CFI Inactivation of Human Plasma

人血浆的 CFI 灭活

• 批准号: 7675084
• 负责人: TREVOR P. CASTOR
• 金额: $ 34.43万
• 依托单位: APHIOS CORPORATION
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7675084
• 关键词: Acquired Immunodeficiency Syndrome; American; Animal Viruses; Area; Biocompatible Materials; Biological; Bioterrorism; Blood Circulation; Capsid Proteins; Chemicals; Collaborations; Community Healthcare; Country; Cyclic GMP; Decontamination; Deforestation; Department of Defense; Depreciation; Detergents; Disease Outbreaks; Drops; England; Enzymes; Epidemic; Equipment; Europe; Excision; Exhibits; Ficusin; Fresh Frozen Plasmas; Gases; HIV; Habitats; Heating; Hepatitis A; Human; Human Parvovirus B19; Influenza; Influenza A Virus, H5N1 Subtype; Injection of therapeutic agent; Institutes; International; Laboratories; Life Cycle Stages; Lipids; Liquid substance; London; Methods; Modeling; Mutation; National Heart, Lung, and Blood Institute; Operative Surgical Procedures; Ozone; Penetration; Pharmacologic Substance; Pharmacology; Phase; Phase I Clinical Trials; Plasma; Plasma Proteins; Poliomyelitis; Process; Production; Property; Proteins; Psoralens; Red Cross; Research; Rupture; Safety; Serum; Severe Acute Respiratory Syndrome; Smallpox; Solvents; Staging; Sterilization for infection control; System; Techniques; Technology; Testing; Time; Travel; Urbanization; Vesicular stomatitis Indiana virus; Viral; Virion; Virus; Virus Inactivation; West Nile virus; Work; authority; base; clinical toxicology; cost; density; design; design and construction; fetal bovine serum; irradiation; mathematical model; model design; pandemic disease; particle; pasteurization; pathogen; pre-clinical; pressure; programs; prototype; public health relevance; scale up; therapeutic protein; ultraviolet irradiation

DESCRIPTION (provided by applicant): The worldwide AIDS epidemic, the periodic emergence of Ebola and SARS, and the recent outbreaks of potentially pandemic strains of influenza such as H5N1 have highlighted a persistent concern in the health- care community -- the need for effective sterilization techniques for human blood plasma and plasma-derived products. There are also a number of emerging viruses such as West Nile and a number of potential bioterrorism pathogens such as smallpox that are of concern to the safety of the human plasma supply. The causes of the more rapid emergence and spread of these "killer" viruses are not entirely known, but are thought to be caused by some combination of deforestation with urbanization of wild virus habitats, evolutionary mutations, and rapid travel between countries around the globe that can facilitate the spread of both natural and bioterrorism pathogens. A number of approaches have been employed for the inactivation or removal of viruses in human plasma and therapeutic proteins derived from human plasma, including: heating or pasteurization; solvent-detergent technique; Ultra Violet (UV) irradiation; chemical inactivation utilizing hydrolyzable compounds such as 2- proprionolactone and ozone; and photochemical decontamination using synthetic psoralens. Thus, current approaches are not always effective against a wide spectrum of human and animal viruses, are sometimes encumbered by process-specific deficiencies, and often result in denaturation of the biologicals that they are designed to protect. Some of the commercially available methods are effective in inactivating enveloped viruses, such as HIV, but are not very effective against nonenveloped viruses such as Hepatitis A (HAV) and parvovirus B19. We propose to develop a safe and cost-effective universal process and equipment for the inactivation of non- enveloped and enveloped viruses in pooled as well as single units of human plasma. We propose to do so by further advancing a unique, rapid and generally applicable virus inactivation technique based on supercritical and near-critical fluids (SuperFluids" or SFS) technology. SuperFluids" are normally gases which, when compressed, exhibit enhanced solvation, penetration and expansion properties. These gases are used to permeate and inflate the virus particles. The overfilled particles are then decompressed and, as a result of rapid phase conversion, rupture at their weakest points. The process is