Birinapant

Manufactured by Selleck Chemicals

Sourced in United States

Birinapant is a small molecule compound that functions as an inhibitor of Inhibitor of Apoptosis Proteins (IAPs). IAPs are a family of proteins that play a role in regulating cell death and survival pathways.

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Lab products found in correlation

33 protocols using birinapant

Etanercept and Birinapant Inhibit Tumor Metastasis

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Mice were injected with the BSVTK MDA-MB-231 population (as described above), and the primary tumors were resected when they reached 100 mm³ before the treatment. For the etanercept experiment (two independent experiments), the control group received saline as vehicle; the treated group received etanercept (Enbrel) (10 mg/kg), three times weekly until the end of the experiment. For the birinapant experiment (one experiment), the treated group received birinapant (#S7015, Selleck Chemicals) at the dose of 30 mg/kg, three times weekly until the end of the experiment. The mice were euthanized when the control groups died from metastatic disease. Metastases were analyzed by imaging and flow cytometry, as described below.

Berthelet J., Wimmer V.C., Whitfield H.J., Serrano A., Boudier T., Mangiola S., Merdas M., El-Saafin F., Baloyan D., Wilcox J., Wilcox S., Parslow A.C., Papenfuss A.T., Yeo B., Ernst M., Pal B., Anderson R.L., Davis M.J., Rogers K.L., Hollande F, & Merino D. (2021). The site of breast cancer metastases dictates their clonal composition and reversible transcriptomic profile. Science Advances, 7(28), eabf4408.

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Cytarabine and Birinapant Cytotoxicity Assays

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1 × 10⁴ U937 CUX1^−/− or wild-type cells were dispensed into 96 well plates in triplicate. Cytarabine (Apexbio) was dissolved in water. Drug or vehicle were added to cells to achieve the desired concentrations as indicated. Cells were incubated for 48 h before cell viability was determined using CellTiter 96 AQ_ueous One reagent (Promega). For birinapant treatment of murine cKit⁺ cells, 2 × 10⁴ cells were plated in 96 well plates containing RPMI supplemented with 10% FBS, glutamine, antibiotics and cytokines (mouse IL-3, 10 ng/ml; mouse IL-6, 10 ng/ml; and mouse SCF, 25 ng/ml; Peprotech) in triplicate from three mice per genotype. birinapant (Selleckchem) or DMSO vehicle was added to achieve desired concentrations and a final well volume of 200 µl before incubating for a further 48 h. Proliferation assays using murine cKit⁺ cells infected with Cflar shRNA vectors were performed similarly using 2 × 10⁴ cells plated in 96 well plates in triplicate from two mice per genotype with doxycycline (Sigma) or drug vehicle added to 0.1 µg/ml. Apoptosis was determined by Annexin-V and DAPI staining. Necrostatin-1 (20 µM, Sigma) or zVAD (10 µM, Selleckchem) was added at the same time as birinapant or doxycycline where indicated in the text.

Supper E., Rudat S., Iyer V., Droop A., Wong K., Spinella J.F., Thomas P., Sauvageau G., Adams D.J, & Wong C.C. (2021). Cut-like homeobox 1 (CUX1) tumor suppressor gene haploinsufficiency induces apoptosis evasion to sustain myeloid leukemia. Nature Communications, 12, 2482.

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Breast Cancer Cell Line Maintenance Protocol

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Human breast cancer cell lines MDA-MB-231, MDA-436, MDA-415, AU565 were purchased from Cell Bank of the Chinese Academy of Sciences Shanghai Institute of Cell Biology (Shanghai, China) and maintained in RPMI 1640 (HyClone/Thermo Fisher Scientific, Beijing, China) supplemented with 10% heat-inactivated fetal bovine serum (Hangzhou Sijiqing Biological Engineering Materials Co., Ltd, Hangzhou, China). Cells were incubated in a humidified 5% CO2 atmosphere at 37°C. Birinapant and Z-IETD-FMK were purchased from Selleck (Shanghai, China) and was dissolved in DMSO with a stock concentration of 1 and 5 mmol/L and stored at −20°C. NCTD was purchased from Nanjing Zelang Medical Technology Co., Ltd (Nanjing, China), and was dissolved in distill water with a stock concentration of 2 mM.

Zhao L., Yang G., Bai H., Zhang M, & Mou D. (2017). NCTD promotes Birinapant-mediated anticancer activity in breast cancer cells by downregulation of c-FLIP. Oncotarget, 8(16), 26886-26895.

