Dooku1
(Synonyms: 2-((2,6-二氯苄基)硫基)-5-(1H-吡咯-2-基)-1,3,4-恶二唑) 目录号 : GC38553
Dooku1,作为可逆的Yoda1拮抗剂,对 Yoda1诱导的Ca2+进入HUVEC中具有浓度依赖性抑制作用,IC50为1.49µM。
Cas No.:2253744-54-4
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
Quality Control & SDS
- View current batch:
- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
|
Cell lines |
Rat podocyte cell line C7 |
|
Preparation Method |
Cells were pretreated with Dooku1(10µM) for 30min or Rac inhibitor EHT 1864 (50µM) for 2h, prior to Yoda1 (3µM) or vehicle stimulation. |
|
Reaction Conditions |
10µM Dooku1 for 30min |
|
Applications |
Yoda1-induced upregulation of Pai1, Sgk1, and Mcp1 was significantly inhibited by Dooku1 via Yoda1-Piezo1 signaling. |
| Animal experiment [2]: | |
|
Animal models |
Male adult C57BL/6 mice (aged 8-10 weeks, weighing 22-30g) |
|
Preparation Method |
Dooku1 was dissolved in dimethyl sulfoxide to a final concentration of less than 0.1%. Dooku1 (5, 10, 20mg/kg) was intraperitoneally administered at 24 and 72 hours after intracerebral hemorrhage (ICH) to evaluate the effects of Dooku1 on neurological function. |
|
Dosage form |
5, 10, 20mg/kg twice, intraperitoneal injection |
|
Applications |
In the modified Garcia test, treatment with 5mg/kg Dooku1 did not affect the score at 24 and 72 hours after ICH, treatment with 10 and 20 mg/kg Dooku1 significantly improved the score at 72 hours after ICH, but only 10 mg/kg Dooku1 significantly improved the score 24 hours after ICH. |
|
References: [1]. Ogino S, Yoshikawa K, Nagase T, Mikami K, Nagase M. Roles of the mechanosensitive ion channel Piezo1 in the renal podocyte injury of experimental hypertensive nephropathy.Hypertens Res. 2024;47(3):747-59. Epub 20231225. doi: 10.1038/s41440-023-01536-z. PubMed PMID: 38145990. | |
Dooku1, a reversible Yoda1 antagonist, has a concentration-dependent inhibitory effect against Yoda1-induced Ca2+ entry in HUVECs, acting with an IC50 of 1.49µM. Dooku1 can antagonize Yoda1‐evoked activation of Piezo1 and aortic relaxation and be used for research related to hypertensive nephropathy[1].
Dooku1 pretreatment (5µM) in rat cauda epididymal epithelial cells significantly suppressed the Yoda1-stimulated Ca2+ response. The Yoda1-stimulated Ca2+ response was abolished when ambient Ca2+ was removed [2]. Dooku1 inhibited 2µM Yoda1‐induced Ca2+‐entry with IC50 of 1.3µM in HEK 293 cells [1].
Dooku1 treatment (10 mg/kg) significantly increased the right turn percentage 72 hours after intracerebral hemorrhage (ICH) among Male adult C57BL/6 mice in the corner turn test. Compared with the ICH +vehicle group, Dooku1 treatment (10 mg/kg) significantly decreased the brain water content at 24 and 72 hours after ICH. A moderate dose (10 mg/kg) of the Piezo1 antagonist Dooku1 could relieve neurological impairment and myelin damage among C57BL/6 mice[3].
References:
[1]. Evans EL, Cuthbertson K, Endesh N, Rode B, Blythe NM, Hyman AJ, et al. Yoda1 analogue (Dooku1) which antagonizes Yoda1-evoked activation of Piezo1 and aortic relaxation. Br J Pharmacol. 2018;175(10):1744-59. Epub 20180406. doi: 10.1111/bph.14188. PubMed PMID: 29498036.
