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Study of bioavailability and biodistribution of Ambbyomin-X. 

Boufleur P1, Sciani JM2, Chudzinski-Tavassi AM2

1 Programa de Pós- graduação em Biologia Molecular, Escola Paulista de Medicina Universidade Federal de São Paulo, Brasil; 2 Laboratório de Bioquímica e Biofísica, Instituto Butantan, Brasil

Introduction:  Amblyomin-X is a 12.2 kDa recombinant protein, characterized as Kunitz-type inhibitor. This protein causes inhibition of Factor Xa and apoptosis in tumor cells, beside tumor regression in mouse models. Objectives: To study the initial parameters of bioavailability and biodistribution, to verify and quantify the presence and the maximum concentration of the protein in organs and plasma after different times of administration.  Methods: Amblyomin-X was injected intravenously (100 mg/kg) in female mice (CEUAIB n°1251/14) and after different times (10, 45 min, 1 h, 2 h, 6 h e 24 horas), plasma, urine and organs were collected. The organs were immersed in appropriate buffer and sonicated to extract proteins. These samples and plasma were subjected to a polyacrylamide gel and the band corresponding to the Amblyomin-X was extracted and analyzed by liquid chromatography coupled to mass spectrometry (LC-MS and MALDI-TOF). Collected urine was treated for the extraction of proteins and peptides also analyzed by LC-MS and MALDI-TOF. 
  Results and Discussion: Amblyomin-X was found in the plasma immediately after application and up to 2 hours. In organs, it was identified after 10 min in the thymus, lung and liver; 45 min in the heart; 1h in the thymus and heart; 2h in the heart; 6h in the thymus, heart, spleen and kidney and 24 hours only in kidney. Amblyomin-X was identified in the urine after 24 hours of application, indicating it´s been eliminated after that. 


2 - BIOCHEMISTRY 21th Annual Scientific Meeting of Instituto Butantan.