Amino acid ester prodrugs of 2-bromo-5,6-dichloro-1-(β-dribofuranosyl) benzimidazole enhance metabolic stability in vitro and in vivo

Philip L. Lorenzi, Christopher P. Landowski, Xueqin Song, Katherine Z. Borysko, Julie M. Breitenbach, Seung Kim Jae, John M. Hilfinger, Leroy B. Townsend, John C. Drach, Gordon L. Amidon

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

2-Bromo-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole (BDCRB) is a potent and selective inhibitor of human cytomegalovirus (HCMV), but it lacks clinical utility due to rapid in vivo metabolism. We hypothesized that amino acid ester prodrugs of BDCRB may enhance both in vitro potency and systemic exposure of BDCRB through evasion of BDCRB-metabolizing enzymes. To this end, eight different amino acid prodrugs of BDCRB were tested for N-glycosidic bond stability, ester bond stability, Caco-2 cell uptake, antiviral activity, and cytotoxicity. The prodrugs were resistant to metabolism by BDCRB-metabolizing enzymes, and ester bond cleavage was rate-limiting in metabolite formation from prodrug. Thus, BDCRB metabolism could be controlled by the selection of promoiety. In HCMV plaque-formation assays, L-Asp-BDCRB exhibited 3-fold greater selectivity than BDCRB for inhibition of HCMV replication. This potent and selective antiviral activity in addition to favorable stability profile made L-Asp-BDCRB an excellent candidate for in vivo assessment and pharmacokinetic comparison with BDCRB. In addition to rapid absorption and sufficient prodrug activation after oral administration to mice, L-Asp-BDCRB exhibited a 5-fold greater half-life than BDCRB. Furthermore, the sum of area under the concentration-time profile (AUC)BDCRB and AUCprodrug after L-Asp-BDCRB administration was roughly 3-fold greater than AUCBDCRB after BDCRB administration, suggesting that a reservoir of prodrug was delivered in addition to parent drug. Overall, these findings demonstrate that amino acid prodrugs of BDCRB exhibit evasion of metabolizing enzymes (i.e., bioevasion) in vitro and provide a modular approach for translating this in vitro stability into enhanced in vivo delivery of BDCRB.

Original languageEnglish
Pages (from-to)883-890
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume314
Issue number2
DOIs
Publication statusPublished - 1 Aug 2005
MoE publication typeA1 Journal article-refereed

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Prodrugs
Esters
Amino Acids
Cytomegalovirus
Antiviral Agents
Enzymes
Caco-2 Cells
In Vitro Techniques
benzimidazole
Oral Administration
Half-Life
Pharmacokinetics
Pharmaceutical Preparations

Cite this

Lorenzi, Philip L. ; Landowski, Christopher P. ; Song, Xueqin ; Borysko, Katherine Z. ; Breitenbach, Julie M. ; Jae, Seung Kim ; Hilfinger, John M. ; Townsend, Leroy B. ; Drach, John C. ; Amidon, Gordon L. / Amino acid ester prodrugs of 2-bromo-5,6-dichloro-1-(β-dribofuranosyl) benzimidazole enhance metabolic stability in vitro and in vivo. In: Journal of Pharmacology and Experimental Therapeutics. 2005 ; Vol. 314, No. 2. pp. 883-890.
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abstract = "2-Bromo-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole (BDCRB) is a potent and selective inhibitor of human cytomegalovirus (HCMV), but it lacks clinical utility due to rapid in vivo metabolism. We hypothesized that amino acid ester prodrugs of BDCRB may enhance both in vitro potency and systemic exposure of BDCRB through evasion of BDCRB-metabolizing enzymes. To this end, eight different amino acid prodrugs of BDCRB were tested for N-glycosidic bond stability, ester bond stability, Caco-2 cell uptake, antiviral activity, and cytotoxicity. The prodrugs were resistant to metabolism by BDCRB-metabolizing enzymes, and ester bond cleavage was rate-limiting in metabolite formation from prodrug. Thus, BDCRB metabolism could be controlled by the selection of promoiety. In HCMV plaque-formation assays, L-Asp-BDCRB exhibited 3-fold greater selectivity than BDCRB for inhibition of HCMV replication. This potent and selective antiviral activity in addition to favorable stability profile made L-Asp-BDCRB an excellent candidate for in vivo assessment and pharmacokinetic comparison with BDCRB. In addition to rapid absorption and sufficient prodrug activation after oral administration to mice, L-Asp-BDCRB exhibited a 5-fold greater half-life than BDCRB. Furthermore, the sum of area under the concentration-time profile (AUC)BDCRB and AUCprodrug after L-Asp-BDCRB administration was roughly 3-fold greater than AUCBDCRB after BDCRB administration, suggesting that a reservoir of prodrug was delivered in addition to parent drug. Overall, these findings demonstrate that amino acid prodrugs of BDCRB exhibit evasion of metabolizing enzymes (i.e., bioevasion) in vitro and provide a modular approach for translating this in vitro stability into enhanced in vivo delivery of BDCRB.",
author = "Lorenzi, {Philip L.} and Landowski, {Christopher P.} and Xueqin Song and Borysko, {Katherine Z.} and Breitenbach, {Julie M.} and Jae, {Seung Kim} and Hilfinger, {John M.} and Townsend, {Leroy B.} and Drach, {John C.} and Amidon, {Gordon L.}",
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Amino acid ester prodrugs of 2-bromo-5,6-dichloro-1-(β-dribofuranosyl) benzimidazole enhance metabolic stability in vitro and in vivo. / Lorenzi, Philip L.; Landowski, Christopher P.; Song, Xueqin; Borysko, Katherine Z.; Breitenbach, Julie M.; Jae, Seung Kim; Hilfinger, John M.; Townsend, Leroy B.; Drach, John C.; Amidon, Gordon L.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 314, No. 2, 01.08.2005, p. 883-890.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Amino acid ester prodrugs of 2-bromo-5,6-dichloro-1-(β-dribofuranosyl) benzimidazole enhance metabolic stability in vitro and in vivo

