Obatoclax Mesylate (GX15-070)

Catalog No.S1057

Obatoclax Mesylate (GX15-070) is an antagonist of Bcl-2 with Ki of 0.22 μM in a cell-free assay, can assist in overcoming MCL-1 mediated resistance to apoptosis. Phase 3.

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Obatoclax Mesylate (GX15-070) Chemical Structure

Obatoclax Mesylate (GX15-070) Chemical Structure
Molecular Weight: 413.49

Validation & Quality Control

Customer Product Validation(9)

Quality Control & MSDS

Related Compound Libraries

Obatoclax Mesylate (GX15-070) is available in the following compound libraries:

Bcl-2 Inhibitors with Unique Features

  • Pan Bcl-2 Inhibitor

    ABT-737 Bcl-xL, EC50=78.7 nM; Bcl-2, EC50=30.3 nM; Bcl-w, EC50=197.8 nM.

  • Most Potent Bcl-2 Inhibitor

    ABT-263 (Navitoclax) Bcl-xL, Ki≤ 0.5 nM; Bcl-2, Ki≤ 1 nM; Bcl-w, Ki≤ 1 nM.

  • Newest Bcl-2 Family Activator

    BAM7 Direct and selective activator of proapoptotic Bax with EC50 of 3.3 μM.

  • Classic Bcl-2 Inhibitor

    ABT-737 BH3-mimetic inhibitor, Bcl-xL, EC50=78.7 nM; Bcl-2, EC50=30.3 nM; Bcl-w, EC50=197.8 nM.

Product Information

  • Compare Bcl-2 Inhibitors
    Compare Bcl-2 Products
  • Research Area

Product Description

Biological Activity

Description Obatoclax Mesylate (GX15-070) is an antagonist of Bcl-2 with Ki of 0.22 μM in a cell-free assay, can assist in overcoming MCL-1 mediated resistance to apoptosis. Phase 3.
Targets Bcl-2 [1]
(Cell-free assay)
IC50 0.22 μM(Ki)
In vitro Obatoclax completely inhibits Bak recovery of Mcl-1 at 5 μM in SK-Mel5 cells and overcomes resistance to ABT-373-induced apoptosis conferred by Mcl-1 in KB/Bcl-2 cells. [1] Obatoclax is a BH3 mimetic which binds to a broad spectrum of Bcl-2 family members, including Bcl-2, Bcl-xL, and Mcl-1. Obatoclax uniquely displaces BH3 domains by activation of the pocket of Mcl-1 followed by a triggering of apoptosis mediated by oligomerization of Bak and release of cytochrome c. Obatoclax is sensitive to Bcl-xL in cell lines lacking it or showing low expression. It shows low cytotoxicity to Mcl-1, Bcl-2, and Bcl-xL in all the strongly-expressed cell lines. Obatoclax inhibits multiple myeloma (MM) cell lines (KMS12PE, KMS18, MY5, etc.) with IC50 values ranging from 52 to 1100 nM and the inhibition is umimpaired even in the presence of IL-6 or IGF-1, which are resistance to cytotoxic agents, at a concentration of 150 nM. Obatoclax enhances the antimyeloma activity induced by melphalan, dexamethasone, or bortezomib. [2] Obatoclax potentiates TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-mediated apoptosis by unsequestering Bak and Bim from Bcl-2/Bcl-xL or Mcl-1 proteins in PANC-1 and BxPC-3 cells. [3]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity DescriptionPMID
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NCI-H23NVvpc5h[T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=MVrJR|UxRTFyLk[0OFIh|ryPMkDzV2FPT0WU
UACC-812NVT1Wo41T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NEDtTYZKSzVyPUGxMlcyPyEQvF2=NXn1WJQ3W0GQR1XS
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... Click to View More Cell Line Experimental Data

In vivo Obatoclax also exhibits high antitumor activity in SCID mice bearing human C33A cerivical carcinoma tumors at a dose of 0.5 mg/kg. [1]
Features Potential 1st-in-class small molecule antagonist of Bcl-2 designed to inhibit all relevant Bcl-2 family members, including Mcl-1.

Protocol(Only for Reference)

Kinase Assay: [1]

Bcl-2 Binding affinity calculation A predicted binding affinity for Obatoclax binding to BCL-2 is calculated using the SIE scoring function. [4] As a control in determining the reliability of the calculation, predicted binding affinities Ki) are calculated for a set of 12 small molecules with experimentally measured binding affinities to BCL-2.

