Highly Selective Pathogen Inactivators For Treatment of Convalescent Transfusion Plasma

摘要 目前，输注恢复期血浆(CP)正被开发为新冠肺炎患者的一种治疗方法 并作为高危人群的避孕药。此外，用血浆或中和抗体治疗 康复期患者的制剂可能是治疗新发传染病的唯一方法， 可能没有其他治疗方法可用。同时，CP输血会使接受者面临风险 输血传播疾病(TTD)的风险，这一风险因受感染的人而进一步加剧 危重病人的免疫状况。目前有限的TTD血液检测不能提供 为获得充分保护，请限制严重受限的捐赠者池，并可能在某些地区不可用。病原体 灭活可以提供解决方案。不幸的是，目前使用的减少病原体的治疗方法 血浆(溶剂洗涤剂、干热巴氏杀菌、紫外线或伽马辐射)是非选择性的，可以 影响血浆中和抗体或其他保护性蛋白因子的质量。我们在Zata 制药公司已开发出一类基于天然多胺的新型病原体灭活剂(ZPI) 支架，在灭活病原体基因组分子方面具有真正的选择性，同时节省血浆蛋白。 我们的初步结果表明，ZPI对核酸具有很高的反应性，并且不修改模型 蛋白质(Cyt-C、RSV融合蛋白)和动物血清生长因子。使用它们，我们灭活了不同类型的 病原体(G+和G-细菌、支原体、真菌、原生动物)和包膜或包膜的高效价制剂 无包膜病毒。目前我们正在开发ZPI用于减少输血红细胞中的病原体， 研究由NIH SBIR拨款资助(R44 HL145783)。在本应用程序中，我们建议将新的 用于治疗恢复期血浆(CP)的病原体灭活方法：(1)使用6种病毒 选择人血浆中病原菌灭活的最佳ZPI和条件；(2)使用 已经开发了分析方法，以建立完全中和和/或去除 处理后血浆中残留的灭活剂；(3)使用针对4种病毒的特异性抗体 经酶联免疫吸附试验证实，病毒灭活处理对抗体结合无影响 (4)用抗SARS-CoV-2 S蛋白的中和抗体显示保存 血浆处理后抗体的病毒中和特性；(5)重复使用自体 输注处理后的血浆，建立其在兔体内的安全性模型。 在实现这些初步目标后，我们将申请资金，包括完成SBIR资金， 与纽约血液中心合作，进行临床前评估，并启动I期人体试验，或 或者，将许可治疗程序，以完成其开发和营销。最终， 这项建议将导致安全和高质量的恢复期人体血浆用于治疗或预防 COVID19或其他目前没有其他有效治疗方法的致命疾病。

ABSTRACT Presently, convalescent plasma (CP) transfusion is being developed as a therapy for COVID-19 patients and as a prophylactic for high risk individuals. In addition, treatment with plasma or neutralizing antibody preparations from convalescent patients could be the only treatment for emerging infectious diseases, for which no other treatments may be available. At the same time, CP transfusion exposes the recipient to the risk of transfusion transmitted diseases (TTD), a risk which is additionally exacerbated by the compromised immune conditions of the critically ill patients. The limited number of current TTD blood tests does not provide for full protection, restrict the critically limited donor’s pool and may not be available in some areas. Pathogen Inactivation can provide the solution. Unfortunately, the currently utilized treatments for pathogen reduction in plasma (solvent-detergent, pasteurization of dry heat, UV or gamma irradiations) are non-selective and can compromise the quality of plasma’s neutralizing antibodies or other protective protein factors. We at ZATA Pharmaceuticals have developed a new class of pathogen inactivators (ZPI) based on the natural polyamines scaffold, which are truly selective in inactivating pathogens genomic molecules while sparing plasma proteins. Our preliminary results show that ZPI have high reactivity toward nucleic acids and do not modify model proteins (Cyt-C, RSV fusion protein) and animal sera growth factors. Using them, we inactivated different types of pathogens (G+ and G- bacteria, mycoplasma, fungi, protozoa) and high titer preparations of enveloped or non-enveloped viruses. Currently we are developing ZPI for pathogens reduction in transfusion red blood cells, research funded by NIH SBIR grant (R44 HL145783). In this application we propose to adapt the new pathogen inactivation process for treatment of convalescent plasma (CP) by: (1) using 6 virus species in human plasma to select the optimal ZPI and conditions for pathogen inactivation in human plasma; (2) using already developed analytical methods to establish conditions for complete neutralization and/or removal of the residual inactivator from the treated plasma; (3) using specific antibodies against 4 virus species to demonstrate by ELISA that the virus inactivation treatment has no effect on the binding of the antibodies to their targets; (4) using neutralizing antibodies against SARS-CoV-2 S protein to demonstrate preservation of the virus neutralizing properties of the antibodies after plasma treatment; (5) using repeated autologous infusion of treated plasma to establish its in vivo the safety in the rabbit models. After accomplishment of those initial goals we will apply for funding, including SBIR funding to complete, in collaboration with New York Blood Center, its pre-clinical evaluation and to initiate phase I human trials, or alternatively, will license the treatment procedure for completion of its development and marketing. Ultimately, this proposal will lead to a safe and high quality convalescent human plasma for treatment or prophylactics of COVID19 or other deadly diseases for which no other effective treatment is currently available.