| 名稱 | Navitoclax |
| 描述 | Navitoclax (ABT-263) is a Bcl-2 inhibitor that binds to Bcl-xL, Bcl-2, and Bcl-w proteins (Ki<1 nM) with potent and oral activity. Navitoclax has antitumor activity and induces apoptosis. |
| 細(xì)胞實(shí)驗 | Human tumor cell lines were maintained at 37°C containing 5% CO2. SCLC cell lines were cultured in RPMI 1640 with 10% fetal bovine serum (FBS), 1% sodium pyruvate, 25 mmol/L HEPES, 4.5 g/L glucose, and 1% penicillin/streptomycin. Leukemia and lymphoma cell lines were cultured in RPMI 1640 supplemented with 10% FBS and 1% penicillin/streptomycin. Cells (1 × 10^4–5 × 10^4) were treated for 48 h in 96-well culture plates in a final volume of 100 μL and cytotoxicity was assessed with the CellTiter Glo assay [1]. |
| 激酶實(shí)驗 | ABT-737 and ABT-263 were synthesized as previously described. The enantiomer and BH3-only peptides were synthesized at Abbott. Binding affinities (Ki or IC50) were determined with competitive fluorescence polarization assays. The following peptide probe/protein pairs were used: f-bad (1 nmol/L) and Bcl-xL (6 nmol/L), f-Bax (1 nmol/L) and Bcl-2 (10 nmol/L), f-Bax (1 nmol/L) and Bcl-w (40 nmol/L), f-Noxa (2 nmol/L) and Mcl-1 (40 nmol/L), and f-Bax (1 nmol/L) and Bcl-2-A1 (15 nmol/L). Binding affinities for Bcl-xL were also determined using a time-resolved fluorescence resonance energy transfer assay. Bcl-xL (1 nmol/L, His tagged) was mixed with 200 nmol/L f-Bak, 1 nmol/L Tb-labeled anti-His antibody, and compound at room temperature for 30 min. Fluorescence was measured on an Envision plate reader using a 340/35 nm excitation filter and 520/525 (f-Bak) and 495/510 nm (Tb-labeled anti-His antibody) emission filters. Dissociation constants (Ki) were determined using Wang's equation [1]. |
| 動物實(shí)驗 | C.B.-17 scid-bg or C.B.-17 scid mice were implanted with 5 × 10^6 (1 × 10^6 for DoHH2) cells in 0.2 mL 50% Matrigel s.c. into the right flank. Tumor-bearing mice were size matched (～235 mm3; day 0) into treatment and control groups, ear tagged, and monitored individually. Tumor volume was measured two to three times weekly by electronic calipers (volume = length × width2 / 2). Tumor growth inhibition was calculated based on the difference in mean tumor volumes between treated and appropriate vehicle control groups. Partial response (PR) is defined as ≥50% tumor growth inhibition, and complete response (CR) is defined as nonpalpable tumor. All studies used 8 to 10 mice per group. ABT-263 was formulated in 10% ethanol, 30% polyethylene glycol 400, and 60% Phosal 50 PG and administered p.o. The other agents used [rituximab, doxorubicin, cyclophosphamide, vincristine, bortezomib, and prednisone] were administered i.p., p.o., or i.v. and formulated according to the manufacturers' recommendations. For combination studies, ABT-263 was given ～2 h before the other agents, except bortezomib, which was given ～4 h before ABT-263 [1]. |
| 體外活性 | 方法：鼠原 B 淋巴細(xì)胞 FL5.12/Bcl-xL 和 FL5.12/Bcl-2 用 Navitoclax (0.001-1000 nmol/L) 處理 48 h，使用 CellTiter Glo 方法檢測細(xì)胞活力。
結(jié)果：Navitoclax 逆轉(zhuǎn)了 Bcl-2 或 Bcl-xL 的過表達(dá)所提供的保護(hù) (EC50 分別為 60 和 20 nmol/L)。在 IL-3 存在的情況下，在 FL5.12 細(xì)胞不受促凋亡刺激的情況下，Navitoclax 誘導(dǎo)細(xì)胞死亡是無效的。[1]
方法：HCC 細(xì)胞 PLC/PRF/5、Hep3B、HepG2 和 Huh7 用 Navitoclax (5 μM) 處理 18 h，使用 Western Blot 方法檢測靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：在用 Navitoclax 治療后，所有 HCC 細(xì)胞系中的 Mcl-1 水平顯著增加，但 Bcl-2 和B cl-xL 水平?jīng)]有顯著變化。[2] |
| 體內(nèi)活性 | 方法：為檢測體內(nèi)抗腫瘤活性，將 Navitoclax (100 mg/kg in 10% ethanol+30% polyethylene glycol 400+60% Phosal 50 PG) 口服給藥給攜帶人 SCLC 和 ALL 異種移植物的 scid 小鼠，每天一次，持續(xù)二十一天。
結(jié)果：口服 Navitoclax 導(dǎo)致體內(nèi) SCLC 和 ALL 異種移植物腫瘤消退。[1]
方法：為檢測體內(nèi)抗腫瘤活性，將 Navitoclax (50-100 mg/kg in 10% ethanol+30% polyethylene glycol 400+60% Phosal 50 PG) 單劑量口服給藥給攜帶人 SCLC 腫瘤 H146 的 scid 小鼠。
結(jié)果：單劑量 Navitoclax 治療的 H146 腫瘤顯示出大量的死亡和垂死細(xì)胞，包括血管化良好的腫瘤周圍。[3] |
| 存儲條件 | store at low temperature | 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)
DMSO : 50 mg/mL (51.3 mM), Sonication is recommended.
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | inhibit | Inhibitor | Bcl-2 Family | Navitoclax | ABT 263 | ABT263 |
| 相關(guān)產(chǎn)品 | Diphenyl disulfide | Venetoclax | Thymoquinone | Gossypol (acetic acid) | (S)-(+)-Ibuprofen | Sonrotoclax TFA | Rasagiline | Gossypol | UMI-77 | (E)-Ferulic acid | Lisaftoclax | Allethrin |
| 相關(guān)庫 | 抑制劑庫 | 抗癌活性化合物庫 | 經(jīng)典已知活性庫 | 已知活性化合物庫 | 自噬庫 | 線粒體靶向庫 | 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 |