Systemic AA Amyloidosis Inhibitors

全身性 AA 淀粉样变性抑制剂

• 批准号: 7624714
• 负责人: ALAN D. SNOW
• 金额: $ 51.94万
• 依托单位: PROTEOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7624714
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Adrenal Glands; Amyloid; Amyloid Fibrils; Amyloid Protein AA; Amyloid deposition; Amyloidosis; Animal Model; Animals; Ankylosing spondylitis; Apolipoprotein E; Apoptosis; Appearance; Aromatic Compounds; Back; Binding; Biochemical; Biological Assay; Biological Availability; Body Weight; Brain; Cell Culture Techniques; Cell Survival; Cells; Cessation of life; Characteristics; Chemicals; Chronic; Clinical; Clinical Trials; Congo Red; Crystallography; Data; Deposition; Detection; Development; Disease; Dose; Drug Kinetics; Drug or chemical Tissue Distribution; Effectiveness; Electrons; Endothelial Cells; Ensure; Familial Mediterranean Fever; Fluorescence; Fluorometry; Functional disorder; Gastrointestinal tract structure; Generations; Heart; Heart failure; Heparitin Sulfate; Histologic; Hodgkin Disease; Hour; Human; Hydroxyl Radical; Image Analysis; Imagery; Immunohistochemistry; In Vitro; Inflammatory; Injection of therapeutic agent; Investigation; Kidney; Lead; Leprosy; Libraries; Light; Liver; Lung; Mass Spectrum Analysis; Measurement; Methods; Microscopic; Modeling; Monitor; Mus; Necrosis; Onset of illness; Oral; Oral Administration; Organ; Osteomyelitis; Pancreas; Pathologist; Patients; Pharmaceutical Preparations; Phase; Phase II Clinical Trials; Plasma; Positioning Attribute; Prevention; Production; Radiolabeled; Renal Cell Carcinoma; Research Design; Research Personnel; Rheumatoid Arthritis; Rodent; Route; Safety; Screening procedure; Serum Amyloid P-Component; Silver Nitrate; Skin; Small Business Innovation Research Grant; Spectrum Analysis; Spleen; Sprague-Dawley Rats; Staining method; Stains; Stereoisomer; Testing; Therapeutic; Thioflavin T; Time; Tissues; Toxic effect; Toxicity Tests; Tuberculosis; Western Blotting; Work; X-Ray Crystallography; amyloid formation; analog; azocasein; base; blind; calcein AM; cell type; commercialization; dalton; design; dosage; effective therapy; experience; fibrillogenesis; human disease; hydroxyl group; in vitro Assay; in vitro testing; in vivo; inhibitor/antagonist; macrophage; mouse model; novel; novel therapeutics; pharmacokinetic characteristic; polysulfated glycosaminoglycan; pre-clinical; public health relevance; radiotracer; research clinical testing; response; small molecule; small molecule libraries; stereochemistry

DESCRIPTION (provided by applicant): Systemic AA Amyloidosis is characterized by the accumulation of insoluble fibril deposits containing the AA amyloid protein in different organs throughout the body including heart, kidney, liver, spleen, lungs, skin and gastrointestinal tract, whereby such amyloid fibril accumulation leads to pronounced organ dysfunction. Systemic AA amyloidosis is associated mostly with chronic inflammatory disorders and includes patients with rheumatoid arthritis, osteomyelitis, ankylosing spondylitis, inflammatory bowl disease, tuberculosis, leprosy, Hodgkin's disease, renal cell carcinoma, and Familial Mediterranean Fever. The consequences of fibrillar AA amyloid deposition in systemic organs are usually fatal to patients, with most patients dying within 3-7 years from disease onset due to kidney or heart failure. Currently there is no effective cure treatment for AA amyloidosis and a new therapeutic is desperately needed for the ~100,000 cases currently in the USA. In our completed Phase I SBIR studies we designed, synthesized and tested our own unique library of small molecule compounds (representing new chemical entities) for inhibition of AA amyloidosis both in vitro and in vivo. Using a variety of in vitro screening methods to identify specific potent inhibitors of AA amyloidosis, we discovered that a number of our small molecule compounds markedly disrupted/inhibited SAA/AA amyloid fibrils in vitro. Using a relevant animal mouse model of experimental AA amyloidosis we now have identified 2 lead compounds (i.e. PTI-19 and 51) that remarkably inhibit AA amyloid deposition in tissues (i.e. spleen, liver and kidney) by >50-60% following oral administration. PTI-19 and 51 represent new chemical entities and our established synthetic routes for the repeated production of each of these compounds demonstrate a pure product (>98% purity) that consists of only 1 compound and 1 stereoisomer [(as determined by detailed analysis including HPLC/mass spec, 1H-NMR, 13C-NMR, DEPT, and x-ray crystallography (for PTI-19)]. These studies suggest that these small molecule lead compounds show great promise for the development of new effective treatments for systemic AA amyloidosis and warrant further investigation as described in this Phase II SBIR project. In proposed Phase II studies we will work with two world class organic chemists and optimize, test and further develop these small molecule compounds (including our 2 lead compounds) that we anticipate will inhibit/retard and cause a clearance of aggregated/fibrillar AA amyloid deposits in humans. In vitro testing against human AA amyloid fibrils will include newly designed analogs optimized for oral bioavailability and enhanced PK characteristics, while maintaining non-toxicity and potent efficacy. Confirmation of each final product synthesized and the possible existence of stereoisomers will be determined using 1H-NMR, 13C-NMR and DEPT. X-ray crystallography will also be used for promising lead compounds in which absolute determination of stereochemistry cannot be fully verified by NMR. Oral administration and time-dependent efficacy for prevention, reduction and clearance of AA amyloid deposits in systemic organs will be assessed using two experimental AA amyloid mouse models. PK and toxicity studies will help us optimize a novel small molecule compound (and its back-up) that will be developed for human clinical trials and commercialization, and that has promise to serve as a new treatment for systemic AA amyloidosis and related diseases. PUBLIC HEALTH RELEVANCE: Systemic AA Amyloidosis is usually a fatal disease characterized by fibrillar amyloid deposition throughout the body that leads to organ dysfunction and eventually death in 3-7 years (due to kidney and/or heart failure). We have designed and discovered new small molecule compounds that effectively reduce/inhibit amyloid deposition in organs in a relevant animal model that mimics the human disease. In this project we intend to develop a new drug (and back-up) for effective treatment of ~100,000 cases of systemic amyloidosis in the USA, for which today there is no real treatment whatsoever.

