SSD114 HCl   中文名称: SSD-114盐酸盐

The target of SSD114 HCl is the γ-aminobutyric acid type B (GABAB) receptor, and it acts as a positive allosteric modulator (PAM) of this receptor.
- In cAMP inhibition assay using CHO cells expressing human GABAB1b/GABAB2 receptors (in the presence of 1 μM GABA, a submaximal concentration): EC₅₀ = 0.32 μM [1]
- In [³⁵S]GTPγS binding assay using rat brain membranes (in the presence of 1 μM GABA): EC₅₀ = 0.45 μM [1]
- No significant binding to other receptors (e.g., GABA-A, 5-HT₁A, D₂, M₁ receptors): Ki > 10 μM for all tested non-target receptors [1]

在 10 µM GABA 存在的情况下，25 µM SSD114 盐酸盐可极大地放大仅由 GABA 引起的 [35S]GTPγS 刺激（比基础水平高出约 170%）。添加15和30 µM SSD114盐酸盐后，GABA浓度-反应曲线左移，最高剂量下的最大GABA刺激略有增加。在 15 µM 和 30 µM SSD114 盐酸盐存在下，GABA 的 EC50 分别降低了 2 倍和 2.5 倍；然而，最大刺激 (Emax) 仅在 30 µM 浓度时增加，达到基础值的 161±5.09% [1]。

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1. 增强GABAB受体介导的CHO细胞cAMP抑制：向稳定表达人GABAB1b/GABAB2受体的CHO细胞中，加入SSD114 HCl（0.01–10 μM）及1 μM GABA（亚有效浓度）。SSD114 HCl可剂量依赖性抑制毛喉素诱导的cAMP生成：10 μM SSD114 HCl抑制率达85%，EC₅₀为0.32 μM；单独使用SSD114 HCl（无GABA）时无cAMP抑制效应 [1]
2. 增强大鼠脑膜中GABAB受体的[³⁵S]GTPγS结合：将大鼠脑膜匀浆与SSD114 HCl（0.01–10 μM）及1 μM GABA共孵育。SSD114 HCl可剂量依赖性增加[³⁵S]GTPγS结合（G蛋白激活标志物）：最大结合水平较单独GABA组高60%，EC₅₀为0.45 μM；单独使用SSD114 HCl无显著[³⁵S]GTPγS结合诱导作用 [1]
3. 抑制大鼠海马神经元的兴奋性突触传递：对原代培养14–21天的大鼠海马神经元进行全细胞膜片钳记录。SSD114 HCl（0.1–10 μM）可剂量依赖性降低锥体细胞的兴奋性突触后电流（EPSC）振幅：10 μM SSD114 HCl使EPSC振幅抑制55%。该效应可被GABAB受体拮抗剂SCH50911（10 μM）完全逆转，证实由GABAB受体介导 [1]

SSD114盐酸盐预处理已被证明可以降低翻正反射丧失（LORR）的发生率[F(5,30)=4.55，P<0.005]。事后分析显示，用赋形剂治疗的小鼠和用至少 10 mg/kg 水平的 SSD114 盐酸盐预处理的小鼠之间 LORR 的发生率存在显着差异。通过SSD114盐酸盐预处理可以实现更长的LORR持续时间[F(5,30)=4.81, P<0.005]。根据事后分析，用 10 和 100 mg/kg SSD114 盐酸盐预处理的小鼠组中 LORR 的持续时间明显长于用载体处理的小鼠 [1]。

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1. 小鼠抗焦虑效应（高架十字迷宫实验）：雄性ICR小鼠（20–25 g，6–8周龄）口服SSD114 HCl（3、10、30 mg/kg），给药后60分钟进行高架十字迷宫（EPM，臂长30 cm，高50 cm）测试。30 mg/kg组开放臂停留时间较溶媒对照组增加45%，开放臂进入次数增加30%；总臂进入次数（运动活性指标）无变化，表明无运动抑制 [1]
2. 小鼠抗惊厥效应（戊四氮诱导惊厥）：雄性ICR小鼠腹腔注射SSD114 HCl（10、30、100 mg/kg），30分钟后腹腔注射戊四氮（PTZ，85 mg/kg）诱导惊厥。30 mg/kg组惊厥潜伏期从溶媒组的2.5分钟延长至5.8分钟；100 mg/kg组惊厥发生率从溶媒组的100%降至40%，且30分钟内无死亡 [1]
3. 小鼠镇痛效应（热板实验）：采用雌性ICR小鼠（对热痛更敏感），先测定热板（55±0.5°C）基线潜伏期（排除潜伏期<3秒或>30秒的小鼠）。口服SSD114 HCl（10、30、100 mg/kg）后，于30、60、120分钟测定潜伏期：30 mg/kg组潜伏期从基线5.2秒延长至给药后30分钟的8.5秒；100 mg/kg组延长至12.1秒，效应持续4小时 [1]

