CFI Pathogen Inactivation Technology

CFI病原体灭活技术

• 批准号: 7933943
• 负责人: TREVOR P. CASTOR
• 金额: $ 42.05万
• 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: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7933943
• 关键词: Acquired Immunodeficiency Syndrome; American; Animal Viruses; Antibodies; Applications Grants; Babesia; Bacteria; Biological; Bioterrorism; Blood; Blood donor screening; Chemicals; Clinical Trials; Collaborations; Community Healthcare; Conduct Clinical Trials; Cyclic GMP; Deforestation; Department of Defense; Detergents; Development; Disease; Disease Outbreaks; England; Ensure; Enzyme Immunoassay; Enzymes; Epidemic; Escherichia coli; Eukaryota; Europe; Evaluation; Excision; Exhibits; Factor VIII; Fibrinogen; Gases; Grant; HIV; Habitats; Heating; Human; Human Parvovirus B19; Individual; Infection; Infectious Agent; Influenza; Influenza A Virus, H5N1 Subtype; Institutes; International; Laboratories; Lead; Liquid substance; London; Manufacturer Name; Methods; Mexican; Modeling; Morbidity - disease rate; Mutation; National Heart, Lung, and Blood Institute; Parasites; Penetration; Pharmacologic Substance; Pharmacology; Phase; Plasma; Plasma Proteins; Plasmodium; Poliomyelitis; Process; Prokaryotic Cells; Property; Proteins; Recombinants; Recovery; Red Cross; Research; Residual state; Risk; Rupture; Saccharomyces cerevisiae; Safety; Screening procedure; Severe Acute Respiratory Syndrome; Small Business Innovation Research Grant; Smallpox; Solvents; Staging; Techniques; Technology; Testing; Therapeutic Monoclonal Antibodies; Thick; Transfusion; Transgenic Organisms; Travel; United States National Institutes of Health; Urbanization; Vaccines; Vascular blood supply; Vesicular stomatitis Indiana virus; Viral; Viral Antigens; Virus; Virus Inactivation; West Nile virus; Work; authority; base; clinical toxicology; design; design and construction; healthy volunteer; irradiation; meetings; microorganism; mortality; operation; pandemic disease; particle; pasteurization; pathogen; pathogenic bacteria; pre-clinical; preclinical study; programs; prototype; public health relevance; research and development; response; serological marker; swine flu; ultraviolet irradiation

DESCRIPTION (provided by applicant): This project entitled "CFI Pathogen Inactivation Technology," is in response to the challenge of developing Enabling Technologies to "Ensure a safe and adequate blood supply through the development of new processing technologies" Topic 06-HL-106, p. 94 of the National Institutes of Health American Recovery and Reinvestment Act of 2009 Challenge Grant Omnibus Solicitation RFA-OD-09-003. 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 healthcare community -- the need for effective pathogen inactivation and removal techniques for human blood plasma and plasma-derived products. There are also a number of emerging viruses such as West Nile and the breaking Mexican swine flu, and a number of potential bioterrorism pathogens such as smallpox that are of concern to the safety of the human plasma supply chain. In addition to viruses, bacteria and parasites such as Babesia spp. and Plasmodium spp. are major threats of spreading diseases through transfusion. The causes of the more rapid emergence and spread of these "killer" viruses and pathogens 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 global travel. Annually, an estimated 3.8 million Americans are transfused with 28.2 million blood components derived from 12.8 million units of blood donated by apparently healthy volunteers. A rigorous scrutiny of blood donors and the screening of donated blood for various serological markers have significantly reduced the mortality and morbidity due to transfusion-associated infectious agents. Some enzyme immunoassays used for routine screening may detect viral antigens or antibodies, but not the infectious agents themselves. Thus, there could be an asymptomatic window period of infectivity responsible for a residual risk of post-transfusion infection. Current approaches such as pasteurization; solvent-detergent; UV irradiation; and chemical and photochemical inactivation not always effective against a wide spectrum of pathogens, are sometimes encumbered by process-specific deficiencies, and often result in denaturation of the biologics that they are designed to protect. We plan to develop a physical pathogen inactivation process for non-enveloped and enveloped viruses as well as pathogenic bacteria and parasites in human plasma and plasma protein products. The process utilizes supercritical and near-critical fluids (SuperFluids(tm) or SFS). SuperFluids(tm) are normally gases which, when compressed, exhibit enhanced solvation, penetration and expansion