MMP-9-related brain hemorrhage

依达拉奉对谷氨酸毒性具有治疗和预防作用。依达拉奉预处理降低了谷氨酸对 SGN 的毒性。依达拉奉可减少谷氨酸引起的坏死和细胞凋亡。通过 500 μM 依达拉奉预处理，这些改变恢复到接近正常水平。 Edaravone 对谷氨酸诱导的 SGN 细胞凋亡的保护作用与 Bcl-2 蛋白家族和 PI3K/Akt 通路相关 [4]。

在脑缺血时，依达拉奉可减少神经元损伤并防止内皮损伤，从而发挥神经保护作用。当 eNOS 存在时，可以挽救缺血性中风的有用 NOS 会增加，但当依达拉奉存在时，nNOS 和 iNOS（有毒的 NOS）会减少。依达拉奉预处理可减少溶栓治疗引起的出血事件和再灌注后脑水肿[1]。依达拉奉大大减少了梗死面积；依达拉奉组大鼠的平均梗死面积为227.6 mm3，明显小于对照组大鼠（264.0 mm3）。此外，依达拉奉治疗可减少缺血后出血量（依达拉奉治疗组大鼠为 53.4 mm3，对照组为 53.4 mm3）。 176.4 毫米）。此外，与未治疗的大鼠 (63.2%) 相比，接受依达拉奉治疗的大鼠出血量与梗死体积的比率 (23.5%) 降低 [2]。给予依达拉奉（20 mg/kg）的大鼠的胼胝体、生发基质和大脑皮层显示星形胶质细胞活性（胶质纤维酸性蛋白）和凋亡细胞（caspase-3）降低[3]。

Ho.33342和碘化丙啶（PI）双染色法检测细胞凋亡和坏死[4]
SGNs与谷氨酸和依达拉奉（500 μM）。对照细胞未经任何处理。用PBS洗涤细胞两次，用95%乙醇固定10 分钟，然后用Ho.33342染色（10 mg/mL）和PI（50 mg/mL）在37°C下保持30 min。在绿光（515–560）下通过荧光显微镜检查形态学变化 nm）和紫外线（340–380 nm）。每组在5个随机选择的场中计数至少500个细胞。所有治疗重复三次

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分光光度计检测GSH含量、SOD活性和MDA水平[4]
SGN与2 mM谷氨酸10 min，预处理或不预处理500 μM依达拉奉2 h在前面。对照细胞未经任何处理。然后用冰冷的PBS洗涤细胞两次，超声处理，并收获用于以下测定。根据制造商的说明，通过商业检测试剂盒测量所有组的细胞内GSH含量、SOD活性和MDA水平。使用分光光度计测量最佳波长下的OD值，并计算与对照细胞相比的相对水平。所有实验重复三次。

药物治疗[4]
在96孔或24孔板中传代培养的SGNs（1.0×105/mL）用2 mM谷氨酸盐10分钟。然后用正常DMEM代替培养基。将不同浓度的依达拉奉添加到培养基中20 分钟之前或2 h、 6 h、 和12 谷氨酸处理后h。所有的剂量和时间点都是通过初步实验确定的（数据未显示）

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MTT和台盼蓝染色法评估细胞活力[4]
细胞活力通过MTT法和台盼蓝染色进行定量。MTT（5 mg/mL，20 μL）加入每个孔中并孵育4 如上所述药物处理后，在37°C下放置h。除去培养基并将细胞沉淀溶解在DMSO中。然后，在570测量光密度（OD）值 nm，使用ELISA读取器。所有实验重复三次。根据以下公式计算细胞相对活力：
单间牢房  相对的  生存能力  （%）=ODexperimentODcontrol×100%
ODblank被用作零
在台盼蓝染色中，用0.4%台盼蓝将SGNs染色5 分钟。显微镜下拍照，然后计数台盼蓝阳性和阴性细胞。细胞存活率定义为阴性细胞的百分比。

