ETA/endothelin A receptor

克拉生坦是一种选择性内皮素a受体拮抗剂，用于预防和治疗蛛网膜下腔出血后血管痉挛。根据临床前数据，它是有机阴离子转运多肽1B1/1B3的底物。目前的数据表明，克唑生坦是ET(a)受体介导的由ET-1及其前体大ET-1引起的脑血管收缩的一种有效的竞争性拮抗剂。这些功能数据也可用于确定具有高临床疗效可能性的ET受体拮抗剂的体外轮廓。［3］

Clazosentan的药代动力学特征为：中间清除率、分布体积与细胞外液体积相似、剂量比例暴露、不依赖于药物代谢酶的消除，以及主要依赖于肝摄取转运体有机阴离子转运多肽1B1/1B3的处置。在健康受试者中，克唑生坦可导致ET-1浓度升高，防止ET-1输注介导的心脏和肾脏效应。在患者中，它显著降低了中度或重度血管痉挛的发生率以及asah后血管痉挛相关的发病率和死亡率。在预期治疗剂量为15mg /h时，克唑生坦耐受性良好，在aSAH患者中，肺并发症、低血压和贫血是克唑生坦治疗后比安慰剂更常见的不良事件。综上所述，克唑生坦具有药代动力学、药效学和安全性特征，适合成为预防asah诱导的脑血管痉挛的治疗方式的宝贵资产。[1]

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脑血管痉挛和晚期脑缺血（LCI）仍然是蛛网膜下腔出血（SAH）患者死亡的主要原因。这通常发生在初次出血后3至4天，并在5至7天达到高峰。其潜在的病理生理学仍然知之甚少。由于SAH与内皮素-1 （ET-1）水平升高有关，人们一直关注用受体拮抗剂如clazosentan来对抗内皮素受体的激活，然而，临床试验的结果不佳。我们假设抑制细胞内转录信号将是预防LCI的有效方法。在这里，我们比较了克拉生坦和MEK1/2阻滞剂U0126在SAH大鼠模型中的作用。虽然clazosentan直接抑制体内对ET-1的收缩反应，但它并不能阻止sah诱导的脑动脉ET受体的上调，也没有显示出对神经系统预后的有益影响。U0126本身没有血管舒缩作用，但可以抵消SAH诱导的脑动脉受体上调，改善SAH后的预后。我们认为，由于SAH诱导几种收缩受体亚型的表达升高，仅阻断其中一种受体（ET受体）是不够的，抑制mek1 /2介导的几种收缩受体的转录上调可能是缓解LCI的可行方法。[2]

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这项随机、双盲、两期交叉研究调查了在静脉注射安慰剂或利福平（600mg / 100ml， 30分钟）后静脉输注clazosentan （15mg /h，持续3小时）的药代动力学、安全性和耐受性。共有14名健康男性参与者参加，其中13人完成了这项研究。血浆浓度-时间曲线下面积的几何平均比值（90%置信区间）为3.88(3.24-4.65)，表明在有机阴离子运输多肽1B1/1B3抑制后，Clazosentan暴露量增加了3 -4倍。间隙和分布体积也有类似程度的下降。消去半衰期不受影响。与安慰剂相比，克唑生坦与利福平同时使用时，观察到类似的模式，但不良事件的发生率和频率更高。［5］

Isometric force measurements were performed in rat basilar artery (BA) ring segments with (E+) and without (E-) endothelial function. Concentration effect curves (CECs) were constructed by cumulative application of ET-1 or big ET-1 in the absence or presence of clazosentan (10(-9), 10(-8), and 10(-7) M). The inhibitory potency of clazosentan was determined by the value of the affinity constant (pA2). The CECs for contraction induced by ET-1 and big ET-1 were shifted to the right in the presence of clazosentan in a parallel dose-dependent manner, which indicates competitive antagonism. The pA2 values for ET-1 were 7.8 (E+) and 8.6 (E-) and the corresponding values for big ET-1 were 8.6 (E+) and 8.3 (E-). [3]

[1]. Clinical Pharmacology of Clazosentan, a Selective Endothelin A Receptor Antagonist for the Prevention and Treatment of aSAH-Related Cerebral Vasospasm. Front Pharmacol. 2021 Feb 4;11:628956.

[2]. MEK1/2 inhibitor U0126 but not endothelin receptor antagonist clazosentan reduces upregulation of cerebrovascular contractile receptors and delayed cerebral ischemia, and improves outcome after subarachnoid hemorrhage in rats. J Cereb Blood Flow Metab. 2015 Feb;35(2):329-37.

