| 名称 | BI-D1870 |
| 描述 | BI-D1870 is a cell-permeable, ATP-competitive inhibitor of ribosomal S6 kinases (RSKs; IC50s: 31/24/18/15 nM for RSK1/2/3/4). |
| 细胞实验 | The rat embryo fibroblast cell line, Rat-2 cells were cultured on 10 cm-diameter dishes in Dulbecco's Modified Eagle's medium supplemented with 10% (v/v) FBS. HEK-293 cells were cultured on 10 cm-diameter dishes in Dulbecco's Modified Eagle's medium supplemented with 10% FBS and 1×antimycotic/antibiotic solution. Prior to stimulation, cells were cultured in the absence of serum for 16 h. Inhibitors were dissolved in DMSO at a 1000-fold higher concentration than they were used at. These inhibitors, or the equivalent volume of DMSO as a control, were added to the tissue culture medium 30 min prior to stimulation unless indicated otherwise. The final concentration of DMSO in the culture medium was 0.1% and had no effect on agonist-induced activation or phosphorylation of any of the substrates examined. The cells were stimulated with the indicated agonists and lysed in 1 ml of ice-cold Lysis Buffer and centrifuged at 16000 g at 4 °C for 5 min. The supernatants were frozen in liquid nitrogen and stored at ?80 °C until use. Protein concentrations were determined using the Bradford method with BSA as the standard [1]. |
| 激酶实验 | Purified His6–RSK1, His6–RSK2 or GST–RSK21–389:S381E (1–2 units/ml) were assayed for 10 min at 30 °C in a 50 μl assay mixture in Buffer A containing 30 μM substrate peptide (KEAKEKRQEQIAKRRRLSSLRASTSKSGGSQK), 10 mM magnesium acetate and 100 μM of [γ-32P]ATP. Reactions were terminated and analyzed as described previously. The amount of enzyme that catalyzed the phosphorylation of 1 nmol of substrate peptide in 1 min was termed one unit. In order to assay RSK and MSK1 in HEK-293 or Rat-2 cell lysates, these kinases were immunoprecipitated from the cell lysates (0.1 mg of lysate protein for RSK and 0.3 mg for MSK1) and assayed as described previously, except that for RSK assays the immunoprecipitates were washed twice with Buffer A containing 1 mM ATP and twice with Buffer A prior to the assay, as a precaution to ensure dissociation of BI-D1870 from the RSK isoforms [1]. |
| 动物实验 | Myelin oligodendrocyte glycoprotein (MOG) peptide 35–55. (MEVGWYRSPFSRVVHLYRNGK) (BEX) was used to induce EAE in C57/BL6J mice. Mice were injecteds.c. with 200 g of MOG peptide in100 L of PBS emulsified in 100 L complete Freund's adjuvant (CFA) that was further supplemented with five mg mL?1 Mycobacterium tuberculosis (H37Ra). In addition, 500 ng pertussis toxin was injected i.p. on days zero and two. The RSK inhibitor (BI-D1870; 0.5 mg kg?1) was injected i.p. into mice two days after immunization with MOG peptide, and injection was repeated every other day for 11 days. Mice that received only dimethyl sulfoxide (DMSO) solution were used as controls. Paralysis was evaluated according to the following scale: zero, no disease; one, tail limpness; two, hind limb weakness; three, hind limb paralysis; four, forelimb weakness; five, quadriplegia; six, death. For histological analysis, CNS samples were fixed with 4% paraformaldehyde and sliced at 4 m, and then hematoxylin & eosin (H & E) staining was performed [4]. |
| 体外活性 | BI-D1870能够穿透细胞并阻止在HEK 293细胞和Rat-2细胞中由RSK介导的phorbol酯和EGF（表皮生长因子）引发的GSK-3β和LKB1的磷酸化。相比之下，BI-D1870并不影响由其他六种AGC激酶触发的底物磷酸化。此外，BI-D1870也不抑制phorbol酯或EGF诱导的CREB（cAMP反应元素结合蛋白）的磷酸化[1]。在LN-229细胞中，1μM的BI-D1870对p70S6K的磷酸化和rpS6的减少几乎无影响。然而，10μM的BI-D1870在孵育90分钟后强烈诱导p70S6K激活。在LN-18细胞中，10μM的BI-D1870刺激了rpS6和p70S6K的磷酸化，峰值出现在90分钟[2]。BI-D1870对OSCC细胞展示出剂量依赖的抗增殖效应，同时相对保护正常的人类口腔角质形成细胞。该化合物通过抑制下游RSK靶标YB-1并诱导细胞凋亡来发挥作用。此外，BI-D1870还通过调节p21等细胞周期调节器的表达来诱导G2/M阻滞，并展示出新发现的抗癌特性，包括产生反应性氧种、增加内质网应激和自噬[3]。 |
| 体内活性 | BI-D1870 (0.5 mg/kg) 的给药能够保护小鼠免受实验性自身免疫性脑炎（EAE）的影响，其机制是通过减少TH1和TH17细胞向中枢神经系统（CNS）的浸润，并降低TH17细胞中Ccr6的mRNA水平[4]。 |
| 存储条件 | 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 : 6.88 mg/mL (17.56 mM), Sonication is recommended.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 7.2 mg/mL (18.39 mM), Suspension. Working solution is recommended to be prepared and used immediately.
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| 关键字 | BI-D 1870 | BI-D1870 | inhibit | Ribosomal S6 Kinase (RSK) | S6K | Inhibitor | BI-D-1870 | Autophagy | BI D1870 | BID1870 |
| 相关产品 | Oxyresveratrol | Guanidine hydrochloride | Naringin | Taurine | Gefitinib | Xylitol | Hydroxychloroquine | Curcumin | Stavudine | Myricetin | Paeonol | Sodium 4-phenylbutyrate |
| 相关库 | 抑制剂库 | 经典已知活性库 | 已知活性化合物库 | 抗结直肠癌化合物库 | 激酶抑制剂库 | 抗衰老化合物库 | 谷氨酰胺代谢化合物库 | 抗肝癌化合物库 | 疼痛相关化合物库 | 表型筛选靶点鉴定库 |