LY-404187   中文名称: N-[2-(4'-氰基联苯-4-基)丙基]-2-丙烷磺酰胺

AMPA receptors (EC50 = 0.15, 1.44, 1.66, 0.21 µM, 5.65 for GluR2i, GluR2o, GluR3i, GluR4i, GluR1i, respectively)

转染人 GluR4 的 HEK293 细胞对 LY-404187 (3-10 nM) 表现出增强的谷氨酸诱发内向电流 [2]。在急性分离的锥体神经元中，LY-404187 (0.03-10 µM) 选择性增加通过 AMPA 受体/通道的谷氨酸诱发电流，具有显着的效力 (EC50=1.3±0.3 µM) 和功效（Emax = 45.3±8.0 倍增加）[3 ]。剂量为 10 µM 时，LY-404187 对前额皮质 (PFC) 锥体神经元全细胞 K+ 或 Na+ 电流的幅度或时间过程没有影响 [3]。

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与NMDA受体活性的募集一致，LY404187在需要不同记忆过程的认知功能动物模型中被证明可以提高表现。这些数据表明，AMPA受体增强剂可能对治疗多种疾病的认知缺陷有益，特别是那些与谷氨酸能信号减少相关的疾病，如精神分裂症。此外，LY404187已被证明在行为绝望的动物模型中有效，对抗抑郁药物活性具有相当大的预测有效性。尽管AMPA受体增强剂在这些疾病和其他疾病中的治疗效果最终将在临床中确定，但有证据表明，这些化合物的益处将由多种作用机制介导。这些机制包括AMPA受体功能的直接增强，细胞内信号级联的二次动员，以及基因表达的长期调节。[1]

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近年来，随着α -氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）受体分子生物学和药理学的发展，发现了选择性、强效和系统活性的AMPA受体增强剂。这些分子增强突触传递，证据表明它们在可塑性和认知过程中发挥重要作用。AMPA受体的激活也会增加神经元的激活和活动依赖的信号，这可能会增加脑源性神经营养因子（BDNF）的表达并促进脑内细胞的增殖。因此，我们假设AMPA受体增强剂可能在帕金森病的啮齿动物模型中提供神经营养作用。在本研究中，我们报道了强效和选择性的AMPA受体增强剂R，S-N-2-（4-（4-氰苯基）苯基）丙基2-丙磺酸酰胺（LY404187），对大鼠单侧向黑质或纹状体输注6-羟多巴胺提供功能、神经化学和组织学保护。该化合物还降低了1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）对小鼠的毒性。有趣的是，当我们将治疗延迟到细胞死亡后（6-羟多巴胺输注后3或6天），我们也能够观察到巨大的功能和组织学影响，支持神经营养作用机制。此外，LY404187提供了纹状体中生长相关蛋白43表达的剂量依赖性增加。因此，我们提出AMPA受体增强剂提供了一种潜在的新疗法来阻止帕金森病的进展，并可能修复帕金森病的退化。[4]

皮下暴露于 0.5 mg/kg LY-404187 11 天的小鼠可免受 MPTP 诱导的神经毒性 [4]。当以 0.5 mg/kg 的剂量皮下注射 28 天时，LY-404187 显着防止酪氨酸羟化酶阳性黑质细胞体的损失，并减少阿扑吗啡诱导的反向旋转[4]。 LY-404187（0.1 或 0.5 mg/kg；皮下注射 14 天）可对注射 6-羟基多巴胺的大鼠黑质进行功能、神经化学和组织学保护[4]。延迟治疗后皮下注射 LY-404187 (0.5 mg/kg) 14 天，观察到功能和组织学的改善。这表明细胞死亡后给予该药物可能会产生营养影响[4]。当以 0.1 和 0.5 mg/kg 皮下给药 14 天时，LY-404187 以剂量依赖性方式提高纹状体中的 GAP-43 免疫反应性 [4]。

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LY404187是一种选择性、强效且具有中枢活性的AMPA受体正向变构调节剂。LY404187优先作用于重组人同聚体GluR2和GluR4 AMPA受体，而非GluR1和GluR3。此外，与flop剪接变体相比，LY404187对这些AMPA受体的flip剪接变体具有更强的增强作用。在重组和天然AMPA受体中，LY404187的增强作用表现出独特的时间依赖性增长，这似乎涉及对这些离子通道脱敏过程的抑制。LY404187已被证明能在体外和体内增强谷氨酸能突触传递。这种突触活动的增强源于对AMPA受体功能的直接增强，以及电压依赖性NMDA受体活性的间接募集。通过NMDA受体增强钙内流被认为是启动突触功能长期改变（如长时程增强，LTP）的关键步骤。这些突触功能的改变可能是某些记忆编码形式的基础。[1]

