METALLOPEPTIDES AS THERAPEUTIC AGENTS

描述（由申请人提供）：药物发现仍然是医学科学的重中之重。耐药现象使人们更加需要新的药物种类和新的作用方式。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一个蛋白质识别结构域和一个金属结合结构域，后者介导治疗靶点的不可逆失活。失活是催化和多重翻转，而结合和催化中心的结合提供了双重过滤机制，以提高目标选择性和降低剂量。该资助期的具体目标将集中于改善对心血管靶点具有潜在治疗价值的铅金属肽，在动物模型中证明其疗效，并阐明这些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)设计并评价针对3种关键心血管酶（ACE、ECE-1和NEP）的三作用金属药物。(2)通过调整多肽序列和氨基酸构型来优化体内稳定性和有效性。(3)评价蛋白靶点催化失活的机制。优化催化金属药物对金属介导的目标分子降解的固有反应性将需要对催化裂解机制的深入了解（通过使用各种动力学和质谱方法来实现）。这些研究将结合ACE、ECE-1和NEP金属肽失活的荧光活性分析，以及自发性高血压大鼠模型的动物研究。质谱分析修饰蛋白和蛋白水解酶解将揭示催化失活背后侧链修饰的细节。将研究L-取代d -氨基酸对金属药物活性、靶标识别和体内稳定性的影响。动物研究也将用于验证最初的观察结果，即仅递送无金属肽就足以产生活性，在摄取后从细胞环境中招募金属辅助因子。公共卫生相关性：药物发现仍然是医学科学的重中之重。耐药现象使人们更加需要新的药物种类和新的作用方式。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。这一概念允许提高靶标选择性和降低剂量要求，并将在溶液研究和动物模型中进一步发展针对心血管蛋白靶标。

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.