ODQ

sGC/soluble guanylyl cyclase (nitric oxide-activated enzyme)
Rat cardiomyoblasts are exposed to H2O2 in vitro for 4 hours, which reduces mitochondrial respiration because it produces hydroxyl radicals. ODQ pretreatment of the cells does not prevent this cell damage. Additionally, superoxide anions are not scavenged by ODQ. [1]

在30和50 μM时，ODQ会显着着诱导NCI-H2452细胞，分别将丙酮水平提高12倍和15倍。当浓度为10μM时，浓度低于ODQ诱导细胞的阈值，ODQ与顺铂联合使用可增强（实际上是双倍）1 μM 顺铂的促进丙酮作用[1]。

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在大鼠心室肌母细胞（H9c2细胞）中，预孵育 ODQ（0.1 μM 至 1 mM）未能减轻由 H₂O₂（1 mM，4 小时）引起的线粒体呼吸功能下降。[1]
在无细胞的次黄嘌呤/黄嘌呤氧化酶测定系统中，ODQ（0.1 μM 至 1 mM）未能清除超氧阴离子，该结果通过化学发光法测定。[1]

ODQ (2 mg/kg；ip) 减少麻醉体内由革兰氏急性或革兰氏阴性菌壁废物引起的多器官损伤和功能障碍[2]。 动物模型：麻醉的雄性 Wistar 大鼠[2]剂量：2 mg/kg 给药方式：腹腔注射 结果：减轻脂磷壁酸/肽聚糖或脂多糖引起的肾功能障碍、肺损伤和肝细胞损伤。

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在体内，给予脂磷壁酸/肽聚糖或脂多糖会在6小时内导致低血压、急性肾功能不全、肝细胞损伤和肺损伤。用ODQ预处理大鼠可减轻脂磷壁酸/肽聚糖或脂多糖引起的肾功能障碍、肺损伤和肝细胞损伤。在体外，对大鼠心肌母细胞施用H2O2（4小时）可降低线粒体呼吸，这是由于羟基自由基的产生。用ODQ预处理细胞并不能消除这种细胞损伤。此外，ODQ不清除超氧阴离子。 结论：这些结果表明，鸟苷酸环化酶抑制剂ODQ可以减轻麻醉大鼠中革兰氏阳性或革兰氏阴性细菌壁碎片引起的多器官损伤和功能障碍。观察到的ODQ的保护作用不是由于ODQ减少超氧阴离子或羟基自由基的形成或影响的能力[1]。

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在大鼠中，预处理 ODQ（2 mg/kg，腹腔注射，诱导休克前 2 小时）可减轻由脂磷壁酸 + 肽聚糖（革兰氏阳性休克）或脂多糖（革兰氏阴性休克）引起的肾功能障碍（降低血浆尿素和肌酐水平）和肝细胞损伤（降低血浆AST和ALT水平）。[1]
在革兰氏阳性和革兰氏阴性休克模型中，预处理 ODQ 可减轻肺、肾和肝的组织学损伤（炎性浸润、细胞坏死）。[1]
预处理 ODQ 可显著降低休克大鼠肺、肾和肝组织中硝基酪氨酸的免疫染色（硝基酪氨酸是过氧亚硝酸盐/活性氧介导的蛋白质硝化的指标）。[1]
ODQ 未能预防由脂磷壁酸/肽聚糖或脂多糖引起的低血压，尽管它减轻了器官损伤。[1]

评估ODQ对超氧阴离子生成的影响。[1]
如前所述，我们使用次黄嘌呤/黄嘌呤氧化酶测定法产生超氧阴离子。简而言之，以黄嘌呤氧化酶水溶液（2单位/mL）和次黄嘌呤（0.7 mmol/L）为底物。通过在磷酸盐缓冲盐水中搅拌等重量（10mg/mL）的牛血清白蛋白和鲁米诺过夜来制备鲁米诺结合白蛋白。然后将溶液加热至40°C以增加鲁米诺的溶解度。溶液通过0.22μm过滤器过滤，得到澄清溶液，冷冻保存。根据Trevisthick等人的研究，使用DMSO稳定化学发光。将溶液与ODQ（0.1μM至1 mM）一起孵育，并在测量期前加入黄嘌呤氧化酶/次黄嘌呤。用市售计数器每13秒记录一次化学发光。所有实验均进行了两次n=4的观察。

