文章：

rAAV2-sTRAIL的临床前研究：药物疗效、生物分布和动物安全性

Preclinical study of rAAV2-sTRAIL: pharmaceutical efficacy, biodistribution and safety in animals

原文发布日期：2017-04-21 

英文摘要：

The recombinant sTRAIL has been in clinical trial for various human malignancies. However, the half-life time of sTRAIL is very short, which might be an important factor influencing its clinical efficacy for cancer therapy. We previously reported the recombinant adeno-associated virus (AAV)-encoding sTRAIL95–281-mediated sTRAIL expression in vivo up to 8 months and suppressed tumor growth markedly in mouse xenografts. In the present study, we further evaluated the clinical potency for cancer gene therapy and the safety in mouse and non-human primates. The mouse models with HCT-116, NCI-H460 and BEL-7402 cancers were injected intraperitoneally with a single dose of 1.0 × 1011, 1.0 × 1010 and 1.0 × 109 vg of rAAV2-sTRAIL95–281 virus, respectively. The cynomolgus monkeys were injected (i.m.) with a single dose of rAAV2-sTRAIL95–281 of 1 × 1011, 3 × 1011 and 1 × 1012 vg, corresponding to 6-, 20- and 60-fold of intended use dosage for humans, respectively. The efficacy, pharmacology and toxicity of rAAV-sTRAIL in the animals were analyzed accordingly. The tumor inhibitory rates reached 44–76%, 48–52% and 55–74% in the three tumor models, respectively, and they had no influence on mouse spontaneous activity. Administration (s.c.) of a single dose of rAAV2-sTRAIL95–281 virus of 1.0 × 109 or 1.0 × 1010 vg in mice with implanted tumor led to mainly distribution in the spleen, liver, implanted tumor, blood, injected site of muscle and bone marrow. Two weeks later, there was no rAAV2-sTRAIL95–281 detected in blood and bone marrow, and it significantly decreased in other tissues and organs and then gradually cleared away in 4–12 weeks after administration. There was no rAAV2-sTRAIL accumulation in the animal’s body and no influence on the body weights. Administration (i.v.) did not cause animal death, and no dose-related abnormal clinical symptoms were found in the mice. There were no abnormal tissue and organ found in all animals. Long-term toxicity test in cynomolgus monkeys did not cause rAAV2-sTRAIL95–281-related toxic and side effects, except that anti-AAV and anti-sTRAIL antibodies were generated. In conclusion, these data demonstrated that administration of rAAV2-sTRAIL95–281 in mice and in cynomolgus monkeys is safe without obvious toxic and side effects to the animals, and throw light on pharmacokinetics and safety in human clinical trials for cancer gene therapy. 

摘要翻译： 

重组sTRAIL蛋白已针对多种人类恶性肿瘤开展临床试验。然而sTRAIL的半衰期极短，这可能是影响其癌症治疗临床疗效的重要因素。我们既往报道过重组腺相关病毒（AAV）编码的sTRAIL95-281可在体内持续表达sTRAIL达8个月之久，并能显著抑制小鼠移植瘤的生长。本研究进一步评估了其用于癌症基因治疗的临床潜力及在小鼠和非人灵长类动物中的安全性。分别向HCT-116、NCI-H460和BEL-7402癌症小鼠模型腹腔注射单剂量1.0×10¹¹、1.0×10¹⁰和1.0×10⁹ vg的rAAV2-sTRAIL95-281病毒。食蟹猴肌肉注射单剂量1×10¹¹、3×10¹¹和1×10¹² vg的rAAV2-sTRAIL95-281（分别相当于拟用于人体剂量的6倍、20倍和60倍），并相应分析rAAV-sTRAIL在动物体内的疗效、药理学和毒性作用。三种肿瘤模型的抑瘤率分别达到44-76%、48-52%和55-74%，且对小鼠自主活动无影响。给荷瘤小鼠皮下注射单剂量1.0×10⁹或1.0×10¹⁰ vg rAAV2-sTRAIL95-281病毒后，主要分布于脾脏、肝脏、移植瘤、血液、注射部位肌肉和骨髓。两周后血液和骨髓中未检测到rAAV2-sTRAIL95-281，其他组织器官中的病毒量显著降低，并在给药后4-12周内逐渐清除。未发现rAAV2-sTRAIL在动物体内蓄积，对体重也无影响。静脉给药未导致动物死亡，小鼠未出现剂量相关的异常临床症状。所有动物均未发现异常组织和器官。食蟹猴长期毒性试验未引起rAAV2-sTRAIL95-281相关的毒副作用，但产生了抗AAV和抗sTRAIL抗体。综上所述，这些数据表明rAAV2-sTRAIL95-281在小鼠和食蟹猴中的应用是安全的，对动物无明显毒副作用，并为癌症基因治疗人体临床试验的药代动力学和安全性研究提供了重要依据。

原文链接：

Preclinical study of rAAV2-sTRAIL: pharmaceutical efficacy, biodistribution and safety in animals