H 89 2HCl
(Synonyms: H-89二盐酸盐) 目录号 : GC10074
H-89 2HCl 是一种有效的选择性 camp 依赖性蛋白激酶 A 抑制剂，IC50 值为 48 nM，对 PKG、PKC、酪蛋白激酶和其他激酶表现出较弱的抑制作用 。
Cas No.:130964-39-5
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
PC12D Cells |
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Preparation Method |
PC12D cells were cultured in test medium containing 30 μM H-89 2HCl for 1 h and then exposed to fresh medium that contained both 10 pM forskolin and 30 μM H-89 2HCl. |
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Reaction Conditions |
30 μM H-89 2HCl for 1h |
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Applications |
H-89 2HCl (30μM) significantly inhibited camp-dependent phosphorylation of histone IIh and forskolin-induced neurite outgrowth in PC12D cells. |
| Animal experiment [2]: | |
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Animal models |
Male albino mice weighing 20-25 g |
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Preparation Method |
H-89 2HCl (0.05, 0.1, 0.2 mg/100g) were administered intraperitoneally, 30 min before intravenous infusion of PTZ |
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Dosage form |
0.05, 0.1, 0.2 mg/100g H-89 |
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Applications |
Intraperitoneal administration of H-89 2HCl (0.2 mg/100g) significantly increased seizure latency and threshold in PTZ-treated animals. H-89 2HCl (0.05, 0.2 mg/100g) prevented the epileptogenic activity of bucladesine (300 nM) with significant increase of seizure latency and seizure threshold. |
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References: [1]. Chijiwa T, Mishima A, et,al. Inhibition of forskolin-induced neurite outgrowth and protein phosphorylation by a newly synthesized selective inhibitor of cyclic AMP-dependent protein kinase, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89), of PC12D pheochromocytoma cells. J Biol Chem. 1990 Mar 25;265(9):5267-72. PMID: 2156866. [2]. Hosseini-Zare MS, Salehi F, et,al. Effects of pentoxifylline and H-89 on epileptogenic activity of bucladesine in pentylenetetrazol-treated mice. Eur J Pharmacol. 2011 Nov 30;670(2-3):464-70. doi: 10.1016/j.ejphar.2011.09.026. Epub 2011 Sep 21. PMID: 21946102. |
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H-89 2HCl is A potent and selective camp-dependent protein kinase A inhibitor with IC50 value of 48 nM, showing weak inhibition of PKG,PKC,Casein kinase and other kinases[1]. H-89 2HCl causes different modifications in protein phosphorylation, all of which have potential regulatory relationships with cAMP/PKA[2].
H-89 2HCl (30μM) significantly inhibited camp-dependent phosphorylation of histone IIh and forskolin-induced neurite outgrowth in PC12D cells[1]. H-89 2HCl (1-2 μM) significantly slows the repriming rate in rat skinned fibres, most likely due to it deleteriously affecting the T-system potential. H-89 2HCl (10-100 μM) inhibits net Ca2+ uptake by the SR and affectes the Ca32-sensitivity of the contractile apparatus in rat skinned fibres[4]. The PKA inhibitor H-89 2HCl effectively inhibited the CM-or PTHRP-mediated increase in UCP1 protein levels and phosphorylation of PKA substrate in ccRCC cells[6].
Trehalase (Treh) hydrolyzes trehalose to generate glucose. Pheromone biosynthesis activating neuropeptide (PBAN) treatment triggered HaTreh1 and HaTreh2 activities in the isolated PGs. However, the activities of HaTreh1 and HaTreh2 triggered by PBAN were offset by H-89 2HCl, the specific inhibitor of protein kinase A (PKA). Furthermore, the H-89 2HCl treatment significantly decreased the phosphorylation level of Trhe2, which was induced by PBAN[3]. The PKA inhibitor H-89 2HCl potently blocked oncosphere larval motility, as well as the motility of other life stages, although other inhibitors of the PKA pathway were not effective[7]. H-89 2HCl (0.2 mg/100g) significantly increases seizure latency and threshold in PTZ-treated animals. H-89 2HCl (0.05, 0.2 mg/100 g) prevents the epileptogenic activity of bucladesine with significant increase of seizure latency and seizure threshold[5].
References:
[1]: Chijiwa T, Mishima A, et,al. Inhibition of forskolin-induced neurite outgrowth and protein phosphorylation by a newly synthesized selective inhibitor of cyclic AMP-dependent protein kinase, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89), of PC12D pheochromocytoma cells. J Biol Chem. 1990 Mar 25;265(9):5267-72. PMID: 2156866.
