Dopamine Receptor; Adenosine A2A receptor (Ki = 2.3 nM) [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。

- 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]。

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]

- 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]。

- 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.

- 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]

- 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.

[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.

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。