Improvement of L-dopa absorption by dipeptidyl derivation, utilizing peptide transporter PepT1.

In the present study, possible enhancement of intestinal absorption of L-dopa by utilizing intestinal peptide transporter was examined using Caco-2 cells and Xenopus oocytes expressing human peptide transporter (hPepT1). To see whether this peptide transporter could be utilized for the improvement of L-dopa absorption, we employed a dipeptide-mimetic derivative of L-dopa, L-dopa-L-Phe. L-Dopa-L-Phe inhibited the uptake of [14C]Gly-Sar, but not that of L-[3H]-dopa by Caco-2 cells. Uptake of L-dopa-L-Phe was increased by expression of hPepT1 in Xenopus oocytes. The appearance of L-dopa and its metabolite, dopamine, on the basolateral side of Caco-2 cells was significantly higher after addition of L-dopa-L-Phe than after that of L-dopa and was reduced by the presence of Gly-Sar on the apical side. These results indicate that the L-dopa-L-Phe is absorbed more efficiently than L-dopa and is taken up via the peptide transporter, but not via the amino acid transporter, demonstrating the possibility of targeting the peptide transporter as a means for improving intestinal absorption of peptide-like drugs.

[1]  S. Shirazi-Beechey,et al.  Glycyl-L-proline transport in rabbit enterocyte basolateral-membrane vesicles. , 1990, The Biochemical journal.

[2]  T. Nakanishi,et al.  Immunolocalization and pharmacological relevance of oligopeptide transporter PepT1 in intestinal absorption of β‐lactam antibiotics , 1996, FEBS letters.

[3]  H. Yamamoto,et al.  Sequence, tissue distribution and developmental changes in rat intestinal oligopeptide transporter. , 1996, Biochimica et biophysica acta.

[4]  A. Asatoor,et al.  Intestinal absorption of stereoisomers of dipeptides in the rat. , 1973, Clinical science and molecular medicine.

[5]  D. Matthews,et al.  Rates of absorption of a dipeptide and the equivalent free amino acid in various mammalian species. , 1971, Biochimica et biophysica acta.

[6]  J. Moss,et al.  Involvement of N-Myristoylation in Monoclonal Antibody Recognition Sites on Chimeric G Protein α Subunits (*) , 1995, The Journal of Biological Chemistry.

[7]  L. Johnson,et al.  Physiology of the gastrointestinal tract , 2012 .

[8]  M. Hediger,et al.  Human Intestinal H+/Peptide Cotransporter , 1995, The Journal of Biological Chemistry.

[9]  D. Matthews,et al.  Observations on the intestinal absorption of some oligopeptides of methionine and glycine in the rat. , 1969, Clinical science.

[10]  T. Orfeo,et al.  One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. , 1977, Journal of the National Cancer Institute.

[11]  D. Perrett,et al.  Absorption of two tripeptides by the human small intestine: a study using a perfusion technique. , 1974, Clinical science and molecular medicine.

[12]  H. Saito,et al.  Dipeptide transporters in apical and basolateral membranes of the human intestinal cell line Caco-2. , 1993, The American journal of physiology.

[13]  S. Takenoshita,et al.  Immuno-localization of H+/peptide cotransporter in rat digestive tract. , 1996, Biochemical and biophysical research communications.

[14]  M. Romero,et al.  Expression cloning of a mammalian proton-coupled oligopeptide transporter , 1994, Nature.

[15]  K. Sasahara,et al.  Dosage form design for improvement of bioavailability of levodopa II: bioavailability of marketed levodopa preparations in dogs and parkinsonian patients. , 1980, Journal of pharmaceutical sciences.

[16]  D. Perrett,et al.  Dipeptide absorption in man , 1972, Gut.

[17]  G. Amidon,et al.  Passive and carrier-mediated intestinal absorption components of captopril. , 1988, Journal of pharmaceutical sciences.

[18]  H. Higashida,et al.  Functional expression of transporter for beta-lactam antibiotics and dipeptides in Xenopus laevis oocytes injected with messenger RNA from human, rat and rabbit small intestines. , 1995, The Journal of pharmacology and experimental therapeutics.

[19]  P. Artursson,et al.  Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells. , 1991, Biochemical and biophysical research communications.

[20]  H. Saito,et al.  Cloning and characterization of a rat H+/peptide cotransporter mediating absorption of beta-lactam antibiotics in the intestine and kidney. , 1995, The Journal of pharmacology and experimental therapeutics.

[21]  H. Kuo,et al.  Intestinal Absorption Studies on Peptide Mimetic α‐Methyldopa Prodrugs , 1996 .

[22]  J. Morris,et al.  Active Transport of L-Dopa in the Intestine , 1973, Nature.

[23]  D. Matthews,et al.  Uptake of dipeptides containing basic and acidic amino acids by rat small intestine in vitro. , 1972, Clinical science.

[24]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[25]  R. Borchardt,et al.  Transport of a large neutral amino acid in a human intestinal epithelial cell line (Caco-2): uptake and efflux of phenylalanine. , 1992, Biochimica et biophysica acta.

[26]  D. Matthews,et al.  Absorption of glycine and glycine peptides from the small intestine of the rat. , 1968, Clinical science.

[27]  T. Nakanishi,et al.  The Predominant Contribution of Oligopeptide Transporter PepT1 to Intestinal Absorption of β‐Lactam Antibiotics in the Rat Small Intestine , 1997, The Journal of pharmacy and pharmacology.

[28]  A. Tsuji,et al.  Kinetics and mechanism of in vitro uptake of amino-beta-lactam antibiotics by rat small intestine and relation to the intact-peptide transport system. , 1984, Biochemical pharmacology.