DEVELOPMENT OF UNIFIED MODELS OF CCK RECEPTOR SUBTYPE

CCK受体亚型统一模型的开发

• 批准号: 2392175
• 负责人: GREGORY V NIKIFOROVICH
• 金额: $ 15.61万
• 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/2392175
• 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-A 肽药效团，之前通过与亲本比较获得 肽及其肽类似物，以及新开发的 3D 模型 CCK-B和CCK-A受体本身，设计非肽激动剂和 拮抗剂可能与 CCK-B 和 CCK-A 的相同位点结合 受体。 我们在这个项目中要遵循的策略是 基于合理的设计。 根据之前CCK的结果 项目中，我们将在以下方向扩展未来的研究： (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。 我们将通过两种收敛的方法来设计这些化合物 路径：（i）通过刚性化所提出的受体结合构象异构体（因此 进一步完善模型），并且，（ii）通过修改，根据 模型，已知非肽CCK-B拮抗剂以获得非肽激动剂 （目前尚未发现CCK-B非肽激动剂）。 这些研究 将开辟先导化合物的路线，可将其开发用于 药品水平。 （2）。 我们将进一步完善模型 CCK-A受体结合构象异构体通过分子建模、合成和 构象受限的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 和对手。 我们还将研究构象关系 肽（激动剂和拮抗剂）和非肽拮抗剂之间 （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 根据我们可用的独特计算机程序提出建议 实验室。 这些程序包括确定跨膜末端 蛋白质序列中的螺旋片段，通过以下方式将这些片段包装在一起 特殊分子识别算法，恢复螺旋间 通过残基-残基接触矩阵技术形成环。 (4) 我们将使用 获得了 CCK-B 和 CCK-A 肽药效团以及 CCK-B 的模型 和CCK-A受体用于肽和非肽配体的国家设计 对于两种受体。 我们将合成设计的化合物并将 将它们提交用于体外以及细胞测试系统中的生物学研究 具有克隆和表达的突变体和嵌合 CCK 受体。