Ligand with Two Modes of Interaction with the Dopamine D(2) Receptor-An Induced-Fit Mechanism of Insurmountable Antagonism.
Ligand with Two Modes of Interaction with the Dopamine D(2) Receptor-An Induced-Fit Mechanism of Insurmountable Antagonism.
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DOI：
10.1021/acschemneuro.0c00477
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发表时间：
2020-10-07
影响因子：
5
通讯作者：
Sahlholm K
中科院分区：
文献类型：
--
作者：
Ågren R;Zeberg H;Stępniewski TM;Free RB;Reilly SW;Luedtke RR;Århem P;Ciruela F;Sibley DR;Mach RH;Selent J;Nilsson J;Sahlholm K
对配体结合机制的深入理解对于合理设计靶向多巴胺D2受体（D2 R）的治疗药物至关重要。在这里，我们使用G蛋白偶联内向整流钾（GIRK）通道激活非洲爪蟾卵母细胞来测量一系列阿立哌唑类似物的D2 R拮抗作用的动力学，以及洗脱后多巴胺（DA）响应的恢复。阿立哌唑类似物包含正构和二级药效团，并且不同之处在于连接这两个药效团的饱和碳接头的长度。在1或100 μM DA存在下，含有3-和5-碳连接体的两种化合物允许从拮抗作用中恢复相似程度（分别为对照的> 25%和>90%），而在洗脱4-碳接头化合物SV-III-130后，回收率不太显著（约20%），延长与SV-III-130的共孵育时间进一步降低了回收率。曲线移动实验与SV-III-130和DA之间的竞争一致。D2 R的二级结合口袋中的两个突变（V91 A和E95 A）降低了拮抗效力并增加了从SV-III-130拮抗中的恢复，而第三个突变（L94 A）仅增加恢复。我们的研究结果表明，二级结合口袋的影响，从研究的阿立哌唑类似物的抑制恢复。我们提出了一种机制，通过计算机模拟支持，其中SV-III-130最初可逆地结合到D2 R，之后药物-受体复合物经历缓慢过渡到第二配体结合状态，这取决于二级结合口袋的完整性，并且在我们实验的时间范围内不可逆。
A solid understanding of the mechanisms governing ligand binding is crucial for rational design of therapeutics targeting the dopamine D2 receptor (D2R). Here, we use G protein-coupled inward rectifier potassium (GIRK) channel activation in Xenopus oocytes to measure the kinetics of D2R antagonism by a series of aripiprazole analogues, as well as the recovery of dopamine (DA) responsivity upon washout. The aripiprazole analogues comprise an orthosteric and a secondary pharmacophore and differ by the length of the saturated carbon linker joining these two pharmacophores. Two compounds containing 3- and 5-carbon linkers allowed for a similar extent of recovery from antagonism in the presence of 1 or 100 μM DA (>25 and >90% of control, respectively), whereas recovery was less prominent (∼20%) upon washout of the 4-carbon linker compound, SV-III-130, both with 1 and 100 μM DA. Prolonging the coincubation time with SV-III-130 further diminished recovery. Curve-shift experiments were consistent with competition between SV-III-130 and DA. Two mutations in the secondary binding pocket (V91A and E95A) of D2R decreased antagonistic potency and increased recovery from SV-III-130 antagonism, whereas a third mutation (L94A) only increased recovery. Our results suggest that the secondary binding pocket influences recovery from inhibition by the studied aripiprazole analogues. We propose a mechanism, supported by in silico modeling, whereby SV-III-130 initially binds reversibly to the D2R, after which the drug-receptor complex undergoes a slow transition to a second ligand-bound state, which is dependent on secondary binding pocket integrity and irreversible during the time frame of our experiments.
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影响因子：
3.3
作者：
Klauda, Jeffery B.;Venable, Richard M.;Freites, J. Alfredo;O'Connor, Joseph W.;Tobias, Douglas J.;Mondragon-Ramirez, Carlos;Vorobyov, Igor;MacKerell, Alexander D., Jr.;Pastor, Richard W.
通讯作者：
Pastor, Richard W.
β-甲蛋白偏置的配体的结构启发的设计。
影响因子：
14.8
作者：
McCorvy JD;Butler KV;Kelly B;Rechsteiner K;Karpiak J;Betz RM;Kormos BL;Shoichet BK;Dror RO;Jin J;Roth BL
通讯作者：
Roth BL
影响因子：
5.5
作者：
Leduc-Nadeau, Alexandre;Lahjouji, Karim;Bichet, Daniel G.
通讯作者：
Bichet, Daniel G.
影响因子：
2.8
作者：
Manuel Ramirez-Anguita, Juan;Rodriguez-Espigares, Ismael;Selent, Jana
通讯作者：
Selent, Jana
影响因子：
4.8
作者：
Agren, Richard;Arhem, Peter;Sahlholm, Kristoffer
通讯作者：
Sahlholm, Kristoffer