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本研究描述了两个新型强效选择性AMPA受体增强分子LY392098和LY404187的活性。LY392098和LY404187能增强谷氨酸（100 μM）刺激的通过重组人同聚体AMPA受体离子通道（GluR1-4）的离子内流，其EC50估计值分别为：LY392098对GluR1i（1.77 μM）、GluR2i（0.22 μM）、GluR2o（0.56 μM）、GluR3i（1.89 μM）和GluR4i（0.20 μM）；LY404187对GluR1i（5.65 μM）、GluR2i（0.15 μM）、GluR2o（1.44 μM）、GluR3i（1.66 μM）和GluR4i（0.21 μM）。在没有谷氨酸的情况下，这两种化合物都不会影响未转染HEK293细胞或GluR转染细胞的离子内流。两种化合物对AMPA受体都具有选择性活性，对人重组红藻氨酸受体无活性。电生理记录表明，LY392098和LY404187在低浓度（3-10 nM）下能增强谷氨酸（1 mM）在人GluR4转染HEK293细胞中诱发的内向电流。此外，这两种化合物都能消除重组GluR4 AMPA受体的谷氨酸依赖性脱敏。这些研究表明，LY392098和LY404187能在体外变构增强HEK293细胞表达的人AMPA受体离子通道介导的反应。[2]

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谷氨酸α-氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）受体的正向调节剂可增强多种物种的认知功能。本实验比较了新型联芳基丙基磺酰胺化合物LY404187与原型苯甲酰哌啶化合物1-(喹喔啉-6-羰基)-哌啶（CX516）对前额叶皮层（PFC）锥体神经元AMPA受体的作用。LY404187（0.03-10 μM）选择性增强急性分离锥体神经元AMPA受体/通道的谷氨酸诱发电流，其效价（EC50=1.3±0.3 μM）和效能（Emax=45.3±8.0倍增加）显著高于CX516（EC50=2.8±0.9 mM；Emax=4.8±1.4倍增加）。LY404187和CX516分别使谷氨酸浓度-反应曲线的效价提高了6倍和3倍。快速灌流实验表明，LY404187显著抑制受体脱敏的幅度但不改变其动力学，而CX516对脱敏程度影响较小，仅适度减缓脱敏过程。在前额叶皮层切片中，纳摩尔浓度的LY404187能增强自发的和刺激诱发的AMPA受体介导的兴奋性突触后电位。电压敏感的N-甲基-D-天冬氨酸（NMDA）受体依赖性突触反应也因突触后去极化增强而间接增强。与体外数据一致，LY404187在体内增强前额叶皮层神经元对海马谷氨酸能传入刺激放电概率的效力比CX516强1000倍。这种LY404187的增强作用可被选择性AMPA受体拮抗剂（LY300168，1 mg/kg，静脉注射）和NMDA受体拮抗剂（LY235959，5 mg/kg，静脉注射）减弱。总之，这些结果表明LY404187是一种极其强效且具有中枢活性的天然AMPA受体增强剂，具有独特的作用机制。本文还讨论了AMPA受体增强剂的治疗意义。[3]

钙内流测量[2]
制备了含有稳定表达人AMPA受体的HEK293细胞的融合单层96孔板。细胞在缓冲液中孵育(10 mM葡萄糖，138 mM氯化钠，1 mM氯化镁，5 mM氯化钾，5 mM氯化钙，10 mM n -2-羟乙基哌嗪- n -2-乙磺酸，到pH 7.1至7.3)，含20 μM Fluo3-AM染料60分钟。用缓冲液洗涤细胞，使用FLUOROSKAN II荧光仪进行荧光测量，表明在环噻嗪（100 μM）或化合物存在的情况下，谷氨酸（100 μM）刺激钙流入细胞时，荧光变化。化合物应用在谷氨酸添加前5分钟，在谷氨酸添加前和谷氨酸添加后3分钟进行荧光测量。数据相对于环噻嗪（100 μM）对GluR1-4 (i)产生的荧光变化表示，对于GluR2 flop数据相对于LY392098或LY404187获得的最大荧光表示。

Animal/Disease Models: Male C57BL/6J mice (20-25 g) were challenged with MPTP on day 8 [4]
Doses: 0.5 mg/kg
Route of Administration: Sc; twice (two times) daily on weekdays and one time/day on weekends for 11 days
Experimental Results: Attenuated loss of tyrosine hydroxylase immunoreactivity in the substantia nigra. There were no significant changes in dorsal and ventral striatal tyrosine hydroxylase immunoreactivity.

