Carboxymethyl-starch excipients for gastrointestinal stable oral protein formulations containing protease inhibitors.
OBJECTIVE
The aim of this study was to develop a formulation for bioactive compounds using Carboxymethyl Starch (CMS) as excipient containing protease inhibitors. This formulation provided gastro protection and enhanced stability against pancreatic enzymes. Such stability is needed for formulation of oral vaccines with specific antigens.


METHODS
CMS was synthesized by treatment of starch with monochloroacetic acid in conditions leading to a substitution degree of about 1 meq/g and used as excipient for monolithic devices (300 mg tablets). Pefabloc SC and Aprotinin inhibitors were tested in dissolution media and in formulation to prevent the degradation of released bioactive materials. To evaluate the structural integrity and biological stability of plant proteins in the CMS formulation, albumin and lipase were added to the plant protein extract as protein and respectively as enzyme markers. The amounts of released and recovered proteins were evaluated by SDS-PAGE and densitometric analysis.


RESULTS
It was found that 1.6 % (w/w) of Pefabloc SC provides 98 % protection of the released plant proteins for formulations of 30 % alfalfa protein extract (APE) with CMS. In addition, when bovine serum albumin (BSA) added to the plant protein extract as a marker, 90 % protection of the released BSA was observed. Furthermore, a much higher lipase activity was found in the releasing media when the formulations contained Pefabloc SC.


CONCLUSIONS
Formulations with CM-Starch excipients and containing protease inhibitors prevent protein degradation and protect lipase activity, showing a marked potential to use for orally administered bioactive peptides and therapeutic enzymes.
De Koninck
Patrick
P
Department of Chemistry, Université du Québec à Montréal, Montréal (Québec) Canada.
Archambault
Denis
D
Hamel
Francine
F
Sarhan
Fathey
F
Mateescu
Mircea Alexandru
MA
D000284:Administration, Oral; D000818:Animals; D007611:Aprotinin / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D002417:Cattle; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D005079:Excipients / Q000737:chemistry*; D005750:Gastric Juice / Q000378:metabolism; D007419:Intestinal Secretions / Q000378:metabolism; D008049:Lipase / Q000378:metabolism; D000455:Medicago sativa / Q000737:chemistry; D010936:Plant Extracts / Q000378:metabolism; D055550:Protein Stability; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013213:Starch / Q000031:analogs & derivatives* / Q000737:chemistry; D013450:Sulfones / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D013607:Tablets; D014361:Trypsin Inhibitors / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology
D005079:Excipients; D010936:Plant Extracts; D013450:Sulfones; D013607:Tablets; D014361:Trypsin Inhibitors; D012710:Serum Albumin, Bovine; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; D013213:Starch; C034848:carboxymethyl starch; D007611:Aprotinin; D008049:Lipase