H2O2 是研究活性氧如何影响各种器官的有用工具。它很容易渗透穿过细胞膜，在过渡金属存在下，在细胞内发生芬顿反应，转化为有毒的羟基自由基。使用 ODQ（0.1 mM 至 1 mM）、盐水或 DMSO（含有 10% DMSO 的培养基）在 37°C 下将细胞预孵育两小时。在 37°C 下暴露于介质或 H2O2 (1 mM) 4 小时后，测量细胞的损伤程度。每个实验使用 n = 4 个观察值运行两次。

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ODQ对H2O2引起的大鼠心肌母细胞损伤的影响评估：实验设计。[1]
H2O2是研究活性氧对不同器官影响的有用工具。它可以很容易地扩散穿过细胞膜（36），并在过渡金属存在下通过芬顿反应在细胞内位点转化为有毒羟基自由基。细胞用a）ODQ（0.1μM至1 mM）、b）生理盐水或C）DMSO（含10%DMSO的培养基）预孵育（2小时，37°C）。然后将细胞暴露于a）培养基或b）H2O2（1 mM，37°C下4小时），之后评估细胞损伤（见前面的描述）。所有实验均进行了两次n=4的观察。

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大鼠心室肌母细胞（H9c2细胞）在补充有L-谷氨酰胺和胎牛血清的DMEM培养基中培养，并于96孔板中生长。细胞与 ODQ（0.1 μM 至 1 mM）、生理盐水或DMSO在37°C预孵育2小时，然后暴露于H₂O₂（1 mM）中4小时。通过MTT法评估细胞活力，测量MTT还原为甲臜的量，并在550 nm波长下读取吸光度。[1]
使用无细胞化学发光法评估超氧阴离子清除能力。次黄嘌呤/黄嘌呤氧化酶系统产生超氧阴离子，鲁米诺结合的白蛋白作为检测剂。将 ODQ（0.1 μM 至 1 mM）与系统共孵育，每13秒记录一次化学发光。[1]

Anesthetized, male Wistar rats
2 mg/kg
I.p.

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For the in vivo portion of the study, after surgical preparation, anesthetized rats were observed for 6 hrs. All rats were pretreated and received an intravenous infusion of saline (1.5 mL·kg−1·hr−1), which was maintained throughout the experiment. The rats were assigned to nine groups. Group 1 contained control rats (sham) subjected to 2 mL/kg saline intraperitoneally, 2 hrs before the experiment (n = 7). Group 2 contained control rats (sham) that received 2 mg/kg ODQ intraperitoneally, 2 hrs before the experiment (n = 9). Group 3 contained control rats (sham) that received 2 mL/kg dimethyl sulfoxide, 30% v/v in saline intraperitoneally, as a vehicle for ODQ, 2 hrs before the experiment (n = 6). In group 4 rats, Gram-positive shock was induced by coadministration of lipoteichoic acid (3 mg/kg intravenously) and peptidoglycan (10 mg/kg intravenously) (n = 10). In group 5, rats were pretreated with ODQ (as described previously) before lipoteichoic acid/peptidoglycan (n = 9). In group 6, rats were pretreated with dimethyl sulfoxide (as de- scribed previously) before lipoteichoic acid/peptidoglycan (n = 7). In group 7, Gram-negative shock was induced by lipopolysaccharide (6 mg/kg intravenously) (n = 11). In group 8, rats were pretreated with ODQ (as described previously) before lipopolysaccharide (n = 8). In group 9, rats were pretreated with dimethyl sulfoxide (as described previously) before lipopolysaccharide (n = 8).[1]

