Current Good Manufacturing Practice Production of an Oncolytic Recombinant Vesicular Stomatitis Viral Vector for Cancer Treatment
Current Good Manufacturing Practice Production of an Oncolytic Recombinant Vesicular Stomatitis Viral Vector for Cancer Treatment
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DOI：
10.1089/hum.2010.159
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发表时间：
2011-04-01
影响因子：
4.2
通讯作者：
Couture, L.
中科院分区：
文献类型：
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作者：
Ausubel, L. J.;Meseck, M.;Couture, L.
水泡性口炎病毒（VSV）是一种溶瘤病毒，由于其在癌细胞中选择性复制的能力，目前正被研究作为一种有希望的治疗癌症的工具。为了增强VSV非病理性实验室菌株的溶瘤特性，我们生成了表达鼠γ疱疹病毒M3的重组载体[rVSV（M Delta 51）-M3]，M3是一种分泌型病毒趋化因子结合蛋白，可与多种哺乳动物趋化因子高亲和力结合。如前所述，当rVSV（MD51）-M3用于大鼠肝细胞癌（HCC）的原位模型时，它抑制炎性细胞迁移到病毒感染的肿瘤部位，这使得肿瘤内病毒复制增强，导致肿瘤坏死增加和生存期显著延长。这些令人鼓舞的结果导致了这种载体在HCC患者中的临床转化的发展。然而，尚未描述该载体的可扩展的符合现行药品生产质量管理规范（cGMP）的生产工艺。为了生产临床试验所需的高滴度病毒，开发了一种适合GMP生产和规模化的工艺。我们在此描述了一种大规模（50升）载体生产工艺，其能够在cGMP下实现约10（9）噬斑形成单位（PFU）/ml的粗滴度。该工艺用于生产主病毒种子贮备液和cGMP下临床试验药物的临床批次，感染性病毒滴度约为2 x 10（10）PFU/ml（总产率，1 x 10（13）PFU）。该批次已通过美国食品和药物管理局（FDA）授权的所有放行检测，并将用于晚期HCC患者的1期临床转化试验。
Vesicular stomatitis virus (VSV) is an oncolytic virus currently being investigated as a promising tool to treat cancer because of its ability to selectively replicate in cancer cells. To enhance the oncolytic property of the nonpathologic laboratory strain of VSV, we generated a recombinant vector [rVSV(M Delta 51)-M3] expressing murine gammaherpesvirus M3, a secreted viral chemokine-binding protein that binds to a broad range of mammalian chemokines with high affinity. As previously reported, when rVSV(MD51)-M3 was used in an orthotopic model of hepatocellular carcinoma (HCC) in rats, it suppressed inflammatory cell migration to the virus-infected tumor site, which allowed for enhanced intratumoral virus replication leading to increased tumor necrosis and substantially prolonged survival. These encouraging results led to the development of this vector for clinical translation in patients with HCC. However, a scalable current Good Manufacturing Practice (cGMP)compliant manufacturing process has not been described for this vector. To produce the quantities of high-titer virus required for clinical trials, a process that is amenable to GMP manufacturing and scale-up was developed. We describe here a large-scale (50-liter) vector production process capable of achieving crude titers on the order of 10(9) plaque-forming units (PFU)/ml under cGMP. This process was used to generate a master virus seed stock and a clinical lot of the clinical trial agent under cGMP with an infectious viral titer of approximately 2 x 10(10) PFU/ml (total yield, 1 x 10(13) PFU). The lot has passed all U.S. Food and Drug Administration-mandated release testing and will be used in a phase 1 clinical translational trial in patients with advanced HCC.