Development of a Novel MRI Contrast Agent for Early Detection of Alcoholic Steatohepatitis

摘要 慢性肝病（CLD）是全球发病率和死亡率的主要原因[1-4]。肝纤维化可以 任何类型的慢性肝病（CLD）患者，包括酒精性肝病（ALD），肝炎 丙型肝炎、B型肝炎、非酒精性脂肪性肝病（NAFLD）和自身免疫性肝炎。主要临床 肝硬化的后果是肝功能衰竭和肝细胞癌（HCC），这两者都增加了风险 死亡之酒精性肝病（ALD）是全球肝病和肝脏相关死亡的主要原因， 特别是在发达国家。目前的金标准诊断方法，肝活检是侵入性的， 许多局限性，包括采样误差和高观察者间的变异性，即使对于先进的诊断， 肝纤维化的阶段。作为替代品研究的几种技术都没有提供所需的灵敏度 和纤维化早期检测和分期的特异性。因此，一种创新的，安全的MRI造影剂， 需要克服几个主要的限制，包括对早期纤维化的低灵敏度和特异性， 异质组织中的检测。我们的团队开创了一类新的钆基造影剂 其利用蛋白质支架结合Gd 3+原子。我们已经证明，ProCA32.collagen，一种胶原蛋白1靶向 蛋白质MRI造影剂，表现出对胶原蛋白I的强亲和力，其表达水平和空间模式 取决于纤维化的阶段。ProCA 32.胶原蛋白在1.4 和7.0 T，这使得能够稳健地检测早期酒精诱导的肝纤维化和非酒精性肝纤维化。 通过双对比模式在动物模型中观察脂肪性肝炎。我们的初步研究毒性概况和在体内 我们的化合物在使用非GLP级材料的动物中的稳定性提供了信心， 化合物将在人类临床试验中表现出用于肝纤维化的早期检测的良好灵敏度和特异性。 在本提案的第一阶段，我们寻求及时支持，以开展IND启用CMC研究， GLP/cGMP级ProCA 32胶原蛋白。在第二阶段，我们将进行毒性和安全性研究， GLP/cGMP级产品，以完成IND要求的临床试验研究。

Abstract Chronic liver disease (CLD) is a major cause of morbidity and mortality worldwide[1-4]. Hepatic fibrosis can develop in patients with any type of chronic liver disease (CLD), including alcoholic liver disease (ALD), hepatitis C, hepatitis B, nonalcoholic fatty liver disease (NAFLD) and autoimmune hepatitis. The major clinical consequences of cirrhosis are liver failure and hepatocellular carcinoma (HCC), both of which increase the risk of death. Alcoholic liver disease (ALD) is a leading cause of liver disease and liver-related deaths globally, particularly in developed nations. The current gold standard diagnostic method, liver biopsy is invasive and has many limitations including sampling error and high inter-observer variability even for the diagnosis of advanced stages of liver fibrosis. None of several technologies investigated as alternatives offers the desired sensitivity and specificity for the early detection and staging of fibrosis. Thus, an innovative, safe MRI contrast agent is required to overcome several major limitations including low sensitivity and specificity for early stage fibrosis and detection in heterogeneous tissue. Our team has pioneered a new class of gadolinium based contrast agents which utilize a protein scaffold to bind Gd3+ atoms. We have shown that ProCA32.collagen, a collagen 1 targeted protein MRI contrast agent, exhibits strong affinity to collagen I, whose expression level and spatial pattern depend on the stage of fibrosis. ProCA32.collagen possesses high relaxivities per particle (r1 and r2) at both 1.4 and 7.0 T, which enables the robust detection of early-stage alcohol-induced liver fibrosis and nonalcoholic steatohepatitis in animal models via dual contrast modes. Our preliminary studies on toxicity profiles and in vivo stability of our compound in animals using non-GLP grade materials have provided confidence that the compound will exhibit good sensitivity and specificity for early detection of liver fibrosis in human clinical trials. In Phase I of this proposal, we seek timely support for conducting IND enabled CMC studies for generating GLP/cGMP grade ProCA32.collagen. In Phase II, we will conduct toxicity and safety profile studies using GLP/cGMP grade products to complete IND required studies for clinical trials.