purely physical and does not involve the use of heat, chemicals and/or irradiation, each of which has significant drawbacks in the viral inactivation of human plasma. Our research to date indicates that the SuperFluids" CFI (critical fluid inactivation) process inactivates enveloped viruses such as MuLV, VSV, TGE, BVD, Sindbis and HIV in fetal bovine serum by a lipid solubilization mechanism, similar to the solvent detergent method. Our research also indicates that SFS-CFI inactivates non-enveloped viruses surrounded by a tough protein capsid through rapid expansion of the fluid with concomitant physical disruption of viral particles. SFS-CFI's ability to inactivate non-enveloped viruses such as Polio, Adeno, Reo and EMC in fetal bovine serum, while preserving biological activity of the treated product, has been demonstrated. In a research collaboration with the National Institute of Biological Standards and Control (NIBSC), London, England, we have also demonstrated that SuperFluids" CFI can inactivate more than 4 logs of human Parvovirus B19 in human serum in a two-stage CFI unit in less than one minute. The revised Specific Aims of the proposed Phase I research program are: " Specific Aim 1: Mathematically model and evaluate key process parameters that are critical for scaling-up of the laminar flow CFI unit and the SFS-CFI process. Mathematical modeling and transport phenomena calculations will be conducted to define optimum SFS and plasma parameters such as density, fluid type and droplet size for effective contact of the viral particles by the SFS, penetration, saturation, expansion, disruption and inactivation. " Specific Aim 2: Based on the results of this modeling, design and test injection nozzles and isobaric chamber as well as define operating conditions for achieving > 3 logs of virus inactivation of a prototypical enveloped virus and a prototypical nonenveloped virus in a single laminar flow CFI unit with retention of > 90% of protein integrity. In Phase II, we will scale-up by a factor of 500X then scale-down by a factor of 10X to 100X, a single-stage laminar flow SFS-CFI prototype. We also plan to validate a large-scale SuperFluids" CFI prototype for human plasma under cGMP conditions. In Phase III, with a pharmaceutical/biologics partner such as Baxter International and/or an institutional partner such as the American Red Cross, the Department of Defense or the National Heart, Blood and Lung Institute, we plan to conduct pre-clinical toxicology and pharmacology studies, file an IND with the FDA and conduct Phase I clinical trials. PUBLIC HEALTH RELEVANCE: There are a number of emerging viruses such as West Nile, Ebola, SARS potential pandemic strains of influenza (H5N1) and a number of potential bioterrorism pathogens such as smallpox that are of concern to the safety of the human plasma supply. Current approaches are not always effective against a wide spectrum of human and animal viruses, are sometimes encumbered by process-specific deficiencies, and often result in denaturation of the biologicals that they are designed to protect. We propose to develop a purely physical technique that gives the virus the "bends," inactivating them without damaging proteins and enzymes in medically important fluids such as human plasma. This technique does not involve the use of heat, chemicals and/or irradiation, each of which has significant drawbacks in the viral inactivation of human plasma. Aphios Corporation will focus on developing this physical virus inactivation technique for areas of human use where a virus inactivation step is essential and will remain necessary for the manufacture of safe products. A generally applicable and effective virus inactivation process that increases yield from plasma and reduces processing cost will have a significant impact in the marketplace. We plan to commercialize this technology as an orthogonal virus inactivation technology to techniques such as solvent-detergent (S/D) that is not effective against non-enveloped viruses and passive virus removal techniques such as nanofiltration. This approach is consistent with the regulatory authorities in Europe and the US that require a minimum of two virus inactivation technologies, which work by different mechanisms of action.

