Hemoglobin Modifiers for Sickle Cell Disease Therapy

用于镰状细胞病治疗的血红蛋白调节剂

• 批准号: 9053281
• 负责人: Martin K Safo
• 金额: $ 39.59万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-11 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9053281
• 关键词: Acute; Adverse effects; Adverse event; Affect; Affinity; African American; Aldehydes; Antisickling Agents; Ants; Binding; Biological; Biological Assay; Biological Availability; Biological Markers; Biological Models; Blood Transfusion; Blood flow; Cell Adhesion; Cells; Cellular Morphology; Cytosol; Data; Dehydration; Disease; Disease Pathway; Disease Progression; Dose; Endothelium; Erythrocytes; Event; Exhibits; Functional disorder; Generations; Goals; Health; Hemoglobin; Hemolysis; Hepatic; Human; Hydration status; In Vitro; Inflammation; Inherited; Kinetics; Lead; Mediating; Metabolism; Methodology; Methods; Minority; Modeling; Modification; Monitor; Mus; Oral; Oxidative Stress; Oxygen; Pain; Parents; Pathway interactions; Patients; Personal Satisfaction; Pharmaceutical Preparations; Phase; Polymers; Population; Process; Prodrugs; Property; Rattus; Regimen; Research Personnel; Resources; Series; Serum Albumin; Sickle Cell; Sickle Cell Anemia; Sickle Hemoglobin; Solubility; Structure; Surface; Survival Rate; System; TNFRSF5 gene; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Tissues; Toxicology; Transgenic Mice; Variant; X-Ray Crystallography; adduct; aldehyde dehydrogenases; atomic interactions; base; cytotoxicity; design; drug candidate; experience; health care disparity; health disparity; hydroxyurea; improved; in vivo; insight; insoluble fiber; mouse model; next generation; novel; novel therapeutics; polymerization; preclinical efficacy; premature; prevent; research clinical testing; sickling; vanillin

DESCRIPTION (provided by applicant): Our goal is to design and establish novel therapeutic agents for sickle cell disease (SCD), namely drugs that inhibit the initial sickle hemoglobin (HbS) polymerization and the subsequent pathophysiology. When deoxygenated, HbS polymerizes into long, rigid, and insoluble fibers causing red blood cells (RBCs) to sickle, a process worsened by the unusual low affinity of HbS for oxygen, resulting in premature release of oxygen. Based on several evidence--our preliminary data, studies by others, and the results of a recently completed phase I/II clinical testing of our lead compound, 5-HMF (an allosteric effector of Hb, AEH)-we hypothesize that AEHs, not only prevent HbS polymerization, but also mitigates several secondary sickling related pathological events that include inflammation, oxidative stress/damage, RBC hemolysis, and pain. We also have preliminary evidence that our next generation AEHs (INN- and TD-series) exhibit enhanced potency and improved in-vitro duration of action. These AEHs act via a novel mechanism of action, i.e., destabilize HbS polymer contacts, in addition to increasing Hb affinity for oxygen; providing positive synergistic effects. We propose to test our hypothesis by further investigating candidate drugs from the INN- and TD-series, as well as derivatives of 5-HMF and INN-312 for their pharmacologic properties, focusing on the secondary SCD pathways, as well as the underlying HbS polymerization problem using our model systems. The specific aims are: 1. Design and synthesis of novel allosteric effectors of hemoglobin (AEHs). We will modify our parent compounds and synthesize derivatives with enhanced efficacy and prolonged half-lives. We will also synthesize prod rugs to protect the active aldehyde functional moiety from aldehyde dehydrogenase (ALDH)- mediated metabolism as necessary. 2. Investigate in-vitro functional, ant sickling, and cytotoxicity activities of novel AEHs. We will investigate the AEHs for their in-vitro ant sickling/functional activities (RBC sickling tests, P50 analyses, HbS solubility, and Hb adduct formation), and carefully monitor for adverse effects. 3. Determine in-vivo/in-vitro PK/PD properties, binding and metabolism and evaluate preclinical efficacy of AEHs in SCD Berkeley mice. We will show that serum albumin binding and/or metabolism by ALDH in RBC or hepatic cytosol are not likely to adversely affect in-vivo pharmacologic activity. We will demonstrate using a Berkeley mouse model of SCD transgenic mice that AEHs show potent pharmacologic effects, increase short- and long-term survival rates of mice. We will also study their potential beneficial effects, e.g. amelioration of hemolysis, inflammation, endothelial damage, and overall reversal of the SCD pathophysiology observed in this model. 4. Determine the atomic interactions between AEHs and Hb. X-ray crystallography will be used to validate our hypothesis that AEH potency is directly dependent upon their abilities to bind Hb with their pyridyl substituents toward the surface of the Hb molecule. The structures would provide valuable insight to help guide rational modifications for better pharmacologic properties.

