Mucoadhesive polymers in peroral peptide drug delivery. II. Carbomer and polycarbophil are potent inhibitors of the intestinal proteolytic enzyme trypsin.
OBJECTIVE
The evaluation of the inhibitory action of two mucoadhesive poly(acrylates), polycarbophil and carbomer, registered by the Food and Drug Administration (FDA), on the intestinal proteolytic enzyme trypsin.


METHODS
The effect of the polymers on trypsin activity by measuring the degradation of a trypsin specific substrate. Binding of Ca2+ ions and proteins (125I-BSA) to the poly(acrylates). The influence of the polymers on the secondary trypsin structure by circular dichroism.


RESULTS
Trypsin inhibition was found to be time-dependent upon addition of Ca2+ in the degradation experiment. Only when Ca2+ was added within 10 min after trypsin incubation, recovery of the enzyme could be observed. Both polymers showed a strong Ca2+ binding ability. Carbomer, which had a higher inhibitory effect on trypsin activity, also revealed a higher Ca2+ binding affinity than polycarbophil. The amount of Ca2+ depleted out of the trypsin structure and the reduction of enzyme activity were comparable. Immobilization of trypsin by binding to the polymers could not be observed at pH 6.7. Circular dichroism studies suggested that, under depletion of Ca2+ from trypsin, the secondary structure changed its conformation, followed by an increased autodegradation of the enzyme.


CONCLUSIONS
The poly(acrylates) investigated may have potential to protect peptides from tryptic degradation and may be used to master the peroral delivery of peptide drugs.
Luessen
H L
HL
Leiden/Amsterdam Center for Drug Research, Leiden University, Division of Pharmaceutical Technology, The Netherlands.
Verhoef
J C
JC
Borchard
G
G
Lehr
C M
CM
de Boer
A G
AG
Junginger
H E
HE
D000180:Acrylic Resins / Q000494:pharmacology*; D000284:Administration, Oral; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002942:Circular Dichroism; D016503:Drug Delivery Systems; D066298:In Vitro Techniques; D007422:Intestines / Q000201:enzymology; D010455:Peptides / Q000008:administration & dosage*; D011485:Protein Binding; D014361:Trypsin Inhibitors / Q000494:pharmacology*
D000180:Acrylic Resins; D010455:Peptides; D014361:Trypsin Inhibitors; C006903:carbopol 940; C030312:calcium polycarbophil; D002118:Calcium