title: Development of an enteric-coated formulation containing freeze-dried, viable recombinant Lactococcus lactis for the ileal mucosal delivery of human interleukin-10.
abstract: Recombinant hIL-10 producing Lactococcus lactis (Thy12) looks a promising intestinal mucosal delivery system for treatment of Crohn's disease [L. Steidler, W. Hans, L. Schotte, S. Neirynck, F. Obermeirer, W. Falk, W. Fiers, E. Remaut, Treatment of murine colitis by L. lactis secreting interleukin-10, Science 289 (2000) 1352-1355. L. Steidler, S. Neirynck, N. Huyghebaert, V. Snoeck, A. Vermeire, B.M. Goddeeris, E. Cox, J.P. Remon, and E. Remaut, Biological containment of genetically modified L. lactis for intestinal delivery of human interleukin-10, Nat. Biotechnol. 21 (7) (2003) 785-789]. As the hIL-10 production is strictly related to Thy12's viability and gastric fluid negatively influences this viability, an enteric-coated formulation had to be developed with maintenance of its viability after production and storage. L. lactis MG1363, used for optimization, was grown until stationary phase in milk (glucose/casiton supplemented) and freeze-dried. This resulted in a viability of about 60%. Storage at different conditions showed that viability remained highest at 8 degrees C/N2 atmosphere (32.5% of initial remained viable after 6 months). To increase the concentration of bacteria in the freeze-dried powder, they were concentrated by centrifugation. L. lactis tolerated this procedure. However, the concentration factor was limited to 10. Freeze-dried Thy12 was filled in ready-to-use enteric-coated capsules. Despite the good enteric properties of the capsules, viability of Thy12 dropped to about 43 and 28% after gastric fluid stage, depending on the enteric polymer used. Freeze-dried Thy12 filled in ready-to-use enteric-coated capsules, packed in Alu sachets (sealed at 20% RH) maintained 6.1 and 44.3% of initial viability after storage for 1 year at 8 and -20 degrees C, respectively, as well as its hIL-10 producing capacity.
authors.affiliation: Laboratory of Pharmaceutical Technology, Ghent University, Ghent, Belgium.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Huyghebaert
authors.forename: An
authors.initials: A
authors.lastname: Vermeire
authors.forename: Sabine
authors.initials: S
authors.lastname: Neirynck
authors.forename: Lothar
authors.initials: L
authors.lastname: Steidler
authors.forename: Erik
authors.initials: E
authors.lastname: Remaut
authors.forename: Jean Paul
authors.initials: JP
authors.lastname: Remon
mesh_terms: D004339:Drug Compounding; D016503:Drug Delivery Systems / Q000379:methods*; D004355:Drug Stability; D004356:Drug Storage; D005612:Freeze Drying; D006801:Humans; D007082:Ileum / Q000378:metabolism* / Q000382:microbiology; D016753:Interleukin-10 / Q000008:administration & dosage* / Q000235:genetics; D007413:Intestinal Mucosa / Q000378:metabolism* / Q000382:microbiology; D013294:Lactococcus lactis / Q000235:genetics / Q000254:growth & development*; D011994:Recombinant Proteins / Q000008:administration & dosage* / Q000235:genetics; D013608:Tablets, Enteric-Coated
chemical_list: D011994:Recombinant Proteins; D013608:Tablets, Enteric-Coated; D016753:Interleukin-10