properties. These gases are used to permeate and inflate the virus and pathogen particles. The overfilled particles are then decompressed and, as a result of rapid phase conversion, rupture at their weakest points. Research to date indicates that the SuperFluids(tm) CFI (critical fluid inactivation) process inactivates both enveloped viruses such as MuLV, VSV, TGE, BDVD, Sindbis and HIV and nonenveloped viruses such as Polio, Adeno, Reo, Parvo and EMC while preserving biological activity of the treated product. In a research collaboration with the National Institute of Biological Standards and Control (NIBSC), London, England, we demonstrated that SuperFluids(tm) CFI can inactivate more than 4 logs of human Parvovirus B19 (one of the smallest and toughest viruses) in human plasma in a two-stage CFI unit in less than 20 seconds. We have also demonstrated that SFS can disrupt and inactivate microorganisms such as E. coli, and thick-walled prokaryotes such as B. subtilis and tough eukaryotes such as S. cerevisiae at viral inactivation SFS conditions. CFI can be used with conventional viral reduction methods such as SD and nanofiltration as an orthogonal method of pathogen clearance. Our specific plans for this challenge grant are to: (1) design and construct laboratory-scale prototypes to conduct evaluation and trade-off studies that will lead to the selection of a commercial-scale SFS-CFI design to achieve > 6 logs of inactivation levels of nonenveloped and enveloped viruses with >90% retention of protein (e.g., Factor VIII) integrity; (2) test prototypical and emerging viruses and bacteria in human plasma in extant SFS-CFI units and in laboratory-scale prototypes in order to establish universal operation conditions as well as the universality of the SFS-CFI process for enveloped and nonenveloped viruses and pathogenic bacteria; (3) evaluate the potential of applying SFS-CFI enabling technology to human plasma proteins such as fibrinogen and other components; and (4) evaluate compatibility of SFS-CFI with other enabling pathogen inactivation/ reduction technologies such as nanofiltration to define orthogonal effective approaches to meet manufacturers' specifications and FDA requirements. Subsequently, with a pharmaceutical/biologics partner such as Baxter International and/or an institutional partner such as the American Red Cross, DOD or NHLBI, we plan to conduct pre-clinical studies, file an IND with the FDA and conduct clinical trials on CFI-treated plasma. A generally-applicable physical technology for inactivating viruses and emerging pathogens with high retention of biological activity will help ensure a blood supply that is safe from emerging and unknown pathogens as well as bioterrorism threats. In addition to human plasma and human plasma proteins, the developed technology will also be applicable to recombinant therapeutics, monoclonal antibodies, transgenics and vaccines. PUBLIC HEALTH RELEVANCE: There are a number of emerging viruses such as West Nile, Ebola, SARS, potential pandemic strains of influenza (H5N1), the breaking Mexican swine flu, bacteria, parasites 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. CFI pathogen inactivation technology gives pathogens the "bends," inactivating them without damaging proteins and enzymes in medically important transfusion fluids such as human plasma. This purely physical 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. As such, while CFI is capable of inactivating wide classes of viruses, bacteria and parasites, CFI has negligible negative impact on biological integrity and potency of the treated fluids. We plan to develop this technology as an orthogonal virus inactivation technology to techniques such as solvent-detergent (SD) that is not effective against non-enveloped viruses and passive virus removal techniques such as nanofiltration which does not render viruses inactive. This orthogonal approach is consistent with the regulatory authorities in Europe and the US that require a minimum of two pathogen inactivation technologies, which work by different mechanisms of action. The potential impact of a generally-applicable physical technology for inactivating viruses and emerging pathogens with high retention of biological activity will be very significant. Such a technology, especially when used with conventional virus inactivation or removal methods such as SD or nanofiltration, will help ensure a blood supply that is safe from emerging and unknown pathogens and bioterrorism threats. In addition to human plasma and human plasma proteins such as fibrinogen, the developed technology will also be applicable to recombinant therapeutics, monoclonal antibodies, transgenics and vaccines.

