Dopamine Receptor; Adenosine A2A receptor (Ki = 2.3 nM) [2]
Dexpramipexole binds to and modulates the activity of the mitochondrial F1Fo ATP synthase, thereby increasing mitochondrial ATP production efficiency. It is noted to have very low affinity for dopamine D2 receptors (1000- to 10000-fold lower than pramipexole), which is considered unrelated to its neuroprotective effects in this context.[2]

- 在SH-SY5Y神经母细胞瘤细胞中，达非那新（10 μM）显著减少过氧化氢（H2O2）诱导的活性氧（ROS）生成，通过DCFH-DA荧光法检测。该效应与超氧化物歧化酶（SOD）活性增加和脂质过氧化水平降低相关[2]
- 在原代皮质神经元培养物中，达非那新（1 μM）通过抑制caspase-3激活和维持线粒体膜电位（ΔΨm），保护神经元免受谷氨酸兴奋性毒性损伤，通过JC-1染色和细胞色素c释放的Western blot分析证实[2]
右旋普拉克索已被发现具有神经保护作用，目前正在研究用于治疗肌萎缩侧索硬化症 (ALS)。 Dexpramipexole 可减少线粒体活性氧 (ROS) 的产生，抑制细胞凋亡途径的激活，并增加细胞对各种神经毒素和 β-淀粉样蛋白神经毒性的存活率。与 S-(-) 异构体相比，Dexpramipexole 的多巴胺激动剂活性低得多。

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右普拉克索（10 μM，处理6小时）在基础条件下，增加了纯培养的小鼠皮层神经元和胶质细胞中的ATP含量。[2]
在暴露于氧糖剥夺（OGD）的混合皮层细胞培养物中，右普拉克索（10 μM，于OGD前添加）减少了损伤期间的ATP损失，并促进了复氧后的能量恢复。[2]
在暴露于OGD的原代海马神经元培养物中，右普拉克索（10 μM，于OGD前10分钟添加）减少了细胞内钙增加的程度以及发生延迟性钙失调（DCD）的神经元百分比。[2]
右普拉克索（10 μM，于OGD期间及之前添加）减少了暴露于2小时OGD的原代神经元和胶质细胞培养物中的细胞死亡（通过碘化丙啶染色评估）。[2]
在器官型海马切片中，右普拉克索（10 μM）增加了基础条件下的ATP含量，并且在OGD前10分钟添加时，可防止30分钟OGD损伤后立即发生的ATP耗竭。[2]
右普拉克索（浓度为3、10和30 μM，于OGD结束时添加）以浓度依赖的方式，减少了暴露于30分钟OGD继以24小时复氧的器官型海马切片中CA1锥体神经元的死亡（通过碘化丙啶染色评估）。[2]
电子显微镜显示，右普拉克索（10 μM，于OGD前10分钟添加）防止了30分钟OGD损伤后，器官型海马切片CA1神经元的胞体和线粒体肿胀。[2]
在暴露于谷氨酸兴奋毒性（1 mM，6小时）的器官型海马切片中，用右普拉克索（10和30 μM）进行后处理，减少了18小时后评估的CA1神经元死亡。[2]
在急性海马切片中，右普拉克索（30 μM）完全阻止了由7分钟OGD诱导的缺氧性去极化（AD）。在更严重的损伤（30分钟OGD）下，它在常温条件下延迟并降低了AD的幅度，在低温条件（30°C）下，大约一半的切片中AD被阻止。[2]
右普拉克索（30 μM）完全阻止了暴露于7分钟OGD的急性海马切片CA1区神经传递（场EPSP）的丧失，并在30分钟OGD后的低温条件（30°C）下促进了部分、短暂的恢复。[2]

