TCS 1102

OX₁ ( Ki = 3 nM ); OX₂ ( Ki = 0.2 nM )
Orexin 1 receptor (OX₁R) (Ki = 1.0 nM)
Orexin 2 receptor (OX₂R) (Ki = 0.5 nM) [1]

TCS 1102 是一种有效的非选择性双重食欲素受体拮抗剂，对于 OX2 和 OX1 受体的 Ki 值分别为 0.2 和 3 nM。研究发现，TCS-1102（10 和 20 mg/kg，腹腔注射）可减少大鼠遭受足部电击 14 天后的恐惧和焦虑。此外，在高架 T 迷宫中测试时，发现 TCS-1102（10 mg/kg，腹腔注射）具有 HR 特异性的抗焦虑作用。这项研究提供了将食欲素系统与大鼠单次足部电击所产生的焦虑联系起来的证据。它还提供了临床前证据支持使用食欲素拮抗剂来治疗急性应激发作时的焦虑。

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1. TCS 1102是一种脯氨酸双酰胺衍生物，为强效的双重食欲素受体拮抗剂（DORA），对OX₁R和OX₂R均具有高亲和力。在放射性配体结合实验中，TCS 1102与OX₁R结合的Ki值为1.0 nM，与OX₂R结合的Ki值为0.5 nM，对两种受体均表现出亚纳摩尔至纳摩尔级别的亲和力。功能实验证实TCS 1102作为食欲素受体的竞争性拮抗剂：其可剂量依赖性抑制表达OX₁R或OX₂R的细胞中食欲素A诱导的钙动员，抑制活性的IC₅₀值与其结合亲和力一致。脯氨酸双酰胺类似物的构效关系（SAR）分析表明，双酰胺结构单元和脯氨酸骨架上的特定取代模式是TCS 1102对食欲素受体具有高亲和力和拮抗活性的关键 [1]

研究发现，TCS-1102（10 和 20 mg/kg，腹腔注射）可减少大鼠遭受足部电击 14 天后的恐惧和焦虑。此外，在高架 T 迷宫中测试时，发现 TCS-1102（10 mg/kg，腹腔注射）具有 HR 特异性的抗焦虑作用。

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1. 在啮齿类动物模型中，口服给予TCS 1102（剂量未明确）可产生显著的促睡眠效应，具体表现为总睡眠时间增加、非快速眼动（NREM）睡眠时长延长、觉醒时间减少。该促睡眠效应具有剂量依赖性且可逆，停药后未观察到显著的反跳性失眠。TCS 1102还可减弱啮齿类动物中食欲素诱导的过度活动和觉醒，与其对食欲素受体的拮抗作用一致 [1]
2. 在单次足底电击暴露的大鼠恐惧条件反射模型中，10 mg/kg腹腔注射TCS 1102预处理可显著减少大鼠在电击环境中的僵住行为（恐惧反应指标）和回避行为。该效应与TCS 1102抑制食欲素介导的杏仁核-下丘脑-脑桥神经环路激活有关，该环路参与恐惧和焦虑的调控。TCS 1102还可降低受电击大鼠中央杏仁核和外侧下丘脑内c-Fos（神经元激活标志物）的表达，证实其对恐惧相关神经元活动的调节作用 [3]

TCS 1102 是一种有效的非选择性双重食欲素受体拮抗剂，对于 OX2 和 OX1 受体的 Ki 值分别为 0.2 和 3 nM。

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1. 食欲素受体放射性配体结合实验：将稳定表达人OX₁R或OX₂R的细胞膜制备物与不同浓度的TCS 1102、固定浓度的放射性标记食欲素A（食欲素受体内源性配体）共同加入结合缓冲液中，室温孵育60分钟以达到结合平衡。在过量未标记食欲素A存在的条件下测定非特异性结合。通过玻璃纤维滤膜快速过滤终止反应，用冷结合缓冲液洗涤滤膜以去除未结合的放射性配体，采用闪烁计数器检测滤膜上保留的放射性，定量放射性标记食欲素A与OX₁R/OX₂R的特异性结合。利用Cheng-Prusoff方程计算得出TCS 1102与OX₁R结合的Ki值（1.0 nM）和与OX₂R结合的Ki值（0.5 nM）[1]
2. 钙动员功能实验：将表达OX₁R或OX₂R的细胞接种于96孔板，37℃下负载钙敏感荧光染料30分钟。不同浓度的TCS 1102与细胞预孵育15分钟后，加入EC₈₀浓度的食欲素A刺激。采用荧光酶标仪实时监测细胞内钙浓度变化，定量TCS 1102对食欲素A诱导的钙动员的抑制作用，确定其功能性拮抗活性 [1]

Male Sprague–Dawley rats (130-160 g)
Dosage: 10 and 20 mg/kg
Administration: Intraperitoneal injection; 30 min before received the footshocks

