芳杂基哌嗪基脒类化合物的设计、合成及5-HT和NE重摄取双重抑制活性

本文基于药效团模型的指导和课题组前期的研究结果,设计合成了一类全新结构类型的芳杂基哌嗪基脒类化合物,通过¹H NMR、HRMS对化合物结构进行了确证,并完成了初步的体外药理活性评价。结果表明这些化合物显示不同程度的5-HT和NE重摄取抑制活性,值得进一步研究。     

单胺重摄取抑制剂的研究进展

单胺重摄取抑制剂在临床上被广泛应用于抑郁症的治疗。本文通过文献检索综述了2004～2010年间5-羟色胺重摄取抑制剂和去甲肾上腺素再摄取抑制剂的研究进展,发现具有5-HT、NA重摄取抑制(选择性抑制或双重抑制)活性的新结构及部分构效关系。     

单胺重摄取抑制剂的研究进展

单胺重摄取抑制剂在临床上被广泛应用于抑郁症的治疗.本文通过文献检索综述了2004～2010年间5-羟色胺重摄取抑制剂和去甲肾上腺素再摄取抑制剂的研究进展,发现具有5-HT、NA重摄取抑制(选择性抑制或双重抑制)活性的新结构及部分构效关系.     

间尼索地平对5-羟色胺诱导的大鼠肺动脉平滑肌细胞增殖中Ca2+/CaM/CaN通路的影响

探讨Ca²⁺/CaM/CaN信号通路在5-羟色胺 (5-HT) 诱导的大鼠肺动脉平滑肌细胞 (PASMCs) 增殖中的作用以及间尼索地平 (m-Nis) 对此的影响。采用细胞培养、噻唑蓝 (MTT) 比色检测、激光共聚焦显微镜及反转录聚合酶链反应 (RT-PCR) 等方法, 研究5-HT (1 μmol·L⁻¹) 对大鼠PASMCs细胞增殖的影响以及m-Nis对此的抑制作用, 并通过检测m-Nis对5-HT诱导的大鼠PASMCs增殖中Ca²⁺/CaM/CaN通路的影响深入探讨其作用机制。结果显示, 5-HT (1 μmol·L⁻¹) 可明显诱导大鼠PASMCs增殖 (P < 0.01), m-Nis对此有明显的抑制作用, 并呈现一定的浓度依赖性 (P < 0.05或P < 0.01); 另外, m-Nis还明显抑制了5-HT引起的[Ca²⁺]_i的升高 (P < 0.01)、CaM和CaN mRNA的表达以及CaN活性的升高 (P < 0.05或P < 0.01)。结果表明, Ca²⁺/CaM/CaN信号通路在5-HT诱导的大鼠PASMCs增殖中起重要作用, m-Nis抗5-HT诱导的增殖作用可能与抑制[Ca²⁺]_i增高从而阻断了Ca²⁺/CaM/CaN信号通路有关。     

维拉唑酮：多靶标和新机制抗抑郁药研发的新方向

维拉唑酮是兼有5-羟色胺1A(5-HT_1A)受体部分激动作用和5-HT重摄取抑制作用的双重活性药物。该药物的设计是基于特异性加速5-HT_1A自身受体脱敏的原理,以达到加快起效和增强疗效的目的。临床前研究结果表明,维拉唑酮是重组细胞系和正常组织的5-HT_1A受体的高效的部分激动剂,能占有并功能性阻断5-HT转运蛋白的活性位点,抑制5-HT重摄取;在多个动物模型中具有抗抑郁和抗焦虑活性。维拉唑酮已经被美国食品药品监督管理局批准用于重度抑郁症的治疗。目前尚无将维拉唑酮与其他抗抑郁药进行直接比较的报道,但现有临床研究已表明该药物抗抑郁的治疗效果与其他抗抑郁药相当。与5-HT重摄取抑制剂一样,维拉唑酮也存在胃肠道不良反应,但性功能障碍和体质量增加的不良反应发生率极低,并且起效可能更快速。维拉唑酮全新的抗抑郁机制,为重度抑郁症的治疗提供了新的选择。     