AU - Lorenzi, Philip L.

AU - Landowski, Christopher P.

AU - Song, Xueqin

AU - Borysko, Katherine Z.

AU - Breitenbach, Julie M.

AU - Jae, Seung Kim

AU - Hilfinger, John M.

AU - Townsend, Leroy B.

AU - Drach, John C.

AU - Amidon, Gordon L.

PY - 2005/8/1

Y1 - 2005/8/1

N2 - 2-Bromo-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole (BDCRB) is a potent and selective inhibitor of human cytomegalovirus (HCMV), but it lacks clinical utility due to rapid in vivo metabolism. We hypothesized that amino acid ester prodrugs of BDCRB may enhance both in vitro potency and systemic exposure of BDCRB through evasion of BDCRB-metabolizing enzymes. To this end, eight different amino acid prodrugs of BDCRB were tested for N-glycosidic bond stability, ester bond stability, Caco-2 cell uptake, antiviral activity, and cytotoxicity. The prodrugs were resistant to metabolism by BDCRB-metabolizing enzymes, and ester bond cleavage was rate-limiting in metabolite formation from prodrug. Thus, BDCRB metabolism could be controlled by the selection of promoiety. In HCMV plaque-formation assays, L-Asp-BDCRB exhibited 3-fold greater selectivity than BDCRB for inhibition of HCMV replication. This potent and selective antiviral activity in addition to favorable stability profile made L-Asp-BDCRB an excellent candidate for in vivo assessment and pharmacokinetic comparison with BDCRB. In addition to rapid absorption and sufficient prodrug activation after oral administration to mice, L-Asp-BDCRB exhibited a 5-fold greater half-life than BDCRB. Furthermore, the sum of area under the concentration-time profile (AUC)BDCRB and AUCprodrug after L-Asp-BDCRB administration was roughly 3-fold greater than AUCBDCRB after BDCRB administration, suggesting that a reservoir of prodrug was delivered in addition to parent drug. Overall, these findings demonstrate that amino acid prodrugs of BDCRB exhibit evasion of metabolizing enzymes (i.e., bioevasion) in vitro and provide a modular approach for translating this in vitro stability into enhanced in vivo delivery of BDCRB.

AB - 2-Bromo-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole (BDCRB) is a potent and selective inhibitor of human cytomegalovirus (HCMV), but it lacks clinical utility due to rapid in vivo metabolism. We hypothesized that amino acid ester prodrugs of BDCRB may enhance both in vitro potency and systemic exposure of BDCRB through evasion of BDCRB-metabolizing enzymes. To this end, eight different amino acid prodrugs of BDCRB were tested for N-glycosidic bond stability, ester bond stability, Caco-2 cell uptake, antiviral activity, and cytotoxicity. The prodrugs were resistant to metabolism by BDCRB-metabolizing enzymes, and ester bond cleavage was rate-limiting in metabolite formation from prodrug. Thus, BDCRB metabolism could be controlled by the selection of promoiety. In HCMV plaque-formation assays, L-Asp-BDCRB exhibited 3-fold greater selectivity than BDCRB for inhibition of HCMV replication. This potent and selective antiviral activity in addition to favorable stability profile made L-Asp-BDCRB an excellent candidate for in vivo assessment and pharmacokinetic comparison with BDCRB. In addition to rapid absorption and sufficient prodrug activation after oral administration to mice, L-Asp-BDCRB exhibited a 5-fold greater half-life than BDCRB. Furthermore, the sum of area under the concentration-time profile (AUC)BDCRB and AUCprodrug after L-Asp-BDCRB administration was roughly 3-fold greater than AUCBDCRB after BDCRB administration, suggesting that a reservoir of prodrug was delivered in addition to parent drug. Overall, these findings demonstrate that amino acid prodrugs of BDCRB exhibit evasion of metabolizing enzymes (i.e., bioevasion) in vitro and provide a modular approach for translating this in vitro stability into enhanced in vivo delivery of BDCRB.

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