Phe-Gly Dipeptidomimetics Designed for the Di-/Tripeptide Transporters PEPT1 and PEPT2:  Synthesis and Biological Investigations

Våbenø, Jon; Lejon, Tore; Nielsen, Carsten Uhd; Steffansen, Bente; Chen, Weiqing; Ouyang, Hui; Borchardt, Ronald T.; Luthman, Kristina

doi:10.1021/jm031022+

Cited by 39 publications

(28 citation statements)

References 47 publications

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“…The improved oral bioavailability of these anti-virals has been attributed to their enhanced affinity to transporters [3,4,43–45]. A variety of amino acid, dipeptide and tripeptide have been investigated to improve the oral absorption via intake transporters as a part of prodrug approaches [16,17,22–24,46–49]. We have synthesized mono-amino acid and dipeptide prodrugs of floxuridine, gemcita-bine and 2-bromo-5,6- dichloro-1-(beta-d-ribofuranosyl) benz-imidazole (BDCRM) and characterized their stability and membrane permeability [5,6,9–11,13,14,30,50].…”

Section: Discussionmentioning

confidence: 99%

The development of orally administrable gemcitabine prodrugs with d-enantiomer amino acids: Enhanced membrane permeability and enzymatic stability

Tsume

Incecayir

Song³

et al. 2014

European Journal of Pharmaceutics and Biopharmaceutics

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Gemcitabine prodrugs with D- and L-configuration amino acids were synthesized and their chemical stability in buffers, resistance to glycosidic bond metabolism, enzymatic activation, permeability in Caco-2 cells and mouse intestinal membrane, anti-proliferation activity in cancer cell were determined and compared to that of parent drug, gemcitabine. Prodrugs containing D-configuration amino acids were enzymatically more stable than ones with L-configuration amino acids. The activation of all gemcitabine prodrugs was 1.3–17.6-fold faster in cancer cell homogenate than their hydrolysis in buffer, suggesting enzymatic action. The enzymatic activation of amino acid monoester prodrugs containing D-configuration amino acids in cell homogenates was 2.2–10.9-fold slower than one of amino acid monoester prodrugs with L-configuration amino acids. All prodrugs exhibited enhanced resistance to glycosidic bond metabolism by thymidine phosphorylase compared to parent gemcitabine. Gemcitabine prodrugs showed superior the effective permeability in mouse jejunum to gemcitabine. More importantly, the high plasma concentration of D-amino acid gemcitabine prodrugs was observed more than one of L-amino acid gem-citabine prodrugs. In general, the 5′-mono-amino acid monoester gemcitabine prodrugs exhibited higher permeability and uptake than their parent drug, gemcitabine. Cell proliferation assays in AsPC-1 pancreatic ductal cell line indicated that gemcitabine prodrugs were more potent than their parent drug, gemcitabine. The transport and enzymatic profiles of 5′-D-valyl-gemcitabine and 5′-D-phenylalanyl-gem-citabine suggest their potential for increased oral uptake and delayed enzymatic bioconversion as well as enhanced uptake and cytotoxic activity in cancer cells, would facilitate the development of oral dosage form for anti-cancer agents and, hence, improve the quality of life for the cancer patients.

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Section: Discussionmentioning

confidence: 99%

The development of orally administrable gemcitabine prodrugs with d-enantiomer amino acids: Enhanced membrane permeability and enzymatic stability

Tsume

Incecayir

Song³

et al. 2014

European Journal of Pharmaceutics and Biopharmaceutics

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“…Moreover, the PEPT2 affinity values for kyotorphin, cephalexin, and δ-aminolevulinic acid obtained in this study using the SKPT cells grown in basic culture media are comparable to the values previously reported in the literature when using other assay systems (Table 2). [30][31][32][33][34][35][36] Altogether, SKPT cells grown in basic culture media may serve as a useful standard model for di-/ tripeptide uptake studies.…”

Section: Comparison Of Skpt Cells Grown Under Different Culture Condimentioning

confidence: 99%

Characterization of rPEPT2-Mediated Gly-Sar Transport Parameters in the Rat Kidney Proximal Tubule Cell Line SKPT-0193 cl.2 Cultured in Basic Growth Media