黄酮类化合物对细胞色素P450 CYP1A2的抑制作用及其构效关系研究

利用本实验室已建立的体外肝微粒体模型， 测定36个黄酮类单体化合物对人细胞色素P450 CYP1A2的抑制活性， 并使用三维定量构效关系方法研究化合物的分子结构参数与其抑制活性之间的关系。CoMSIA模型证实黄酮类化合物的结构参数与其CYP1A2抑制活性存在明显的相关性(模型的相关系数R²为0.948)， 且有良好的预测能力(交叉验证相关系数q²为0.630)， 同时使用“留五法”证实模型的稳定性和可靠性。结果表明， 相对于黄酮， α-萘黄酮的π-π共轭体系更有利于提高化合物的抑制活性。根据获得的模型的三维等势图， 在α-萘黄酮基础上， 6、3′、4′位引入带正电基团或是疏水基团，同时5位引入带负电基团，能有效改善化合物的CYP1A2抑制活性。     

苯氧茚酮类衍生物的合成和乙酰胆碱酯酶抑制活性

目的设计、合成新型的AchE抑制剂。方法在K₂CO₃和乙腈存在下，将2-溴-5,6-二甲氧基-1-茚酮与多种胺烷基苯酚缩合得到一系列苯氧茚酮类衍生物，并采用改进的Ellman方法研究它们的体外AchE和BchE抑制活性。结果合成了16个未见文献报道的化合物8a～p，结构经IR，¹H NMR，MS及元素分析确证。初步体外AchE，BchE抑制活性试验表明，绝大部分化合物都显示出良好的抑制AchE活性，其中8h的IC₅₀为50.0 nmol·L^-1，与石杉碱甲相近(IC₅₀＝53.0 nmol·L^-1)；而所有化合物基本都无BchE抑制活性。结论该类衍生物具有较强的AchE抑制活性，有进一步研究的价值。     

查尔酮异甘草素类化合物的合成及其对子宫颈癌细胞的抑制作用

目的 设计并合成查尔酮异甘草素类化合物,并评价其抗子宫颈癌活性。方法 利用Claisen-Schmidt反应原理,通过微波固相合成5种目标化合物,并进行结构表征。以SiHa(人子宫颈鳞癌细胞)和HeLa(子宫颈癌细胞)细胞株作为体外模型,利用MTT法考察目标化合物对宫颈癌细胞增殖的抑制活性,利用流式细胞仪测定目标化合物对宫颈癌细胞促凋亡作用。结果与结论 快速高效合成了5个查尔酮异甘草素类化合物,其结构经¹H-NMR、¹³C-NMR谱确认。目标化合物能有效抑制宫颈癌细胞增殖,促进宫颈癌细胞凋亡。     

苯二醛缩氨基硫脲的合成及其酪氨酸酶抑制活性研究

目的 合成苯二醛单缩和二缩氨基硫脲类化合物,并初步研究其抑制酪氨酸酶的活性和作用机制。方法 以5种苯二醛和氨基硫脲为原料,通过缩合反应合成9个目标化合物;采用蘑菇酪氨酸酶多巴速率氧化法和酶抑制动力学实验,测定目标化合物对酪氨酸酶的抑制活性和作用机制;选择化合物3a和4a进行抑制机制和抑制动力学研究。结果 目标化合物的结构经¹H-NMR、¹³C-NMR及MS确证;所有化合物抑制酪氨酸酶的活性均优于对照药物曲酸;苯二醛二缩氨基硫脲3a~3d的活性明显强于相应的单缩氨基硫脲4a~4d;化合物3a和4a对酪氨酸酶的抑制作用均表现为混合型可逆抑制作用。结论 苯二醛二缩氨基硫脲类化合物具有优异的抑制酪氨酸酶的活性,值得进一步深入研究。     