描述（由申请人提供）：全球流行病，埃博拉病毒和SARS的周期性出现，以及最近在潜在的流感菌株（例如H5N1）爆发的爆发，突显了健康社区中的持续关注 - 对人血浆和血浆血浆和血浆菌的有效灭菌技术的需求。还有许多新兴病毒，例如西尼罗河，以及许多潜在的生物恐怖病原体，例如天花，这些病原体涉及人血浆供应的安全性。这些“杀手”病毒的出现更快出现和传播的原因并不完全清楚，但被认为是由于森林砍伐与野生病毒栖息地，进化突变的城市化，全球各国之间的快速旅行而引起的，可以促进自然和生物恐怖症病原体的传播。已经采用了许多方法来灭活或去除人血浆中的病毒和源自人血浆的治疗蛋白，包括：加热或巴氏杀菌；溶剂少量技术； Ultra Violet（UV）辐射；利用可水解化合物（例如2-丙二酮和臭氧）的化学灭活；和光化学去污染物使用合成的薄膜。因此，当前的方法并不总是有效地针对各种各样的人类和动物病毒，有时会因过程特异性缺陷而抑制，并且通常会导致其旨在保护的生物学变性。某些市售方法可有效地灭活诸如HIV的包膜病毒，但对诸如丙型肝炎A（HAV）和细小病毒B19等非发育病毒的有效性不是很有效。我们建议开发一种安全且具有成本效益的通用过程和设备，以使人类等离子体的合并和单个单元中的非包膜和包裹病毒失活。我们建议通过进一步推进基于超临界和近临界流体（超氟”或SFS）技术的独特，快速且普遍适用的病毒灭活技术来做到这一点。超级流体“通常是气体”，通常是压缩的气体，它表现出增强的溶剂化，穿透性，穿透性和扩张性能。这些气体用于渗透和充气病毒颗粒。然后对过度填充的颗粒进行解压缩，并且由于快速相位转换而在其最弱点处破裂。该过程纯粹是物理的，不涉及使用热，化学物质和/或辐照，每个过程在人血浆的病毒失活中都有显着的缺点。迄今为止，我们的研究表明，超流体“ CFI（临界液体失活）过程失活了诸如MULV，VSV，TGE，BVD，BVD，SINDBIS，SINDBIS，SINDBIS，SINDBIS，SINDBIS和HIV中的包蛋白牛血清中通过脂质溶液机制，类似于我们的研究，该方法与我们的研究相似。通过快速扩张液体与病毒颗粒的物理破坏，可以使胎儿牛血清中的polio，Adeno，Reo和EMC灭活，而在伦敦的生物学中保留了伦敦的合作，则蛋白质的capsid。我们还证明了超级流体“ CFI可以在不到一分钟的时间内在两个阶段的CFI单元中在人类血清中灭活4个以上的人类细胞谷病毒B19。 The revised Specific Aims of the proposed Phase I research program are: " Specific Aim 1: Mathematically model and evaluate key process parameters that are critical for scaling-up of the laminar flow CFI unit and the SFS-CFI process. Mathematical modeling and transport phenomena calculations will be conducted to define optimum SFS and plasma parameters such as density, fluid type and droplet size for effective contact of the viral particles by the SF，渗透，饱和度，膨胀，中断和灭活。 正直。在第二阶段，我们将扩大500倍的比例，然后缩小10倍至100倍，即单级层流SFS-CFI原型。我们还计划在CGMP条件下验证人类血浆的大规模超流体。临床试验。缺乏症，通常会导致它们旨在保护它们的生物学变性。该技术不涉及使用热，化学物质和/或辐照，每种技术在人血浆的病毒灭活中都有显着的缺点。 Aphios Corporation将专注于开发这种物理病毒灭活技术，用于人类使用的领域，在该领域中，病毒失活步骤至关重要，对于生产安全产品仍然是必要的。通常适用且有效的病毒灭活过程，从血浆中增加产量并降低加工成本将对市场产生重大影响。我们计划将这项技术作为正交病毒灭活技术将其商业化为溶剂含量（S/D）等技术，这些技术对非发育的病毒和诸如纳米过滤等非发育病毒和被动病毒去除技术无效。这种方法与欧洲和美国需要至少两种病毒灭活技术的监管机构一致，这些技术通过不同的作用机理起作用。