Bromelain Proteinase Activity in IBD

描述（由申请人提供）：菠萝蛋白酶是一种从菠萝茎中提取的蛋白酶混合物，在保健食品商店中作为“助消化”出售。菠萝蛋白酶在几种人类炎症性疾病和动物炎症模型中也有有益的治疗作用。口服菠萝蛋白酶自我治疗2例常规多药治疗难治性溃疡性结肠炎患者的临床和内镜缓解。我们的对照动物研究表明，在IL-10敲除（IL-10 -/-）炎症性肠病（IBD）小鼠模型中，菠萝蛋白酶治疗降低了自发性结肠炎的发展和已建立结肠炎的严重程度。菠萝蛋白酶的生物学效应似乎取决于其蛋白水解活性，根据其蛋白酶组成的不同，其蛋白水解活性可能有很大差异。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，目前尚不清楚菠萝蛋白酶中的哪些蛋白酶在IBD中具有抗炎活性。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、菠萝蛋白酶和水果菠萝蛋白酶）的蛋白水解活性的方法，以及纯化这些酶的方法。在本研究中，IL-10-/-结肠炎小鼠将口服纯化的

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.