| 名稱 | B-AP15 |
| 描述 | B-AP15 (NSC-687852)(NSC-687852) is a selective inhibitor of the deubiquitinating enzymes Usp14 and UCHL5 of the 26S proteasome. It blocks the deubiquitinating activity of the 26S proteasome. |
| 細胞實驗 | b-AP15 is dissolved in DMSO and stored, and then diluted with appropriate medium before use[2]. Cell viability is monitored by either the fluorometric microculture cytotoxicity assay or the MTT assay. For the MTT assay, cells are seeded into 96-well flat-bottomed plates overnight and exposed to drugs, using DMSO as the control. At the end of incubations, 10 μl of a stock solution of 5 mg/mL MTT is added into each well, and the plates are incubated 4 hours at 37°C. Formazan crystals are dissolved with 100 μL 10% SDS/10 mM HCl solution overnight at 37°C. Absorbance is measured using an enzyme-linked immunosorbent assay (ELISA) plate reader at 590 nm[2]. |
| 激酶實驗 | For deubiquitinase inhibition assays, 19S regulatory particle (5 nM), 26S (5 nM) UCH-L1 (5 nM), UCH-L3 (0.3 nM), USP2CD (5 nM) USP7CD (5 nM) USP8CD (5 nM) or BAP1 (5 nM) is incubated with DMSO or b-AP15 and monitored the cleavage of ubiquitin-AMC (1,000 nM) using a Wallac VICTOR Multilabel counter or a Tecan Infinite M1000 equipped with 380 nm excitation and 460 nm emission filters[1]. |
| 體外活性 | 在HCT-116結(jié)腸癌移植瘤的小鼠中,B-AP15（5 mg/kg）可使腫瘤發(fā)病顯著延緩.對攜帶鱗狀細胞癌移植瘤的重癥聯(lián)合免疫缺陷小鼠,B-AP15（5 mg/kg）可產(chǎn)生顯著的抗腫瘤活性. |
| 體內(nèi)活性 | 作為UPS抑制劑,通過誘導(dǎo)cathepsin-D依賴性的溶酶體凋亡途徑,B-AP15可誘導(dǎo)細胞死亡。與外周血單核細胞或永生化上皮細胞（hTERT-RPE1）相比,B-AP15對HCT-116細胞毒性更大。B-AP15劑量依賴性地使UbG76V-YFP受體累積（IC50：0.8 μM）,表明損傷的蛋白酶體降解受損。作用于人類結(jié)腸癌HCT-116細胞時,B-AP15（1 μM）可使多聚泛素化蛋白快速累積。B-AP15（1 μM）對ATP誘導(dǎo)的IL-1β從脂多糖誘導(dǎo)的腹腔巨噬細胞中釋放有抑制作用。作用于THP-1細胞,B-AP15（1 μM）可使尼日利亞菌素誘導(dǎo)的細胞死亡水平降低。B-AP15（1 μM）處理脂多糖誘導(dǎo)的THP-1細胞,可使尼日利亞菌素處理后形成的ASC斑點數(shù)量顯著減少。作用于HCT-116細胞,B-AP15（1 μM）使細胞周期在G2/M期停滯,并造成細胞周期抑制劑累積。作用于HCT-116細胞, B-AP15（2.2 μM）使細胞周期蛋白依賴性激酶CDKN1A和CDKNIB,及腫瘤抑制基因TP53的數(shù)量劑量依賴性地提高,但不影響鳥氨酸脫羧酶1的數(shù)量。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year |
| 溶解度 | 10% DMSO+90% Saline : < 2.45 mg/mL (5.84 mM), Lower concentrations may be soluble, but exact solubility limit is unknown.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 2.45 mg/mL (5.84 mM), Suspension.
DMSO : 1 mg/mL (2.38 mM), Sonication is recommended.
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| 關(guān)鍵字 | UCH-L5 | NSC-687852 | NSC687852 | Inhibitor | inhibit | DUBs | Deubiquitinase | B-AP-15 | B-AP15 | BAP15 | B AP15 | Apoptosis |
| 相關(guān)產(chǎn)品 | Urea | Formamide | Dimethyl phthalate | Aceglutamide | Alginic acid | Dextran sulfate sodium salt (MW 5000) | Metronidazole | Sildenafil citrate | Citric Acid Triammonium | Stavudine | Tamoxifen | Adenosine |
| 相關(guān)庫 | 已知活性庫Plus |