Coumarinic acid-based cyclic prodrugs of opioid peptides that exhibit metabolic stability to peptidases and excellent cellular permeability.
Coumarinic acid-based cyclic prodrugs of opioid peptides that exhibit metabolic stability to peptidases and excellent cellular permeability.
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基于香豆酸的阿片肽环状前药，对肽酶表现出代谢稳定性和优异的细胞渗透性。
DOI：
10.1023/a:1018828207920
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发表时间：
1999
影响因子：
3.7
通讯作者：
Borchardt,RT
中科院分区：
文献类型：
--
作者：
Gudmundsson,OS;Pauletti,GM;Wang,W;Shan,D;Zhang,H;Wang,B;Borchardt,RT
Purpose. To evaluate the cellular permeation characteristics and the chemical and enzymatic stability of coumarinic acid-based cyclic pro-drugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ of the opioid peptides [Leu5]-enkephalin (H-Tyr-Gly-Gly-Phe-Leu-OH) and DADLE (H-Tyr-D-Ala-Gly-Phe-D-Leu-OH), respectively.Methods. The rates of conversion of the cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ to [Leu5]-enkephalin and DADLE, respectively, in HBSS, pH 7.4 (Caco-2 cell transport buffer) and in various biological media having measurable esterase activity were determined by HPLC. The cell permeation characteristics of [Leu5]-enkephalin, DADLE and cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ were measured using Caco-2 cell monolayers grown onto microporus membranes and monitored by HPLC.Results. In HBSS, pH 7.4, cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ degraded chemically to intermediates that further degraded to [Leu5]-enkephalin and DADLE, respectively, in stoichiometric amounts. In 90% human plasma and rat liver homogenate, the disappearance of cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ was significantly faster than in HBSS, pH 7.4. The half-lives in 90% human plasma and in rat liver homogenate were substantially longer after pretreatment with paraoxon, a known inhibitor of serine-dependent esterases. When applied to the AP side of a Caco-2 cell monolayer, cyclic prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ exhibited significantly greater stability against peptidase metabolism than did [Leu5]-enkephalin. Cyclic pro-drug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ and DADLE exhibited similar stability when applied to the AP side of the Caco-2 cell monolayer. Prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ was 665-fold more able to permeate the Caco-2 cell monolayers than was [Leu5]-enkephalin, in part because of its increased enzymatic stability. Prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ was shown to be approximately 31 fold more able to permeate a Caco-2 cell monolayer than was DADLE.Conclusions. Cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ , prepared with the coumarinic acid promoiety, were substantially more able to permeate Caco-2 cell monolayers than were the corresponding opioid peptides. Prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ exhibited increased stability to peptidase metabolism compared to [Leu5]-enkephalin. In various biological media, the opioid peptides were released from the prodrugs by an esterase-catalyzed reaction, which is sensitive to paraoxon inhibition.
Purpose. To evaluate the cellular permeation characteristics and the chemical and enzymatic stability of coumarinic acid-based cyclic pro-drugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ of the opioid peptides [Leu5]-enkephalin (H-Tyr-Gly-Gly-Phe-Leu-OH) and DADLE (H-Tyr-D-Ala-Gly-Phe-D-Leu-OH), respectively.Methods. The rates of conversion of the cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ to [Leu5]-enkephalin and DADLE, respectively, in HBSS, pH 7.4 (Caco-2 cell transport buffer) and in various biological media having measurable esterase activity were determined by HPLC. The cell permeation characteristics of [Leu5]-enkephalin, DADLE and cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ were measured using Caco-2 cell monolayers grown onto microporus membranes and monitored by HPLC.Results. In HBSS, pH 7.4, cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ degraded chemically to intermediates that further degraded to [Leu5]-enkephalin and DADLE, respectively, in stoichiometric amounts. In 90% human plasma and rat liver homogenate, the disappearance of cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ was significantly faster than in HBSS, pH 7.4. The half-lives in 90% human plasma and in rat liver homogenate were substantially longer after pretreatment with paraoxon, a known inhibitor of serine-dependent esterases. When applied to the AP side of a Caco-2 cell monolayer, cyclic prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ exhibited significantly greater stability against peptidase metabolism than did [Leu5]-enkephalin. Cyclic pro-drug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ and DADLE exhibited similar stability when applied to the AP side of the Caco-2 cell monolayer. Prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ was 665-fold more able to permeate the Caco-2 cell monolayers than was [Leu5]-enkephalin, in part because of its increased enzymatic stability. Prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ was shown to be approximately 31 fold more able to permeate a Caco-2 cell monolayer than was DADLE.Conclusions. Cyclic prodrugs $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ and $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{2} $$ , prepared with the coumarinic acid promoiety, were substantially more able to permeate Caco-2 cell monolayers than were the corresponding opioid peptides. Prodrug $$\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{1} $$ exhibited increased stability to peptidase metabolism compared to [Leu5]-enkephalin. In various biological media, the opioid peptides were released from the prodrugs by an esterase-catalyzed reaction, which is sensitive to paraoxon inhibition.