U46619

Thromboxane A2 (TXA2)

U-46619 (1 nM–10 μM) 对血小板聚集和形状改变的浓度依赖性影响的 EC50 分别为 0.58 μM 和 0.013 μM [1]。 U-46619 (10 nM–10 μM) 以浓度依赖性方式刺激磷酸肌醇 (PI) 水解，并提高内部 Ca2+ 浓度 ([Ca2+]i [1]。血小板膜中的 GTPase 也可通过以下方式激活： U-46619 (3 nM–10 μM) 以浓度依赖性方式[1]。通过激活 p38MAPK 和 ERK1/2 信号通路，U-46619 增强人诱导多能干细胞向内皮细胞的分化效率。 2]。

雄性自发性高血压大鼠 (SHR) 对 U-46619（5 μg/kg；静脉注射）表现出血压升高 [3]。

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在雄性和雌性自发性高血压大鼠（SHR）中研究了U 46619（5微克/千克静脉注射）单独或与乙酰水杨酸（ASA）（100mg/kg口服）联合对平均动脉血压（MABP）的影响。在雄性SHR中，服用U 46619 1分钟后观察到MABP显著增加。与单独使用U 46619相比，用ASA预处理雄性SHR延迟了静脉注射U 46619后MABP的增加。在对照组动物中，静脉注射U 46619 5分钟后，升高的MABP恢复到基线值，而ASA预处理的雄性SHR的MABP仍显著增加约30 mmHg。相比之下，雌性SHR的MABP对单独使用U 46619或U 46619和ASA的组合没有反应。血栓素B2（TXB2）的血管形成进一步显示了性别差异。而在雄性SHR中，TXB2的血管形成增加了U 46619，而雌性SHR的TXB2形成减少了。单独使用U 46619或U 46619和ASA的组合不影响雄性和雌性SHR的6-酮-PGF1α的血管形成。总之，我们的研究结果表明，SHR的血压对TXA2模拟物U 46619的反应在两性之间是不同的。此外，通过调节对TXA2的血压反应，血管活性前列腺素可能显著参与了男性SHR高血压的维持。[3]

1.在洗涤的兔血小板中研究了血栓素A2（TXA2）受体介导的信号转导，以阐明诱导形状变化和聚集的机制。2.TXA2激动剂U46619（1nM至10微M）以浓度依赖的方式引起形状变化和聚集。聚集所需的U46619浓度（EC50为0.58微M）是形状变化所需浓度（EC50=0.013微M）的40倍。聚集仅在存在外部1mM Ca2+时发生，但在没有Ca2+的情况下可能会发生形状变化。3.30 nM的SQ29548和0.3 microM的GR32191B（TXA2受体拮抗剂）以相似的效力竞争性抑制U46619诱导的形状变化和聚集，表明U46619引起的聚集和形状变化都是TXA2受体介导的事件。然而，1 nM的ONO NT-126（另一种TXA2受体拮抗剂）比形状变化更能抑制U46619诱导的聚集，这表明TXA2受体亚型可能存在。4.ONO NT-126（2nM至3微M）本身以浓度依赖的方式引起形状变化而不聚集，与外部Ca2+无关。因此，ONO NT-126是兔血小板TXA2受体的部分激动剂。5.U46619（10nM至10microM）以浓度依赖的方式增加了内部Ca2+浓度（[Ca2+]i），并激活了磷酸肌醇（PI）水解，具有类似的浓度依赖性。6.U46619（3 nM至10微M）也能浓度依赖性地激活血小板膜中的GTP酶。U46619诱导的GTP酶活化被QL（一种针对Gq/11的抗体）处理的膜部分抑制。7.U46619在Ca2+动员（0.15微米）、PI水解（0.20微米）和GTP酶活性增加（0.12微米）方面的EC50值相似，但在形状变化（0.013微米）方面与EC50值不同，表明TXA2受体的激活可能通过未知的机制导致形状变化。8.U46619诱导的形状变化不受钙调素拮抗剂W-7（30微M）或肌球蛋白轻链激酶抑制剂ML-7（30微m）的影响，表明[Ca2+]i的增加可能与形状变化无关。事实上，U46619（10 nM）可以在不影响[Ca2+]i水平的情况下引起形状变化，这是通过同时记录来确定的。9.[3H]-SQ29548和[3H]-U46619分别以14.88 nM的Kd值和106.1 fmol/10（8）血小板的Bmax与血小板在单个位点结合，Kd值为129.8 nM，Bmax为170.4 fmol/108血小板。U46619作为3H配体结合抑制剂的抑制常数Ki值在GTP酶活性、磷酸肌醇水解和Ca2+动员方面与U46619的EC50值相似，但对形状变化的影响存在显著差异（Student's t检验P<0.001）。10.在存在或不存在外部Ca2+和/或异丁基甲基黄嘌呤的情况下，U46619和ONO NT-126均不影响腺苷3'，5'-环单磷酸（环AMP）水平。11.结果表明，TXA2受体刺激会导致磷脂酶C激活，并通过Gq/11家族的G蛋白增加[Ca2+]i，导致在外部Ca2+存在的情况下聚集，并且TXA2受体兴奋诱导的形状变化可能在不涉及Gq磷脂酶C-Ca2+途径的情况下发生[1]。

