Naturally Occurring Agent for Alcohol Liver Diseases

酒精性肝病的天然药物

• 批准号: 6703343
• 负责人: DAVID Yue-Wei LEE
• 金额: $ 22.27万
• 依托单位: NATURAL PHARMACIA INTERNATIONAL, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-18 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6703343
• 关键词: alcoholism /alcohol abuse; biological signal transduction; biotherapeutic agent; cytoprotection; drug design /synthesis /production; genetically modified animals; high performance liquid chromatography; laboratory mouse; laboratory rat; liver cells; liver disorder chemotherapy; liver regeneration; mass spectrometry; medicinal plants; pharmacokinetics; plant extracts

DESCRIPTION (provided by applicant): Chronic heavy ethanol consumption has major adverse effects on the mammalian liver, namely the induction of hepatocellular damage and impairment of regeneration following injury. For a medication development project focusing on alcohol liver diseases, it is essential to understand, at the molecular level, potential alterations in cellular signal transduction pathways that adversely affect the hepatocyte proliferative response, as well as those survival mechanisms critical for liver regeneration. The objective of this STTR Phase I and II fast track application is to develop a naturally occurring agent, namely Silybin (NPI-MK-001), as a hepatoprotective agent for alcohol liver problems. Based on our preliminary studies, NPI-MK-001 is the bioactive component isolated from the seed of Milk thistle (Silybum marianum), the best-studied medicinal plant for the treatment of liver disease. We plan to study the cytoprotective effect of NPIMK-001 on liver injury produced by chronic ethanol consumption in rat hepatocyte culture. We will also evaluate the therapeutic potential of NPI-MK-001 on the hepatic repair process in vivo by analyzing effects on specific growth factor signal transduction cascades associated with hepatocyte DNA synthesis. Our specific aims are: Aim #1 - Determine the in vitro cytoprotective effects of NPI-MK-001 on hepatocyte injury caused by chronic ethanol feeding+ hypoxia; Aim #2 - Explore the in vivo action of NPI-MK-001 on the IRS-1 mediated growth factor signal transduction pathway associated with liver repair in chronic ethanol-fed rats and IRS-1 transgenic mice. In Phase I, we will focus on the standardization of NPI-MK-001, a prerequisite for in vitro and in vivo evaluation. We propose to (1) conduct field inspection of the collection, processing, extraction, and contamination control (insecticides, herbicides, and chemical fertilizers) of the seed of Milk Thistle; (2) optimize the analytical HPLC conditions for qualitative and quantitative analysis of NPI-MK-001; (3) isolate and purify a sufficient quantity of NPI-MK-001 and its diasteric isomers (Silybin A and B) for chemical markers, and for in vitro and in vivo evaluation; and (4) develop a cost-effective separation procedure for large scale purification of NPI-MK-001. In Phase II, we propose to: (1) evaluate relative potency of NPI-MK-001and its diasteric isomers (Silybin A and Silybin B) by in vitro and in vivo models; (2) investigate the mechanism of action of NPI-MK-001 as specified in Aims 1 and 2; (3) Synthesize radio-labeled NPIMK-001 and assess the bioavailability and pharmcokinetics; (4) standardize NPI-MK-001 and corresponding placebo according to FDA guidelines; (5) obtain data on mutagenesis and acute and chronic toxicity; (6) conduct stability testing of the standardized NPI-MK-001; and (7) establish protocols for the capsulation of standardized NPI-MK-001 and placebo under GMP conditions. Natural Pharmacia International, Inc, an established natural products laboratory, has assembled an excellent consortial team to assure that the quality and purity of NPI-MK-001would meet the stringent FDA requirements. The key to the success of this STTR program is not merely the exceptional experience of the team members in carry out the proposed studies in Phases I and II, but also the outstanding business development plan which supports the further development of NPI-MK-001 including filing of a DMF and an IND in Phase III.

描述（由申请人提供）：长期大量饮用乙醇对哺乳动物肝脏有重大不良影响，即诱导肝细胞损伤和损伤后再生障碍。对于酒精性肝病的药物开发项目，必须在分子水平上了解细胞信号转导途径的潜在改变，这些改变会对肝细胞增殖反应产生不利影响，以及对肝再生至关重要的生存机制。STTR I期和II期快速通道申请的目的是开发一种天然存在的药物，即水飞蓟宾（NPI-MK-001），作为酒精性肝脏问题的肝保护剂。根据我们的初步研究，NPI-MK-001是从水飞蓟（Silybum marianum）种子中分离出来的生物活性成分，水飞蓟是研究得最好的治疗肝脏疾病的药用植物。我们计划在大鼠肝细胞培养中研究NPIMK-001对慢性乙醇消耗肝损伤的细胞保护作用。我们还将通过分析与肝细胞DNA合成相关的特定生长因子信号转导级联反应的影响，评估NPI-MK-001在体内肝脏修复过程中的治疗潜力。我们的具体目标是：目的1 -确定NPI-MK-001对慢性乙醇喂养+缺氧引起的肝细胞损伤的体外细胞保护作用；目的2：探讨NPI-MK-001在慢性乙醇喂养大鼠和IRS-1转基因小鼠中对IRS-1介导的与肝脏修复相关的生长因子信号转导通路的体内作用。在I期，我们将重点关注NPI-MK-001的标准化，这是体外和体内评估的先决条件。我们建议(1)