Exploring structure-function relationship of DHODH enzyme in the development of new molecules with trypanocidal and leishmanicidal activities

• Grant number: 11/23504-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Effective date (Start): June 01, 2012
• Effective date (End): June 02, 2016
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Maria Cristina Nonato
• Grantee: Renata Almeida Garcia Reis
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 14/21068-5 - Integrated in vitro and in silico strategies in the search for lead compounds against trypanosomatid-related diseases based on the selective inhibition of dihydroorotate dehydrogenases, BE.EP.DR

Abstract

The parasitic infectious diseases affect millions of people especially in the poorest regions in the world and represent a growing threat worldwide. Considered neglected diseases, parasitic infectious diseases also contribute to the maintenance of social inequality, and represent a strong barrier to the development of various countries. The most of deaths caused by parasitic diseases occur in regions situated below the equator. We know that today, more than a billion people - one seventh of the world population - are affected by tropical diseases. In particular, the diseases caused by parasites of trypanosomatids family as Chagas disease, leishmaniasis and sleeping sickness incapacitate a significant fraction of patients. The need for alternative therapies is clear, but overall investment in research and development (R & D) of drugs and vaccines are extremely insufficient. Recently, diidroorotato dehydrogenase enzyme has been validated as a therapeutic target for the development of drugs with trypanocidal activity. DHODH belongs to the route of de novo synthesis of pyrimidine nucleotides and catalyzes the diidroorotato L-orotate oxidation, through a mechanism dependent the FMN and electron acceptors. The present project aims use structural biology techniques and molecular design based on protein structure to search for compounds that have selective inhibitory activity against DHODH of trypanosomatids. According to this modern approach, the steps involved in the development of the project are: (1) complete mapping of DHODH enzyme catalytic mechanism, combining techniques of molecular biology, biochemistry and structural biology, (2) identification of new interaction sites that can be exploited in designing new ligands for DHODH, (3) identification of molecules with potential inhibitory activity against DHODH enzyme through virtual screening of compounds databases, testing "in vitro" in the presence of synthetic and natural products, with subsequent determination of the inhibitory constant through enzyme kinetics assays in the presence of the compounds identified by the search, (4) structural characterization of the interaction between the enzyme and inhibitors by techniques of single-crystal X-ray diffraction, (5) proposals in chemical modifications in compounds with an inhibitory effect against DHODH, identified by the research and proven by enzyme kinetics assays, in order to maximize the selectivity and inhibitory activity against the parasite enzyme.

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

REIS, RENATA A. G.; CALIL, FELIPE ANTUNES; FELICIANO, PATRICIA ROSA; PINHEIRO, MATHEUS PINTO; NONATO, M. CRISTINA. The dihydroorotate dehydrogenases: Past and present. Archives of Biochemistry and Biophysics, v. 632, n. SI, p. 175-191, OCT 15 2017. (11/23504-9, 07/08703-0, 12/25075-0)

PADUA, RICARDO A. P.; TOMALERI, GIOVANI P.; REIS, RENATA A. G.; DAVID, JULIANA S.; SILVA, VALERIA C.; PINHEIRO, MATHEUS P.; NONATO, MARIA CRISTINA. ThermoFMN - a thermofluor assay developed for ligand-screening as an alternative strategy for drug discovery. Journal of the Brazilian Chemical Society, v. 25, n. 10, p. 1864+, OCT 2014. (11/23504-9, 07/08703-0, 08/11644-8, 11/14269-6, 12/25075-0)

REIS, RENATA A. G.; CALIL, FELIPE ANTUNES; FELICIANO, PATRICIA ROSA; PINHEIRO, MATHEUS PINTO; NONATO, M. CRISTINA. The dihydroorotate dehydrogenases: Past and present. Archives of Biochemistry and Biophysics, v. 632, p. 17-pg., 2017-10-15. (12/25075-0, 11/23504-9, 07/08703-0)

GARCIA REIS, RENATA ALMEIDA; LORENZATO, EDER, JR.; SILVA, VALERIA CRISTINA; NONATO, MARIA CRISTINA. Recombinant production, crystallization and crystal structure determination of dihydroorotate dehydrogenase from Leishmania (Viannia) braziliensis. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 71, p. 6-pg., 2015-05-01. (11/23504-9, 14/01257-8, 12/25075-0)

REIS, RENATA A. G.; FERREIRA, PATRICIA; MEDINA, MILAGROS; NONATO, M. CRISTINA. The mechanistic study of Leishmania major dihydro-orotate dehydrogenase based on steady- and pre-steady-state kinetic analysis. Biochemical Journal, v. 473, n. 5, p. 651-660, MAR 1 2016. (12/25075-0, 11/23504-9)

GARCIA REIS, RENATA ALMEIDA; LORENZATO, JR., EDER; SILVA, VALERIA CRISTINA; NONATO, MARIA CRISTINA. Recombinant production, crystallization and crystal structure determination of dihydroorotate dehydrogenase from Leishmania (Viannia) braziliensis. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 71, n. 5, SI, p. 547-552, MAY 2015. (12/25075-0, 14/01257-8, 11/23504-9)

REIS, RENATA A. G.; FERREIRA, PATRICIA; MEDINA, MILAGROS; NONATO, M. CRISTINA. The mechanistic study of Leishmania major dihydro-orotate dehydrogenase based on steady- and pre-steady-state kinetic analysis. Biochemical Journal, v. 473, p. 10-pg., 2016-03-01. (12/25075-0, 11/23504-9)