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WAM Essentials, Inc.
Systemic Enzyme Therapy
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Transport of Proteolytic Enzymes
Across Caco-2 Cell Monolayers
Bock U.,1 Kolac
C.,1 Borchard G.,1 KochK.,1 Fuchs
R.,1 Streichhan P.,2 and
Lehr C.-M.1
1 Department of Biopharmaceutics and Pharmaceutical
Technology, University of Saarland, Saarbrücken, Germany
2 Mucos Pharma GmbH, Geretsried, Germany
Pharmaceutical Research 1998, Vol. 15, No. 9, pp.
1393-1400.
SO 119 (18-09-2) |
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Summary
Purpose. To investigate the mechanisms
by which proteolytic enzymes. such as (trypsin. chymotrypsin.
papain. and bromelain, are able to cross the intestinal mucosal
barrier after oral administration to man. Methods. Filter-grown
Caco-2 cell monolayers were incubated with proteolytic enzymes
and then the transepithelial electrical resistance (TEER)
and the transport of the paracellular marker fluorescein
were monitored. The effects of (he enzymes on the cells were
investigated by light microscopy and by biochemical assays.
Transport of intact proteases across the cells was verified
by monitoring the proteolytic activity and MALDI-TOF mass
spectroscopic identification of undegraded trypsin.
Results. Depending on time. concentration.
and side of exposure to Caco-2 cell monolayers. all proleases
decreased the TEER and increased the transport of fluorescein.
Some morphological and metabolic changes were observed. The
effects were reversible, but until 24 hours after removal
of the proteases. Under the conditions of this in-vitro model,
approximately 10 % of the apically applied dose reached the
basolateral compartment as biologic. ally active, non-degraded
molecules.
Conclusions. Proteolytic enzymes
were found to exert considerable effects on the barrier function
of Caco-2 monolayers. facilitating the transport of normally
non-absorbable compounds. This suggests the also reported,
but so far unexplained systemic absorption of proteolytic
enzymes after oral administration in vivo may occur by self-enhanced
paracellular transport. |
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