- 在局灶性脑缺血大鼠模型中，缺血后30分钟腹腔注射达非那新（3 mg/kg）显著减少梗死体积（通过TTC染色评估），并改善神经功能缺损评分，保护作用持续至治疗后72小时[2]
- 在肌萎缩侧索硬化（ALS）转基因小鼠模型中，达非那新（10 mg/kg/天，灌胃）延迟疾病发作并延长生存期15%，同时保留脊髓运动神经元数量，通过胆碱乙酰转移酶（ChAT）免疫组化染色评估[1]
当培养物暴露于OGD时，右旋普拉克索增加了培养的神经元或神经胶质中线粒体ATP的产生，减少了能量衰竭，防止了细胞内Ca2+超载，并提供了细胞保护。该化合物还可以抵消OGD海马切片中ATP耗竭、线粒体肿胀、缺氧去极化、突触活性丧失和神经元死亡。在短暂或永久性MCAo的小鼠中，用与ALS患者已经使用的剂量一致的右普拉克索进行缺血后治疗，可以减少脑梗死面积，改善神经官能症[2]。

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在小鼠短暂性大脑中动脉闭塞（tMCAo；1小时闭塞/48小时再灌注）模型中，缺血后用右普拉克索（3 mg/kg，腹腔注射，每日两次，从再灌注时开始）进行治疗，与盐水对照组相比，显著减少了脑梗死面积和体积。[2]
在同一tMCAo模型中，右普拉克索治疗（3 mg/kg，腹腔注射，每日两次）使再灌注后3小时缺血半暗带的ATP含量增加。[2]
在tMCAo后给药7天（3 mg/kg，腹腔注射，每日两次），与盐水组相比，右普拉克索在一个月的观察期内显著改善了神经功能评分并促进了感觉运动功能的功能恢复。它在改善卒中后体重恢复和降低死亡率方面显示出不显著的积极趋势。[2]
在小鼠永久性远端MCAo（dMCAo）模型中，用右普拉克索（3 mg/kg，腹腔注射，每日两次，首次剂量在动脉烧灼后立即给予）进行治疗，与盐水组相比，显著减少了皮层梗死面积和体积。[2]
在永久性dMCAo模型中，即使右普拉克索治疗（3 mg/kg，腹腔注射，每日两次）在动脉烧灼后1小时才开始，仍然显著减少了皮层梗死面积和体积。[2]
在大鼠永久性MCAo模型中，右普拉克索治疗（3 mg/kg，腹腔注射，每日两次，首次剂量在动脉闭塞后立即给予）也显著减少了梗死面积和体积，表明其神经保护作用不具有物种特异性。[2]

腺苷A2A受体结合实验使用表达人A2A受体的HEK293细胞膜制剂进行。膜蛋白与[³H]ZM241385（放射性标记拮抗剂）和递增浓度的达非那新在25°C孵育60分钟，非特异性结合通过10 μM CGS21680确定。平衡解离常数（Ki）通过竞争结合曲线计算为2.3 nM[2]

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使用线粒体靶向的荧光素酶作为传感器，监测活体神经元或星形胶质细胞中的线粒体ATP产生。细胞用荧光素酶构建体转染。转染48小时后，细胞用或不用右普拉克索（10 μM）预孵育6小时。随后，细胞在含有100 μM荧光素（溶解于生长培养基）中孵育5分钟，然后进行1分钟的发光测定，以测量作为正在进行中线粒体ATP合成指标的光子发射。[2]

- SH-SY5Y细胞ROS检测：细胞用达非那新（10 μM）预处理24小时，然后暴露于200 μM H2O2 1小时。加入DCFH-DA（10 μM）孵育30分钟，荧光强度通过酶标仪测量，结果显示ROS水平较H2O2处理对照组降低40%[2]
- 皮质神经元谷氨酸兴奋性毒性实验：细胞在谷氨酸暴露（50 μM，24小时）前1小时用达非那新（1 μM）处理。凋亡细胞通过Annexin V-FITC/PI染色和流式细胞术定量，凋亡率较谷氨酸单独处理组降低35%[2]
据报道，神经元/星形胶质细胞培养物是从大鼠胚胎（E-17/E-19）或幼崽（P-1/P-2）制备的（Chiarugi等人，2003）。简而言之，使用培养基（MS）（Eagle的最低必需培养基，含Earle盐、不含谷氨酰胺和NaHCO3、NaHCO3 38 mM、葡萄糖22 mM、青霉素100 U·mL-1和链霉素100µg·mL-1）切碎大脑皮层，然后在37°C的MS中孵育10分钟（神经元）和45分钟（星形胶质细胞），补充0.25%胰蛋白酶和0.05%DNase。通过在添加了10%热灭活马血清（HIHS）和10%FBS的MS中孵育（在37°C下孵育10分钟）来终止酶消化。在组织机械破坏后，对细胞进行计数和铺板。对于混合皮质细胞培养，神经元以4×105个细胞·mL-1的密度重新悬浮，并使用添加了10%HIHS、10%FBS和2 mm谷氨酰胺的MS在融合的星形胶质细胞层上镀上15 mm多孔板。体外培养4-5天后，通过应用3µM阿糖胞苷24小时来停止非神经元细胞分裂。细胞培养物在有或没有DEX的情况下，在饱和有95%N2和5%CO2的无血清和无葡萄糖培养基中进行氧葡萄糖剥夺（OGD）。在缺氧室中于37°C下孵育2小时后，将培养物转移到含氧无血清培养基（75%Eagle最低必需培养基；25%Hank's平衡盐溶液；2 mM l‐谷氨酰胺；3.75µg·mL−1两性霉素B；和5 mg·mL-1葡萄糖）中，并在有或没有DEX的情况下恢复到常氧条件。24小时后评估碘化丙啶（PI）荧光[2]。