[2]. Gao DD, Huang JH, Ding N, Deng WJ, Li PL, Mai YN, et al. Mechanosensitive Piezo1 channel in rat epididymal epithelial cells promotes transepithelial K(+) secretion. Cell Calcium. 2022;104:102571. Epub 20220315. doi:10.1016/j.ceca.2022.102571. PubMed PMID: 35314382.
[3]. Qu J, Zong HF, Shan Y, Zhang SC, Guan WP, Yang Y, et al. Piezo1 suppression reduces demyelination after intracerebral hemorrhage. Neural Regen Res. 2023;18(8):1750-6. doi: 10.4103/1673-5374.361531.PubMed PMID: 36751801; PubMed Central PMCID: PMCPMC10154511.
Dooku1,作为可逆的Yoda1拮抗剂,对 Yoda1诱导的Ca2+进入HUVEC中具有浓度依赖性抑制作用,IC50为1.49µM。Dooku1可以拮抗Yoda1诱发的Piezo1激活和主动脉舒张并且可用于高血压肾病相关研究[1]。
Dooku1预处理(5µM)大鼠附睾上皮细胞显著抑制Yoda1刺激的Ca2+反应。当环境中Ca2+被去除时,Yoda1刺激的Ca2+反应被消除[2]。Dooku1在HEK 293细胞中抑制 2µM Yoda1 诱导的Ca2+ 内流,IC50为1.3µM[1]。
Dooku1治疗 (10 mg/kg) 显著增加了转弯试验中雄性成年 C57BL/6 小鼠脑出血(ICH) 后 72小时的右转率。与ICH +对照组相比,Dooku1治疗(10 mg/kg)显著降低了ICH后24小时和72小时的脑含水量。中等剂量(10 mg/kg)的 Piezo1 拮抗剂 Dooku1可以缓解C57BL/6 小鼠神经损伤和髓鞘损伤[3]。
| Cas No. | 2253744-54-4 | SDF | |
| 别名 | 2-((2,6-二氯苄基)硫基)-5-(1H-吡咯-2-基)-1,3,4-恶二唑 | ||
| Canonical SMILES | ClC1=C(C(Cl)=CC=C1)CSC2=NN=C(C3=CC=CN3)O2 | ||
| 分子式 | C13H9Cl2N3OS | 分子量 | 326.2 |
| 溶解度 | DMSO: 125 mg/mL (383.20 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
||
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 3.0656 mL | 15.328 mL | 30.656 mL |
| 5 mM | 0.6131 mL | 3.0656 mL | 6.1312 mL |
| 10 mM | 0.3066 mL | 1.5328 mL | 3.0656 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Yoda1 analogue (Dooku1) which antagonizes Yoda1-evoked activation of Piezo1 and aortic relaxation
Br J Pharmacol 2018 May;175(10):1744-1759.PMID:29498036DOI:10.1111/bph.14188.
Background and purpose: The mechanosensitive Piezo1 channel has important roles in vascular physiology and disease. Yoda1 is a small-molecule agonist, but the pharmacology of these channels is otherwise limited. Experimental approach: Yoda1 analogues were generated by synthetic chemistry. Intracellular Ca2+ and Tl+ measurements were made in HEK 293 or CHO cell lines overexpressing channel subunits and in HUVECs, which natively express Piezo1. Isometric tension recordings were made from rings of mouse thoracic aorta. Key results: Modification of the pyrazine ring of Yoda1 yielded an analogue, which lacked agonist activity but reversibly antagonized Yoda1. The analogue is referred to as Dooku1. Dooku1 inhibited 2 μM Yoda1-induced Ca2+ -entry with IC50 s of 1.3 μM (HEK 293 cells) and 1.5 μM (HUVECs) yet failed to inhibit constitutive Piezo1 channel activity. It had no effect on endogenous ATP-evoked Ca2+ elevation or store-operated Ca2+ entry in HEK 293 cells or Ca2+ entry through TRPV4 or TRPC4 channels overexpressed in CHO and HEK 293 cells. Yoda1 caused dose-dependent relaxation of aortic rings, which was mediated by an endothelium- and NO-dependent mechanism and which was antagonized by Dooku1 and analogues of Dooku1. Conclusion and implications: Chemical antagonism of Yoda1-evoked Piezo1 channel activity is possible, and the existence of a specific chemical interaction site is suggested with distinct binding and efficacy domains.