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Cell Lines and Reagents for Signaling Studies

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REH (#ACC-22), TF-1 (#ACC334) and L929 (#ACC-2) cells were obtained from the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany), KYM-1 cells were kindly provided by Harald Wajant (University of Wuerzburg). All cell lines were grown RPMI 1640 medium (PAN Biotech, Aidenbach, Germany) supplemented with 10% (v/v) fetal calf serum (Sigma, Steinheim, Germany). TF-1 cells were additionally supplemented with 5 ng/ml human granulocyte–macrophage colony-stimulating factor (Immunotools, Friesoythe, Germany). Antibodies used in the study: TNF #3707, p38 #9212, phospho-p38 #9211, ERK #4695, phospho-ERK #4370 (Cell Signaling, Beverly, MA, USA); tubulin #MS-581: Dunnlab (Asbach, Germany); NFAT5 #PA1-023: Thermo Fisher Scientific (Waltham, MA, USA). Chemicals: MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide): Biomol (Hamburg, Germany); zVAD-fmk (carbobenzoxy-valyl-alanyl-aspartyl-(Omethyl)-fluoromethylketone): Bachem (Bubendorf, Switzerland); BIRB796, PH797804, SB203580, BV6, LCL161 and birinapant: Selleck Chemicals (Houston, TX, USA). Necrostatin-1 (Nec-1): Stress-Marq (Victoria, Canada); Etanercept was obtained from Pfizer (Berlin, Germany).

Bittner S., Knoll G, & Ehrenschwender M. (2017). Hyperosmotic stress enhances cytotoxicity of SMAC mimetics. Cell Death & Disease, 8(8), e2967-.

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Characterization of Burkitt Lymphoma Cell Lines

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Human BL cell lines RAMOS, DG-75, RAJI, BJAB and DAUDI were obtained from the German Collection of Microorganisms and Cell cultures (DSMZ, Braunschweig, Germany) and BL-2, BL-30, Seraphine, BL-60, BL-70 and Salina cells were kindly provided by T. Oellerich, Department of Medicine II, Hematology/Oncology, University Hospital Frankfurt, Germany. All cell lines were authenticated by STR profiling and continuously monitored for mycoplasma contamination. Cells were cultured in RPMI 1640 (Life Technologies), supplemented with 10% or 20% fetal calf serum (FCS) and 1% penicillin/streptomycin (Invitrogen). The bivalent Smac mimetic BV6 was kindly provided by Genentech. The broad-range caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethylketone (zVAD.fmk) was purchased from Bachem, Necrosulfonamide (NSA) from Toronto Research Chemicals Inc., GSK’872 and Necrostatin-1s (Nec-1s) from Merck and Dabrafenib from Selleck Chemicals. Recombinant human TRAIL was obtained from R&D Systems, human recombinant TNFα from PeproTech and human multimeric FASL from AdipoGen. Doxycycline hydrochloride (DOX) was purchased from Sigma-Aldrich. LCL-161 was purchased from Novartis and AT-406, Birinapant and SGI-110 (Guadecitabine) were obtained from Selleck Chemicals. All other chemicals were purchased from Sigma-Aldrich or Carl Roth unless indicated otherwise.

Koch A., Jeiler B., Roedig J., van Wijk S.J., Dolgikh N, & Fulda S. (2021). Smac mimetics and TRAIL cooperate to induce MLKL-dependent necroptosis in Burkitt's lymphoma cell lines. Neoplasia (New York, N.Y.), 23(5), 539-550.

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Apoptosis-Inducing Compound Screening

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UMI-77 (APExBIO);^{8 (link)} Bax-inhibiting peptide V5 (BIP-V5; Sigma Aldrich); MG132 (Enzo Life Sciences), etoposide (Cell Signaling Technologies); PAC1, Bam7, Birinapant, Lexibulin (CYT997), Nutlin-3a, YM155, A1210477, Navitoclax (ABT-263), Venetoclax (ABT-199) and Obatoclax mesylate were all purchased from Selleckchem. All inhibitors and stressors were dissolved in DMSO; control conditions were all treated with the appropriate amount of DMSO.

Robinson E.J., Aguiar S.P., Kouwenhoven W.M., Starmans D.S., von Oerthel L., Smidt M.P, & van der Heide L.P. (2018). Survival of midbrain dopamine neurons depends on the Bcl2 factor Mcl1. Cell Death Discovery, 4, 107.