使用U-46619上调p38丝裂原活化蛋白激酶（p38MAPK）和细胞外信号调节激酶1/2（ERK1/2）信号传导，这使两个已建立的hiPSC系的分化效率（通过CD31表达测量）提高到高达89%。分化的细胞表达动静脉标志物，但不表达淋巴标志物；体外形成管状结构和EC腔；其群体倍增时间明显短于单层分化的hiPSC EC；并在小鼠后肢缺血模型中恢复灌注和血管。在三个来源于与内皮功能障碍相关的疾病或疾病症状患者的hiPSC系中，分化效率也>85%[2]。

Animal/Disease Models: 12-15 weeks old male and female SHR [2]
Doses: 5 μg/kg
Route of Administration: intravenous (iv) (iv)injection
Experimental Results: A significant increase in MABP was induced in male SHR after 1 minute.

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Pressure Myography [4]
Vasodilation was determined using a pressure myography system as previously described.20,46 Briefly, rats were sacrificed under anesthesia, and the brains were rapidly harvested and placed in precooled physiological salt dissolution (PSS, composition in mmol/L: NaCl 118, KCl 4.7, CaCl2 1.6, KH2PO4 1.2, MgSO4 1.2, NaHCO3 25, EDTA 0.026, glucose 5.5, pH 7.4) bubbled with 95% O2 + 5% CO2. CBA was carefully isolated from the brain and cut into an unbranched artery segment of 3 mm in length. The artery segment was inserted with glass micropipettes at both ends and fixed in a perfusion chamber of a DMT-114P Pressure Myograph System, which was filled with PSS aerated with 95% O2 + 5% CO2 at 37 °C. The lumen of the segment was perfused with the same aerated PPS. After 60 min of equilibrium, 100 nmol/L U46619 or 30 mmol/L KCl was added to the luminal superfusate to induce stable vasocontraction. Vasodilation was subsequently caused by cumulatively adding ACh or NaHS. The diameter of the artery segment was continually measured by Pressure Myograph System software. Vasodilation was expressed as the percentage of the maximum diameter using the following formula:
where Dmax is the initial diameter of the artery segment at equilibration for 60 min, Dmin is the stable diameter after adding KCl or U46619, and D is the diameter after adding ACh or NaHS. [4]

[1]. Thromboxane A2-mediated shape change: independent of Gq-phospholipase C--Ca2+ pathway in rabbit platelets. Br J Pharmacol. 1996 Mar;117(6):1095-104.

