Targeting MMP9 to Improve Outcomes in Serious Influenza Infections

描述(由申请人提供)：尽管有疫苗和全球监测，流感感染仍然是一个巨大的公共卫生问题。甲型流感病毒(IAV)的高突变率使其能够逃避自然和疫苗介导的免疫。此外，当前的抗病毒疗法不能在延迟开始治疗时预防与IAV相关的死亡。因此，治疗流感疾病的新疗法是迫在眉睫的公共卫生优先事项。我们的初步研究确定了基质金属蛋白酶-9是一个有吸引力的IAV治疗靶点。在感染季节性和甲型H1N1禽流感病毒的人的血浆样本和感染H1N1禽流感病毒的小鼠的肺样本中，基质金属蛋白酶-9显著上调。在小鼠中，基质金属蛋白酶-9增加了IAV相关死亡率、晚期肺部炎症和晚期肺部病毒负荷。我们建议在三个整合和高度协作的目标下，测试一种“重新定位”的候选治疗药物(ADZ1236)的治疗效果，该候选药物在两个物种的IAV感染中选择性和有效地抑制基质金属蛋白酶-9活性。目的1检测AZD1236对感染BL2 H1N1禽流感病毒的小鼠的治疗效果。AZD1236将单独在小鼠身上进行测试，并与临床使用的抗病毒药物(神经氨酸酶抑制剂奥司他韦)联合进行测试。将对延迟启动的治疗方法进行优化，以模拟严重IAV感染的“真实世界”治疗情景。目的2确定基质金属蛋白酶-9促进(和AZD1236限制)小鼠严重IAV感染的机制。我们将研究H1N1感染的WT与MMP9-/-小鼠和MMP9骨髓嵌合小鼠，并使用体外方法来验证我们的假设，即MMP9通过裂解宿主或病毒蛋白而促进IAV疾病的不利结果。这些基质金属蛋白酶-9的底物将被鉴定。目的3将测试AZD1236对感染BL2 H1N1和BL3高致病性禽流感H5N1毒株的雪貂的疗效(如果该毒株在资助期内出现流行病或大流行毒株，则测试该毒株对感染H7N9 IAV的雪貂的疗效)。我们的研究将确定AZD1236介导的基质金属蛋白酶-9抑制对小动物和大动物的IAV是否具有足够的治疗效果，从而满足FDA为证明疗效而申请FDA批准的“两个动物规则”。本文建议的工作的成功完成可能会为严重流感引起的肺部疾病提供一种“一流”的治疗干预。

DESCRIPTION (provided by applicant): Influenza infection remains an enormous public health concern despite the availability of vaccines and worldwide surveillance. The high mutation rates of influenza A viruses (IAVs) enable the viruses to evade natural and vaccine-mediated immunity. Also, current anti-viral therapies do not prevent IAV-related deaths when there is a delay in initiating treatment. Thus, new therapeutic options for treating influenza disease are an immediate public health priority. Our pilot studies identify MMP-9 as an attractive IAV therapeutic target. MMP- 9 is strikingly upregulated in plasma samples from human subjects infected with seasonal and H1N1 IAV and in lungs from mice infected with H1N1 IAV. In mice, Mmp-9 increases IAV-associated mortality and late-stage lung inflammation and late-stage lung viral burdens. We proposed to test the therapeutic efficacy of a "re- purposed" therapeutic candidate (ADZ1236) that selectively and potently inhibits MMP-9 activity in IAV infections in two species in three integrated and highly collaborative aims. Aim 1 will test the therapeutic efficacy of AZD1236 therapy in mice infected with BL2 H1N1 IAV. AZD1236 will be tested in mice alone and in combination with a clinically-used antiviral agent (the neuraminidase inhibitor, oseltamivir). A delayed initiation of treatment approach will be optimized to model "rea-world" treatment scenarios for serious IAV infections. Aim 2 will identify the mechanisms by which Mmp-9 promotes (and AZD1236 limits) serious IAV infections in mice. We will study H1N1-infected WT vs. Mmp-9-/- mice and Mmp-9 bone marrow chimeric mice and use in vitro approaches to test our hypothesis that Mmp-9 promotes adverse outcomes in IAV disease by cleaving host or viral proteins. These Mmp-9 substrates will be identified. Aim 3 will test the therapeutic efficacy of AZD1236 in ferrets infected with BL2 H1N1 and the BL3 highly pathogenic avian influenza (HPAI) H5N1 strain (and in H7N9 IAV-infected ferrets if this strain emerges as an epidemic or pandemic strain during the funding period). Our studies will determine whether AZD1236-mediated MMP-9 inhibition has therapeutic efficacy against IAV in both small and large animals sufficient to thereby satisfy the FDA "Two Animal Rule" required to demonstrate efficacy to apply for FDA approval. Successful completion of the work proposed herein may provide a "first in class" therapeutic intervention for serious influenza-mediated lung disease.