Highly Selective Pathogen Inactivators For Treatment of Convalescent Transfusion Plasma

摘要 目前，恢复期血浆（CP）输注正在开发作为COVID-19患者的治疗方法 并作为高危个体的预防剂。此外，用血浆或中和抗体治疗 康复期病人的制剂可能是治疗新发传染病的唯一方法， 没有其他治疗方法可用。同时，CP输血使接受者面临风险 输血传播疾病（TTD）的风险，这是额外加剧了妥协 重症患者的免疫状况。目前TTD血液测试数量有限，无法提供 为了得到充分保护，限制极为有限的捐助者库，在某些地区可能无法获得。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 等离子体（溶剂-洗涤剂、干热的巴氏灭菌、UV或γ辐射）是非选择性的， 损害血浆中和抗体或其他保护性蛋白质因子的质量。我们在ZATA 制药业已经开发出一类基于天然多胺的新型病原体灭活剂（ZPI） 支架，这是真正的选择性灭活病原体基因组分子，同时节省血浆蛋白。 我们的初步结果表明，ZPI对核酸具有高反应性，并且不改变模型 蛋白质（Cyt-C，RSV融合蛋白）和动物血清生长因子。利用它们，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 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， 由NIH SBIR资助的研究（R44 HL 145783）。在本申请中，我们建议采用新的 一种用于处理恢复期血浆（CP）的病原体灭活方法，包括：（1）使用6种病毒， 人血浆中选择最佳ZPI和人血浆中病原体灭活的条件;（2）使用 已经开发的分析方法，以建立完全中和和/或去除 从处理的血浆中残留灭活剂;（3）使用针对4种病毒的特异性抗体， 通过ELISA证明病毒灭活处理对抗体与 （4）使用针对SARS-CoV-2S蛋白的中和抗体来证明SARS-CoV-2S蛋白的保留， 血浆处理后抗体的病毒中和特性;（5）使用重复的自体 输注处理后的血浆，以确定其在兔模型中的体内安全性。 完成这些初始目标后，我们将申请资金，包括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，将许可完成其开发和销售的治疗程序。最后， 该建议将产生安全和高质量的恢复期人血浆，用于治疗或预防 COVID 19或其他致命疾病，目前没有其他有效的治疗方法。

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.