Identification of Piezo1 channels in perivascular adipose tissue (PVAT) and their potential role in vascular function
Pharmacol Res 2022 Jan;175:105995.PMID:34818570DOI:10.1016/j.phrs.2021.105995.
The vasculature constantly experiences distension/pressure exerted by blood flow and responds to maintain homeostasis. We hypothesized that activation of the stretch sensitive, non-selective cation channel Piezo1 would directly increase vascular contraction in a way that might be modified by perivascular adipose tissue (PVAT). The presence and function of Piezo1 was investigated by RT-PCR, immunohistochemistry, and isolated tissue bath contractility. Superior and mesenteric resistance arteries, aortae, and their PVATs from male Sprague Dawley rats were used. Piezo1 mRNA was detected in aortic vessels, aortic PVAT, mesenteric vessels, and mesenteric PVAT. Both adipocytes and stromal vascular fraction of mesenteric PVAT expressed Piezo1 mRNA. In PVAT, expression of Piezo1 mRNA was greater in magnitude than that of Piezo2, transient receptor potential cation channel, subfamily V, member 4 (TRPV4), anoctamin 1, calcium activated chloride channel (TMEM16), and Pannexin1 (Panx1). Piezo1 protein was present in endothelium and PVAT of rat aortic and in PVAT of mesenteric artery. The Piezo1 agonists Yoda1 and Jedi2 (1 nM - 10 µM) did not stimulate aortic contraction [max < 10% phenylephrine (PE) 10 µM contraction] or relaxation in tissues + or -PVAT. Depolarizing the aorta by modestly elevated extracellular K+ did not unmask aortic contraction to Yoda1 (max <10% PE 10 µM contraction). Finally, the Piezo1 antagonist Dooku1 did not modify PE-induced aorta contraction + or -PVAT. Surprisingly, Dooku1 directly caused aortic contraction in the absence (Dooku1 =26 ± 11; Vehicle = 11 ± 11%PE contraction) but not in the presence of PVAT (Dooku1 = 2 ± 1; Vehicle = 8 ± 5% PE contraction). Thus, Piezo1 is present and functional in the isolated rat aorta but does not serve direct vascular contraction with or without PVAT. We reaffirmed the isolated mouse aorta relaxation to Yoda1, indicating a species difference in Piezo1 activity between mouse and rat.
Mechanosensitive Piezo1 channel in rat epididymal epithelial cells promotes transepithelial K+ secretion
Cell Calcium 2022 Jun;104:102571.PMID:35314382DOI:10.1016/j.ceca.2022.102571.
The Piezo1 channel, a mechanosensitive channel that exhibit a preference for Ca2+, play multifarious physiological and pathological roles in the endothelium and epithelium of various tissues. However, the functional expression of Piezo1 channel in the epithelium of the male reproductive tract remains unknown. In the present study, the expression of Piezo1 channel in the rat epididymis was determined by real-time quantitative PCR, western blot and immunohistochemical analysis. Our data revealed that Piezo1 channel was located in the epithelial layer of the rat epididymis, with higher expression levels in the corpus and cauda regions. The pro-secretion function of Piezo1 channel was then investigated using short circuit current (ISC) and intracellular Ca2+ imaging techniques. Application of Yoda1, a selective Piezo1 channel activator, stimulated a remarkable decrease in the ISC of the epididymal epithelium. Pharmacological experiments revealed that the ISC response induced by Piezo1 channel activation was abolished by pretreating epithelial cells with the Yoda1 analogue, Dooku1, the selective mechanosensitive cation channel blocker, GsMTx4, or removal of basolateral K+. Meanwhile, we demonstrated that activation of Piezo1 channel triggered a robust Ca2+ influx in epididymal epithelial cells. The possible involvement of Ca2+- activated K+channels (KCa) in transepithelial K+ secretion was then evaluated. And that big conductance KCa (BK), but not small conductance or intermediate conductance KCa, mediated Piezo1-elicited transepithelial K+ secretion. Moreover, we demonstrated that NKCC and NKA were responsible for supplying substrate K+ during transepithelial K+ secretion. These data demonstrate that the activation of Piezo1 channel promotes BK-mediated transepithelial K+ secretion, and thus may plays an important role in the formation of a high K+ concentration in epididymal intraluminal fluid.