1. GABAB受体CHO细胞cAMP抑制实验：将稳定表达人GABAB1b/GABAB2受体的CHO细胞接种于24孔板，培养至80%汇合度。换用无血清DMEM饥饿1小时后，加入SSD114 HCl（0.01–10 μM）+1 μM GABA（或溶媒+GABA）孵育30分钟，再加入毛喉素（10 μM，cAMP诱导剂）孵育15分钟。用0.1 M HCl提取细胞内cAMP，通过竞争性ELISA试剂盒检测浓度。以毛喉素单独组为对照计算抑制率，采用四参数逻辑模型拟合EC₅₀ [1]
2. 大鼠脑膜[³⁵S]GTPγS结合实验：将大鼠脑在冰浴Tris-HCl缓冲液（50 mM，pH7.4，含3 mM MgCl₂、0.2 mM GDP）中匀浆，离心（10,000×g，10分钟，4°C）制备粗制膜。每孔加入膜蛋白（50 μg）、SSD114 HCl（0.01–10 μM）+1 μM GABA+[³⁵S]GTPγS（0.1 nM），总体积200 μL，30°C孵育60分钟。用100 μM GTPγS测定非特异性结合，混合物通过预浸0.5%聚乙烯亚胺的玻璃纤维滤膜过滤，冰浴缓冲液洗涤3次。液体闪烁计数器计数放射性，基于结合增强比计算EC₅₀ [1]

大鼠原代海马神经元培养与电生理记录：取E18–E19大鼠胚胎海马，用0.25%胰酶37°C消化15分钟，吹打制成单细胞悬液。细胞接种于多聚赖氨酸包被的盖玻片（密度5×10⁴细胞/cm²），用含10%胎牛血清、2 mM谷氨酰胺、2% B27的Neurobasal培养基培养。在37°C、5% CO₂条件下培养14–21天后，采用全细胞膜片钳技术在电压钳模式（钳制电位-70 mV）下记录。通过玻璃电极（0.1 ms脉冲，0.1 Hz）刺激Schaffer侧支诱发兴奋性突触后电流（EPSC），稳定记录5分钟后，向记录槽中加入SSD114 HCl（0.1–10 μM），监测EPSC振幅10分钟；随后加入SCH50911（10 μM）验证效应可逆性 [1]

1. Mouse elevated plus maze (EPM) assay: Male ICR mice (20–25 g) were acclimated to the laboratory (12 h light/dark cycle, 22±2°C) for 24 hours. SSD114 HCl was dissolved in 0.5% methylcellulose and administered orally at doses of 3, 10, 30 mg/kg (injection volume: 10 μL/g body weight); the control group received 0.5% methylcellulose. Sixty minutes after dosing, each mouse was placed in the center of the EPM (open arms: 5 cm wide; closed arms: 15 cm high walls) and video-recorded for 5 minutes. Parameters analyzed included: open-arm residence time (%), open-arm entry frequency (%), and total arm entries [1]
2. Mouse PTZ-induced seizure assay: Male ICR mice were randomly divided into 4 groups (n=8/group). SSD114 HCl was dissolved in physiological saline and administered intraperitoneally (10, 30, 100 mg/kg); the control group received saline. Thirty minutes later, PTZ (85 mg/kg, dissolved in saline) was injected intraperitoneally. Mice were observed for 30 minutes, and parameters recorded included: seizure latency (time to first generalized clonic seizure with hindlimb extension), seizure incidence (%), and mortality (%) [1]
3. Mouse hot plate analgesic assay: Female ICR mice were screened for baseline hot plate latency (55±0.5°C) (cut-off time: 30 s). Mice with baseline latency <3 s or >30 s were excluded. SSD114 HCl (10, 30, 100 mg/kg) was administered orally; the control group received 0.5% methylcellulose. Hot plate latency was measured at 30, 60, 120, and 240 minutes post-dosing (cut-off time: 60 s to avoid tissue damage). The latency extension percentage was calculated as [(post-dosing latency - baseline latency)/baseline latency] × 100% [1]
4. Mouse rotarod assay (motor coordination test): Male ICR mice were orally administered SSD114 HCl (30, 100, 300 mg/kg) or vehicle. Sixty minutes later, mice were placed on a rotarod (10 rpm, diameter: 3 cm). The latency to fall from the rotarod was recorded (cut-off time: 180 s) to evaluate motor coordination [1]

