovery, AP.CEPID

Abstract

Malaria is a deadly parasitic disease with a global prevalence, and despite considerable efforts in the search for new treatments, a high mortality rate still persists. In the search for new antimalarial drugs, targeting the glycolytic pathway has been explored to inhibit parasite development and combat the infection. Among the enzymes in the glycolytic pathway, Plasmodium falciparum pyruvate kinase II (PfPKII) has emerged as an attractive molecular target. PfPKII catalyzes the conversion of phosphoenolpyruvate (PEP) and ADP to pyruvate and ATP in the parasite's apicoplast. It plays a crucial role in the functioning of this organelle and is involved in the biosynthesis of type II fatty acids. Given the significance of glycolytic enzymes in the parasite's metabolism and the need for validated drug targets in P. falciparum, this proposal aims to discover inhibitors of PfPKII from chemical libraries provided by the not-for-profit organization "Medicines for Malaria Venture" (MMV). Promising preliminary results indicated that the standardized protocols have been successful in obtaining soluble amounts of PfPKII with high degree of purity, suitable for kinetic assays investigation and biological screenings. Once potential inhibitors are identified, their inhibitory properties, including IC50 and Ki values, and mechanism of inhibition, will be thoroughly investigated for confirmation and validation as hits for the discovery of new lead compounds. Therefore, this research proposal will contribute to the fight against malaria and the development of effective treatments for this devastating disease.