[3]. Cerebrovascular characterization of clazosentan, the first nonpeptide endothelin receptor antagonist clinically effective for the treatment of cerebral vasospasm. Part I: inhibitory effect on endothelin(A) receptor-mediated contraction. J Neurosurg. 2005 Jun;102(6):1101-7.

[4]. IL-33 mediates antigen-induced cutaneous and articular hypernociception in mice. Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2723-8.

[5]. Influence of Rifampin-Mediated Organic Anion-Transporting Polypeptide 1B1/1B3 Inhibition on the Pharmacokinetics of Clazosentan. Clin Transl Sci. 2019 Sep;12(5):440-444.

CLAZOSENTAN is a small molecule drug with a maximum clinical trial phase of III (across all indications) and has 2 investigational indications.

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The central role of endothelin (ET)-1 in the development of cerebral vasospasm after subarachnoid hemorrhage is indicated by the successful treatment of this vasospasm in several animal models by using selective ET(A) receptor antagonists. Clazosentan is a selective ET(A) receptor antagonist that provides for the first time clinical proof that ET-1 is involved in the pathogenesis of cerebral vasospasm. The aim of the present investigation was, therefore, to define the pharmacological properties of clazosentan that affect ET(A) receptor-mediated contraction in the cerebrovasculature.[3]

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IL-33, a new member of the IL-1 family, signals through its receptor ST2 and induces T helper 2 (Th2) cytokine synthesis and mediates inflammatory response. We have investigated the role of IL-33 in antigen-induced hypernociception. Recombinant IL-33 induced cutaneous and articular mechanical hypernociception in a time- and dose-dependent manner. The hypernociception was inhibited by soluble (s) ST2 (a decoy receptor of IL-33), IL-1 receptor antagonist (IL-1ra), bosentan [a dual endothelin (ET)(A)/ET(B) receptor antagonist], clazosentan (an ET(A) receptor antagonist), or indomethacin (a cyclooxygenase inhibitor). IL-33 induced hypernociception in IL-18(-/-) mice but not in TNFR1(-/-) or IFNgamma(-/-) mice. The IL-33-induced hypernociception was not affected by blocking IL-15 or sympathetic amines (guanethidine). Furthermore, methylated BSA (mBSA)-induced cutaneous and articular mechanical hypernociception depended on TNFR1 and IFNgamma and was blocked by sST2, IL-1ra, bosentan, clazosentan, and indomethacin. mBSA also induced significant IL-33 and ST2 mRNA expression. Importantly, we showed that mBSA induced hypernociception via the IL-33 --> TNFalpha --> IL-1beta --> IFNgamma --> ET-1 --> PGE(2) signaling cascade. These results therefore demonstrate that IL-33 is a key mediator of immune inflammatory hypernociception normally associated with a Th1 type of response, revealing a hitherto unrecognized function of IL-33 in a key immune pharmacological pathway that may be amenable to therapeutic intervention.[4]

C25H23N9NA2O6S

623.55

621.113

C, 48.31; H, 3.41; N, 20.28; Na, 7.40; O, 15.44; S, 5.16

503271-02-1

180384-56-9; 503271-02-1 (sodium)

51346597

Typically exists as solid at room temperature

3.186

178.55

1

15

11

43

911

0

S(C1C=CC(C)=CN=1)([N-]C1C(=C(N=C(C2C=CN=C(C3=NN=N[N-]3)C=2)N=1)OCCO)OC1C=CC=CC=1OC)(=O)=O.[Na+].[Na+]

PZNSONUYVNYXJZ-UHFFFAOYSA-N

InChI=1S/C25H21N9O6S.2Na/c1-15-7-8-20(27-14-15)41(36,37)32-24-21(40-19-6-4-3-5-18(19)38-2)25(39-12-11-35)29-22(28-24)16-9-10-26-17(13-16)23-30-33-34-31-23;;/h3-10,13-14,35H,11-12H2,1-2H3;;/q-2;2*+1

disodium;[6-(2-hydroxyethoxy)-5-(2-methoxyphenoxy)-2-[2-(1,2,3-triaza-4-azanidacyclopenta-2,5-dien-5-yl)pyridin-4-yl]pyrimidin-4-yl]-(5-methylpyridin-2-yl)sulfonylazanide

Ro 61-1790 disodium; VML 588 disodium; Clazosentan sodium; Clazosentan disodium salt; Clazosentan disodium; Clazosentan sodium [JAN]; Clazosentan disodium salt [MI]; 180384-56-9; AXV-034343; VML-588; Ro-61-1790; AXV-343434; Ro 61-1790; ACT-108475; AXV-034343 disodium

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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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。