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Evaluation of Apoptosis Induction Inhibitors

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Birinapant, GDC-0152, embelin, and vorinostat (suberoylanilide
hydroxamic acid) were purchased from Selleck Chemicals. Concentrations of
drugs used were based upon previously published whole blood
C_max and
C_min data for each drug (Amaravadi et al., 2015 (link); Flygare et al., 2012 (link); Zutshi et al., 1990 (link)) and were prepared in
dimethyl sulfoxide. Corresponding volumes of DMSO
(5μL DMSO per mL) were used for the vehicle in
each experiment. Staurosporine was purchased from Sigma and was used at 1
μM. For inhibitor experiments, 20
μM z-VAD-FMK, 200 nM wortmannin, 10
μM chloroquine, 10 nM bafilomycin A₁,
or 10 μM necrostatin-1 (all Sigma) were added for 1
h prior to, and for the duration of the stimulation. The human TNF
neutralizing antibody (D1B4) was purchased from Cell Signaling, and was
added 2 h prior to SM treatment at 1 μg
mL⁻¹.

Campbell G.R., Bruckman R.S., Chu Y.L., Trout R.N, & Spector S.A. (2018). SMAC mimetics induce autophagy-dependent apoptosis of HIV-1-infected resting memory CD4+ T cells. Cell host & microbe, 24(5), 689-702.e7.

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TRAIL-induced cell viability assay

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Cells were seeded at 1 × 10⁴ cells per well in 96-well format. After 24 h cells were stimulated with 50 ng/ml (Colo357) or 100 ng/ml (MDA-MB-231) TRAIL for 24 h. In some experiments cells were incubated with either U0126, MK-2206 (see above), Birinapant (see above), Navitoclax or Venetoclax (both 5 μM, Selleck Chemicals) prior to (for 2 h) and concomitant with TRAIL (for 24 h). Cell viability was assayed by crystal violet staining as described previously (Trauzold et al., 2006 (link)).

Rambow A.C., Aschenbach I., Hagelund S., Tawfik D., Gundlach J.P., Weiße S., Maass N, & Trauzold A. (2022). Endogenous TRAIL-R4 critically impacts apoptotic and non-apoptotic TRAIL-induced signaling in cancer cells. Frontiers in Cell and Developmental Biology, 10, 942718.

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Necroptosis Induction and Blockade

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Necroptosis was induced in vitro by pre-treatment 30min with Q-VD-OPh (R&D Systems, 10 μM) and Birinapant (Selleckchem, 5 μM) prior stimulation with TNFα (R&D Systems, 10 ng.mL-1) for 48h. Necroptosis was blocked using Necrostatin Nec (Selleckchem, 20 μM).

André-Grégoire G., Maghe C., Douanne T., Rosińska S., Spinelli F., Thys A., Trillet K., Jacobs K.A., Ballu C., Dupont A., Lyne A.M., Cavalli F.M., Busnelli I., Hyenne V., Goetz J.G., Bidère N, & Gavard J. (2022). Inhibition of the pseudokinase MLKL alters extracellular vesicle release and reduces tumor growth in glioblastoma. iScience, 25(10), 105118.

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Inhibitor Procurement and Utilization

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Inhibitors were purchased from commercial suppliers as listed below, or were provided (Compound 29, (Patch et al., 2011 (link))) by the Donald McDonnell Laboratory (Duke University). ABT199 (Selleckchem #S8048), ABT263 (Selleckchem #S1001), LCL161 (Selleckchem #S7009), Birinapant (Selleckchem #S7015), Fomepizole (Selleckchem #S1717), CHR2797 (Tocris #3595), XCT790 (Sigma Aldrich #X4753), Topiramate (Selleckchem #S1438), ACET (Tocris #2728), Almorexant (Selleckchem #S2160), SB334867 (R&D Systems, #1960), anti-IL1R1 (R&D Systems, #AF269), anti-IL9 (R&D Systems, #AF209), anti-ITGAV (Abcam, #ab16821), Cilengitide (Selleckchem, #S7077), Guanidine HCl (Sigma Aldrich, #G3272), Dalfampridine (Sigma Aldrich, #275875), BEZ235 (Selleckchem, #S1009), Apitolisib (Selleckchem, #S2696), S17092 (Sigma Aldrich, #SML0181), WNK463 (Selleckchem, #S8358).

Kishton R.J., Patel S.J., Decker A.E., Vodnala S.K., Cam M., Yamamoto T.N., Patel Y., Sukumar M., Yu Z., Ji M., Henning A.N., Gurusamy D., Palmer D.C., Stefanescu R.A., Girvin A.T., Lo W., Pasetto A., Malekzadeh P., Deniger D.C., Wood K.C., Sanjana N.E, & Restifo N.P. (2022). Cancer genes disfavoring T cell immunity identified via integrated systems approach. Cell reports, 40(5), 111153.

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