描述（由申请人提供）：该项目名为“CFI病原体灭活技术”，是为了响应开发使能技术的挑战，以“通过开发新的处理技术确保安全和充足的血液供应”，主题06-HL-106，第94页，美国国立卫生研究院美国复苏和再投资法2009年挑战拨款综合招标RFA-OD-09-003。世界范围内的艾滋病流行、埃博拉和非典型肺炎的周期性出现，以及最近爆发的H5N1等潜在的大流行流感毒株，都突出了医疗界持续关注的一个问题——需要有效的人血浆和血浆衍生产品的病原体灭活和清除技术。还有一些新出现的病毒，如西尼罗河病毒和正在爆发的墨西哥猪流感，以及一些潜在的生物恐怖主义病原体，如天花，它们对人类血浆供应链的安全构成了威胁。除病毒外，细菌和寄生虫，如巴贝斯虫和疟原虫，也是通过输血传播疾病的主要威胁。这些“杀手”病毒和病原体更迅速出现和传播的原因尚不完全清楚，但据认为是森林砍伐与野生病毒栖息地城市化、进化突变和快速全球旅行的某种结合造成的。据估计，每年有380万美国人接受2820万份血液成分的输血，这些血液成分来自表面上健康的志愿者捐献的1280万份血液。对献血者的严格审查和对捐献的血液进行各种血清学标志物的筛查，大大降低了因输血相关感染因子引起的死亡率和发病率。一些用于常规筛选的酶免疫测定法可以检测病毒抗原或抗体，但不能检测感染因子本身。因此，可能存在一个无症状的感染性窗口期，导致输血后感染的残留风险。目前的方法，如巴氏灭菌法；solvent-detergent;紫外线照射;化学和光化学失活并不总是对广泛的病原体有效，有时会受到特定工艺缺陷的阻碍，并经常导致它们旨在保护的生物制剂变性。我们计划对人类血浆和血浆蛋白制品中的非包膜病毒和包膜病毒以及致病菌和寄生虫开发一种物理病原体灭活工艺。该工艺利用超临界和近临界流体（超流体（tm）或SFS）。超流体（tm）通常是气体，当被压缩时，表现出增强的溶剂化、渗透和膨胀特性。这些气体被用来渗透和膨胀病毒和病原体颗粒。然后，过度填充的颗粒被减压，由于快速的相变，在它们最薄弱的地方破裂。迄今为止的研究表明，superfluid (tm) CFI（临界流体失活）工艺既灭活了包膜病毒，如MuLV、VSV、TGE、BDVD、Sindbis和HIV，也灭活了非包膜病毒，如脊髓灰质炎、腺病毒、Reo、Parvo和EMC，同时保留了处理产物的生物活性。在与英国伦敦国家生物标准与控制研究所（NIBSC）合作的一项研究中，我们证明了superfluid (tm) CFI可以在不到20秒的时间内在两阶段CFI装置中灭活人体血浆中4个以上的人细小病毒B19（最小和最顽强的病毒之一）。我们还证明，在病毒灭活SFS条件下，SFS可以破坏和灭活微生物，如大肠杆菌、厚壁原核生物，如枯草芽孢杆菌和坚韧的真核生物，如葡萄球菌。CFI可与常规的病毒还原方法（如SD和纳滤）作为病原体清除的正交方法。我们对这项挑战赠款的具体计划是：(1)设计和构建实验室规模的原型，以进行评估和权衡研究，从而选择商业规模的SFS-CFI设计，以实现无包膜和包膜病毒失活水平的bbbb6日志，并保留b> 90%的蛋白质（例如，因子VIII）完整性；(2)在现有的SFS-CFI装置和实验室规模的原型机中测试人血浆中的原型和新出现的病毒和细菌，以建立通用的操作条件，以及