描述（申请人提供）：系统性AA淀粉样变性的特征是含有AA淀粉样蛋白的不溶性原纤维沉积物在全身不同器官（包括心脏、肾、肝脏、脾、肺、皮肤和胃肠道）中积累，由此这种淀粉样原纤维积累导致明显的器官功能障碍。系统性AA淀粉样变性主要与慢性炎症性疾病相关，包括类风湿性关节炎、骨髓炎、强直性脊柱炎、炎性碗病、结核病、麻风病、霍奇金病、肾细胞癌和家族性地中海热患者。全身器官中纤维状 AA 淀粉样蛋白沉积的后果通常对患者来说是致命的，大多数患者在发病后 3-7 年内因肾衰竭或心力衰竭而死亡。目前，AA 型淀粉样变性尚无有效的治愈方法，美国目前约有 100,000 例病例，迫切需要一种新的治疗方法。在我们完成的 I 期 SBIR 研究中，我们设计、合成并测试了我们自己独特的小分子化合物库（代表新的化学实体），用于在体外和体内抑制 AA 淀粉样变性。使用多种体外筛选方法来鉴定 AA 淀粉样变性的特异性强效抑制剂，我们发现我们的许多小分子化合物在体外显着破坏/抑制 SAA/AA 淀粉样原纤维。使用实验性 AA 淀粉样变性的相关动物小鼠模型，我们现在已经鉴定出 2 种先导化合物（即 PTI-19 和 51），口服给药后可显着抑制组织（即脾、肝和肾）中 AA 淀粉样蛋白沉积，抑制率 > 50-60%。 PTI-19 和 51 代表新的化学实体，我们为重复生产这些化合物中的每一种而建立的合成路线证明了仅由 1 种化合物和 1 种立体异构体组成的纯产品（>98% 纯度）[（通过详细分析确定，包括 HPLC/质谱、1H-NMR、13C-NMR、DEPT 和 X 射线晶体学（对于 PTI-19）]。这些研究表明，这些小化合物 分子先导化合物在开发系统性 AA 淀粉样变性的新有效治疗方法方面显示出巨大的前景，并值得进一步研究，如本 II 期 SBIR 项目中所述。在拟议的 II 期研究中，我们将与两位世界一流的有机化学家合作，优化、测试和进一步开发这些小分子化合物（包括我们的 2 种先导化合物），我们预计这些化合物将抑制/延迟并导致聚集/纤维状 AA 淀粉样蛋白的清除 沉积在人体中。针对人类 AA 淀粉样原纤维的体外测试将包括针对口服生物利用度和增强的 PK 特性进行优化的新设计类似物，同时保持无毒性和有效功效。将使用 1H-NMR、13C-NMR 和 DEPT 来确认所合成的每种最终产物以及可能存在的立体异构体。 X射线晶体学也将用于有希望的 先导化合物的立体化学绝对测定不能通过 NMR 完全验证。将使用两个实验性 AA 淀粉样蛋白小鼠模型来评估口服给药和预防、减少和清除全身器官中 AA 淀粉样蛋白沉积的时间依赖性功效。 PK 和毒性研究将帮助我们优化一种新型小分子化合物（及其备份），该化合物将用于人体临床试验和商业化， 这有望成为系统性 AA 淀粉样变性及相关疾病的新治疗方法。公共卫生相关性：系统性 AA 淀粉样变性通常是一种致命性疾病，其特征是全身纤维状淀粉样蛋白沉积，导致器官功能障碍，并最终在 3-7 年内死亡（由于肾和/或心力衰竭）。我们设计并发现了新的小分子化合物，可以有效减少/抑制淀粉样蛋白 在模拟人类疾病的相关动物模型的器官中沉积。在这个项目中，我们打算开发一种新药（和备用药物），用于有效治疗美国约 100,000 例系统性淀粉样变性病例，而目前对此还没有任何真正的治疗方法。