Henry S P, Templin M V, Gillett N, Rojko J, Levin A A
Isis Pharmaceuticals, Inc., Carlsbad, California 92008, USA.
Toxicol Pathol. 1999 Jan-Feb;27(1):95-100. doi: 10.1177/019262339902700117.
ISIS 2302 is a phosphorothioate oligodeoxynucleotide with a sequence complementary to the mRNA of human intercellular adhesion molecule 1 (ICAM-1). Hybridization of ISIS 2302 to the mRNA inhibits expression of the ICAM-1 protein in response to inflammatory stimuli. A murine active antisense oligonucleotide, ISIS 3082, has been used for in vivo pharmacology studies and has anti-inflammatory activity in models of organ transplant rejection, ulcerative colitis, and collagen-induced arthritis at doses ranging from 0.03 to 5 mg/kg. The safety assessment for ISIS 2302 includes general toxicity studies up to 6 mo in duration in mice and monkeys, genetic toxicity studies, and reproductive/fertility studies. ISIS 3082 was examined in parallel with ISIS 2302 in mouse toxicity and reproductive studies. The toxicities observed following systemic administration of ISIS 2302 and ISIS 3082 were similar and consistent with those observed for other compounds in this chemical class and, therefore, are independent of the suppression of ICAM-1 expression. Toxicokinetic evaluation demonstrated that toxicities occurred in organs containing the highest concentrations of ISIS 2302. Evidence of immune stimulation. including dose-dependent splenomegaly, lymphoid hyperplasia, and multiorgan mixed mononuclear cell infiltrates, was the most common finding in rodent studies. Monkeys were much less sensitive than mice to immune stimulation. Kidney contained the highest concentrations of ISIS 2302. Morphologic changes observed in kidney included atrophic and regenerative changes in proximal tubular epithelium; however, there was no evidence of functional abnormalities. Additional histologic changes noted in proximal tubular epithelium included basophilic granules, which were reflective of oligonucleotide distribution and uptake in these cells. Liver also contained high concentrations of oligonucleotide, which were associated with Kupffer cell hypertrophy in mice. Changes in serum transaminases, cholesterol, and triglycerides were reflective of hepatic alterations. In monkeys, high concentrations of oligonucleotide caused a transient increase in clotting times and activation of the alternative complement pathway. All toxicities associated with ISIS 2302 were reversible and occurred at doses well above those required for pharmacologic activity or currently used in clinical trials. In addition, there has been no evidence of genetic toxicity associated with ISIS 2302, and no changes in reproductive performance, fertility, or fetal development have been noted in animals treated with ISIS 2302 or ISIS 3082.
ISIS 2302是一种硫代磷酸酯寡脱氧核苷酸，其序列与人细胞间黏附分子1（ICAM-1）的mRNA互补。ISIS 2302与mRNA杂交可抑制ICAM-1蛋白在炎症刺激下的表达。一种小鼠活性反义寡核苷酸ISIS 3082已用于体内药理学研究，在器官移植排斥、溃疡性结肠炎和胶原诱导性关节炎模型中，剂量范围为0.03至5 mg/kg时具有抗炎活性。ISIS 2302的安全性评估包括在小鼠和猴子中进行长达6个月的一般毒性研究、遗传毒性研究以及生殖/生育研究。在小鼠毒性和生殖研究中，ISIS 3082与ISIS 2302并行进行检测。全身给药ISIS 2302和ISIS 3082后观察到的毒性相似，与该化学类别中其他化合物观察到的毒性一致，因此与ICAM-1表达的抑制无关。毒代动力学评估表明，毒性发生在含有最高浓度ISIS 2302的器官中。免疫刺激的证据，包括剂量依赖性脾肿大、淋巴组织增生和多器官混合单核细胞浸润，是啮齿动物研究中最常见的发现。猴子对免疫刺激的敏感性远低于小鼠。肾脏中ISIS 2302的浓度最高。在肾脏中观察到的形态学变化包括近端肾小管上皮的萎缩和再生变化；然而，没有功能异常的证据。在近端肾小管上皮中观察到的其他组织学变化包括嗜碱性颗粒，这反映了寡核苷酸在这些细胞中的分布和摄取。肝脏中也含有高浓度的寡核苷酸，这与小鼠库普弗细胞肥大有关。血清转氨酶、胆固醇和甘油三酯的变化反映了肝脏的改变。在猴子中，高浓度的寡核苷酸导致凝血时间短暂延长和替代补体途径激活。与ISIS 2302相关的所有毒性都是可逆的，且发生在远高于药理活性所需剂量或目前临床试验中使用剂量的情况下。此外，没有证据表明ISIS 2302具有遗传毒性，在用ISIS 2302或ISIS 3082治疗的动物中，未观察到生殖性能、生育能力或胎儿发育的变化。
