A novel cytarabine crystalline lipid prodrug: Hexadecyloxypropyl cytarabine 3′,5′-cyclic monophosphate for proliferative vitreoretinopathy
A novel cytarabine crystalline lipid prodrug: Hexadecyloxypropyl cytarabine 3′,5′-cyclic monophosphate for proliferative vitreoretinopathy
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发表时间：
2012-07
期刊：
影响因子：
2.2
通讯作者：
J. S. Kim;J. Beadle;W. Freeman;K. Hostetler;K. Hartmann;Nadejda Valiaeva;I. Kozák;Laura Conner;Julissa Trahan;K. Aldern;Lingyun Cheng
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作者：
J. S. Kim;J. Beadle;W. Freeman;K. Hostetler;K. Hartmann;Nadejda Valiaeva;I. Kozák;Laura Conner;Julissa Trahan;K. Aldern;Lingyun Cheng
目的本研究的目的是合成和表征两种类型的阿糖胞苷（Ara-C）脂质前药，并评估前药通过玻璃体内注射给药后持续眼内递送的情况。方法以阿糖胞苷（1-β-D-阿拉伯呋喃糖基胞嘧啶）为原料，合成十六烷氧基丙基阿糖胞苷5'-单磷酸（HDP-P-Ara-C）和十六烷氧基丙基阿糖胞苷3',5'-环单磷酸（HDP-cP-Ara-C）。使用定制的溶出室模拟它们的玻璃体清除率，使用细胞增殖测定评估体外细胞毒性，并使用生物显微镜、间接检眼镜、眼压测量、视网膜电图和组织学评估大鼠和兔眼的体内眼部特性。结果 HDP-P-Ara-C 在 7 天内从溶出室（流速 2 µL/min）中清除。相比之下，HDP-cP-Ara-C（一种更难溶的前药）在溶解过程开始 36 天后仍然可检测到。 HDP-P-Ara-C 在人视网膜色素上皮 (ARPE-19) 中的 50% 细胞毒性浓度为 52±2.6 µM，在大鼠 Müller 细胞系 rMC-1 中的 50% 细胞毒性浓度为 32±2.2 µM。 ARPE-19 和 rMC-1 细胞中 HDP-cP-Ara-C 的 50% 细胞毒性浓度值分别为 50 µM 和 25 µM。玻璃体内注射最高剂量（228 µg/大鼠眼）后 2 周未检测到 HDP-P-Ara-C，且未发现眼部毒性。使用 HDP-cP-Ara-C，单次玻璃体内注射（800 µg/兔眼）后，药库在 26 周内可见。对于这两种化合物，视网膜电图、眼压和其他毒性研究均呈阴性，但最高剂量的 HDP-cP-Ara-C（800 µg/眼）除外，其因直接接触视网膜而产生局部毒性，并减少暗适应 a 波和闪烁视网膜电图振幅（广义估计方程，p=0.039 和 0.01）。结论 环状单磷酸酯前药 HDP-cP-Ara-C 具有更适合将阿糖胞苷持续递送至眼后段的理化特性。这些特性包括有限的水溶性、体外抗增殖活性以及注射到兔眼后良好的耐受性。
Purpose The objectives of this study were to synthesize and characterize two types of cytarabine (Ara-C) lipid produgs and evaluate the prodrugs for sustained intraocular delivery after administration by intravitreal injection. Methods Hexadecyloxypropyl cytarabine 5′-monophosphate (HDP-P-Ara-C) and hexadecyloxypropyl cytarabine 3′,5′-cyclic monophosphate (HDP-cP-Ara-C) were synthesized starting from cytarabine (1-β-D-arabinofuranosylcytosine). Their vitreal clearance profile was simulated using a custom dissolution chamber, in vitro cytotoxicity was evaluated using cell proliferation assays, and in vivo ocular properties in rat and rabbit eyes were assessed using biomicroscopy, indirect ophthalmoscopy, tonometry, electroretinography, and histology. Results HDP-P-Ara-C was cleared from the dissolution chamber (flow rate 2 µL/min) within 7 days. In contrast, HDP-cP-Ara-C, a much more insoluble prodrug, was still detectable 36 days after the dissolution process was started. HDP-P-Ara-C had a 50% cytotoxicity concentration of 52±2.6 μM in human retinal pigment epithelium (ARPE-19) and 32±2.2 µM in a rat Müller cell line, rMC-1. The 50% cytotoxicity concentration values for HDP-cP-Ara-C in ARPE-19 and rMC-1 cells were 50 µM and 25 µM, respectively. HDP-P-Ara-C was not detectable 2 weeks after the highest intravitreal dose (228 µg/rat eye) was injected, and no ocular toxicity was found. With HDP-cP-Ara-C, the drug depot was visible for 26 weeks following a single intravitreal injection (800 µg/rabbit eye). For both compounds, the electroretinogram, intraocular pressure, and other toxicity studies were negative except for the highest dose of HDP-cP-Ara-C (800 µg/eye), which had focal toxicity from the direct touch of the retina and decreased dark adapted a-waves and decreased flicker electroretinogram amplitudes (generalized estimating equations, p=0.039 and 0.01). Conclusions The cyclic monophosphate prodrug, HDP-cP-Ara-C, was found to have physiochemical properties better suited for sustained delivery of cytarabine to posterior segments of the eye. These properties included limited aqueous solubility, in vitro antiproliferative activity, and good tolerability after injection into rabbit eyes.