[2]. The prostaglandin H2 analog U-46619 improves the differentiation efficiency of human induced pluripotent stem cells into endothelial cells by activating both p38MAPK and ERK1/2 signaling pathways. Stem Cell Res Ther. 2018 Nov 15;9(1):313.

[3]. U 46619 induces different blood pressure effects in male and female spontaneously hypertensive rats (SHR). Prostaglandins Leukot Essent Fatty Acids. 1993 Jun;48(6):469-73.

[4]. Roles of the RhoA-ROCK Signaling Pathway in the Endothelial H2S Production and Vasodilation in Rat Cerebral Arteries. ACS Omega. 2022 May 20;7(22):18498-18508.

[5]. Thromboxane A2 receptors in prostate carcinoma: expression and its role in regulating cell motility via small GTPase Rho. Cancer Res. 2008 Jan 1;68(1):115-21.

A stable prostaglandin endoperoxide analog which serves as a thromboxane mimetic. Its actions include mimicking the hydro-osmotic effect of VASOPRESSIN and activation of TYPE C PHOSPHOLIPASES. (From J Pharmacol Exp Ther 1983;224(1): 108-117; Biochem J 1984;222(1):103-110)

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Background: We have shown that the differentiation of human-induced pluripotent stem cells (hiPSCs) into endothelial cells (ECs) is more efficient when performed with a 3-dimensional (3D) scaffold of biomaterial than in monolayers. The current study aims to further increase hiPSC-EC differentiation efficiency by deciphering the signaling pathways in 3D scaffolds. Methods and results: We modified our 3D protocol by using U-46619 to upregulate both p38 mitogen-activated protein kinase (p38MAPK) and extracellular signal-regulated kinase 1/2 (ERK1/2) signaling, which increased the differentiation efficiency (as measured by CD31 expression) to as high as 89% in two established hiPSC lines. The differentiated cells expressed arteriovenous, but not lymphatic, markers; formed tubular structures and EC lumen in vitro; had significantly shorter population-doubling times than monolayer-differentiated hiPSC-ECs; and restored perfusion and vascularity in a murine hind limb ischemia model. The differentiation efficiency was also > 85% in three hiPSC lines that had been derived from patients with diseases or disease symptoms that have been linked to endothelial dysfunction. Conclusions: These observations demonstrate that activating both p38MAPK and ERK1/2 signaling pathways with U-46619 improves the efficiency of arteriovenous hiPSC-EC differentiation and produces cells with greater proliferative capacity. Keywords: Endothelial differentiation; Human-induced pluripotent stem cells; Signaling pathways. PubMed Disclaimer[2]

C21H34O4

350.49226

350.245

C, 71.96; H, 9.78; O, 18.26

56985-40-1

56985-40-1;

5311493

Colorless to light yellow liquids

1.1±0.1 g/cm3

519.7±30.0 °C at 760 mmHg

176.1±18.1 °C

0.0±3.1 mmHg at 25°C

1.548

3.9

66.76

2

4

12

25

457

5

CCCCC[C@@H](/C=C/[C@H]1C2OCC(C2)[C@@H]1C/C=C/CCCC(=O)O)O

LQANGKSBLPMBTJ-BRSNVKEHSA-N

InChI=1S/C21H34O4/c1-2-3-6-9-17(22)12-13-19-18(16-14-20(19)25-15-16)10-7-4-5-8-11-21(23)24/h4,7,12-13,16-20,22H,2-3,5-6,8-11,14-15H2,1H3,(H,23,24)/b7-4-,13-12+/t16-,17+,18+,19-,20-/m1/s1

(Z)-7-[(1R,4S,5S,6R)-6-[(E,3S)-3-hydroxyoct-1-enyl]-2-oxabicyclo[2.2.1]heptan-5-yl]hept-5-enoic acid

U-46619; U46619; U 46619; 9,11-Methanoepoxy PGH2; 11alpha,9alpha-epoxymethano-PGH2; MLS000028857; CHEMBL521784;

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));
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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；
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