查耳酮类化合物合成及抗乳腺癌活性研究

目的 设计、合成并筛选具有抗乳腺癌活性的查耳酮类化合物。方法 利用Claisen-Schmidt 羟醛缩合反应,合成了一系列的查耳酮类化合物,并通过MTT法测试化合物抗乳腺癌活性。结果 合成了36个查耳酮类化合物,其结构均通过¹H-NMR和¹³C-NMR进行了表征。初步生物活性结果表明大部分目标分子查耳酮对MCF-7有较强的抑制活性,对MDA-MB-231也展现出了中等的抑制活性,其中化合物27(IC₅₀=11.3 μmol·L^-1)对MCF-7的抑制活性是他莫昔芬的1.5倍,同时这些查尔酮类化合物对正常细胞均没有毒性。结论 本研究为开发具有抗乳腺癌活性的查耳酮类化合物提供了参考和依据。     

5-HT reuptake inhibitors with 5-HT(1B/1D) antagonistic activity: a new approach toward efficient antidepressants

As part of our research program toward new, potential antidepressants, a series of unsymmetrical ureas has been prepared and evaluated as 5-HT reuptake inhibitors with 5-HT(1B/1D) antagonistic activities. The design of these compounds was based on coupling of various indole derivatives, previously shown to inhibit 5-HT reuptake, to three different aniline moieties, which are part of known 5-HT(1B/1D) ligands. Binding experiments in rat frontal cortex using [(125)I]iodocyanopindolol, in calf striatum using [(3)H]5-HT, and in rat hippocampus using [(3)H]8-OH-DPAT as radioligands, respectively, revealed significantly higher affinity at the 5-HT(1B) receptor as compared to the affinities for the 5-HT(1A) and 5-HT(1D) receptors for a number of compounds, among them 4-(5-fluoro-1H-indol-3-yl)piperidine-1-carboxylic acid [4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]amide (5), the corresponding 4-fluoro-1H-indol-3-yl analogue 21a, and the corresponding 6-fluoro-1H-indol-3-yl analogue 21b. Conformational restriction of the aniline moiety in 5 only slightly enhanced the 5-HT(1B) affinity, whereas introduction of an aniline moiety with higher conformational flexibility resulted in a less potent 5-HT(1B) receptor ligand as compared to 5. The functional 5-HT(1B/1D) antagonistic activity was investigated using the rabbit saphenous vein model as well as the [(3)H]5-HT release from guinea pig cortical slices. All new compounds tested in the rabbit saphenous vein model were shown to antagonize the sumatriptan-evoked contractile responses with pA(2) values ranging from 7.3 to 8.7. These observations were consistent with the results of the cortical slice model, in which the ureas were found to block the sumatriptan-induced inhibition of potassium-evoked [(3)H]5-HT release. The 5-HT reuptake inhibition of the ureas determined in rat brain synaptosomes was found to be either increased or decreased as compared to the uncoupled indole derivatives indicating that the reuptake inhibition shown by the ureas is not only due to the indole part but also affected by the aniline moiety of the molecule. Among this series of compounds described the ureas 5, 21a, and 21b seem to be the most interesting candidates showing both 5-HT reuptake inhibition and 5-HT(1B/1D) antagonism in vitro. This dual pharmacological profile should in theory lead to a pronounced enhancement in serotonergic neurotransmission and consequently to a more efficient treatment of depression.     

The partial NMDA agonist D-cycloserine stimulates LH secretion in healthy volunteers

 Because of clinical interest in the effects of antidepressant drugs that exert their effects on multiple neurotransmitter systems, the behavioral effects produced by combined treatment with an SSRI (fluoxetine) with a selective norepinephrine (NE; desipramine) or dopamine (DA) reuptake inhibitor (buproprion) were examined in the forced swimming test (FST), a behavioral test in rodents that predicts the clinical activity of antidepressants. Three additional compounds with mixed activity as NE-5-HT reuptake inhibitors, milnacipran, duloxetine and venlafaxine, were also examined. Desipramine and fluoxetine both reduced immobility in the FST, but desipramine increased only climbing behavior, whereas fluoxetine increased only swimming behavior. The combination of fluoxetine with desipramine or bupropion increased both climbing and swimming behaviors at certain doses, but higher doses of desipramine when combined with fluoxetine replaced swimming behavior with climbing behavior. The mixed NE-5-HT reuptake inhibitors milnacipran and duloxetine reduced immobility and increased climbing behavior, but did not alter swimming. Venlafaxine reduced immobility and increased swimming behavior, except at the highest dose tested (80mg/kg), which increased both swimming and climbing behaviors. Thus, combining certain doses of pharmacologically selective monoamine reuptake inhibitors, or the mixed reuptake inhibitor venlafaxine, produced a pattern of mixed active behaviors in the FST (climbing and swimming) that may reflect the activity of multiple neurotransmitters, especially the combination of enhanced 5-HT and DA activity. The combination of higher doses of desipramine with fluoxetine, or compounds with mixed activity at inhibiting NE and 5-HT reuptake, demonstrated effects similar to those of desipramine alone and may reflect inhibition of the expression of serotonergic antidepressant behavioral effects by selective NE reuptake inhibitors. Received: 10 June 1997/Final version: 19 August 1997     

