DEVELOPMENT OF UNIFIED MODELS OF CCK RECEPTOR SUBTYPE

CCK受体亚型统一模型的开发

• 批准号: 2685005
• 负责人: GREGORY V NIKIFOROVICH
• 金额: $ 16.24万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2685005
• 关键词: chemical models; cholecystokinin; drug design /synthesis /production; guinea pigs; hormone receptor; laboratory rabbit; laboratory rat; ligands; model design /development; neuropeptide receptor; neurotransmitter agonist; neurotransmitter antagonist; neurotransmitter receptor; protein structure function; receptor binding; site directed mutagenesis; synthetic protein

The general goal of the present study is to use 3D models of the CCK-B and CCK-A peptide pharmacophores, previously obtained by comparison of a parent peptide and its peptide analogs, as well as newly developed 3D models for CCK-B and CCK-A receptors themselves, to design non-peptide agonists and antagonists presumably binding to the same sites of the CCK-B and CCK-A receptors. The strategy we are going to follow in this project will be based on a rational design. Based on the results of the previous CCK project, we will extend the future studies in the following directions: (1) We will use the developed model for the CCK-B receptor-bound conformation for rational design of non-peptide agonists and antagonists with CCK-B selectivity. We will design these compounds by two convergent paths: (i) by rigidifying the proposed receptor-bound conformer (thus refining the model further), and, (ii) by modifying, in accordance with the model, known non-peptide CCK-B antagonists to obtain non-peptide agonists (there is no CCK-B non-peptide agonists discovered so far). These studies will open the route to lead compounds, which could be developed to the level of pharmaceuticals. (2). We will refine further the model of the CCK-A receptor-bound conformer by molecular modeling, synthesis and biological testing of conformationally constrained cyclic compounds. Then, this model will be used for design of CCK-A selective non-peptide agonists and antagonists. We will study also the conformational relationships between peptide (agonists and antagonists) and non-peptide antagonists (again, non-peptide agonists are not known) with CCK-A selectivity. (3) We will develop further the initial 3D models for CCK-B and CCK-A receptors suggested on the basis of the unique computer procedures available in our lab. These procedures include determining tof the ends for transmembrane helical segments in protein sequences, packing these segments together by special molecular recognition algorithm, and restoring the inter helical loops by residue-residue contact matrix technique. (4) We will use the obtained models for CCK-B and CCK-A peptide pharmacophores and for CCK-B and CCK-A receptors for national design of peptide and non-peptide ligands for both receptors. We will synthesize the designed compounds and will submit them for biological studies in vitro as well as in cell test systems with cloned and expressed mutants and chimeric CCK receptors.

本研究的总体目标是使用CCK-B的3D模型和 CCK-一种多肽药效团，以前通过与亲本比较而获得 多肽及其类似物，以及新开发的三维模型 CCK-B和CCK-A受体本身，设计非肽激动剂和 推测拮抗剂与CCK-B和CCK-A的相同位点结合 感受器。我们在这个项目中要遵循的策略是 基于合理的设计。根据前一次CCK的结果 随着项目的实施，我们将从以下几个方面拓展未来的研究： (1)我们将使用开发的CCK-B受体结合模型 非肽类激动剂和拮抗剂合理设计的构象 具有CCK-B选择性。我们将通过两个收敛的方法来设计这些化合物 途径：(I)通过使所建议的受体结合构象刚性(因此 进一步完善模型)，以及，(Ii)根据 模型，已知的非肽CCK-B拮抗剂获得非肽激动剂 (到目前为止还没有发现CCK-B非肽激动剂)。这些研究 将打开先导化合物的道路，这可能会发展成 药品的水平。(2)。我们将进一步完善这一模型 CCK-A受体结合构象的分子模拟、合成和鉴定 构象受限环状化合物的生物测试。然后, 该模型将用于CCK-A选择性非肽激动剂的设计 和敌手。我们还将研究构象关系 多肽(激动剂和拮抗剂)和非多肽拮抗剂之间 (同样，非肽激动剂未知)具有CCK-A选择性。(3) 我们将进一步开发CCK-B和CCK-A受体的初始3D模型 建议的依据是我们的 实验室。这些程序包括确定跨膜末端的tOf。 蛋白质序列中的螺旋片段，通过以下方式将这些片段打包在一起 特殊的分子识别算法，并恢复螺旋间 利用残基-残基接触矩阵技术进行环路。(4)我们将使用 CCK-B和CCK-A多肽药效团及CCK-B模型的建立 和CCK-A受体用于肽和非肽配体的国产化设计 对两种受体都有影响。我们将合成设计的化合物，并将 在体外和细胞测试系统中提交它们进行生物学研究 利用克隆和表达的突变体和嵌合的CCK受体。