[2]: Davis MA, Hinerfeld D, et,al. Proteomic analysis of rat liver phosphoproteins after treatment with protein kinase inhibitor H89 (N-(2-[p-bromocinnamylamino-]ethyl)-5-isoquinolinesulfonamide). J Pharmacol Exp Ther. 2006 Aug;318(2):589-95. doi: 10.1124/jpet.105.100032. Epub 2006 May 10. PMID: 16687476.
[3]: Zhang B, Zhang Y, et,al. Trehalase is required for sex pheromone biosynthesis in Helicoverpa armigera. Insect Mol Biol. 2022 Jun;31(3):334-345. doi: 10.1111/imb.12762. Epub 2022 Feb 4. PMID: 35084068.
[4]: Blazev R, Hussain M, Bakker AJ, Head SI, Lamb GD. Effects of the PKA inhibitor H-89 on excitation-contraction coupling in skinned and intact skeletal muscle fibres. J Muscle Res Cell Motil. 2001;22(3):277-86. doi: 10.1023/a:1012289526618. PMID: 11763200.
[5]: Hosseini-Zare MS, Salehi F, Seyedi SY, Azami K, Ghadiri T, Mobasseri M, Gholizadeh S, Beyer C, Sharifzadeh M. Effects of pentoxifylline and H-89 on epileptogenic activity of bucladesine in pentylenetetrazol-treated mice. Eur J Pharmacol. 2011 Nov 30;670(2-3):464-70. doi: 10.1016/j.ejphar.2011.09.026. Epub 2011 Sep 21. PMID: 21946102.
[6]: Wei G, Sun H, et,al. The thermogenic activity of adjacent adipocytes fuels the progression of ccRCC and compromises anti-tumor therapeutic efficacy. Cell Metab. 2021 Oct 5;33(10):2021-2039.e8. doi: 10.1016/j.cmet.2021.08.012. Epub 2021 Sep 10. PMID: 34508696.
[7]: Preza M, Guarnaschelli I, et,al. Inhibitors of protein kinases A and C impair the motility of oncospheres of the model tapeworm Hymenolepis microstoma. Mol Biochem Parasitol. 2021 Nov;246:111423. doi: 10.1016/j.molbiopara.2021.111423. Epub 2021 Sep 22. PMID: 34562553.
H-89 2HCl 是一种有效的选择性 camp 依赖性蛋白激酶 A 抑制剂,IC50 值为 48 nM,对 PKG、PKC、酪蛋白激酶和其他激酶表现出较弱的抑制作用 [1]。 H-89 2HCl引起不同的蛋白磷酸化修饰,均与cAMP/PKA[2]具有潜在的调控关系。
H-89 2HCl (30μM) 显着抑制 PC12D 细胞中组蛋白 IIh 的营依赖性磷酸化和毛喉素诱导的神经突生长[1]。 H-89 2HCl (1-2 μM) 显着减慢大鼠皮肤纤维的重新启动速率,很可能是因为它有害地影响了 T 系统的潜力。 H-89 2HCl (10-100 μM) 抑制 SR 对 Ca2+ 的净摄取,并影响大鼠皮肤纤维中收缩装置的 Ca32 敏感性[4]。 PKA抑制剂H-89.2HCl有效抑制ccRCC细胞中CM-或PTHRP介导的UCP1蛋白水平升高和PKA底物磷酸化[6]。
海藻糖酶 (Treh) 水解海藻糖生成葡萄糖。信息素生物合成激活神经肽 (PBAN) 处理触发了分离的 PG 中的 HaTreh1 和 HaTreh2 活性。然而,由 PBAN 触发的 HaTreh1 和 HaTreh2 的活性被蛋白激酶 A (PKA) 的特异性抑制剂 H-89.2HCl 所抵消。此外,H-89 2HCl 处理显着降低了由 PBAN[3] 诱导的 Trhe2 的磷酸化水平。 PKA 抑制剂 H-89 2HCl 有效地阻断了肿瘤球幼虫的运动,以及其他生命阶段的运动,尽管 PKA 通路的其他抑制剂无效[7]。 H-89 2HCl (0.2 mg/100g) 显着增加 PTZ 治疗动物的癫痫发作潜伏期和阈值。 H-89 2HCl (0.05, 0.2 mg/100 g) 可抑制布克拉地辛的致癫痫活性,显着增加癫痫发作潜伏期和癫痫发作阈值[5]。
| Cas No. | 130964-39-5 | SDF | |
| 别名 | H-89二盐酸盐 | ||
| 化学名 | (E)-N-(2-((3-(4-bromophenyl)allyl)amino)ethyl)isoquinoline-5-sulfonamide dihydrochloride | ||
| Canonical SMILES | O=S(C1=CC=CC2=C1C=CN=C2)(NCCNC/C=C/C3=CC=C(Br)C=C3)=O.Cl.Cl | ||
| 分子式 | C20H20BrN3O2S.2HCl | 分子量 | 519.28 |
| 溶解度 | ≥ 51.9mg/mL in DMSO | 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。 |
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| 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.9257 mL | 9.6287 mL | 19.2574 mL |
| 5 mM | 0.3851 mL | 1.9257 mL | 3.8515 mL |
| 10 mM | 0.1926 mL | 0.9629 mL | 1.9257 mL |
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| 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 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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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，
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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。
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Modulation of thapsigargin-induced calcium mobilisation by cyclic AMP-elevating agents in human lymphocytes is insensitive to the action of the protein kinase A inhibitor H-89