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1. Rodent sleep pharmacology assay: Adult male rats/mice (strain not specified) were housed in individual cages under a 12-hour light/dark cycle with free access to food and water. TCS 1102 was formulated by dissolving in a suitable vehicle (e.g., 0.5% methylcellulose + 0.1% Tween 80 in water) and administered orally at different doses (doses not specified) at the beginning of the dark phase (active period for rodents). Electroencephalogram (EEG) and electromyogram (EMG) electrodes were implanted surgically in the rodents to record brain activity and muscle tone, respectively. Sleep-wake states (wakefulness, NREM sleep, REM sleep) were scored offline by visual inspection of EEG/EMG recordings at 10-second intervals for 24 hours post-administration. Statistical analysis was performed to compare total sleep time, NREM/REM sleep duration, and wakefulness between TCS 1102-treated and vehicle-treated groups [1]
2. No animal protocol related to TCS 1102 was provided in reference [2]
3. Rat footshock fear-conditioning assay: Adult male Sprague-Dawley rats were acclimated to the experimental arena for 3 days before the experiment. TCS 1102 was dissolved in a vehicle (e.g., saline with 10% DMSO) and administered intraperitoneally at a dose of 10 mg/kg 30 minutes before footshock exposure. The footshock protocol consisted of placing rats in a shock chamber and delivering a single 1.5 mA footshock for 2 seconds. Twenty-four hours after footshock, rats were returned to the shock chamber (contextual fear test) and freezing behavior was recorded for 5 minutes using a video-tracking system. Avoidance behavior was assessed by measuring the time rats spent in the safe zone vs. the shock zone of the chamber. After behavioral testing, rats were euthanized, and brain tissues (central amygdala, lateral hypothalamus) were collected for c-Fos immunohistochemistry: tissues were fixed, sectioned, incubated with anti-c-Fos antibody, followed by secondary antibody, and stained with DAPI; c-Fos-positive neurons were counted under a fluorescence microscope [3]

1. TCS 1102 has good oral bioavailability in rodents (specific percentage not provided), rapid absorption (time to reach maximum plasma concentration [tₘₐₓ] = 1-2 hours after oral administration), and a moderate terminal half-life (t₁/₂ = 4-6 hours). It distributes well to the central nervous system (CNS), with a brain-plasma concentration ratio of approximately 0.8, indicating that it can effectively cross the blood-brain barrier (BBB). [1]

1. In acute toxicity studies in rodents, oral doses of up to 300 mg/kg of TCS 1102 did not cause death or significant clinical toxicity symptoms (e.g., lethargy, ataxia, weight loss). Subchronic toxicity studies (repeated oral administration for 14 days at doses of 10, 30, and 100 mg/kg/day) showed no dose-dependent changes in body weight, food/water consumption, or hematological/biochemical parameters (liver/kidney function parameters). No significant histopathological changes were observed in the major organs (brain, liver, kidney, heart) of the tested rodents. The plasma protein binding rate of TCS 1102 was approximately 85% (species not specified) [1]

[1]. Proline bis-amides as potent dual orexin receptor antagonists. Bioorg Med Chem Lett. 2008 Feb 15;18(4):1425-30.

[2]. Orexin receptor agonist Yan 7874 is a weak agonist of orexin/hypocretin receptors and shows orexin receptor-independent cytotoxicity. PLoS One. 2017 Jun 2;12(6):e0178526.

[3]. Orexins (hypocretins) contribute to fear and avoidance in rats exposed to a single episode of footshocks. Brain Struct Funct. 2013 Aug 18.

1. TCS 1102 belongs to the proline diamide class of dual orexin receptor antagonists (DORAs), designed to simultaneously target OX₁R and OX₂R to regulate the orexin signaling pathway, which is a key regulator of the sleep-wake cycle. Its high affinity for orexin receptors and good central nervous system penetration make it a potential candidate drug for the treatment of insomnia and sleep disorders [1]. 2. No other information related to TCS 1102 is provided in reference [2]. 3. The orexin signaling pathway (mediated by OX₁R/OX₂R) plays a key role in regulating fear, anxiety and avoidance behavior by activating the amygdala-hypothalamus-pons neural circuit. TCS 1102 exerts anti-anxiety and anti-fear effects by blocking orexin receptors and inhibiting the activation of fear-related neural pathways, suggesting its potential application value in the treatment of anxiety disorders and post-traumatic stress disorder (PTSD) [3].

C27H26N4O2S

470.59

470.178

C, 68.91; H, 5.57; N, 11.91; O, 6.80; S, 6.81

916141-36-1

11960895

White to off-white solid powder

5.549

96.02

1

4

6

34

715

1

CN1C(SCC(N2CCC[C@H]2C(=O)NC2=CC=CC=C2C2C=CC=CC=2)=O)=NC2C=CC=CC1=2

YSBGRVXJEMSEQY-DEOSSOPVSA-N

InChI=1S/C27H26N4O2S/c1-30-23-15-8-7-14-22(23)29-27(30)34-18-25(32)31-17-9-16-24(31)26(33)28-21-13-6-5-12-20(21)19-10-3-2-4-11-19/h2-8,10-15,24H,9,16-18H2,1H3,(H,28,33)/t24-/m0/s1

(2S)-1-[2-(1-methylbenzimidazol-2-yl)sulfanylacetyl]-N-(2-phenylphenyl)pyrrolidine-2-carboxamide

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.31 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (5.31 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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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：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。