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.

BCL-W:<1 nM (Ki, cell free), BCL2:<1 nM (Ki, cell free), BCL-XL:0.4 nM (Ki, cell free)

方法：鼠原 B 淋巴细胞 FL5.12/Bcl-xL 和 FL5.12/Bcl-2 用 Navitoclax (0.001-1000 nmol/L) 处理 48 h，使用 CellTiter Glo 方法检测细胞活力。
结果：Navitoclax 逆转了 Bcl-2 或 Bcl-xL 的过表达所提供的保护 (EC50 分别为 60 和 20 nmol/L)。在 IL-3 存在的情况下，在 FL5.12 细胞不受促凋亡刺激的情况下，Navitoclax 诱导细胞死亡是无效的。[1]
方法：HCC 细胞 PLC/PRF/5、Hep3B、HepG2 和 Huh7 用 Navitoclax (5 μM) 处理 18 h，使用 Western Blot 方法检测靶点蛋白表达水平。
结果：在用 Navitoclax 治疗后，所有 HCC 细胞系中的 Mcl-1 水平显著增加，但 Bcl-2 和B cl-xL 水平没有显著变化。[2]

方法：为检测体内抗肿瘤活性，将 Navitoclax (100 mg/kg in 10% ethanol+30% polyethylene glycol 400+60% Phosal 50 PG) 口服给药给携带人 SCLC 和 ALL 异种移植物的 scid 小鼠，每天一次，持续二十一天。
结果：口服 Navitoclax 导致体内 SCLC 和 ALL 异种移植物肿瘤消退。[1]
方法：为检测体内抗肿瘤活性，将 Navitoclax (50-100 mg/kg in 10% ethanol+30% polyethylene glycol 400+60% Phosal 50 PG) 单剂量口服给药给携带人 SCLC 肿瘤 H146 的 scid 小鼠。
结果：单剂量 Navitoclax 治疗的 H146 肿瘤显示出大量的死亡和垂死细胞，包括血管化良好的肿瘤周围。[3]

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].

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].

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].