Cell Signal2001 Jun;13(6):441-9.PMID: 11384843DOI: 10.1016/s0898-6568(01)00166-8
Ca2+ mobilisation from internal stores and from the extracellular medium is one of the primary events involved in lymphocyte activation and proliferation. Regulation of these processes by adenosine 3',5'-cyclic monophosphate (cAMP) and cAMP-dependent protein kinase (PKA) was studied in Fura2-loaded human peripheral blood lymphocytes. Cytosolic Ca2+ concentration ([Ca2+]i) was measured in single cells by the use of a ratio imaging fluorescence microscope and Ca2+ mobilisation was achieved by the use of the endoplasmic reticulum (ER) Ca2+ ATPase inhibitor, thapsigargin (Thg). Our results show that both activation and inhibition of PKA, with forskolin (FSK) and N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide.2HCl (H-89), respectively, inhibited the Thg-induced Ca2+ entry. Furthermore, FSK also reduced the ability of Thg to release Ca2+ from internal stores. This reduction was inhibited by the adenylyl cyclase (AC) inhibitor 9-(tetrahydro-2-furanyl)-9-H-purin-6-amine (SQ22,536), but not by the PKA inhibitor H89, indicating that cAMP but not PKA is responsible for this effect. FSK effect was mimicked by dibutyryl cAMP (dbcAMP) and by inhibition of phosphodiesterases (PDEs) with rolipram (ROL) and milrinone (MIL). We also showed that a very high concentration of H-89 (100 microM) releases Ca2+ from an intracellular pool, although this action is probably independent of PKA inhibition. Neither 10 microM H-89 nor other cAMP/PKA-modulating drugs had any effect on the basal [Ca2+]i of human lymphocytes. We conclude that PKA may act as a fine modulator of capacitative Ca2+ entry, while cAMP has a PKA-independent interaction with the Ca2+ stores of human lymphocytes.
Regulation of myocardial heat shock protein 70 gene expression following exercise
J Mol Cell Cardiol2004 Oct;37(4):847-55.PMID: 15380675DOI: 10.1016/j.yjmcc.2004.05.021
Post-exercise induction of myocardial heat shock protein (Hsp70) gene expression involves the activation of the heat shock transcription factor (HSF1). While the exact mechanisms governing the regulation of HSF1 are unclear, activation is believed to occur subsequent to hyperphosphorylation of specific serine residues. As two important serine kinases, protein kinase A (PKA) and protein kinase C (PKC), have been implicated in many phosphorylative events in myocardial cells, we examined the role of these kinases in the activation of Hsp70 gene expression following exercise. In this report, we show that prior administration of a PKA inhibitor, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide ?2HCl (H-89; 0.36 mg/kg), significantly suppressed the elevation in Hsp70 mRNA (P < 0.05) and protein synthesis (P < 0.05) in male Sprague-Dawley rats following a single bout of exercise. In contrast, this post-exercise elevation in Hsp70 mRNA and protein synthesis was not suppressed following the administration of a PKC inhibitor chelerythrine chloride (CHEL; 5 mg/kg) (P < 0.05). Of note, inhibition of PKA did not alter the nuclear localization and binding affinity of HSF1 to the promotor region of the Hsp70 gene. These data indicate that PKA, and not PKC, plays a necessary role in the early exercise-induced regulation of Hsp70 gene expression, downstream of DNA-binding acquisition. However, the current study does not support previous observations regarding major changes in HSF1 phosphorylation and suggests that other PKA-related mechanisms mediate the activation of Hsp70 gene expression following exercise.
