| 名稱 | GS-444217 |
| 描述 | GS-444217 is a selective ATP-competitive inhibitor of apoptosis signal-regulating kinase 1 (ASK1, IC50: 2.87 nM). |
| 細(xì)胞實驗 | Human embryonic kidney cells (HEK293T) were infected with full-length human ASK1 adenovirus or with an inactive truncated ASK1 adenovirus (K709R mutant with N-terminally truncated protein) as a negative control using the following conditions: (a) dose response: cells were infected for 24 hours followed by incubation for 2 hours with 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μM GS-444217; (b) kinetics: after cells were infected for 24 hours, 1 μM GS-444217 was added to cells for 1, 5, 10, or 30 minutes or 1, 2, or 4 hours; (c) off-rate kinetics: cells were infected for 24 hours followed by incubation with GS-444217 for 30 minutes. After 30 minutes, medium was replaced with serum-free medium without compound and incubated for 0, 10, or 30 minutes or 1, 2, or 4 hours [1]. |
| 動物實驗 | Male Sprague-Dawley rats (176–200 g, 7–8 weeks old) were randomly assigned to weight-matched treatment groups: (a) sham procedure, n = 8; (b) ischemia 30 minutes, n = 8; (c) ischemia 30 minutes plus GS-444217 (30 mg/kg, p.o.), n = 8. Bilateral renal occlusion was performed on anesthetized rats held at 37°C for 30 minutes. After recovering from anesthesia, rats were placed in metabolic cages for a 24-hour collection of urine. Sham rats underwent midline incision with surgery duration of 30 minutes but were not subjected to occlusion. Necropsy was performed on all rats 24 hours after surgery to collect kidneys and blood. Renal I/R studies were performed at Physiogenix Inc. Serum was analyzed for creatinine and blood urea nitrogen concentrations on a clinical chemistry analyzer. The right kidney was fixed in formalin and stained with H&E to assess tubular necrosis by pathology and with TUNEL to detect apoptotic cells [1]. |
| 體外活性 | GS-444217 對 ASK1 的結(jié)合具有高選擇性，與面板中的其他激酶相比。GS-444217 對 ASK1 的親和力（KD = 4.1 nM）是對 DYRK1A（KD = 220 nM）的 53 倍，是對 RSK4（KD = 430 nM）的 104 倍。GS-444217 的處理降低了 ASK1 的磷酸化，并防止了在 0.3 μM 及以上濃度下 MKK3/6、MKK4、p38 和 JNK 的磷酸化，1 μM 時完全抑制了 ASK1 活性。GS-444217 降低 ASK1 活性，在加入培養(yǎng)物后 5 分鐘內(nèi)達(dá)到，并在 30 分鐘內(nèi)達(dá)到最大抑制水平。從培養(yǎng)物中移除 GS-444217 后，ASK1 自磷酸化在 10 分鐘內(nèi)重新激活，并在化合物清洗后 2 小時內(nèi)近乎完全恢復(fù)[1]。 |
| 體內(nèi)活性 | 使用GS-444217處理能夠抑制糖尿病腎臟中p38 MAPK的激活，但對Nos3(-/-)小鼠的高血壓無影響。GS-444217的早期干預(yù)顯著抑制了糖尿病性腎小球硬化，減少了腎功能障礙，但對白蛋白尿的發(fā)展無效。GS-444217的晚期干預(yù)改善了腎功能，阻止了腎小球硬化、腎臟炎癥和腎小管損傷的進(jìn)展，盡管對已確立的白蛋白尿無影響[2]。GS-444217（30 mg/kg）單劑量給藥，在給予金剛烷胺（30 mg/kg）前30分鐘，能夠抑制腎皮質(zhì)中ASK1、p38和JNK的激活。金剛烷胺的給藥引起了炎癥細(xì)胞因子（Il1b、Ccl2和Cxcl2）mRNA表達(dá)的增加，并在腎臟中增加了caspase活性。這些ASK1激活的下游效應(yīng)被GS-444217所抑制。通過比較GS-444217的血漿濃度與腎臟中相應(yīng)的磷酸化p38（p-p38）信號，GS-444217對抑制嚙齒動物腎臟中的ASK1通路具有大約1.6 μM的體內(nèi)EC50[1]。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 55 mg/mL (133.67 mM)
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| 關(guān)鍵字 | Apoptosis | inhibit | MAP kinase kinase kinase, MEKK, MAPKKK | MAP3K | GS-444217 | Inhibitor |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Metronidazole | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Myricetin | Sorafenib | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 細(xì)胞焦亡化合物庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗糖尿病庫 | 抗肥胖化合物庫 | 疼痛相關(guān)化合物庫 | 細(xì)胞重編程化合物庫 |