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Evaluation of the Effects of ODQon Circulatory Failure and Multiple Organ Dysfunction Syndrome: Experimental Design.[1]
Nine experimental groups were studied. After an injection of drugs (saline, 2 mL/kg intraperitoneally; ODQ, 2 mg/kg intraperitoneally; or dimethyl sulfoxide [DMSO] as vehicle for ODQ, 2 mL/kg DMSO 30% v/v in saline intraperitoneally), rats were connected to an infusion of saline (1.5 mL·kg−1·hr−1 intravenously), which was maintained throughout the experiment. The nine groups were as follows:
Control rats were treated with saline 2 hrs before the experiment (sham saline, n = 6).
Control rats were treated with ODQ 2 hrs before the experiment (sham ODQ, n = 7).
Control rats were treated with vehicle for ODQ 2 hrs before the experiment (sham DMSO, n = 6).
Rats were treated with saline 2 hrs before they were subjected to Gram-positive shock: LTA (Staphylococcus aureus, 3 mg/kg intravenously) was given over 1 min followed by PepG (S. aureus, 10 mg/kg intravenously) over 15 mins (LTA/PepG, n = 8).
Rats were treated with ODQ 2 hrs before they were subjected to Gram-positive shock (as described previously) (ODQ + LTA/PepG, n = 8).
Rats were treated with vehicle for ODQ (DMSO, as described previously) 2 hrs before they were subjected to Gram-positive shock (as described previously) (DMSO + LTA/PepG, n = 6).
Rats were treated with saline 2 hrs before they were subjected to Gram-negative shock: LPS (Escherichia coli, 6 mg/kg intravenously) was given over 15 mins (LPS, n = 8).
Rats were treated with ODQ 2 hrs before they were subjected to Gram-negative shock (as described previously; ODQ + LPS, n = 8).
Rats were treated with vehicle for ODQ (DMSO) 2 hrs before they were subjected to Gram-negative shock (as described previously; DMSO + LPS, n = 6).[1]

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Male Wistar rats were anesthetized with thiopentone sodium. The trachea, carotid artery, jugular vein, and bladder were cannulated. Rats received a continuous intravenous infusion of saline (1.5 mL·kg⁻¹·hr⁻¹). ODQ (2 mg/kg) or vehicle (DMSO in saline) was administered intraperitoneally 2 hours before shock induction. Gram-positive shock was induced by intravenous coadministration of lipoteichoic acid (3 mg/kg) and peptidoglycan (10 mg/kg). Gram-negative shock was induced by intravenous lipopolysaccharide (6 mg/kg). Hemodynamics were monitored for 6 hours, after which blood and organ samples were collected for analysis of biochemical markers, histology, and immunohistochemistry. [1]

[1]. Crit Care Med . 2001 Aug;29(8):1599-608.

1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one is a member of the class of oxadiazoloquinoxalines that is 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline substituted at position 1 by an oxo group. It has a role as an EC 4.6.1.2 (guanylate cyclase) inhibitor.

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ODQ is described as a selective inhibitor of soluble guanylate cyclase. Its beneficial effects in shock models are not due to scavenging of superoxide anions or hydroxyl radicals. [1]
The study contrasts ODQ with methylene blue, another guanylate cyclase inhibitor that has non-specific effects such as inhibition of nitric oxide synthase and generation of reactive oxygen species. [1]
ODQ reduced protein nitration in shocked organs, suggesting it may modulate peroxynitrite or reactive nitrogen species formation indirectly via guanylate cyclase inhibition. [1]

C9H5N3O2

187.15

187.038

C, 57.76; H, 2.69; N, 22.45; O, 17.10

41443-28-1

1456

Off-white to yellow solid powder

1.6±0.1 g/cm3

321.3±25.0 °C at 760 mmHg

160-170 °C

148.1±23.2 °C

0.0±0.7 mmHg at 25°C

1.781

0.28

60.4

0

4

0

14

337

0

O1C(N2C(C([H])=NC3=C([H])C([H])=C([H])C([H])=C23)=N1)=O

LZMHWZHOZLVYDL-UHFFFAOYSA-N

InChI=1S/C9H5N3O2/c13-9-12-7-4-2-1-3-6(7)10-5-8(12)11-14-9/h1-5H

[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one

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: ≥ 5 mg/mL (26.72 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 50.0 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。