Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
J Pharm Pharm Sci. 2013;16(2):279-88. doi: 10.18433/j3s898.
We have previously shown favorable in vitro gut permeability for three novel di-peptide esters of glucosamine (GlcN) likely facilitated by the peptide transporter 1 (PepT1). Herein, we report the development of a novel assay for the determination of bioavailability of the peptide ester of interest, the anti-inflammatory properties of a glycine-valine ester derivative of GlcN (GVG) as well as its pharmacokinetics under healthy and inflammatory conditions.
A pre-column derivatization (with 9-fluorenylmethoxycarbonyl) HPLC assay was developed to study bioavailability of GVG, GlcN or cleaved GlcN in the rats that were cannulated in their right jugular vein for blood collection. The compounds of interest were orally administered to both healthy and arthritic rats. Serial blood samples and urine were collected and assayed for the compounds. The stability of the GVG was also tested after incubation with the rat feces. Efficacy of GVG was tested in inflamed rats (injection of 0.2 mL of Mycobacterium butyricum in squalene) following GVG (20 and 30 mg/kg/day GlcN equivalent) or GlcN (20 and 90 mg/kg/day) administration. Arthritis index was calculated at the end of the experiment.
The assay was linear (ranged between 0.05-20 μg/mL) and reproducible (intra- and inter-day<10%). Among the tested compounds, only GVG showed a significantly higher plasma concentrations and urinary excretion than GlcN (≈3-fold increase). GVG showed a favorable stability in the rat feces. Adjuvant arthritis was completely prevented with doses greater than 20 mg/kg/day with GVG being 3-fold more potent than GlcN.
The examined glycine-valine-GlcN di-peptide aminosugar is a potent anti-inflammatory compound due to its favorable properties to deliver GlcN into the systemic circulation.
我们之前已经证明了三种新型氨基葡萄糖二肽酯（GlcN）具有有利的体外肠道通透性，这可能是由于肽转运蛋白 1（PepT1）的作用。在此，我们报告了一种新型测定法的开发，用于测定所关注的肽酯的生物利用度，即 GlcN 的甘氨酸-缬氨酸酯衍生物（GVG）的抗炎特性，以及在健康和炎症条件下的药代动力学。
建立了一种预柱衍生（9-芴甲氧羰基）HPLC 测定法，用于研究 GVG、GlcN 或裂解的 GlcN 在右侧颈静脉插管采集血液的大鼠中的生物利用度。将感兴趣的化合物口服给予健康和关节炎大鼠。收集并测定化合物的系列血样和尿样。还测试了 GVG 在与大鼠粪便孵育后的稳定性。在注射 0.2 mL 鲨烯中的丁酸分枝杆菌后，用 GVG（20 和 30 mg/kg/天 GlcN 当量）或 GlcN（20 和 90 mg/kg/天）对 GVG 进行治疗，测试 GVG 的疗效。在实验结束时计算关节炎指数。
该测定法呈线性（范围在 0.05-20 μg/mL 之间）且重现性好（日内和日间<10%）。在所测试的化合物中，只有 GVG 显示出明显高于 GlcN 的血浆浓度和尿排泄（约 3 倍增加）。GVG 在大鼠粪便中表现出良好的稳定性。GVG 的剂量大于 20 mg/kg/天时可完全预防佐剂性关节炎，其效力是 GlcN 的 3 倍。
所检查的甘氨酸-缬氨酸-GlcN 二肽氨基糖是一种有效的抗炎化合物，因为它具有将 GlcN 递送到全身循环的有利特性。