Synthesis and pharmacological characterization of a series of geometrically constrained 5-HT(2A/2C) receptor ligands

In studies of the SAR of phenethylamine-type serotonin 5-HT(2A) receptor agonists, substituted conformationally constrained tetrahydronaphthofurans were designed to investigate the optimal conformation of the 2-aminoethyl moiety. These compounds were tested using in vitro assays for affinity at 5-HT(1A), 5-HT(2A), and 5-HT(2C) receptors. The benzofuran-containing analogues, 6a and 6b, had significantly higher affinity for the 5-HT receptors tested than did the benzodihydrofuran-containing compounds, 4a, 4b, 5a, and 5b. The most potent compound (8-bromo-6-methoxy-4,5-dihydro-3H-naphtho[1,8-bc]furan-5-yl)aminomethane, 6b, had K(i) values for displacement of [(125)I]-DOI from 5-HT(2A) and 5-HT(2C) cloned rat receptors of 2.6 and 1.1 nM, respectively. Despite their high affinity, the compounds of this naphthofuran series lacked high intrinsic activity at the 5-HT(2A) receptor as measured using the phosphoinositide hydrolysis assay. The most potent compound in vitro, 6b, was tested in the two-lever drug discrimination assay in rats trained to discriminate LSD from saline, and failed to substitute, a result typical for compounds with low intrinsic activity. Thus, although conformational constraint has led to high-affinity 5-HT(2A) ligands with partial agonist activity, all of the spatial and steric properties of the ligand necessary for full receptor activation have not yet been identified.     

芳烷醇哌啶类化合物的合成及抗抑郁活性研究

为寻找新型具有抗抑郁活性的单胺递质再摄取抑制剂, 设计合成了15个芳烷醇哌啶类未见文献报道的新化合物, 结构经¹H NMR及HR-MS分析确证。大鼠脑突触体对5-HT、NA和DA再摄取抑制作用体外活性测试结果表明, 化合物4、5和8对5-HT、NA和DA再摄取抑制作用较强。小鼠强迫游泳实验发现, 化合物4、5和13具有显著的体内抗抑郁效果, 化合物4和5值得进一步研究。     

Pyrroloisoquinoline antidepressants. 3. A focus on serotonin

A collection of hexahydropyrroloisoquinoline derivatives (1-22), which represent a class of compounds that inhibit the neuronal uptake of dopamine (DA), norepinephrine (NE), and serotonin (5-HT), was investigated in vivo for serotonin-potentiating properties in the mouse head-twitch and rat serotonin syndrome assays. The p-methylthio compound 3b (McN-5652-Z) was found to possess exceptional activity in these assays, and the activity was attributable almost exclusively to the (+)-6S,10bR enantiomer. Ten closely related analogues were synthesized, tested, and compared among themselves and with some previously prepared compounds, both in vivo and in vitro. Several trans diastereomers exhibited strong inhibition of 5-HT uptake and substantial potentiation of 5-HT, while the cis diastereomers (3a, 4a, and 10a) tested were virtually devoid of such activity. Although 3b was only moderately selective in inhibiting the uptake of 5-HT vs NE, its 10-substituted analogues 4b, 7b-9b had improved 5-HT selectivity relative to NE, to the extent of 20-25 times (150-200 times relative to DA). Of these more selective compounds (in vitro), only 4b and 7b had substantial activity in vivo. Sulfoxide 11b appeared to function as a prodrug of 3b in vivo.     

