A modified coumarinic acid-based cyclic prodrug of an opioid peptide: its enzymatic and chemical stability and cell permeation characteristics.
A modified coumarinic acid-based cyclic prodrug of an opioid peptide: its enzymatic and chemical stability and cell permeation characteristics.
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阿片肽的改良香豆酸基环状前药：其酶促和化学稳定性以及细胞渗透特性。
DOI：
10.1023/a:1016148631055
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发表时间：
2002
影响因子：
3.7
通讯作者：
Borchardt,RonaldT
中科院分区：
文献类型：
--
作者：
Ouyang,Hui;Tang,Fuxing;Siahaan,TerunaJ;Borchardt,RonaldT
目的。以香豆素酸为基，在肽的酚基和羧基之间插入醛（氧甲基），评价其化学/酶稳定性和细胞渗透特性。采用高效液相色谱法测定了氧甲基修饰的前药2转化为DADLE的化学/酶促速率。利用Caco-2细胞、野生型Madin-Darby犬肾细胞（MDCK- wt）、转染人mdr1基因的MDCK细胞（MDCK- mdr1）和转染人mrp2基因的MDCK细胞（MDCK- mrp2）生长在微孔膜上，采用高效液相色谱法测定了DADLE及其氧甲基修饰前药2的细胞渗透特性。以氧甲基修饰香豆酸为基础的环前药2以ph依赖性的方式化学降解为DADLE，即转化率随ph的增加而增加。前药2在大鼠血浆（t1/2= 39 min）和大鼠肝脏浆液（t1/2= 59.2 min）中降解迅速，但在cco -2细胞浆液（t1/2= 678.7 min）和人血浆（t1/2= 264.3 min）中降解较慢。在用于转运研究的所有四种细胞系中，基侧（BL）到根尖（AP）方向（Papp BL-to-AP）的氧甲基前药2的通量率显著大于AP-to-BL方向（Papp AP-to-BL）的通量率。在Caco-2、MDCK-MDR1、MDCK-MRP2和MDCK-WT细胞中，Papp BL-to-AP/Papp ap -to- blp比值分别为0.116、35.1、21.2和12.6。GF120918 （P-gp抑制剂）和环孢素A （P-gp和MRP2抑制剂）可以抑制修饰前药的外排。以氧甲基修饰香豆酸为基础的DADLE环前药2在化学和酶促介质中均可转化为DADLE。然而，前药是P-gp和MRP2的良好底物，这表明其通过肠黏膜和血脑屏障的渗透将受到显著限制。
Purpose. To evaluate the chemical/enzymatic stability and the cell permeation characteristics of the modified coumarinic acid-based cyclic prodrug2of DADLE (H-Tyr-D-Ala-Gly-Phe-D-Leu-OH), which has an aldehyde equivalent (oxymethyl) inserted between the phenolic group of the promoiety and the carboxylic acid group of the peptide.Methods. The rates of the chemical/enzymatic conversion of the oxymethyl-modified prodrug2to DADLE were measured by HPLC. The cellular permeation characteristics of DADLE and its oxymethyl-modified prodrug2were measured by HPLC using Caco-2 cells, wild type Madin-Darby Canine Kidney cells (MDCK-WT), MDCK cells transfected with humanMDR1gene (MDCK-MDR1), and MDCK cells transfected with humanMRP2gene (MDCK-MRP2) grown onto microporous membranes.Results. The oxymethyl-modified coumarinic acid-based cyclic prodrug2degraded chemically to DADLE in a pH-dependent manner, i.e., rates of conversion increased with increasing pH. The prodrug2degraded rapidly in rat plasma (t1/2= 39 min) and rat liver homogenate (t1/2= 59.2 min), but much slower in Caco-2 cell homogenate (t1/2= 678.7 min) and human plasma (t1/2= 264.3 min). In all four cell lines used for transport studies, the flux rates of the oxymethyl prodrug2in the basolateral (BL)-to-apical (AP) direction (Papp BL-to-AP) were significantly greater than the flux rates in the AP-to-BL direction (Papp AP-to-BL). The Papp BL-to-AP/Papp AP-to-BLratios were >116, 35.1, 21.2, and 12.6 in Caco-2, MDCK-MDR1, MDCK-MRP2, and MDCK-WT cells, respectively. The efflux of the modified prodrug could be inhibited by GF120918 (an inhibitor for P-gp) and cyclosporin A (an inhibitor for P-gp and MRP2).Conclusions. The oxymethyl-modified coumarinic acid-based cyclic prodrug2of DADLE could be converted to DADLE in both chemical and enzymatic media. However, the prodrug was a good substrate for both P-gp and MRP2 suggesting that its permeation across intestinal mucosa and blood-brain barrier would be significantly restricted.