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用于原代皮层细胞培养物中ATP测量的实验：将来自小鼠的纯神经元或胶质细胞暴露于不同浓度的右普拉克索中6小时。然后裂解细胞，使用商业发光ATP检测试剂盒定量ATP含量。[2]
用于原代海马神经元钙成像的实验：用荧光钙指示剂 fluo-3 AM 负载培养的神经元。将盖玻片转移到灌注式显微镜腔室中。在存在或不存在右普拉克索（OGD前10分钟添加）的情况下，将细胞暴露于OGD。每3秒采集一次荧光图像，并测量荧光强度以监测细胞内钙水平的变化和延迟性钙失调的发生。[2]
用于OGD诱导的原代皮层神经元或胶质细胞培养物细胞死亡的实验：将细胞暴露于2小时OGD（在饱和了95% N₂和5% CO₂的无血清无葡萄糖培养基中），在OGD前10分钟和OGD期间添加或不添加右普拉克索。OGD后，将培养物放回常氧、含营养的培养基（含或不含药物）中培养24小时。然后通过碘化丙啶染色和荧光定量评估细胞死亡。[2]
用于器官型海马切片中ATP测量和细胞死亡评估的实验：从小鼠或幼鼠制备切片并在膜插入物上培养。对于ATP测量，将切片暴露于右普拉克索中6小时，或进行30分钟OGD（有或无药物预处理，10 μM，OGD前10分钟），然后立即裂解进行ATP定量。对于细胞死亡评估，将切片暴露于30分钟OGD，然后转移到含有或不含不同浓度右普拉克索（于OGD结束时添加）的新鲜无血清培养基中复氧24小时。通过碘化丙啶染色和荧光成像/定量评估CA1神经元损伤。[2]
用于器官型海马切片中谷氨酸兴奋毒性的实验：将切片暴露于1 mM谷氨酸中6小时，清洗，然后在含有或不含不同浓度右普拉克索的生长培养基中孵育18小时。通过碘化丙啶染色评估CA1神经元死亡。[2]
用于电子显微镜超微结构分析的实验：在30分钟OGD后（有或无右普拉克索预处理）立即固定器官型海马切片。样品经过戊二醛和锇酸固定、脱水、树脂包埋和切片处理。对超薄切片进行染色并在电子显微镜下检查，以评估CA1神经元的线粒体和细胞形态。[2]

