METALLOPEPTIDES AS THERAPEUTIC AGENTS

金属肽作为治疗剂

• 批准号: 7739431
• 负责人: JAMES A COWAN
• 金额: $ 18.53万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7739431
• 关键词: Address; Amino Acid Sequence; Amino Acids; Angiotensins; Animal Model; Animals; Antihypertensive Agents; Binding; Biological Assay; Cardiovascular system; Clinical Trials; Dose; Drug Design; Drug effect disorder; Drug resistance; Effectiveness; Endothelin-converting enzyme 1; Environment; Enzymes; Evaluation; Experimental Designs; Funding; Goals; Human; Inbred SHR Rats; Investigation; Kinetics; Lead; Mediating; Medical; Metals; Methods; Modeling; Modification; Neprilysin; Oral Administration; Outcome; Peptides; Pharmacologic Substance; Principal Investigator; Proteins; Rattus; Reactive Oxygen Species; Relative (related person); Science; Side; Solutions; Testing; Therapeutic; Therapeutic Agents; Thermodynamics; Toxic effect; Work; candidate identification; catalyst; cofactor; design; dosage; drug discovery; endothelin-converting enzyme; hypertension treatment; improved; in vivo; novel; oxidation; oxidative damage; programs; protein aminoacid sequence; public health relevance; therapeutic target; uptake

DESCRIPTION (provided by applicant): Drug discovery remains a top priority in medical science. The phenomenon of drug resistance has heightened the need for both new classes of pharmaceutical as well as novel modes of action. In recent years we have worked to develop a distinct approach to drug design that involves both recognition and subsequent irreversible inactivation of therapeutic targets. The basic drug design strategy incorporates a protein recognition domain and a metal binding domain, where the latter mediates irreversible inactivation of the therapeutic target. Inactivation is both catalytic and multiturnover, while the incorporation of both binding and catalytic centers provides a double-filter mechanism for improved target selectivity and lower dosing. The specific aims for the proposed funding period will focus on improving on lead metallopeptides of potential therapeutic value against cardiovascular targets, the demonstration of efficacy in animal models, and elaborating the mechanism of action of such metallodrugs against protein targets. Our overall goals can be summarized as follows. (1) Design and evaluate triple-action metallodrugs that target three key cardiovascular enzymes (ACE, ECE-1 and NEP). (2) Optimization of in vivo stability and efficacy through tuning of peptide sequence and amino acid configuration. (3) Evaluate the mechanism of catalytic inactivation of protein targets. Optimization of the intrinsic reactivity of the catalytic metallodrugs toward metal-mediated degradation of target molecules will require an advanced understanding of the mechanism of catalytic cleavage (to be achieved by use of a variety of kinetic and mass spectrometric methods). These studies will combine fluorogenic activity assays of metallopeptide inactivation of ACE, ECE-1 and NEP, as well as animal studies of a spontaneously hypertensive rat model. Mass spectrometric analysis of modified proteins and proteolytic digests will reveal details of side-chain modification that underlie catalytic inactivation. The impact of substituting L- for D-amino acids on metallodrug activity, target recognition, and in vivo stability will be investigated. The animal studies will also serve to validate initial observations that delivery of the metal-free peptide alone is sufficient for activity, with recruitment of metal cofactor from the cellular environment following uptake. PUBLIC HEALTH RELEVANCE: Drug discovery remains a top priority in medical science. The phenomenon of drug resistance has heightened the need for both new classes of pharmaceutical as well as novel modes of action. In recent years we have worked to develop a distinct approach to drug design that involves both recognition and subsequent irreversible inactivation of therapeutic targets. This concept allows for improved target selectivity and lower dosage requirements and will be further developed against cardiovascular protein targets in both solution studies and animal models.

描述（由申请人提供）：药物发现仍然是医学科学的首要任务。耐药性现象已经提高了对新的药物类别以及新的作用模式的需求。近年来，我们一直致力于开发一种独特的药物设计方法，包括识别和随后的治疗靶点的不可逆失活。基本的药物设计策略包括蛋白质识别结构域和金属结合结构域，其中后者介导治疗靶点的不可逆失活。失活是催化和多周转，而结合和催化中心的结合提供了一个双过滤器机制，提高目标选择性和较低的剂量。拟议资助期的具体目标将集中在改善对心血管靶点具有潜在治疗价值的铅金属肽，在动物模型中证明疗效，并阐述此类金属药物对蛋白质靶点的作用机制。我们的总体目标可以概括如下。(1)设计和评估针对三种关键心血管酶（ACE，ECE-1和NEP）的三重作用金属药物。(2)通过调整肽序列和氨基酸构型优化体内稳定性和功效。(3)评价蛋白质靶点的催化失活机制。催化金属药物对金属介导的靶分子降解的固有反应性的优化将需要对催化裂解机制的深入理解（通过使用各种动力学和质谱方法来实现）。这些研究将结合联合收割机的荧光活性测定的金属肽失活的ACE，ECE-1和NEP，以及动物研究的自发性高血压大鼠模型。质谱分析修饰的蛋白质和蛋白水解酶将揭示细节的侧链修饰的催化失活的基础。将研究用L-取代D-氨基酸对金属药物活性、靶向识别和体内稳定性的影响。动物研究还将用于验证最初的观察结果，即单独递送无金属肽足以产生活性，在摄取后从细胞环境中募集金属辅因子。公共卫生相关性：药物发现仍然是医学科学的首要任务。耐药性现象已经提高了对新的药物类别以及新的作用模式的需求。近年来，我们一直致力于开发一种独特的药物设计方法，包括识别和随后的治疗靶点的不可逆失活。这一概念允许改进的靶点选择性和较低的剂量要求，并将在溶液研究和动物模型中针对心血管蛋白靶点进一步开发。