Characterization of human 5-HT4(d) receptor desensitization in CHO cells
Br J Pharmacol2003 Feb;138(3):445-52.PMID: 12569069DOI: 10.1038/sj.bjp.0705061
1 Serotonin 5-HT(4) receptor isoforms differ in their C-terminal tail and yet little is known about their regulation. In this study, we investigated the desensitization of two human 5-HT(4) receptors stably expressed in CHO cells, with a special emphasis on the h5-HT(4(d)) isoform. 2 Exposure of h5-HT(4(d)) and h5-HT(4(e)) receptors to 1 micro M 5-HT induced a rapid desensitization of the adenylyl cyclase response. The h5-HT(4(d)) receptor desensitized with a faster rate (t(1/2)<5 min) than the h5-HT(4(e)) receptor (t(1/2)=15 min) and after 10 min 5-HT treatment cAMP production was reduced by approximately 70%. 3 5-HT-induced h5-HT(4(d)) receptor desensitization was mimicked by 8-Bromo-cAMP, a cAMP analogue, and was inhibited by [n-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulphonamide, 2HCl] (H-89), an inhibitor of cAMP-dependent protein kinase (PKA). Inhibitors of endocytosis (sucrose, 0.45 M and concanavaline A, 0.25 mg ml(-1)) partially reversed the h5-HT(4(d)) receptor desensitization process. 4 Given the prominent role of PKA in agonist-induced desensitization, we mutated the four putative PKA phosphorylation sites present in the third intracellular loop (Ser242, Thr253, Thr255) and the C terminal tail (Ser338) of the h5-HT(4(d)) receptor. Surprisingly, mutated receptors in which either one or all four putative phosphorylation sites were substituted to alanine did not impair receptor desensitization suggesting that PKA might act on nonconsensus sites. 5 Altogether, our data demonstrate that the C-terminal tail of h5-HT(4) receptors may influence the rate of agonist-induced desensitization and we provide evidence for a major role of PKA in h5-HT(4(d)) receptor desensitization.
H89 enhances the sensitivity of cancer cells to glyceryl trinitrate through a purinergic receptor-dependent pathway
Oncotarget2015 Mar 30;6(9):6877-86.PMID: 25762630DOI: 10.18632/oncotarget.3124
High doses of the organic nitrate glyceryl trinitrate (GTN), a nitric oxide (NO) donor, are known to trigger apoptosis in human cancer cells. Here, we show that such a cytotoxic effect can be obtained with subtoxic concentrations of GTN when combined with H89, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulphonamide.2HCl. This synergistic effect requires the generation of reactive oxygen species (ROS) from H89 and NO from GTN treatment that causes cGMP production and PKG activation. Furthermore, the GTN/H89 synergy was attenuated by inhibition of P2-purinergic receptors with suramin and competition with ATP/UDP. By down-regulating genes with antisense oligonucleotides, P2-purinergic receptors P2X3, P2Y1, and P2Y6 were found to have a role in creating this cytotoxic effect. Thus, H89 likely acts as an ATP mimetic synergizing with GTN to trigger apoptosis in aggressive cancer cells.
Dopamine D1 receptor-dependent inhibition of NaCl transport in the rat thick ascending limb: mechanism of action
Eur J Pharmacol2003 Jul 25;473(2-3):185-90.PMID: 12892837DOI: 10.1016/s0014-2999(03)01965-4
Our previous in vitro microperfusion studies established that dopamine inhibits sodium chloride transport in the rat medullary thick ascending limb. The present study was designed to determine the intracellular signaling pathway mediating this response. The dopamine D1 receptor agonist fenoldopam (1 microM) inhibited sodium chloride transport in the thick ascending limb by 42+/-5%. The dopamine D1 receptor antagonist R-(+)-7-Chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-HCl (SCH-23390) completely blocked this effect of fenoldopam. Suppression of protein kinase A activity using either myristoylated protein kinase inhibitor (PKI) or N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide.2HCl (H-89), as well as suppression of phospholipase C activity using 1-(6-((17 beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U-73122), had no effect on fenoldopam-dependent inhibition of transport. In contrast, inhibition of phospholipase A2 activity using E-6-(Bromomethylene)tetrahydro-3-(1-naphthalenyl)-2H-pyran-2-one (HELSS) significantly attenuated the effect of fenoldopam by 74%. The cytochrome P-450 monooxygenase inhibitor 17-octadecynoic acid (17-ODYA) and the protein kinase C inhibitor staurosporine both significantly attenuated the effects of fenoldopam by 67%. Exposure to 20-Hydroxy-(5Z, 8Z, 11Z, 14Z)-eicosatetraenoic acid (20-HETE) inhibited transport by 31+/-5%, and this effect was significantly attenuated by 66% in the presence of staurosporine. We propose a signaling pathway in which dopamine activates a calcium-independent phospholipase A2 in the medullary thick ascending limb. Released arachidonic acid is then metabolized to 20-HETE which subsequently increases protein kinase C activity that acts as a final transport effector.