Development of a Novel Anti-Inflammatory Therapeutic Based on Antithrombin

描述（由申请人提供）：感染性休克是败血症的可怕结果，是由感染引发的急性全身性炎症状态。在美国，每年发生超过75万例败血症，死亡率达到30%。这种破坏性结果的高流行率突出了新型靶向抗炎治疗的巨大影响潜力。其中一种候选药物是抗凝血酶（AT）。AT是一种天然血浆蛋白，被认为是最重要的凝血级联抑制剂。然而，AT也表现出不同的抗炎信号活动。因此，治疗性给予AT具有改善许多炎症性疾病的潜力。不幸的是，AT的临床应用受到AT抗凝血活性的限制，这会产生不良的出血事件。为了规避这一限制，正在开发一种缺乏抗凝血活性但保留抗炎活性的突变形式的AT。为了支持这一应用，已经产生并建立了以下方法：(1)ATRCL，一种突变的AT形式，具有最小的抗凝血活性，但保持了阻断NF-:B激活的能力（在体外试验中）；(2)脓毒性休克小鼠模型，在该模型中，野生型AT治疗可改善脂多糖（LPS）诱导的心源性休克。根据其体外抗炎活性（Aim 1）生成ATRCL，并对其剂量进行标准化。将使用凝血试验（Aim 2）对ATRCL的体内和体外安全性进行首次评估。最后，将在LPS诱导的脓毒性休克小鼠模型中测试其疗效（Aim 3）。肝素对安全性和有效性的影响也将被监测。所描述的实验将确定一个极有前途的化合物是否在体内执行其概念和证明的体外潜力。如果成功，ATRCL将成为具有广泛和有价值的适用性和突出的安全性的治疗方法。

DESCRIPTION (provided by applicant): Septic shock is a dire outcome of sepsis, an acute systemic inflammatory state triggered by an infection. In the USA, over 750,000 episodes of sepsis occur each year and mortality reaches 30%. The high prevalence of such devastating outcomes highlights the potential for enormous impact by novel targeted anti-inflammatory therapeutics. One such candidate agent is antithrombin (AT). AT is a natural plasma protein that is considered to be the most important inhibitor of the blood clotting cascade. However, AT also exhibits distinct anti-inflammatory signaling activities. Thus, therapeutic administration of AT has the potential to ameliorate numerous inflammatory diseases. Unfortunately, clinical deployment of AT has been limited by AT's anticoagulant activity, which produces adverse bleeding events. To circumvent this limitation, a mutated form of AT that lacks anticoagulant activity but retains anti-inflammatory activity is being developed. In support of this application the following have been generated and established: (1) ATRCL, a mutated AT form with minimal anticoagulant activity but which maintains the ability to block activation of NF-:B (in an in vitro assay) and (2) a mouse model of septic shock, in which treatment with wild-type AT ameliorates lipopolysaccharide (LPS) induced cardiogenic shock. ATRCL will be generated and its dosage standardized based on its in vitro anti-inflammatory activity (Aim 1). The first assessments of ATRCL's in vivo and in vitro safety using coagulation assays (Aim 2) will be conducted. Finally, efficacy will be tested in mouse models of LPS induced septic shock (Aim 3). The effect of heparin on safety and efficacy will also be monitored. The experiments described will determine whether an extremely promising compound performs in vivo to its conceptual and demonstrated in vitro potential. If successful, ATRCL will become a therapeutic with broad and valuable applicability and an outstanding safety profile. PUBLIC HEALTH RELEVANCE: Septic shock kills 150,000 people per year in the US alone. The process is complex, but is driven by inflammation. Antithrombin (AT) is a potent natural anti-inflammatory agent, but it often leads to serious bleeding. We have a new form of AT which imparts anti-inflammatory benefits without the bleeding risks. The overall goal of this project is to develop this drug to help humans.