et al. 2005

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The rat proximal kidney tubule cell line SKPT-0193 cl.2 (SKPT) expresses the di-/tripeptide transporter PEPT2 (rPEPT2) and has been used to study PEPT2-mediated transport. Traditionally, SKPT cells have been cultured in growth media supplemented with epidermal growth factor (EGF), apotransferrin, dexamethasone, and insulin. It was recently demonstrated that omission of EGF from the culture media caused a drastic increase in the expression of rPEPT2. The hypothesis was therefore that the SKPT cell line might be able to differentiate and express rPEPT2 in the absence of the four agonists traditionally added. The aim of the study was thus to characterize Gly-Sar transport parameters in SKPT cells cultured in basic growth media (conventional media without added agonists). Morphology was studied using confocal laser scanning microscopy (CLSM) and immunohistochemistry. Monolayer integrity was evaluated using transepithelial electrical resistance (TEER) measurements and [(3)H]-mannitol permeabilities. Di-/tripeptide transporter activity was studied using [(14)C]-glycylsarcosine ([(14)C]-Gly-Sar). SKPT cells grown in basic media for 4 days formed confluent monolayers with a TEER of 5.03 +/- 0.33 kOmega.cm(2) (n = 5). Apical Gly-Sar uptake peaked after 3-6 days in culture. Uptake at day 4 was 5.89 +/- 0.30 pmol.cm(-2).min(-1) (n = 3). Di-/tripeptide uptake displayed an optimum at approximately pH 6. Affinity values for cephalexin, kyotorphin, and delta-aminolevulinic acid were comparable to those obtained in other PEPT2-expressing model systems. It can be concluded that SKPT cells grown in the absence of the agonists traditionally added to the culture media retain all necessary properties for PEPT2-mediated peptide uptake studies. Furthermore, the absence of the agonists might facilitate studies of hormonal regulation of PEPT2 expression and transport activity.

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“…6,18,22,35 Therefore, dipeptide prodrugs may be delivered more to a target site by carrier mediated transporters, and extra amino acid attached prodrugs could be more suitable for specific enzymatic activation at a target site than mono amino acid ester prodrugs. Several studies on amino acid modifications to increase specific transporter affinity have been conducted.…”

Section: Introductionmentioning

confidence: 99%

Enhanced Cancer Cell Growth Inhibition by Dipeptide Prodrugs of Floxuridine: Increased Transporter Affinity and Metabolic Stability

2008

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Dipeptide monoester prodrugs of floxuridine were synthesized, and their chemical stability in buffers, resistance to glycosidic bond metabolism, affinity for PEPT1, enzymatic activation and permeability in cancer cells were determined and compared to those of mono amino acid monoester floxuridine prodrugs. Prodrugs containing glycyl moieties were the least stable in pH 7.4 buffer (t1/2 < 100 min). The activation of all floxuridine prodrugs was 2- to 30-fold faster in cell homogenates than their hydrolysis in buffer, suggesting enzymatic action. The enzymatic activation of dipeptide monoester prodrugs containing aromatic promoieties in cell homogenates was 5- to 20-fold slower than that of other dipeptide and most mono amino acid monoester prodrugs (t1/2 ∼ 40 to 100 min). All prodrugs exhibited enhanced resistance to glycosidic bond metabolism by thymidine phosphorylase compared to parent floxuridine. In general, the 5′-O-dipeptide monoester floxuridine prodrugs exhibited higher affinity for PEPT1 than the corresponding 5′-O-mono amino acid ester prodrugs. The permeability of dipeptide monoester prodrugs across Caco-2 and Capan-2 monolayers was 2- to 4-fold higher than the corresponding mono amino acid ester prodrug. Cell proliferation assays in AsPC-1 and Capan-2 pancreatic ductal cell lines indicated that the dipeptide monoester prodrugs were equally as potent as mono amino acid prodrugs. The transport and enzymatic profiles of 5′-l-phenylalanyl-l-tyrosyl-floxuridine, 5′-l-phenylalanyl-l-glycyl-floxuridine, and 5′-l-isoleucyl-l-glycyl-floxuridine suggest their potential for increased oral uptake, delayed enzymatic bioconversion and enhanced resistance to metabolism to 5-fluorouracil, as well as enhanced uptake and cytotoxic activity in cancer cells, attributes that would facilitate prolonged systemic circulation for enhanced therapeutic action.

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Phe-Gly Dipeptidomimetics Designed for the Di-/Tripeptide Transporters PEPT1 and PEPT2: Synthesis and Biological Investigations

Cited by 39 publications

References 47 publications

The development of orally administrable gemcitabine prodrugs with d-enantiomer amino acids: Enhanced membrane permeability and enzymatic stability

The development of orally administrable gemcitabine prodrugs with d-enantiomer amino acids: Enhanced membrane permeability and enzymatic stability

Characterization of rPEPT2-Mediated Gly-Sar Transport Parameters in the Rat Kidney Proximal Tubule Cell Line SKPT-0193 cl.2 Cultured in Basic Growth Media

Enhanced Cancer Cell Growth Inhibition by Dipeptide Prodrugs of Floxuridine: Increased Transporter Affinity and Metabolic Stability

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