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MPTP neurotoxicity in mice [4]
Male C57BL/6J mice weighing 20–25 g were used. They were housed in groups of five mice per cage under a 12:12-h light/dark cycle (lights on 7:00 a.m. to 7:00 p.m.) with food and water available ad libitum. LY404187 was administered at 0.5 mg/kg s.c for 11 days. On day 8 the animals received 4×20 mg/kg MPTP at 2 h intervals.

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Drug studies [4]
For all studies, LY404187 was dissolved in 12.5% β-cyclodextrin and sonicated prior to administration and was administered subcutaneously (s.c.) twice daily on weekdays and once daily at weekends.
For studies using the striatal lesion model, LY404187 was administered for 28 days at 0.5 mg/kg s.c. starting 1 day after 6-hydroxydopamine lesion.
For nigral lesion studies, LY404187 was administered for 14 days at either 0.1 or 0.5 mg/kg s.c. starting 1 day after 6-hydroxydopamine lesion. In additional studies treatment with LY404187 was delayed until 3, or 6 days after 6-hydroxydopamine infusion.

[1]. LY404187: a novel positive allosteric modulator of AMPA receptors. CNS Drug Rev. Fall 2002; 8(3): 255-82.

[2]. Novel AMPA receptor potentiators LY392098 and LY404187: effects on recombinant human AMPA receptors in vitro. Neuropharmacology. 2001 Jun; 40(8): 976-83.

[3]. Positive modulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors in prefrontal cortical pyramidal neurons by a novel allosteric potentiator. J Pharmacol Exp Ther. 2001 Jul; 298(1): 86-102.

[4]. Neurotrophic actions of the novel AMPA receptor potentiator, LY404187, in rodent models of Parkinson's disease. Eur J Pharmacol. 2004 Feb 20; 486(2): 163-74.

Multiple intracellular signaling pathways regulate brain-derived neurotrophic factor (BDNF) levels. Antidepressants that increase norepinephrine release may enhance BDNF levels through the CREB pathway. Activation of the MAP kinase pathway can also increase BDNF mRNA levels. Lyn, a member of the Src family of protein tyrosine kinases, can activate the MAP kinase pathway. Studies have shown that Lyn physically interacts with the AMPA receptor GluR2 and GluR3 subunits and can be activated by potent stimulation of the AMPA receptor. Furthermore, primary cerebellar culture studies have shown that cyclothiazide alone has no effect on Lyn activation, but its activation is enhanced when used in combination with AMPA. Therefore, it is hypothesized that Lyn activation can increase BDNF levels by enhancing AMPA receptor activity. Neuron culture studies have confirmed this hypothesis, showing that adding LY392098 in the presence of AMPA or glutamate increases BDNF mRNA and protein levels. In summary, these data suggest that positively modulating AMPA receptors by increasing BDNF levels through the MAP kinase pathway may be beneficial for the treatment of depression. We have preliminarily explored the application of AMPA receptor enhancers in the treatment of depression. The activity of LY404187 was tested in a forced swimming test model, which can be used to detect antidepressant compounds (P. Skolnick, personal communication). The results showed that LY404187 reduced immobility time in mice and rats during the forced swimming test, with minimum effective doses (MEDs) of 0.05 and 0.025 mgu0001kg (oral), respectively (Figure 13). In this study, imipramine (15 mgu0001kg, intraperitoneal injection) served as a positive control. LY404187 did not affect motor activity, indicating that the efficacy of this compound in the forced swimming test is independent of its motor excitatory effect. Similar results were reported for LY392098. Furthermore, systemic administration of LY300168 blocked the activity of LY392098 but not the activity of imipramine, indicating that the action of this enhancer is mediated by AMPA receptors. In summary, these findings suggest that positive regulators of AMPA receptors are active in animal models capable of detecting clinically effective antidepressants. This supports the hypothesis that such compounds may represent a new class of antidepressants. [1]

