VII Simposio Internacional de Ciencias Farmacéuticas 2019 "VII SICF" -VII Simposio "Diseño, Obtención y Desarrollo de Fármacos"

VII Simposio Internacional de Ciencias Farmacéuticas 2019

VII SICF

VII Simposio "Diseño, Obtención y Desarrollo de Fármacos"

High-Throughput Screen to Identify LAPTc Inhibitors as in vitro Antichagasic Agents Using RapidFire Mass Spectrometry

Resumen [ES]

The American trypanosomiasis better known as Chagas disease is a systemic infection caused by the protozoan Trypanosoma cruzi. It is estimated 56,000 new cases and 12,000 deaths annually in the Americas region in 21 countries. The chemotherapy of the disease is still inadequate. Several proteases involving in vital processes for the survival of parasites such as invasion, migration, nutrition, immune response evasion and immunomodulation have been described. The presence of the leucyl aminopeptidase (LAPTc) in the three life stages of the parasite indicates its importance for one or several physiological processes. The objective of this work is to develop LAPTc enzymatic assays for screening compounds. Using water soluble peptide substrate Leu-Ser-Thr-Val-Ile-Val-Arg (LSTVIVR) as substrate, the RapidFire Mass Spectrometry assay provides higher signal to blank ratio, better assay performance than biochemical assay with chromogenic Leucine-p-nitroanilide (Leu-pNA) substrate and fluorogenic Leucine-7-amino-4-methylcoumarin (Leu-AMC) substrate. With this method was identified a tetrazole-peptidomimetic as a micromolar LAPTc inhibitor, with potentialities as an in vitro antichagasic agent.

Resumen [EN]

The American trypanosomiasis better known as Chagas disease is a systemic infection caused by the protozoan Trypanosoma cruzi. It is estimated 56,000 new cases and 12,000 deaths annually in the Americas region in 21 countries. The chemotherapy of the disease is still inadequate. Several proteases involving in vital processes for the survival of parasites such as invasion, migration, nutrition, immune response evasion and immunomodulation have been described. The presence of the leucyl aminopeptidase (LAPTc) in the three life stages of the parasite indicates its importance for one or several physiological processes. The objective of this work is to develop LAPTc enzymatic assays for screening compounds. Using water soluble peptide substrate Leu-Ser-Thr-Val-Ile-Val-Arg (LSTVIVR) as substrate, the RapidFire Mass Spectrometry assay provides higher signal to blank ratio, better assay performance than biochemical assay with chromogenic Leucine-p-nitroanilide (Leu-pNA) substrate and fluorogenic Leucine-7-amino-4-methylcoumarin (Leu-AMC) substrate. With this method was identified a tetrazole-peptidomimetic as a micromolar LAPTc inhibitor, with potentialities as an in vitro antichagasic agent.

Sobre el ponente

Mirtha Elisa Aguado Casas

Lic. Mirtha Elisa Aguado Casas

CEP,Facultad de Biologia,Universidad de la Habana Flag of Cuba
Información Práctica
Póster digital
English (US)
No definido
30 minutos
No definido
Autores
Martin Zoltner
Anira Méndez
Lauren Webster
Jorge González Bacerio
Tony Hope
Daniel G. Rivera
Mark Field
Lic. Mirtha Elisa Aguado Casas
Maikel Izquierdo. Sandra O’ Neill
Palabras clave
laptc inhibitor
leucyl aminopeptidase
rapidfire mass spectrometry assay
trypanosoma cruzi