ABT199 是一种高效，口服有效的选择性BCL-2抑制剂，可诱导自噬，Ki小于0.01 nM。

ABT-199 is a selective inhibitor of Bcl-2 (Ki< 0.010 nM), binding over 3 orders of magnitude less avidly to Bcl-xL, and Bcl-W (Kis = 48 and 245 nM, respectively).

Venetoclax (ABT-199) has a subnanomolar affinity for BCL-2 (Ki< 0.010 nM) and bound over three orders of magnitude less avidly to BCL-XL (Ki = 48 nM) and BCL-W (Ki = 245 nM). ABT-199 has no measurable binding to MCL-1 (Ki >444 nM). Although ABT-199 potently killed FL5.12–BCL-2 cells (EC50 = 4 nM), it showed much weaker activity against FL5.12–BCL-XL cells (EC50 = 261 nM) [1]. The T-ALL cell line LOUCY, which shows a transcriptional program related to immature T-ALL, exhibited high in vitro and in vivo sensitivity for ABT-199 in correspondence with high levels of BCL-2 [2].

After a single oral dose of 12.5 mg/kg in xenografts derived from RS4;11 cells (ALL), ABT-199 caused a maximal tumor growth inhibition (TGImax) of 47% and tumor growth delay (TGD) of 26%. The magnitude and durability of the response increased in a dose-dependent manner, with the highest dose of 100 mg per kg body weight conferring a TGImax of 95% and a TGD of 152% [1]. Some immature, TLX3- or HOXA-positive primary T-ALLs are highly sensitive to BCL-2 inhibition, whereas TAL1 driven tumors mostly showed poor ABT-199 responses [2].

The equilibrium binding experiments of fluorescent peptides to Bcl-xL protein were performed in an Analyst 96-well plate reader under the following conditions: each individual well in a 96-well assay plate contained 5 μl DMSO, 15 nM fluorescent peptide, and increasing concentrations (from 0 to 2.24 μM) of Bcl-xL protein in assay buffer in a final volume of 125 μl. The plate was mixed on a shaker for 1 min and incubated at room temperature for an additional 15 min. The polarization in millipolarization units (mP) was measured at room temperature with an excitation wavelength at 485 nm and an emission wavelength at 530 nm. For assay stability testing, a plate containing a binding experiment was measured at different times over a 24-h period. Between each reading, the plate was covered with parafilm to prevent any solution evaporation. To determine the effect of DMSO on the assay, binding experiments were performed under conditions similar to those described above except that the amount of DMSO was varied from 0 to 4 to 8%. All experimental data were analyzed using Prism 3.0 software and Kd values were generated by fitting the experimental data using a sigmoidal dose-response nonlinear regression model [1].

RS4;11 cells were seeded at 50,000 per well in 96-well plates and treated with compounds diluted in half-log steps starting at 1 μM and ending at 0.00005 μM. All other leukemia and lymphoma cell lines were seeded at 15,000–20,000 cells per well in the appropriate medium and incubated with ABT-199 or navitoclax for 48 h. Effects on proliferation were determined using Cell TiterGlo reagent. EC50 values were determined by nonlinear regression analysis of the concentration-response data. Mouse FL5.12–BCL-2 and FL5.12–BCL-XL cells were propagated and assessed as described previously. Bak?/? Bax?/? double knockout mouse embryonic fibroblasts were seeded into 96-well microtiter plates at 5,000 cells per well in DMEM supplemented with 10% FBS. ABT-199 in the same culture medium was added in half-log dilutions starting at 5 μM. The cells were then incubated at 37 °C (5% CO2) for 48 h, and the effects on proliferation were determined using Cell TiterGlo reagent according to the manufacturer's instructions [1].

Female C.B-17 SCID mice (DoHH2 and Granta-519 xenografts) and female C.B-17 SCID-beige mice (RS4;11 and Toledo xenografts) were inoculated with 1 × 10^6 (DoHH2) or 5 × 10^6 (Granta-519, Toledo and RS4;11) cells subcutaneously in the right flank. The inoculation volume (0.2 ml) comprised a 50:50 mixture of cells in growth media and Matrigel. Electronic calipers were used to measure the length and width of each tumor 2–3 times per week. Tumor volume was estimated by applying the following equation: volume = length × width2/2. When tumors reached approximately 220 mm3, mice were size matched (day 0) into treatment and control groups. All xenograft trials were conducted using ten mice per group, and all mice were ear tagged and monitored individually throughout the studies. ABT-199 was formulated for oral dosing in 60% phosal 50 propylene glycol (PG), 30% polyethylene glycol (PEG) 400 and 10% ethanol, and bendamustine and rituximab were formulated in accordance with the manufacturer's instructions. ABT-199 was delivered approximately 2 h before bendamustine or bendamustine plus rituximab. TGImax was calculated as the greatest treatment response using the following equation: TGImax = (1 ? mean tumor volume of the treated group/mean tumor volume of the vehicle control group) × 100. The TGD (%) was determined as the percentage increase of the median time period for the treatment group to reach an arbitrary tumor volume of 1,000 mm3 relative to the vehicle control group. A complete tumor regression response was the portion of the population with tumors ≤25 mm3 for at least three consecutive measurements [1].

1257044-40-8

C45H50ClN7O7S

868.45

ABT 199;维奈妥拉;GDC-0199;ABT199;ABT-199

Ethanol:<1 mgml
H2O:<1 mgml
DMSO:100mg/mL
Powder: -20°C for 3 years
In solvent: -80°C for 2 years