Novel 4-alkyl-1-arylpiperazines and 1,2,3,4-tetrahydroisoquinolines containing diphenylmethylamino or diphenylmethoxy fragment with differentiated 5-HT1A/5-HT2A/D2 receptor activity

Two series of 4-alkyl-1-arylpiperazines (1-4) and 1,2,3,4-tetrahydroiso-quinolines (5, 6) with diphenylmethylamino (series a) or diphenylmethoxy (series b) fragment were synthesized in order to obtain potential ligands of 5-HT1A and/or 5-HT2A and dopamine D2 receptors. Four new arylpiperazines (1a, 3a, 1b, 3b) were found to demonstrate high 5-HT1A receptor affinity (Ki = 1.5-35 nM); among them, 3a exhibited satisfactory 5-HT2A receptor affinity (Ki = 74 nM). Only compounds 1b and 2b showed moderate affinity for D2 receptor sites (Ki = 123 and 128 nM, respectively). Compounds 1a, 3a, 1b and 3b were investigated in vivo to determine their functional activity at 5-HT1A receptors; additionally, 3a was tested for 5-HT2A receptor activity. Derivatives 1a, 1b and 3b produced effects characteristic of antagonists of postsynaptic 5-HT1A receptors. Moreover, 1b exhibited features of an agonist of presynaptic 5-HT1A receptors, while 3a behaved like a partial agonist of postsynaptic 5-HT1A sites. The latter derivative may also be classified as a 5-HT2A receptor antagonist. Thus, novel potent 5-HT1A receptor ligands were successfully obtained, and the most promising compound 3a showed mixed 5-HT1A/5-HT2A receptor activity in in vitro and in vivo tests.     

New 1-aryl-3-(4-arylpiperazin-1-yl)propane derivatives, with dual action at 5-HT1A serotonin receptors and serotonin transporter, as a new class of antidepressants.

In a search toward new and efficient antidepressants, 1-aryl-3-(4-arylpiperazin-1-yl)propane derivatives were designed, synthesized, and evaluated for 5-HT reuptake inhibition and 5-HT1A receptor antagonism. This dual pharmacological profile should lead, in principle, to a rapid and pronounced enhancement in serotoninergic neurotransmission and consequently to a more efficacious treatment of depression. The design was based on coupling structural moieties related to inhibition of serotonin reuptake, such as gamma-phenoxypropylamines, to arylpiperazines, typical 5-HT1A ligands. In binding studies, several compounds showed affinity at the 5-HT transporter and 5-HT1A receptors. Antidepressant-like activity was initially assayed in the forced swimming test with those compounds with Ki < 200 nM in both binding studies. Functional characterization was performed by measuring the intrinsic effect on rectal temperature in mice and also the antagonism to 8-OH-DPAT-induced hypothermia. The most efficacious compounds (12f, 23gE, 28a, and 28b) were further explored for their ability to antagonize 8-OH-DPAT-induced inhibition of forskolin-stimulated cAMP formation in a cell line expressing the 5-HT1A receptor. Furthermore, the antidepressant-like properties of 12f, 28a, and 28b, which exhibited 5-HT1A receptor antagonistic property in the latter study, were also evaluated in the learned helplessness test in rats. Among these three compounds, 28b (1-benzo[b]thiophene-3-yl)-3-[4-(2-methoxyphenyl)-1-ylpropan-1-ol) showed the higher affinity at both the 5-HT transporter and 5-HT1A receptors (Ki = 20 nM in both cases) and was also active in the other pharmacological tests. Such a pharmacological profile could lead to a new class of antidepressants with a dual mechanism of action and a faster onset of action.     