Piezo1 suppression reduces demyelination after intracerebral hemorrhage
Neural Regen Res 2023 Aug;18(8):1750-1756.PMID:36751801DOI:10.4103/1673-5374.361531.
Piezo1 is a mechanically-gated calcium channel. Recent studies have shown that Piezo1, a mechanically-gated calcium channel, can attenuate both psychosine- and lipopolysaccharide-induced demyelination. Because oligodendrocyte damage and demyelination occur in intracerebral hemorrhage, in this study, we investigated the role of Piezo1 in intracerebral hemorrhage. We established a mouse model of cerebral hemorrhage by injecting autologous blood into the right basal ganglia and found that Piezo1 was largely expressed soon (within 48 hours) after intracerebral hemorrhage, primarily in oligodendrocytes. Intraperitoneal injection of Dooku1 to inhibit Piezo1 resulted in marked alleviation of brain edema, myelin sheath loss, and degeneration in injured tissue, a substantial reduction in oligodendrocyte apoptosis, and a significant improvement in neurological function. In addition, we found that Dooku1-mediated Piezo1 suppression reduced intracellular endoplasmic reticulum stress and cell apoptosis through the PERK-ATF4-CHOP and inositol-requiring enzyme 1 signaling pathway. These findings suggest that Piezo1 is a potential therapeutic target for intracerebral hemorrhage, as its suppression reduces intracellular endoplasmic reticulum stress and cell apoptosis and protects the myelin sheath, thereby improving neuronal function after intracerebral hemorrhage.
The mechanosensitive Piezo1 channels contribute to the arterial medial calcification
Front Physiol 2022 Nov 10;13:1037230.PMID:36439266DOI:10.3389/fphys.2022.1037230.
Vascular calcification (VC) is associated with a number of cardiovascular diseases, as well as chronic kidney disease. The role of smooth muscle cells (SMC) has already been widely explored in VC, as has the role of intracellular Ca2+ in regulating SMC function. Increased intracellular calcium concentration ([Ca2+]i) in vascular SMC has been proposed to stimulate VC. However, the contribution of the non-selective Piezo1 mechanosensitive cation channels to the elevation of [Ca2+]i, and consequently to the process of VC has never been examined. In this work the essential contribution of Piezo1 channels to arterial medial calcification is demonstrated. The presence of Piezo1 was proved on human aortic smooth muscle samples using immunohistochemistry. Quantitative PCR and Western blot analysis confirmed the expression of the channel on the human aortic smooth muscle cell line (HAoSMC). Functional measurements were done on HAoSMC under control and calcifying condition. Calcification was induced by supplementing the growth medium with inorganic phosphate (1.5 mmol/L, pH 7.4) and calcium (CaCl2, 0.6 mmol/L) for 7 days. Measurement of [Ca2+]i using fluorescent Fura-2 dye upon stimulation of Piezo1 channels (either by hypoosmolarity, or Yoda1) demonstrated significantly higher calcium transients in calcified as compared to control HAoSMCs. The expression of mechanosensitive Piezo1 channel is augmented in calcified arterial SMCs leading to a higher calcium influx upon stimulation. Activation of the channel by Yoda1 (10 μmol/L) enhanced calcification of HAoSMCs, while Dooku1, which antagonizes the effect of Yoda1, reduced this amplification. Application of Dooku1 alone inhibited the calcification. Knockdown of Piezo1 by siRNA suppressed the calcification evoked by Yoda1 under calcifying conditions. Our results demonstrate the pivotal role of Piezo1 channels in arterial medial calcification.