Absorption, Distribution and Excretion
One study investigated the absorption of edaravone in healthy adults, who either received a single oral (105 mg/mL) or intravenous (60 mg/60 min) dose. The mean Cmax (CV%) and Tmax were 1656 (44.3) ng/mL and 0.5 hours, respectively, following oral administration. The absolute oral bioavailability is about 57% because of first-pass metabolism. The mean Cmax (CV%) and Tmax were 1253 (18.3) ng/mL and one hour, respectively, following intravenous administration. When intravenously administered, the maximum plasma concentration (Cmax) of edaravone was reached by the end of infusion. The Cmax and area under the concentration-time curve (AUC) of edaravone increases more than dose-proportional over the dose range of 30 to 300 mg. Edaravone does not accumulate in plasma with once-daily or multiple-dose administration. The Cmax and AUC decreased when the oral suspension formulation of edaravone was administered with a high-fat meal.
In Japanese and Caucasian healthy volunteer studies, edaravone was excreted mainly in the urine as its glucuronide conjugate (60-80% of the dose up to 48 hours). Approximately 6-8% of the dose was recovered in the urine as the sulfate conjugate, and <1% of the dose was recovered in the urine as the unchanged drug. _In vitro_ studies suggest that the sulfate conjugate of edaravone is hydrolyzed back to edaravone, which is then converted to the glucuronide conjugate in the kidney before excretion into the urine.
After intravenous administration, edaravone has a mean volume of distribution of 63.1 L, suggesting substantial tissue distribution. Edaravone has an apparent volume of distribution of 164 L following oral administration. Edaravone readily crosses the blood-brain barrier.
Following intravenous administration, the total clearance of edaravone is estimated to be 35.9 L/h. The apparent total clearance of edaravone is estimated to be 67.9L/h following oral administration.

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Metabolism / Metabolites
The metabolites of edaravone have not been fully characterized. Edaravone is metabolized to a sulfate conjugate and a glucuronide conjugate, which are not pharmacologically active. The glucuronide conjugation of edaravone involves multiple uridine diphosphate glucuronosyltransferase (UGT) isoforms (UGT1A1, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B7, and UGT2B17). In human plasma, edaravone is mainly detected as the sulfate conjugate, which is presumed to be formed by sulfotransferases. Oral edaravone results in 1.3- and 1.7-fold higher exposures for both sulfate and glucuronide metabolites, respectively, when compared to intravenously-administered edaravone because of first-pass metabolism.

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Biological Half-Life
The mean terminal elimination half-life of edaravone is approximately 4.5 to nine hours. The half-lives of its metabolites range from three to six hours.

Hepatotoxicity
Serum aminotransferase elevations occur in a small proportion of patients on edaravone therapy, but the frequency, timing of onset, duration and severity of these elevations has not been defined. The rates of abnormal liver tests during edaravone therapy were said to be similar to those during placebo treatment. Most elevations resolved spontaneously, and there were no reports of drug discontinuation for serum enzyme elevations. Clinically apparent liver injury due to edaravone was not reported in the prelicensure trials and has not been reported with subsequent clinical use of edaravone, but the numbers of patients treated have been few. Thus, clinically apparent liver injury from edaravone must be rare if it occurs at all.
Likelihood score: E (unlikely cause of clinically apparent liver injury).

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Protein Binding
Edaravone is 92% bound to human serum proteins, mainly to albumin, with no concentration dependence in the range of 0.1 to 50 micromol/L.

[1]. Neuroprotective effects of edaravone: a novel free radical scavenger in cerebrovascular injury. CNS Drug Rev, 2006. 12(1): p. 9-20.

[2]. Edaravone, a free radical scavenger, attenuates cerebral infarction and hemorrhagic infarction in rats with hyperglycemia. Neurol Res, 2013.

[3]. Edaravone reduces astrogliosis and apoptosis in young rats with kaolin-induced hydrocephalus. Childs Nerv Syst. 2016 Dec 17. [Epub ahead of print].

[4]. Protective Effect of Edaravone on Glutamate-Induced Neurotoxicity in Spiral Ganglion Neurons. Neural Plast. 2016;2016:4034218. Epub 2016 Nov 10.