Synthesis and molecular modeling of new 1-aryl-3-[4-arylpiperazin-1-yl]-1-propane derivatives with high affinity at the serotonin transporter and at 5-HT(1A) receptors

It has been proposed that 5-HT(1A) receptor antagonists augment the antidepressant efficacy of selective serotonin (5-HT) reuptake inhibitors. In a search toward new and efficient antidepressants, 1-(aryl)-3-[4-arylpiperazin-1-yl]-1-propane molecular hybrids were designed, synthesized, and evaluated for 5-HT reuptake inhibition and 5-HT(1A) receptor affinity. The design was based in coupling structural moieties related to inhibition of serotonin reuptake, such as benzo[b]thiophene derivatives to arylpiperazines, typical 5-HT(1A) receptor ligands. In binding studies, several compounds showed affinity at the 5-HT transporter and at 5-HT(1A) receptors. Molecular modeling studies predicted the pharmacophore elements required for high affinity binding and the features that enable to discriminate between agonist, partial agonist, or antagonist action at 5-HT(1A) receptors and 5-HT transporter inhibition. Solvent interactions in desolvation prior to the binding step along with enthalpy and enthropy compensations might be responsible to explain agonist, partial agonist, and antagonist character. Hydrogen-bonding capability seems to be important to break hydrogen interhelical hydrogen bonds or alternatively to form other bonds upon ligand binding. Partial agonists and antagonists are unable to do this as the full agonist, which interacts closely by long-range forces or directly. The compounds showing the higher affinity at both the 5-HT transporter (K(i) < 50 nM) and the 5-HT(1A) receptors (K(i) < 20 nM) were further explored for their ability to stimulate [(35)S]GTPgammaS binding or to antagonize 8-hydroxy-2-di-n-propylamino-tetralin (8-OH-DPAT)-stimulated [(35)]GTPgammaS binding to rat hippocampal membranes, an index of agonist/antagonist action at 5-HT(1A) receptors, respectively. Compound 8g exhibited agonist activity (EC(50) = 30 nM) in this assay, whereas compounds 7g and 8h,i behaved as weak partial agonists and 7h-j and 8j,l antagonized the R(+)-8-OH-DPAT-stimulated GTPgammaS binding. Functional characterization was performed by measuring the antagonism to 8-OH-DPAT-induced hypothermia in mice.     

New 4-[omega-(diarylmethylamino)alkyl]- and 4-[omega-(diarylmethoxy)alkyl]-1-arylpiperazines as selective 5-HT1A/5-HT2A receptor ligands with differentiated in vivo activity

Two series of novel 4-ethyl- or 4-propyl-1-arylpiperazines (5-12) with the 4,4'-disubstituted diphenylmethylamino (series a) or the diphenylmethoxy (series b) terminal fragment were synthesized and evaluated for their binding affinity at 5-HT1A and 5-HT2A receptors. The influence of the introduction of 4-methyl, 4-chloro or 4-fluoro substituents at both phenyl rings of that terminal moiety on in vitro and in vivo 5-HT1A receptor activity of those modified compounds was discussed. Compounds 5a, 6a, 9a-12a, 5b, 6b, 9b, 11b and 12b displayed high to fairly high affinity for 5-HT1A receptors (Ki = 2.4-72 nM). Compounds of both series showed low or very low 5-HT2A receptor affinity (Ki = 155-5400 nM). Amines 5a, 6a, 11a, and their ether analogs 5b, 6b and 11b, also possessed high or moderate alpha(1)-adrenoceptor affinity (K(i) = 6-104 nM). The functional activity of compounds 5a, 6a, 9a-12a, 5b, 8b, 9b, 11b and 12b was tested in vivo in the commonly used animal models. The majority of those ligands behaved like 5-HT1A receptor antagonists, their influence on the pre- and/or postsynaptic sites being diverse, though. They exhibited characteristics of partial agonists of postsynaptic 5-HT1A receptors (11a), of weak antagonists of pre- and postsynaptic sites (12a, 9b), of antagonists of presynaptic (5a) or of antagonists of postsynaptic 5-HT(1A) receptors (9a, 10a, 5b, 8b, 11b and 12b) while, 6a was devoid of functional activity at those receptors. The above findings indicate that introduction of 4-methyl, 4-chloro or 4-fluoro substituents to the diphenylmethyl part of the 1-(2-methoxyphenyl)piperazines tested in vivo may modify their 5-HT1A receptor functional activity.