| 名稱 | Delanzomib |
| 描述 | Delanzomib (CEP-18770) is an orally active inhibitor of the chymotrypsin-like activity of proteasome (IC50: 3.8 nM). It only marginally inhibits the tryptic and peptidylglutamyl activities of the proteosome. |
| 細胞實驗 | HMEC and TEC cells are seeded into 24-well plates at a density of 104 cells/well in DMEM supplemented with 5% FCS. After incubation with proteasome inhibitors (48 hours), cells are washed, air dried, and stained with crystal violet as described. Cell number is determined, in duplicate samples, on the basis of a standard curve obtained with known cell numbers. All experiments are performed in triplicate. In vitro formation of capillary-like structures is studied on cells (4 × 104 cells/well in DMEM supplemented with 5% FCS. After incubation with proteasome inhibitors (48 hours), cells are washed (cells/well in 24-well plates) and seeded onto Matrigel-coated wells in DMEM containing 0.25% BSA. HMEC and TEC cells (5 × 103 per well), suspended in 200 μL DMEM with 5% FCS (positive control), serum-free medium (negative control), are layered onto the Matrigel surface in the presence or absence of proteasome inhibitor CEP-18770. Cells are observed with a inverted microscope and experimental results are then recorded after a 6-hour incubation at 37 °C. Data is analyzed, as the mean (× 1 SD) of total length of capillary-like structures, by the Micro-Image system and is expressed as mm/field by the computer analysis system in 5 different fields at 100 × magnification in duplicated wells for 4 different experiments. (Only for Reference) |
| 激酶實驗 | Probing proteasome activity in cell extracts: Human multiple myeloma cells are washed twice with cold phosphate-buffered saline, pelleted and lysed with one volume of glass beads (<106 microns, acid-washed) and an equal volume of homogenization buffer (50 mM Tris (pH 7.4), 1 mM dithiothreitol, 5 mM MgCl2, 2 mM ATP and 250 mM sucrose) by vortexing at high speed for 15-30 min at 4 °C. Beads, membrane fractions, nuclei and cell debris are then removed from the supernatant by centrifugation at 16,000 g for 5 min. The protein content of extracts is quantitated using the Bradford assay. Proteasome activity is assayed as described below. Equal amounts (typically 60 g) of protein are denatured by boiling in reducing sample buffer, separated by 12.5% SDS-PAGE and electrotransferred onto polyvinylidene difluoride (PVDF) membranes. Immunoblotting is performed using a dansyl-sulfonamidohexanoyl polyclonal antibody (1:7,500, rabbit) and horseradish peroxidase–coupled goat or swine anti-rabbit secondary antibody followed by enhanced chemiluminescence. |
| 體外活性 | CEP-18770展現(xiàn)了對蛋白酶體的胰蛋白酶樣和肽酰谷氨?；钚缘挠邢拮钄嗄芰?，其IC50值與bortezomib相當，能在低納摩爾濃度下抑制蛋白酶體的胰蛋白酶樣和類凋亡酶活性。[1] CEP-18770對包括A2780卵巢癌細胞、PC3前列腺癌細胞、H460、LoVo結(jié)腸癌細胞、RPMI8226多發(fā)性骨髓瘤和HS-Sultan非霍奇金淋巴瘤等癌癥細胞的IC50值分別為13.7、22.2、34.2、11.3、5.6和8.2 nM。[1] CEP-18770在多發(fā)性骨髓瘤(MM)和慢性髓細胞性白血病細胞系K562中阻斷泛素-蛋白酶體途徑，引發(fā)泛素化蛋白質(zhì)在4到8小時內(nèi)積累，其特征與bortezomib處理后觀察到的相似。[1] 預(yù)處理CEP-18770可完全阻斷IκBα的降解。在RPMI-8226和U266細胞中，CEP-18770顯著抑制高水平的NF-κB活性。CEP-18770對MM細胞系中NF-κB DNA結(jié)合活性的時間和濃度依賴性抑制導致了包括IκBα、X染色體連鎖凋亡抑制蛋白(XIAP)、促炎細胞因子TNF-α和白細胞介素-1β(IL-1β)、細胞間黏附分子(ICAM1)及促血管生成因子血管內(nèi)皮生長因子等數(shù)個由NF-κB調(diào)控、介導腫瘤細胞生長和存活的基因表達的下降。[1] 這些由NF-κB調(diào)控的基因表達及其受bortezomib調(diào)控的方式，凸顯了它們在響應(yīng)CEP-18770暴露方面的潛在預(yù)后價值，從而與此化合物的更佳Clinical反應(yīng)性相關(guān)。 |
| 體內(nèi)活性 | CEP-18770展示了劑量相關(guān)的相對腫瘤重量抑制，并且與硼替佐米比較，以1.2 mg/kg靜脈注射的最大耐受劑量(MTD)實現(xiàn)50%完全緩解(CR)的發(fā)生率相比，CEP-18770引導劑量相關(guān)的完全腫瘤退化。與硼替佐米不同，CEP-18770在完成這些研究（腫瘤移植后120天）時，腫瘤自由小鼠的發(fā)生率呈劑量相關(guān)增加。CEP-18770的口服給藥在120天研究期間顯著減少腫瘤重量，并且完全腫瘤退化的劑量相關(guān)發(fā)生率明顯，同時動物體重變化最小。與硼替佐米比較，等效劑量的CEP-18770顯示出更大且更持久的劑量相關(guān)腫瘤蛋白酶體活性抑制，并且與caspase-3和7活性的最大誘導臨時對應(yīng)。CEP-18770的最大凋亡信號是硼替佐米的2.5倍。相比之下，CEP-18770和硼替佐米在正常的外圍小鼠組織（肝、肺、全血和大腦[無活性]）檢查中的蛋白酶體抑制剖面在其幅度和持續(xù)時間上是可比的。在任何時間點，兩者的大腦組織中都未檢測到蛋白酶體抑制。在MM異種移植模型中，CEP-18770與美法蘭的組合能完全阻止對美法蘭敏感或美法蘭耐藥腫瘤的生長。CEP-18770與硼替佐米的組合導致對硼替佐米敏感的腫瘤完全退化，并與單一化合物治療相比，明顯延緩對硼替佐米耐藥腫瘤的進展。單一化合物CEP-18770 PO在這些異種移植模型中也顯示出顯著的抗MM效應(yīng)。 |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | H2O : < 1 mg/mL (insoluble or slightly soluble)
Ethanol : 77 mg/mL (186.3 mM)
DMSO : 77 mg/mL (186.3 mM)
|
| 關(guān)鍵字 | Inhibitor | CEP 18770 | anti-cancer | Nuclear factor-κB | Nuclear factor-kappaB | chymotrypsin-like | Apoptosis | Delanzomib | antiangiogenic | NF-κB | inhibit | Proteasome | CEP18770 |
| 相關(guān)產(chǎn)品 | L-Glutamic acid | Diallyl disulfide | Metronidazole | 5-Fluorouracil | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | Myricetin | L-Ascorbic acid | Acetylcysteine | Salicylic acid | Sodium 4-phenylbutyrate |
| 相關(guān)庫 | 抑制劑庫 | 抗癌活性化合物庫 | 經(jīng)典已知活性庫 | 抗衰老化合物庫 | 藥物功能重定位化合物庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 |