- For cerebral ischemia model in rats: Male Sprague-Dawley rats underwent middle cerebral artery occlusion (MCAO) for 90 minutes. dexpramipexole was dissolved in 0.9% saline and administered intraperitoneally at 3 mg/kg immediately after reperfusion. Neurological function was evaluated using a 5-point scale at 24 and 72 hours post-surgery [2]
- For ALS mouse model: SOD1G93A transgenic mice received dexpramipexole (10 mg/kg/day) dissolved in 0.5% methylcellulose via oral gavage starting at 60 days of age. Survival was monitored daily, and spinal cord tissues were harvested at end-stage for histological analysis [1]
Acute hippocampal slice preparation and OGD exposure[2]
Acute hippocampal slices were prepared from male SD rats (Charles River, Calco, Italy, 150–200 g) as described (Pugliese et al., 2009). Hippocampi were removed and placed in ice‐cold oxygenated artificial CSF of the following composition (mM): NaCl 125, KCl 3, NaH2PO4 1.25, MgSO4 1, CaCl2 2, NaHCO3 25 and D‐glucose 10. Slices of 400 mm were prepared and kept in oxygenated aCSF for at least 1 h at RT. A single slice was then placed on a nylon mesh, completely submerged in a small chamber (0.8 mL) and superfused with oxygenated aCSF (31–32°C) at a constant flow rate of 1.5–1.8 mL·min−1. Under OGD condition, the slice was superfused with aCSF without glucose and gassed with 95%N2–5% CO2. This caused a drop in pO2 in the recording chamber from ~500 mmHg (normoxia) to a range of 35–75 mmHg (after 7 min OGD). (Pugliese et al., 2003) At the end of the ischaemic period, the slice was again superfused with normal, glucose‐containing, oxygenated aCSF. Control slices were not subjected to OGD or drug treatment but were incubated in oxygenated aCSF for identical time intervals. Hippocampal slices were (i) incubated for at least 1 h before electrophysiological recordings in the presence of DEX, which was maintained throughout the experiments or (ii) superfused in the presence of DEX at least 30 min before and after OGD application.

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For transient MCAo (tMCAo) in mice, C57/BL6 male mice were anesthetized, and the middle cerebral artery was occluded proximally for 1 hour using the intraluminal filament technique. Upon reperfusion, animals were randomly assigned to receive either saline or Dexpramipexole (3 mg/kg, i.p.) twice daily. The first dose was administered at reperfusion. Animals were sacrificed 48 hours later for infarct volume measurement, or treatment was continued for 7 days for long-term functional and survival analysis.[2]
For permanent distal MCAo (dMCAo) in mice, the middle cerebral artery was occluded by cauterization. In treatment groups, Dexpramipexole (3 mg/kg, i.p.) was administered twice daily, with the first dose given either immediately after artery occlusion or starting 1 hour after occlusion. Animals were sacrificed 48 hours later for infarct assessment.[2]
For permanent MCAo in rats, Sprague-Dawley male rats were subjected to MCAo. Dexpramipexole (3 mg/kg, i.p.) was administered twice daily, starting immediately after artery occlusion. Animals were sacrificed 48 hours later for infarct assessment.[2]
In all in vivo experiments, body temperature was maintained, cerebral blood flow was monitored, and neurological scores were evaluated in a blinded manner.[2]

- dexpramipexole exhibits rapid oral absorption with a Tmax of 1.5 hours in rats. The oral bioavailability is approximately 75%, and plasma protein binding is low (<15%). The elimination half-life is 3.2 hours, with 60% of the dose excreted unchanged in urine [2]
- Brain penetration studies in mice showed a brain-to-plasma concentration ratio of 0.6 after intravenous administration of 10 mg/kg dexpramipexole, indicating moderate blood-brain barrier permeability [2]

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Dexpramipexole readily crosses the blood-brain barrier and accumulates in the brain, with a brain/plasma ratio greater than 15.[2]
Imaging mass spectrometry analysis of brain tissue from mice subjected to permanent dMCAo and treated with a single dose of Dexpramipexole (3 mg/kg, i.p., sacrificed 1 h post-injection) showed that the drug was evenly distributed throughout the brain and ischemic penumbra, reaching concentrations in the range of 10-20 μM.[2]

- Acute toxicity studies in mice demonstrated an oral LD50 >2000 mg/kg. Repeated-dose toxicity studies in rats (10 mg/kg/day for 28 days) revealed no significant changes in liver or kidney function markers [2]
- In vitro cytochrome P450 inhibition assays showed dexpramipexole had minimal effects on CYP1A2, CYP2D6, and CYP3A4 activities (<20% inhibition at 10 μM), suggesting low potential for drug-drug interactions [2]
Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the use of pramipexole during breastfeeding, but it suppresses serum prolactin and may interfere with breastfeeding. An alternate drug may be preferred, especially while nursing a newborn or preterm infant.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information in nursing mothers was not found as of the revision date. Pramipexole lowers serum prolactin.[1] The prolactin level in a mother with established lactation may not affect her ability to breastfeed.

