Evaluation of fungi in obtaining chiral active metabolite fexofenadine

Author(s)

Gisele Maria Metta

• Document type: Master's Dissertation
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC)
• Defense date: 2013-12-06

Examining board members

Anderson Rodrigo Moraes de Oliveira; Thiago Barth; Carmen Lúcia Cardoso

• Advisor: Anderson Rodrigo Moraes de Oliveira

Abstract

Fexofenadine (FEX) has been the drug of choice for the symptomatic treatment of allergic manifestations, being an antihistamine H1 receptor 2nd generation non-sedating. It is the active and chiral metabolite of terfenadine (TERF), a drug whose production and marketing was suspended as a result of adverse events. Fungi have been presented as a promising alternative for the production of compounds with biological activity. Thus, the goal of this project was to evaluate the ability of fungi to biotransform asymmetric terfenadine to its active metabolite, fexofenadine using fungi as agents catalysts. For enantioselective analysis of fexofenadine a method for chromatographic separation was developed employing a chiral column Lux® cellulose -1, mobile phase water : methanol (35:65,v/v) + 0.3% triethylamine + 0.4% acetic acid, flow rate of 0.5 mL min-1, with detection at 220nm. Two sample preparation microtechnology were evaluated in the extraction of analytes from the culture medium: the dispersive liquid-liquid microextraction (DLLME) and hollow fiber liquid phase microextraction (HF- LPME). Between the two, the DLLME was the microtechnic chosen because it provided better results such as higher recovery values, chromatograms with no possible interfering peaks, greater speed and ease of sample preparation. The optimized conditions of DLLME were: chloroform (300 ?L) as extractor solvent, isopropanol (300 ?L) as disperser solvent. After the formation of the cloud point, the samples were subjected to agitation by vortexing for 15 seconds and centrifuging for 10 minutes at 3000 rpm. The recoveries were 43 % for both enantiomers. The method was linear in the concentration range from 2.0 -15.0 ?g mL-1 for each enantiomer of FEX (r > 0.990). The limit of quantification was 2 ?g mL-1 for the enantiomers of FEX. Among the seven fungi studied (Papulaspora immersa Hotson SS13, Penicillium crustosum VR4, Mucor rouxii, Nigrospora sphaerica SS67, Fusarium oxysporum SS50, Cunninghamella echinulata var. elegans ATCC 8688A e Cunninghamella elegans NRRL 1393 ATCC 10028B), only the fungi Fusarium oxysporum SS50 e Cunninghamella echinulata var. elegans ATCC 8688A showed potential for biotransformation of terfenadine in fexofenadine in the incubation conditions employed in this work. (AU)

• FAPESP's process: 11/15419-1 - Assessment of Fungi in Obtaining the chiral and active metabolite Fexofenadine.
• Grantee: Gisele Maria Metta
• Support Opportunities: Scholarships in Brazil - Master