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Electrophysiological recordings of glutamate-induced currents in human GluR4-transfected HEK293 cells showed that the detected AMPA receptor enhancers significantly enhanced the inward current. The observed enhancement was reversible, and no effect of the compounds was observed in the absence of agonists. The potency order of LY392098 and LY404187 was similar in both fluorescence assays and electrophysiological studies. Both compounds were more potent in enhancing the glutamate response than cyclothiazide or CX516. As shown by GluR4i (Fig. 3b), the desensitization of the AMPA receptor response to glutamate was rapid. Cyclothiazide significantly reduced the observed desensitization, although the current continued to decrease after sustained application of glutamate. Conversely, in the presence of LY392098 and LY404187, the amplitude of the glutamate-induced current increased during a 10-second glutamate application. The maximum rate of increase in current observed during the 10-second glutamate application appears to depend on the concentration of the AMPA receptor enhancer. The mechanism of this apparent agonist-dependent enhancement is currently under investigation. Shorter glutamate applications lasting 5 milliseconds are closer to the synaptic state (Clements et al., 1992). We investigated recovery after AMPA receptor desensitization using short-duration agonist applications. In this study, the AMPA receptor-mediated response recovery time progression approximates a single exponential time constant t = 48 milliseconds. LY404187 (1 μM), LY392098 (1 μM), and cyclothiazide (10 μM) all accelerated the recovery rate after desensitization. Possible mechanisms for reduced AMPA receptor desensitization include alterations in AMPA receptor inactivation mechanics. Cyclothiazide and aniracetam have both been shown to modulate the inactivation kinetics of recombinant and native AMPA receptors (Hestrin, 1992; Patneau et al., 1993; Partin et al., 1996). In order to study inactivation kinetics, in vitro experiments on the membrane are required. Due to the rapid decrease in current observed in in vitro hGluR receptor membranes, we have been unable to study the potential activity of these compounds in vitro. Current studies have confirmed that LY392098 and LY404187 are potent and selective enhancers of the glutamate response of human AMPA receptors in vitro. It will be important to study the activity of these AMPA receptor enhancers in heterologous oligomeric human recombinant ion channel complexes and neurons. [2] In summary, we have provided strong evidence that AMPA receptor enhancers can provide functional, neurochemical and histological protection in rodent models of Parkinson's disease. Such molecules have been reported to play a positive role in models of depression (Li et al., 2001; Quirk and Nisenbaun, 2002) and cognitive impairment (Staubli et al., 1994; Hampson et al., 1998a; Hampson et al., 1998b; Quirk and Nisenbaun, 2002), both of which are comorbidities in Parkinson's disease. Furthermore, in this study, these protective effects were maintained even after delayed administration until cell death, and the increased expression of GAP-43 in the striatum provided encouraging evidence for neurotrophic effects. These results suggest that LY404187 or its analogues are ideal molecules that could serve as clinical candidates to prevent or potentially reverse the degenerative changes observed in Parkinson's disease. [4]

C19H22N2O2S

342.455183506012

348.187

C, 66.64; H, 6.48; N, 8.18; O, 9.34; S, 9.36

211311-95-4

211311-95-4;

9928016

White to off-white solid powder

1.1±0.1 g/cm3

498.4±47.0 °C at 760 mmHg

255.2±29.3 °C

0.0±1.3 mmHg at 25°C

1.546

4.19

78.34

1

4

6

24

527

0

S(C(C)C)(NCC(C)C1C=CC(C2C=CC(C#N)=CC=2)=CC=1)(=O)=O

HOQAVGZLYRYHSO-UHFFFAOYSA-N

InChI=1S/C19H22N2O2S/c1-14(2)24(22,23)21-13-15(3)17-8-10-19(11-9-17)18-6-4-16(12-20)5-7-18/h4-11,14-15,21H,13H2,1-3H3

2-Propanesulfonamide, N-(2-(4'-cyano(1,1'-biphenyl)-4-yl)propyl)-

LY-404187; LY 404187; 211311-95-4; N-(2-(4'-Cyano-[1,1'-biphenyl]-4-yl)propyl)propane-2-sulfonamide; LY404187; LY-404,187; LY 404,187; N-[2-[4-(4-cyanophenyl)phenyl]propyl]propane-2-sulfonamide; 75W6I8W6OU; CHEMBL435582; LY404187.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~292.00 mM)

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
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注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
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