RS4;11 cells are 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 are 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 are determined using Cell TiterGlo reagent. EC₅₀ values are determined by nonlinear regression analysis of the concentration-response data. Mouse FL5.12-BCL-2 and FL5.12-BCL-XL cells are propagated and assessed. Bak^-/- Bax^-/- double knockout mouse embryonic fibroblasts are 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 is added in half-log dilutions starting at 5 μM. The cells are then incubated at 37°C (5% CO₂) for 48 h, and the effects on proliferation are determined using Cell TiterGlo reagent^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[2]
Nonobese diabetic/severe combined immunodeficient γ (NSG) mice are injected at 6 weeks of age in the tail vein with 150 µL phosphate-buffered saline containing 5×10⁶ luciferase-labeled LOUCY cells. At regular time points, the bioluminescence is measured using the IVIS Lumina II imaging system. At 6 weeks, the cells are engrafted and the mice are randomly divided into 2 groups (with an equal number of males and females in both groups), and the treatment is started on day 0. Mice are treated with 100 mg ABT-199/kg body weight or with vehicle via oral gavage for 4 consecutive days. At days 0, 2, and 4 the bioluminescene is measured. Before imaging, the mice are injected intraperitoneally with 200 µL of a 15 mg/mL firefly D-luciferin potassium salt solution and anesthetized by inhalation of 5% isoflurane. The mice are imaged 10 minutes after luciferin injection. The total bioluminescence signal in each mouse is calculated via the region of interest tool (total counts) in the Living Image software.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Souers AJ, et al. ABT-199, a potent and selective BCL-2 inhibitor, achieves antitumor activity while sparing platelets. Nat Med. 2013 Feb;19(2):202-8.

  [2]. Peirs S, et al. ABT-199 mediated inhibition of BCL-2 as a novel therapeutic strategy in T-cell acute lymphoblastic leukemia. Blood. 2014 Dec 11;124(25):3738-47.

  [3]. Bi C, et al. Inhibition of 4EBP phosphorylation mediates the cytotoxic effect of mechanistic target of rapamycin kinase inhibitors in aggressive B-cell lymphomas. Haematologica. 2017 Apr;102(4):755-764.

868.44

C₄₅H₅₀ClN₇O₇S

1257044-40-8

O=C(NS(=O)(C1=CC=C(NCC2CCOCC2)C([N+]([O-])=O)=C1)=O)C3=CC=C(N4CCN(CC5=C(C6=CC=C(Cl)C=C6)CC(C)(C)CC5)CC4)C=C3OC7=CN=C(NC=C8)C8=C7

Room temperature in continental US; may vary elsewhere