Preclinical Development of PEG-TNF

描述(申请人提供)：未配制的人肿瘤坏死因子(huTNF())在小鼠体内具有抗肿瘤活性，但有效剂量和致死剂量差别不大。未配制的huTNFpha的系统给药已经在人体上进行了测试。所有研究表明，剂量限制性毒性通常为低血压，循环半衰期为20分钟，几乎没有抗肿瘤活性。最近长时间使用huTNFpha隔离肢体灌流的结果表明，这种细胞因子在患有黑色素瘤的人类中具有显著的抗肿瘤活性。这表明，只要其毒性和短暂的循环半衰期能够被克服，huTNFpha可能在治疗上是有用的。 其他具有生物活性的细胞因子和蛋白质与聚乙二醇组分的形成显著提高了它们的安全性，并延长了它们的循环半衰期。例子包括现已获得FDA批准的聚乙二醇α干扰素和聚乙二醇GCSF。 第一阶段SBIR赠款申请建议将huTNFpha与聚乙二醇组成为一体，以延长其循环半衰期并提高其安全性。找到了huTNFpha的最佳配方(命名为huTNFpha-SS PEG20,000 mW)。与未配制的huTNFpha相比，该制剂在小鼠体内的循环半衰期、安全性和有效性均有显著提高。 在IND前与FDA的一次会议上，最终的临床前研究被确定，并成为这一第二阶段SBIR应用的重点。其具体目标是：1)生产cGMP huTNFpha-SS聚乙二醇20,000 mW；2)完成其药理性质；3)按照GLP(良好实验室操作规范)测试其免疫原性和动物安全性。 这些研究的数据将提供最终信息，使IND能够向 FDA将支持人类I期临床测试。

DESCRIPTION (provided by applicant): Unformulated human tumor necrosis factor( (huTNF() has anti-tumor activity in mice, but there is little difference between the effective and lethal doses. Systemic administration of unformulated huTNFalpha has been tested in humans. All studies indicated the dose limiting toxicity was usually hypotension, the circulating half life was < 20 min and little anti-tumor activity was seen. Recent results from isolated limb perfusions with huTNFalpha for prolonged times indicated this cytokine to have marked anti-tumor activity in humans with melanoma. This suggests huTNFalpha may be therapeutically useful, provided the toxicity and short circulating half-life can be overcome. Formulation of other biologically active cytokines and proteins with polyethylene glycol (PEG) has significantly improved their safety and increases their circulating half-life. Examples include PEG-alpha interferon and PEG-GCSF, which are now approved by the FDA. The Phase 1 SBIR grant application proposed formulating huTNFalpha with PEG to increase its circulating half-life and increase its safety. An optimal formulation of huTNFalpha was found (termed huTNFalpha-SS PEG 20,000 mw). The circulating half-life, safety and efficacy of this formulation have been substantially increased compared with unformulated huTNFalpha in mice. In a Pre IND meeting with the FDA the final preclinical studies were identified and are the focus of this Phase 2 SBIR application. The specific aims are to: 1) produce cGMP huTNFalpha-SS PEG 20,000 mw; 2) finalize its pharmacological characterization; and 3) test the immunogenicity and its safety in animals according to GLP (Good Laboratory Practice). The data from these studies will provide the final information enabling the filing of an IND with the FDA to support human Phase I clinical testing.