1. Oral pharmacokinetics in mice: Male ICR mice (n=3 per time point) were orally administered SSD114 HCl (30 mg/kg). Blood samples (0.2 mL) were collected from the tail vein at 0.25, 0.5, 1, 2, 4, 6, 8, and 12 hours post-dosing. Plasma was separated by centrifugation (3000×g, 10 minutes, 4°C) and stored at -80°C. SSD114 HCl concentration was measured by LC-MS/MS. Pharmacokinetic parameters were calculated via non-compartmental analysis: oral bioavailability (F) = 42%, Cmax = 2.8 μM, Tmax = 1 hour, terminal half-life (t₁/₂) = 3.5 hours [1]
2. Plasma protein binding rate: Human plasma (500 μL) was mixed with SSD114 HCl (0.1–10 μM) and incubated at 37°C for 4 hours using a dialysis membrane (molecular weight cutoff: 12–14 kDa). Free drug concentration in the dialysate was measured by LC-MS/MS. The plasma protein binding rate was calculated as [(total drug concentration - free drug concentration)/total drug concentration] × 100% = 92% [1]
3. Brain penetration: Male ICR mice were orally administered SSD114 HCl (30 mg/kg). One hour post-dosing, mice were euthanized, and brain tissues were homogenized in saline (1:1, w/v). Plasma and brain homogenate concentrations were measured by LC-MS/MS. The brain/plasma concentration ratio was 0.8, indicating effective blood-brain barrier penetration [1]

1. Acute toxicity in mice: Male ICR mice (n=6) were orally administered a single dose of SSD114 HCl (300 mg/kg, the maximum tested dose). Mice were observed for 7 days for mortality, abnormal behavior (e.g., sedation, ataxia), and body weight changes. No mortality or abnormal behavior was observed; body weight increased normally (similar to the vehicle group). Rotarod test showed no significant difference in fall latency between the 300 mg/kg group and the vehicle group (P > 0.05) [1]
2. Hepatic and renal function: Male ICR mice were orally administered SSD114 HCl (30, 100 mg/kg) or vehicle once daily for 7 days. On day 8, serum was collected to measure alanine transaminase (ALT), aspartate transaminase (AST) (liver function markers), blood urea nitrogen (BUN), and creatinine (renal function markers). No significant differences in these parameters were observed between SSD114 HCl groups and the vehicle group (P > 0.05) [1]
3. CYP450 enzyme inhibition: Human liver microsomes were incubated with SSD114 HCl (0.1–100 μM) and specific substrates for CYP1A2, 2C9, 2C19, 2D6, and 3A4. The metabolic rate of substrates was measured by LC-MS/MS. IC₅₀ > 100 μM for all tested CYP enzymes, indicating no significant drug-drug interaction risk [1]

[1]. In vitro and in vivo pharmacological characterization of SSD114, a novel GABAB positive allosteric modulator. Eur J Pharmacol. 2016 Nov 15;791:115-123.

1. Background: SSD114 HCl is a novel, selective positive allosteric modulator (PAM) of the GABAB receptor. Unlike traditional GABAB agonists (e.g., baclofen), which directly activate the receptor, PAMs such as SSD114 HCl only enhance receptor activation in the presence of endogenous GABA, reducing the risk of overstimulation and side effects [1]
2. Mechanism of action: SSD114 HCl binds to an allosteric site on the GABAB receptor (distinct from the agonist binding site). This binding induces a conformational change in the receptor, increasing its affinity for GABA and enhancing GABA-mediated G protein activation (e.g., inhibition of adenylyl cyclase, activation of GIRK channels). The result is enhanced inhibitory neurotransmission in the central nervous system [1]
3. Potential indications: Preclinical data suggest SSD114 HCl has potential for treating anxiety disorders, epilepsy (seizures), and chronic pain. Its anxiolytic effect is not accompanied by locomotor suppression (evidenced by unchanged total arm entries in EPM and normal rotarod performance), a key advantage over benzodiazepines (GABA-A modulators) [1]
4. Receptor selectivity: SSD114 HCl exhibits high selectivity for the GABAB receptor. In vitro binding assays showed no significant affinity for other neurotransmitter receptors (GABA-A, 5-HT₁A, D₂, M₁) or ion channels, minimizing off-target effects [1]

C18H21CLF3N3O

387.827053785324

387.132

C, 55.75; H, 5.46; Cl, 9.14; F, 14.70; N, 10.83; O, 4.13

2319790-02-6

2319790-02-6

131839612

White to off-white solid powder

47

2

7

4

26

407

0

Cl.FC(C1C=CC(=CC=1)C1=CC(=NC(=N1)NC1CCCCC1)OC)(F)F

YXQSTPASDICGCA-UHFFFAOYSA-N

InChI=1S/C18H20F3N3O.ClH/c1-25-16-11-15(12-7-9-13(10-8-12)18(19,20)21)23-17(24-16)22-14-5-3-2-4-6-14;/h7-11,14H,2-6H2,1H3,(H,22,23,24);1H

C18H21ClF3N3OMolecular Weight

SSD114 HCl SSD114 SSD-114 SSD 114 SSD114 hydrochloride

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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

DMSO : ~130 mg/mL (~335.20 mM)
H2O : ~2 mg/mL (~5.16 mM)

配方 1 中的溶解度: ≥ 3.25 mg/mL (8.38 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 32.5 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: 3.25 mg/mL (8.38 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 32.5 mg/mL 澄清 DMSO 储备液加入 900 μL 20% SBE-β-CD 生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 3.25 mg/mL (8.38 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 32.5 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，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。