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Virtual screening and experimental validation of casein kinases (CKs) inhibitors from Plasmodium spp. with antimalarial potential

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Author(s):
Kaira Cristina Peralis Tomaz
Total Authors: 1
Document type: Master's Dissertation
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
Defense date:
Examining board members:
Fabio Trindade Maranhão Costa; Maria Aparecida Medeiros Barros do Prado; Marcelo Brocchi
Advisor: Fabio Trindade Maranhão Costa; Gustavo Capatti Cassiano
Abstract

Malaria is an infectious disease that causes thousands of deaths each year worldwide. The use of antimalarials is essential to contain the disease and treat those infected. The recurrent parasitic resistance to current antimalarials, makes it necessary to constantly study for the development of new chemical compounds capable of acting in the death of the parasite through unprecedented modes of action. With the success of kinase inhibitors for the treatment of cancer, the kinome (set of kinases present in the parasite proteome) of Plasmodium spp. started to be investigated in order to find potential therapeutic targets for the treatment of malaria. Through bioinformatics filters, we chose the protein Casein kinase 1 (CK1) and 2 (CK2) from Plasmodium spp. as targets for the selection of chemical compounds for testing.CK1 is conserved among Plasmodium species, highly expressed throughout asexual blood stage development, and has physical interaction with the Rab-5B protein, evidencing being part of the metabolic control. In addition, reverse genetic studies have shown that this protein is essential for the development of the parasite's blood stage. Despite belonging to the same family, CK1 and CK2 are divergent in structure, function, and response to effector molecules. Reverse genetic experiments have shown that P. falciparum CK2? is essential for the survival of the pathogen during its replication cycle within the host's erythrocytes, making, like CK1, CK2? a promising target for the treatment of malaria. With the increasing computational capacity and the improvement of in silico screening, we have performed virtual screening techniques in order to reduce experimental tests and increase assertive capacity. First, we build the 3D structures of the target proteins PvCK1 by homology, and for PfCK2 we use the crystal, and we use two complementary screening methods, the first based on the parasite protein, Structure-Based Virtual Screening (SBVS) to identify CK inhibitors, and the second based on QSAR (Quantitative Structure-Activity Relationship) using the vast information in the literature and databases of active molecules already evaluated in Plasmodium spp., in order to ensure that the selected compounds have activity in vitro against the asexual blood forms of the parasite. After virtual screening, 183 compounds were selected and tested in vitro against P. falciparum, with 11 compounds capable of inhibiting more than 50% of parasitic growth at 1 ?M. These compounds were then evaluated for cytotoxicity in HepG-2 cells, and MCF-7. Finally, 3 compounds were chosen according to structure, cytotoxicity and antiplasmodial activity to produce dose response curves against sensitive and resistant strains of P. falciparum and we obtained 1 compound with a high selectivity index (> 100) (AU)

FAPESP's process: 18/05926-2 - Virtual screening and experimental validation of Plasmodium spp. CK1 (casein kinase 1) inhibitors as potential antimalarial candidates
Grantee:Kaira Cristina Peralis Tomaz
Support Opportunities: Scholarships in Brazil - Master