| 名稱 | CHIR-98014 |
| 描述 | CHIR-98014 (CT98014) is a selective GSK3 inhibitor; potentiate insulin activation of glucose transport and utilization in vitro and in vivo. |
| 細胞實驗 | CHO-IR cells expressing human insulin receptor are grown to 80% con?uence in Hamm’s F12 medium with 10% fetal bovine serum and without hypoxanthine. Trypsinized cells are seeded in 6-well plates at 1 × 106 cells/well in 2 mL of medium without fetal bovine serum. After 24 hours, medium is replaced with 1 mL of serum-free medium containing GSK-3 inhibitor CHIR 98014 or control (?nal DMSO concentration 0.1%) for 30 min at 37 °C. Cells are lysed by freeze/thaw in 50 mM tris (pH 7.8) containing 1 mM EDTA, 1 mM DTT, 100 mM NaF, 1 mM phenylmethylsulfonyl ?uoride, and 25 μg/mL leupeptin (buffer A) and centrifuged 15 min at 4 °C/14000 g. The activity ratio of GS is calculated as the GS activity in the absence of glucose-6-phosphate divided by the activity in the presence of 5 mM glucose-6-phosphate. (Only for Reference) |
| 激酶實驗 | Kinase assays: Polypropylene 96-well plates are ?lled with 300 μL/well buffer (50 mM tris HCl, 10 mM MgCl2, 1 mM EGTA, 1 mM dithiothreitol, 25 mM β-glycerophosphate, 1 mM NaF, 0.01% BSA, pH 7.5) containing kinase, peptide substrate, and any activators. CHIR-98014 or controls are added in 3.5 μL of DMSO, followed by 50 μL of ATP stock to yield a ?nal concentration of 1 μM ATP in all cell-free assays. After incubation, triplicate 100-μL aliquots are transferred to Combiplate eight plates containing 100 μL/well 50 μM ATP and 20 mM EDTA. After 1 hour, the wells are rinsed ?ve times with PBS, ?lled with 200 μL of scintillation ?uid, sealed, left 30 min, and counted in a scintillation counter. All steps are performed at room temperature. |
| 體外活性 | CHIR 98014降低高血糖和改善葡萄糖利用的效果并不限于db/db小鼠和ZDF大鼠,由于觀察到ob/ob小鼠,飲食誘導(dǎo)的糖尿病C57BL/6小鼠和用CHIR-2處理的葡萄糖不耐受的SHHF大鼠出現(xiàn)類似結(jié)果.此外,CHIR-98014降低了出生后大鼠皮質(zhì)和海馬中tau蛋白的磷酸化（Ser396）.GSK-3抑制劑CHIR-98014作用于從瘦弱的患有糖尿病和抗胰島素的ZDF鼠分離的I型骨骼肌,激活GS活性.瘦弱患有糖尿病鼠分離的肌肉中,胰島素和CHIR 98014激活的GS比ZDF鼠分離的肌肉中強很多.CHIR 98014或胰島素作用于這些細胞和肌肉不會改變?nèi)縂S活性. 同時,CHIR 98014不會影響從瘦弱動物分離的肌肉中胰島素劑量反應(yīng). |
| 體內(nèi)活性 | 雖然CHIR-98014作為ATP結(jié)合的簡單競爭性抑制劑,但它對GSK-3顯示出相對于20種其他蛋白激酶（包括Cdc2,ERK2,Tie-2和KDR在內(nèi)）500倍至> 1000倍的選擇性。CHIR 98014抑制人類GSK-3β,Ki值為0.87 nM。CHIR 98014抑制CDC2,IC50為3.7 μM。然而,CHIR 98014對GSK-3高度同源的ɑ和β亞型具有相似效果,但是在GSK-3和最接近的同系物cdc2和erk2間具有明顯區(qū)別。CHIR 98014使CHO-IR和鼠肝細胞達到半數(shù)最大GS刺激反應(yīng)的濃度分別為106和107 nM。將表達胰島素受體的CHO-IR細胞或原代大鼠肝細胞暴露于遞增濃度的抑制劑CHIR98014中導(dǎo)致GS活性比率高于基礎(chǔ)值的兩倍至三倍的刺激。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | Ethanol : < 1 mg/mL (insoluble or slightly soluble)
H2O : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 4.86 mg/mL (10 mM), Sonication is recommended.
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| 關(guān)鍵字 | Glycogen synthase kinase-3 | CT-98014 | GSK-3 | Inhibitor | CHIR-98014 | inhibit | CT 98014 | Glycogen synthase kinase 3 |
| 相關(guān)產(chǎn)品 | Cromolyn sodium | Amlexanox | 4-Chloro-2'-bromoacetophenone | Sodium salicylate | Formononetin | Lenvatinib mesylate | Ferulic Acid | Pazopanib | Nintedanib | Erdafitinib | CHIR-99021 | Ibrutinib |
| 相關(guān)庫 | 神經(jīng)保護化合物庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 神經(jīng)退行性疾病化合物庫 | 膜蛋白靶向化合物庫 | 疼痛相關(guān)化合物庫 | 酪氨酸激酶分子庫 |