Crystalline Endolysin Treatment for Tuberculosis in TB/HIV co-infected patients

描述（由申请人提供）：虽然结核病（TB）通常被认为是过去的疾病，但它仍然是全世界人民的主要杀手。截至2008年底，全球约有20亿人感染结核分枝杆菌（MTb），占世界人口的三分之一。世界卫生组织估计，全世界每年有200万人死于结核病。结核病现在是世界上仅次于人体免疫缺陷病毒/获得性免疫缺陷综合症（艾滋病毒/艾滋病）的第二大常见传染病死亡原因。20世纪80年代中期，美国结核病疫情的重新爆发使人们重新关注结核病。结核病病例增加的主要原因是感染艾滋病毒/艾滋病的人更容易受到感染，以及结核分枝杆菌耐药菌株的发展，结核分枝杆菌可导致人类结核病。世卫组织2010年4月最新报告结果显示，俄罗斯每4名结核病患者中就有1人被诊断为耐多药结核病，而耐多药结核病负担的约50%在中国和印度。2008年，全世界有44万例耐多药结核病例，15万人死亡。根据2010年3月的报告，大约5.4%的耐多药结核病病例是广泛耐药结核病。如果没有新的、更有效的药物、诊断方法和疫苗，消除结核病是不切实际的。世界迫切需要更简单、更快速的治疗药物;更安全、更有效的药物，可以治疗所有形式的结核病，特别是免疫功能低下的艾滋病毒/结核病合并感染者。作为抗生素治疗的替代方案，特别是对于由M.为了治疗结核病，我们建议开发一种酶疗法，其可以与各种制剂条件和递送途径一起使用。酶疗法可能与抗生素疗法一样有效，因为酶是高度特异性的，并且对靶细菌有效。然而，将这些酶递送到感染部位仍然是一个挑战。大多数药物蛋白质在施用部位易于降解，无论是静脉内、皮下、口服、局部或通过任何其他途径施用。我们认为，一个安全有效的输送系统，这种应用可能是通过使用蛋白质晶体，它提供了高度浓缩，稳定和纯净的产品。此外，由于M.结核病存在于巨噬细胞中。细胞穿透肽（CPP），无论是框内工程化的还是非共价地吸附/结合至蛋白质底物的，已被用于将各种各样的分子递送到许多细胞类型中。在该提案中，我们提出开发具有细胞穿透肽（CPP）的结晶配制的重组内溶素疗法，其可以与各种制剂条件和递送途径一起使用。CPP与内溶素偶联的机会是多种多样的，因为它可能能够穿透巨噬细胞，从而能够杀死M。艾滋病毒/结核病合并感染患者的结核病，并可能延长他们的寿命与最小的不适。在第一阶段，我们打算研究使用内溶素的可行性，可溶性和结晶制剂与或不与细胞穿透肽，作为全身递送系统在体内动物模型的M。肺结核感染。如果成功，这种方法将导致引入新的，有效的酶疗法治疗M。肺结核感染。 公共卫生相关性：结核病（TB）是我们这个时代最具破坏性的全球健康问题之一，每年造成约200万人死亡。作为一种预防措施，目前使用卡介苗来避免结核病的严重并发症，并降低儿童结核病的发病率，但它对成人肺结核病几乎没有效果。我们提出了一种新的结晶内溶素，其即使在免疫功能低下的患者中也是纯的、安全的、有效的，并且与需要免疫系统产生抗体的疫苗不同，内溶素可以直接作用于结核分枝杆菌生物体，从而预防和治疗HIV/TB共感染患者中的结核病。

DESCRIPTION (provided by applicant): Though often considered to be a disease of the past, tuberculosis (TB) is still a leading killer of people worldwide. At the end of 2008, approximately 2 billion people one-third of the world's population is infected with Mycobacterium tuberculosis (MTb), the etiologic agent of TB. The World Health Organization estimates that 2 million people worldwide die of TB each year. TB is now the second most common cause of death from infectious disease in the world after human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). In the mid-1980s, a resurgence of outbreaks in the United States brought renewed attention to TB. Main reasons for increase in TB cases are due to higher vulnerability of people infected with HIV/AIDS and development of drug-resistant strains of MTb, which causes TB in human. In the latest 2010 April WHO report results show that one in every 4 TB patients diagnosed with MDR-TB in Russia and about 50% of MDR-TB burden is in China and India. Worldwide there are 440,000 MDR-TB cases and 150,000 deaths occurred in 2008. About 5.4% of MDR-TB cases have XDR-TB according to the report in March 2010.It is impractical to eliminate TB without novel, and more effective drugs, diagnostics and vaccines. The world immediately needs simpler and faster curative drugs; safer and effective drugs that can treat all forms of TB especially in immuno- compromised people with HIV/TB co-infections. As an alternative to antibiotic therapy, especially for systemic lung infections by M. tuberculosis, we are proposing to develop an enzyme therapy that can be used with various formulation conditions and delivery routes. The enzyme therapy may be as effective as antibiotic therapy because the enzyme is highly specific, and potent against the targeted bacteria. However, delivering these enzymes to the site of the infection remains a challenge. Most pharmaceutical proteins are susceptible to degradation at the site of administration, whether administered intravenously, subcutaneously, orally, topically or by any other route. We believe that a safe and effective delivery system for this application may be through the use of protein crystals, which provides highly concentrated, stable, and pure products. In addition, technologies are also needed to deliver endolysin to intracellular compartment since M. tuberculosis resides in macrophages. Cell penetrating peptides (CPP), either engineered in-frame or adsorbed/bound non-covalently to protein substrates, have been used to deliver a wide variety of molecules into a number of cell types. In this proposal we are proposing to develop a crystalline formulated recombinant endolysin therapy with cell-penetrating peptides (CPP) that can be used with various formulation conditions and delivery routes. The opportunities for CPP conjugated to endolysin are multiple since it may be able to penetrate the macrophages thereby able to kill M. tuberculosis in HIV/TB co-infected patients and may extend their life span with minimal discomfort. In Phase I, we intend to study the feasibility of using endolysins, both soluble and crystalline formulations with or without cell penetrating peptides, as a systemic delivery system in an in vivo animal model for M. tuberculosis infection. If successful, this approach will lead to the introduction of novel, efficient enzyme therapy for the treatment of M. tuberculosis infections. PUBLIC HEALTH RELEVANCE: Tuberculosis (TB) is one of the most devastating global health problems of our time, causing approximately 2 million deaths a year. As a preventive measure, currently BCG vaccine is used to avoid serious complications of TB and to reduce the rate of pediatric TB but It has little or no effectiveness for pulmonary TB in adults. We are proposing a novel crystalline endolysin that is pure, safe, effective even in immunocompromised patients, and unlike vaccines which needs immune system to generate the antibodies the endolysin can act directly on the Mycobacterium tuberculosis organism and thus prevent and treat tuberculosis in HIV/TB co-infected patients.