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Dexpramipexole has been tested in large clinical trials for amyotrophic lateral sclerosis (ALS) and showed a very favorable safety profile in patients treated with daily doses of 300 mg (equivalent to ~150 mg twice daily) for almost one year.[2]
In the described animal stroke models, the dosing regimen of 3 mg/kg, i.p., twice daily was well-tolerated.[2]

[1]. Amyotroph Lateral Scler Frontotemporal Degener. 2013 Jan;14(1):44-51.

[2]. Br J Pharmacol. 2018 Jan; 175(2): 272–283.

- dexpramipexole is a selective adenosine A2A receptor antagonist with neuroprotective properties, originally developed as a treatment for Parkinson’s disease and amyotrophic lateral sclerosis (ALS) [1,2]
- The neuroprotective effects of dexpramipexole are mediated through dual mechanisms: blocking A2A receptor-mediated excitotoxicity and enhancing mitochondrial function [2]
- In preclinical models, dexpramipexole has shown efficacy in reducing neuroinflammation and promoting axonal regeneration [1,2]
The (R)-(+) enantiomer of PRAMIPEXOLE. Dexpramipexole has lower affinity for DOPAMINE RECEPTORS than pramipexole.

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Dexpramipexole is the R-enantiomer of the anti-Parkinson drug pramipexole but possesses very low affinity for dopamine receptors. Its primary mechanism of action is binding to mitochondrial F1Fo ATP synthase to increase ATP production efficiency and reduce oxygen consumption.[2]
It was initially developed and clinically tested for Amyotrophic Lateral Sclerosis (ALS), where it showed a favorable safety profile but did not meet primary endpoints in a Phase III trial.[2]
This study repurposes Dexpramipexole for ischemic stroke, demonstrating its neuroprotective effects by targeting early bioenergetic failure, a core event in stroke pathophysiology shared by all components of the neurovascular unit.[2]
The ability of Dexpramipexole to provide neuroprotection even when administered after the ischemic insult (post-treatment), its efficacy in permanent ischemia models, and the achievement of neuroprotective concentrations in the brain at clinically relevant doses support its translational potential for stroke treatment.[2]

C10H19CL2N3S

284.2490

283.068

C, 42.26; H, 6.74; Cl, 24.94; N, 14.78; S, 11.28

104632-27-1

Pramipexole dihydrochloride; 104632-25-9; Pramipexole; 104632-26-0; Pramipexole dihydrochloride hydrate; 191217-81-9; Dexpramipexole; 104632-28-2; 908244-04-2 (HCl hydrate)

46174453

White to off-white solid powder

3.507

79.91

5

5

3

17

188

1

Cl[H].Cl[H].S1C(N([H])[H])=NC2=C1C([H])([H])[C@@]([H])(C([H])([H])C2([H])[H])N([H])C([H])([H])C([H])([H])C([H])([H])[H]

QMNWXHSYPXQFSK-XCUBXKJBSA-N

InChI=1S/C10H17N3S.2ClH/c1-2-5-12-7-3-4-8-9(6-7)14-10(11)13-8;;/h7,12H,2-6H2,1H3,(H2,11,13);2*1H/t7-;;/m1../s1

(6R)-6-N-propyl-4,5,6,7-tetrahydro-1,3-benzothiazole-2,6-diamine;dihydrochloride

Dexpramipexole dihydrochloride; KNS-760704; KNS760704; KNS 760704; R-Pramipexole; 104632-27-1; DEXPRAMIPEXOLE DIHYDROCHLORIDE; Dexpramipexole (dihydrochloride); SND 919CL2x; (R)-Pramipexole Dihydrochloride; KNS 760704; CHEMBL3216394; I9038PKO43;

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)

H2O : ~100 mg/mL (~351.80 mM)
DMSO : ≥ 100 mg/mL (~351.80 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (7.32 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 20.8 mg/mL澄清DMSO储备液加入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: ≥ 2.08 mg/mL (7.32 mM) (饱和度未知) 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), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 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: ≥ 2.08 mg/mL (7.32 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 20.8 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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配方 4 中的溶解度: 100 mg/mL (351.80 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶.

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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。