Bromelain Proteinase Activity in IBD

描述（由申请人提供）：菠萝蛋白酶是一种源自菠萝茎的蛋白酶混合物，在健康食品商店中作为“消化辅助剂”出售。菠萝蛋白酶的有益的治疗效果也已经在几种人类炎性疾病和炎症的动物模型中被提出或证明。口服菠萝蛋白酶的自我治疗诱导了两名溃疡性结肠炎患者的临床和内镜缓解，这些患者的疾病对常规多药药物治疗无效。我们的对照动物研究表明，菠萝蛋白酶治疗降低了炎性肠病（IBD）的IL-10敲除（IL-10 -/-）鼠模型中自发性结肠炎的发展和已建立的结肠炎的严重程度。菠萝蛋白酶的生物学效应似乎取决于其蛋白水解活性，根据其蛋白酶组成，其蛋白水解活性可能因批次而异。用于人体试验的菠萝蛋白酶必须含有适当水平的活性成分，这一点至关重要。然而，目前还不知道菠萝蛋白酶中的哪些蛋白酶导致其在IBD中的抗炎活性。我们已经开发了专门测量3种主要菠萝蛋白酶（茎菠萝蛋白酶、ananain和果实菠萝蛋白酶）的蛋白水解活性的测定以及纯化这些酶的程序。在该研究中，患有已建立的结肠炎的IL-10-/-小鼠将用纯化的抗结肠炎药物口服治疗。 将确定菠萝蛋白酶蛋白酶和对结肠炎症的作用。拟议的研究将明确确定3种主要的菠萝蛋白酶如何有助于菠萝蛋白酶在结肠内的抗炎作用。他们还将开始确定这种情况发生的机制。这些信息将有助于进一步研究所涉及的机制，以及合理设计未来的临床试验，这种有前途的补充结肠炎治疗。因此，它们将加强为补充和替代医学实践提供信息所需的科学知识基础。

DESCRIPTION (provided by applicant): Bromelain is a mixture of proteinases derived from pineapple stem that is sold in health food stores as a "digestive aid". Beneficial therapeutic effects of bromelain have also been suggested or proven in several human inflammatory diseases and animal models of inflammation. Self-treatment with oral bromelain induced clinical and endoscopic remission of ulcerative colitis in two patients whose disease was refractory to conventional multi-agent medical therapy. Our controlled animal studies show that bromelain treatment decreases both the development of spontaneous colitis and the severity of established colitis in the IL-10 knockout (IL-10 -/-) murine model of inflammatory bowel disease (IBD). The biological effects of bromelain appear to depend on its proteolytic activity, which can vary from lot to lot according to its proteinase composition. It is critical that the bromelain to be used in human trials contain appropriate levels of active ingredients. However, it is not currently known which proteinases within bromelain account for its anti-inflammatory activity in IBD. We have developed assays to specifically measure the proteolytic activity of the 3 major bromelain proteinases (stem bromelain, ananain, and fruit bromelain) as well as procedures to purify these enzymes. In this study, IL-10-/- mice with established colitis will be treated orally with purified bromelain proteinases and effects on colonic inflammation will be determined. The proposed studies will definitively determine how the 3 major bromelain proteinases contribute to anti-inflammatory effects of bromelain within the colon. They will also begin to determine the mechanisms by which this occurs. This information will facilitate further investigations of the mechanisms involved as well as the rational design of future clinical trials of this promising complementary colitis therapy. They will thus strengthen the scientific knowledge base needed to inform the practice of complementary and alternative medicine.