title: Enzyme loaded biodegradable microspheres in vitro ex vivo evaluation.
abstract: Prolidase is a naturally occurring enzyme involved in the final stage of protein catabolism. Deficient enzyme activity causes prolidase deficiency (PD), a rare autosomal recessive inherited disorder whose main manifestations are chronic, intractable ulcerations of the skin, particularly of lower limbs. Although several attempts have been made towards the treatment of this pathology, a cure for this disease has yet to be found. The purpose of this work is to evaluate the possibility of enzyme replacement therapy through prolidase microencapsulation in biodegradable microspheres. The poly(D,L-lactide-co-glycolide) (PLGA) prolidase loaded microparticulate systems have been prepared utilizing the w-o-w double emulsion solvent evaporation method. They have been characterized "in vitro" by morphological analysis, total protein content and an in vitro dissolution test of active protein. "Ex vivo" evaluation of prolidase activity from the microspheres has been performed on cellular extracts of cultured skin fibroblasts from healthy subjects (controls) and from patients affected by PD. The results reported in this work on prolidase from pig kidney (available on the market) demonstrate the positive role of microencapsulation as a process of enzymatic activity stabilization inside PLGA microspheres achieving both in vitro and ex vivo active enzyme release. This formulation can be proposed as a parenteral depot drug delivery system.
authors.affiliation: Department of Pharmaceutical Chemistry, A. Castellani, University of Pavia, V. le Taramelli 12, 27100, Pavia, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Genta
authors.forename: P
authors.initials: P
authors.lastname: Perugini
authors.forename: F
authors.initials: F
authors.lastname: Pavanetto
authors.forename: K
authors.initials: K
authors.lastname: Maculotti
authors.forename: T
authors.initials: T
authors.lastname: Modena
authors.forename: B
authors.initials: B
authors.lastname: Casado
authors.forename: A
authors.initials: A
authors.lastname: Lupi
authors.forename: P
authors.initials: P
authors.lastname: Iadarola
authors.forename: B
authors.initials: B
authors.lastname: Conti
mesh_terms: D000328:Adult; D000818:Animals; D001673:Biodegradation, Environmental; D002478:Cells, Cultured; D004150:Dipeptidases / Q000008:administration & dosage / Q000493:pharmacokinetics; D004341:Drug Evaluation / Q000379:methods*; D004789:Enzyme Activation / Q000502:physiology; D004800:Enzymes, Immobilized / Q000008:administration & dosage / Q000493:pharmacokinetics*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D012867:Skin / Q000187:drug effects / Q000201:enzymology; D013552:Swine
chemical_list: D004800:Enzymes, Immobilized; D004150:Dipeptidases; C022449:proline dipeptidase