描述（由申请人提供）：我们的目标是设计和建立镰状细胞病（SCD）的新型治疗药物，即抑制初始镰状血红蛋白（HbS）的药物 聚合和随后的病理生理学。脱氧时，HbS 聚合成又长又硬的不溶性纤维，导致红细胞 (RBC) 镰状化，而 HbS 对氧的亲和力异常低，导致这一过程恶化，导致氧气过早释放。基于多项证据（我们的初步数据、其他人的研究以及最近完成的先导化合物 5-HMF（Hb 的变构效应物，AEH）的 I/II 期临床测试结果），我们假设 AEH 不仅可以防止 HbS 聚合，而且可以减轻一些继发性镰状细胞相关的病理事件，包括炎症、氧化应激/损伤、红细胞溶血和疼痛。我们还有初步证据表明，我们的下一代 AEH（INN 系列和 TD 系列）表现出增强的效力和更长的体外作用持续时间。这些 AEH 通过一种新颖的作用机制发挥作用，即除了增加 Hb 对氧的亲和力外，还会破坏 HbS 聚合物接触的稳定性；提供积极的协同效应。 我们建议通过进一步研究 INN 系列和 TD 系列候选药物以及 5-HMF 和 INN-312 衍生物的药理学特性来检验我们的假设，重点关注次要 SCD 途径以及使用我们的模型系统的潜在 HbS 聚合问题。具体目标是： 1. 新型血红蛋白变构效应物（AEH）的设计和合成。我们将修改我们的母体化合物并合成具有增强功效和延长半衰期的衍生物。我们还将合成前药，以根据需要保护活性醛功能部分免受乙醛脱氢酶（ALDH）介导的代谢影响。 2. 研究新型AEH 的体外功能、蚂蚁镰刀和细胞毒性活性。我们将研究 AEH 的体外蚂蚁镰状/功能活动（红细胞镰状化测试、P50 分析、HbS 溶解度和 Hb 加合物形成），并仔细监测不良影响。 3. 确定体内/体外 PK/PD 特性、结合和代谢，并评估 AEH 在 SCD Berkeley 小鼠中的临床前功效。我们将证明，血清白蛋白结合和/或 RBC 或肝细胞质中 ALDH 的代谢不太可能对体内药理活性产生不利影响。我们将使用 SCD 转基因小鼠的伯克利小鼠模型证明 AEH 显示出有效的药理作用，可提高小鼠的短期和长期存活率。我们还将研究它们潜在的有益影响，例如该模型中观察到的溶血、炎症、内皮损伤的改善以及 SCD 病理生理学的整体逆转。 4. 确定 AEH 和 Hb 之间的原子相互作用。 X 射线晶体学将用于验证我们的假设，即 AEH 效力直接取决于它们通过吡啶基取代基将 Hb 结合到 Hb 分子表面的能力。这些结构将提供有价值的见解，以帮助指导合理的修饰以获得更好的药理特性。