1-phenyl-3-methyl-5-pyrazolone appears as white to off-white powder or crystals. (NTP, 1992)
Edaravone is a pyrazolone that is 2,4-dihydro-3H-pyrazol-3-one which is substituted at positions 2 and 5 by phenyl and methyl groups, respectively. It has a role as a radical scavenger and an antioxidant.
Edaravone is a free radical scavenger and neuroprotective agent with antioxidant properties. It has three tautomers. Edaravone works to scavenge reactive oxygen species, which have been implicated in neurological disorders, such as amyotrophic lateral sclerosis (ALS) and cerebral ischemia. The intravenous formulation of edaravone was first approved in Japan in 2001 for the treatment of acute ischemic stroke. It was later approved for the treatment of amyotrophic lateral sclerosis (ALS) in Japan and South Korea in 2015, followed by the FDA approval in May 2017 and Health Canada approval in October 2018. The oral suspension formulation of edaravone was approved by the FDA in May 2022 and by Health Canada in November 2022. Edaravone was initially granted orphan designation by the European Medicines Agency on June 19, 2015 and was under regulatory review in Europe. However, the drug manufacturer, Mitsubishi Tanabe Pharma, withdrew the Marketing Authorization Application (MAA) for edaravone from the European market on May 24, 2019, in response to the request made by the Committee for Medicinal Products for Human Use (CHMP) for a long-term study demonstrating the long-term efficacy and safety of edaravone. Edaravone was also investigated in other disorders, such as Alzheimer's disease, neuropathic pain, and ischemia-induced nerve injury.
Edaravone is a free radical scavenger and neuroprotective agent used for therapy of amyotrophic lateral sclerosis. Edaravone is associated with a low rate of serum aminotransferase elevations during therapy but has not been linked to instances of clinically apparent, acute liver injury.
Edaravone has been reported in Homo sapiens with data available.
An antipyrine derivative that functions as a free radical scavenger and neuroprotective agent. It is used in the treatment of AMYOTROPHIC LATERAL SCLEROSIS and STROKE.

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Drug Indication
Edaravone is indicated for the treatment of amyotrophic lateral sclerosis (ALS) in the US and Canada. It is also indicated to treat acute ischemic stroke in Japan.
Treatment of amyotrophic lateral sclerosis
Treatment of amyotrophic lateral sclerosis

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Mechanism of Action
Oxidative stress and reactive oxygen species (ROS) production have been implicated in various neurological disorders, such as amyotrophic lateral sclerosis (ALS) and cerebral ischemia. Oxidative stress caused by excess ROS damages endothelial cells in the cerebral vasculature as well as neuronal cell membranes, leading to neuronal cell death. Edaravone is a free radical scavenger that scavenges and suppresses the generation of hydroxyl radicals and peroxynitrite radicals. The exact mechanism of action of edaravone in ALS has not been fully elucidated; however, edaravone is thought to mediate therapeutic effects via its antioxidant properties. Since oxidative stress has been implicated in the pathophysiology of ALS and cerebral ischemia, inhibiting lipid peroxidation, suppressing endothelial cell damage induced by lipid peroxides, and scavenging free radicals may lead to neuroprotective effects. Edaravone has no effect on superoxide production. It is suggested that edaravone may also possess anti-inflammatory properties, as it inhibited neutrophil activation and suppressed inducible nitric oxide synthase (iNOS) and neuronal nitric oxide synthase (nNOS) expression in animal models. It was also shown to ameliorate ROS-induced inflammatory oxidative stress after ischemic brain reperfusion.

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Pharmacodynamics
Edaravone works to delay the disease progression of neurological disorders such as ischemic stroke and ALS by limiting the extent of neuronal damage or death.

C10H10N2O

174.2

174.079

C, 68.95; H, 5.79; N, 16.08; O, 9.18

89-25-8

Edaravone-d5;1228765-67-0

4021

Light yellow to yellow solid

1.2±0.1 g/cm3

333.0±11.0 °C at 760 mmHg

126-128 °C(lit.)

155.2±19.3 °C

0.0±0.7 mmHg at 25°C

1.606

0.44

32.67

0

2

1

13

241

0

O=C1CC(C)=NN1C1C=CC=CC=1

QELUYTUMUWHWMC-UHFFFAOYSA-N

InChI=1S/C10H10N2O/c1-8-7-10(13)12(11-8)9-5-3-2-4-6-9/h2-6H,7H2,1H3

5-methyl-2-phenyl-2,4-dihydro-3H-pyrazol-3-one

MCI-186; NCI-C03952; MCI 186; NSC 12; MCI186; Radicut; trade name: Radicava; Methylphenylpyrazolone; Norantipyrine; Norphenazone; Phenylmethylpyrazolone; Arone

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 (14.35 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 (14.35 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 25.0 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: ≥ 2.5 mg/mL (14.35 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。