旨在抑制巨细胞病毒复制的反义寡核苷酸的抗病毒活性及眼部动力学

Antiviral activity and ocular kinetics of antisense oligonucleotides designed to inhibit CMV replication.

作者信息

Henry S P, Miner R C, Drew W L, Fitchett J, York-Defalco C, Rapp L M, Levin A A

机构信息

Isis Pharmaceuticals, Inc., Carlsbad, California 92008, USA.

出版信息

Invest Ophthalmol Vis Sci. 2001 Oct;42(11):2646-51.

PMID:11581212

Abstract

PURPOSE

To compare the antiviral activity and ocular distribution of first- and second-generation antisense oligonucleotides intended for the treatment of cytomegalovirus (CMV) retinitis.

METHODS

The antiviral activity of ISIS 13312 and ISIS 2922 (Isis Pharmaceuticals, Inc., Carlsbad, CA) against 10 clinical CMV isolates was compared with a plaque-reduction assay. The ocular pharmacokinetics were compared after intravitreal injection in rabbits (36-90 microg) and monkeys (125-500 microg). Vitreous and/or retina were collected after single and multiple injections to characterize ocular distribution, clearance, and accumulation. Oligonucleotide concentrations were measured by capillary gel electrophoresis and immunohistochemical techniques.

RESULTS

ISIS 13312 and ISIS 2922 demonstrated comparable antiviral activity that was consistent among the 10 clinical isolates examined (50% inhibitory concentration [IC(50)], <1 microM). Activity was independent of the resistance of CMV isolates to DNA polymerase inhibitors. After intravitreal injection, the kinetics of ISIS 2922 and ISIS 13312 were characterized by clearance from vitreous and distribution to the retina; however, ISIS 2922 was cleared more quickly from the retina than ISIS 13312. The half-life of ISIS 13312 in the monkey retina was approximately 2 months. Retinal concentrations of ISIS 13312 were dose dependent, with approximately a twofold increase in concentration after once-monthly doses compared with single-dose concentrations. Immunohistochemical analysis indicated that both oligonucleotides were efficiently distributed to numerous ocular tissues, including retina, ciliary body, and optic nerve.

CONCLUSIONS

ISIS 13312 possesses antiviral activity and pharmacokinetic properties that favor its use as a therapeutic agent in treatment of CMV retinitis. The half-life of ISIS 13312 in retina is longer than that of ISIS 2922, potentially allowing for less frequent administration.

摘要

目的

比较用于治疗巨细胞病毒（CMV）视网膜炎的第一代和第二代反义寡核苷酸的抗病毒活性及眼部分布。

方法

采用蚀斑减少试验，比较ISIS 13312和ISIS 2922（Isis制药公司，加利福尼亚州卡尔斯巴德）对10株临床CMV分离株的抗病毒活性。在兔（36 - 90微克）和猴（125 - 500微克）玻璃体内注射后，比较其眼部药代动力学。单次和多次注射后收集玻璃体和/或视网膜，以表征眼部分布、清除和蓄积情况。通过毛细管凝胶电泳和免疫组织化学技术测量寡核苷酸浓度。

结果

ISIS 13312和ISIS 2922表现出相当的抗病毒活性，在所检测的10株临床分离株中一致（50%抑制浓度[IC(50)]，<1微摩尔）。活性与CMV分离株对DNA聚合酶抑制剂的耐药性无关。玻璃体内注射后，ISIS 2922和ISIS 13312的动力学特征为从玻璃体清除并分布到视网膜；然而，ISIS 2922从视网膜清除的速度比ISIS 13312更快。ISIS 13312在猴视网膜中的半衰期约为2个月。ISIS 13312的视网膜浓度呈剂量依赖性，与单剂量浓度相比，每月一次给药后浓度增加约两倍。免疫组织化学分析表明，两种寡核苷酸均有效分布于包括视网膜、睫状体和视神经在内的多种眼部组织。