Isolaton of new phage enzymes to kill B. anthracis

描述（由申请人提供）：我们目前对炭疽杆菌和其他细菌生物武器的防御主要基于使用抗生素治疗。考虑到这种武器中使用的细菌可能对目前的治疗性或预防性抗生素表现出天然或工程抗性的可能性，这一计划是有缺陷的。因此，我们目前正在开发一类新型的抗菌剂。我们的系统是基于使用噬菌体溶素来提供针对感兴趣的细菌病原体（特别是B）的快速和特异性杀伤作用。炭疽病除了提供一种以前无法获得的细菌杀灭方法之外，噬菌体溶素的主要吸引力在于，即使在广泛的尝试之后，也不能检测到细菌对其作用的抗性。我们的实验室是第一个使用这些酶的纯化形式，以杀死粘膜表面和血液中的定植致病菌。所述酶对于所述酶所来源的物种或菌株是特异性的，表明这些酶可用于仅靶向杀死病原性细菌，而对正常植物群细菌几乎没有影响。在这些研究中，我们发现对细胞壁结合具有两种不同特异性的酶（即，酰胺酶和胞壁酶）在它们的杀伤能力方面具有协同效应。在我们用来自γ噬菌体的PlyG噬菌体酶进行的研究中，该酶对B具有特异性。在炭疽杆菌中，我们证明这种酶能够在体外杀死炭疽杆菌，在两分钟内使108个细菌无菌。在体内，我们能够保护动物免受与B密切相关的芽孢杆菌的致命挑战。炭疽菌和B.炭疽病由于这些酶的协同效应，本申请旨在鉴定和开发用于B的酶组合。攻击芽孢杆菌细胞壁中四种不同键的炭疽菌。这将确保更有效的杀灭作用，并降低对这些酶产生抗性的可能性。将从环境中发现的噬菌体和溶原化B的噬菌体中分离噬菌体酶。炭疽病然后将这些酶表征其特异性，纯化并用于体外和体内系统以确定功效。因为这些酶可能是抵抗耐药B攻击的重要防线。炭疽病，有一些酶在我们的处置可能允许更好地决定，如有必要，他们的使用。

DESCRIPTION (provided by applicant): Our current defense against Bacillus anthracis, and other bacterial bioweapons, is based primarily on the use of antibiotic therapies. This plan is flawed considering the possibility that the bacteria used in such weapons may express naturally occurring or engineered resistances to current therapeutic or prophylactic antibiotics. For this reason we are currently developing a novel class of antibacterial agents. Our system is based on the use of phage lysins to provide a rapid and specific killing action against bacterial pathogens of interest, in particular B. anthracis. In addition to offering a previously unavailable method of bacterial killing, phage lysins are primarily attractive in that bacterial resistance to their action cannot be detected, even after extensive attempts. Our laboratory is the first to use these enzymes in their purified form to kill colonizing pathogenic bacteria on mucous membrane surfaces and in blood. The enzymes are specific for the species or strain from which the enzymes were derived, indicating that these enzymes may be used for targeted killing of only the pathogenic bacterium with little to no effect on normal flora bacteria. During these studies we discovered that enzymes with two different specificities for cell wall bonds (i.e., amidase and muramidase) have a synergistic effect in their killing capacity. In our studies with the PlyG phage enzyme from the gamma phage that is specific for B. anthracis, we show that this enzyme is able to kill anthrax bacilli in vitro, reducing 108 bacteria to sterility in two minutes. In vivo, we are able to protect animals from lethal challenge with both a closely related bacillus to B. anthracis as well as B. anthracis. Because of the synergistic effects in these enzymes, this application is designed to identify and develop a combination of enzymes for B. anthracis that attack the four different bonds in the bacillus cell wall. This will ensure a more efficient killing action as well as reduce the possibility of the development of resistance to these enzymes. Phage enzymes will be isolated from phage found in the environment and phage lysogenizing B. anthracis. These enzymes will then be characterized as to their specificity, purified and used in both in vitro and in vivo systems to determine efficacy. Because these enzymes may be an important line of defense against an attack with drug-resistant B. anthracis, having a number of enzymes at our disposal may allow for better decisions as to their use if necessary.