Cell Assay: [2]

Cell lines Human MM cells (HMCLs), peripheral blood lymphocytes (PBLs), bone marrow stromal cells (BMSCs)
Concentrations ~10 μM
Incubation Time 48-72 hours
Method Obatoclax is dissolved in DMSO at a stock concentration of 5 mM. Cell viability is assayed by MTT. Human MM cells (HMCLs), peripheral blood lymphocytes (PBLs), bone marrow stromal cells (BMSCs) are seeded in 96-well plates at a density of 2 × 104 per well for HMCLs or 5~10 × 103 for PBLs. Various concentrations of Obatoclax are added to the cells, with or without IGF-1 (50 ng/mL) or IL-6 (10 ng/mL). Cells are incubated for 48-72 hours and cell viability is determined.

Animal Study: [1]

Animal Models Female BALB/c or CB17 SCID/SCID mice bearing SW480, C33A, PC3, and 4T1 cells are used.
Formulation Obatoclax (tartrate salt) is formulated in 9.6% polyethylene glycol 300, 0.4% polysorbate 20, and 5% dextrose; while for the 4T1 tumor model, Obatoclax is formulated at a concentration of 0.6 mg/mL in 9.48% polyethylene glycol, 0.38% polysorbate 20, 1.2 m
Dosages 0.0313, 0.25, 0.5 and 2 mg/kg
Administration Administered intravenously (tail vein) once a day

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Nguyen M, et al. Proc Natl Acad Sci, 2007, 104(49), 19512-191517.

[2] Trudel S, et al. Blood, 2009, 113(2), 299-305.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2016-04-16)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT01563601 Withdrawn Extensive-stage Small Cell Lung Cancer Cephalon|Teva Pharmaceutical Industries August 2012 Phase 3
NCT01238146 Withdrawn Extranodal Marginal Zone B-cell Lymphoma of Mucosa-associated Lymphoid Tissue|Nodal Marginal Zone B-cell Lymphoma|Recurrent Grade 1 Follicular Lymp  ...more Extranodal Marginal Zone B-cell Lymphoma of Mucosa-associated Lymphoid Tissue|Nodal Marginal Zone B-cell Lymphoma|Recurrent Grade 1 Follicular Lymphoma|Recurrent Grade 2 Follicular Lymphoma|Recurrent Mantle Cell Lymphoma|Recurrent Marginal Zone Lymphoma|Splenic Marginal Zone Lymphoma National Cancer Institute (NCI) October 2010 Phase 1|Phase 2
NCT00933985 Terminated Acute Leukemias of Ambiguous Lineage|Acute Undifferentiated Leukemia|Angioimmunoblastic T-cell Lymphoma|Blastic Phase Chronic Myelogenous Leukemia|  ...more Acute Leukemias of Ambiguous Lineage|Acute Undifferentiated Leukemia|Angioimmunoblastic T-cell Lymphoma|Blastic Phase Chronic Myelogenous Leukemia|Childhood Burkitt Lymphoma|Childhood Chronic Myelogenous Leukemia|Childhood Diffuse Large Cell Lymphoma|Childhood Immunoblastic Large Cell Lymphoma|Childhood Nasal Type Extranodal NK/T-cell Lymphoma|Cutaneous B-cell Non-Hodgkin Lymphoma|Hepatosplenic T-cell Lymphoma|Intraocular Lymphoma|Noncutaneous Extranodal Lymphoma|Peripheral T-cell Lymphoma|Recurrent Childhood Acute Lymphoblastic Leukemia|Recurrent Childhood Acute Myeloid Leukemia|Recurrent Childhood Anaplastic Large Cell Lymphoma|Recurrent Childhood Grade III Lymphomatoid Granulomatosis|Recurrent Childhood Large Cell Lymphoma|Recurrent Childhood Lymphoblastic Lymphoma|Recurrent Childhood Small Noncleaved Cell Lymphoma|Recurrent Cutaneous T-cell Non-Hodgkin Lymphoma|Recurrent Mycosis Fungoides/Sezary Syndrome|Recurrent/Refractory Childhood Hodgkin Lymphoma|Refractory Chronic Lymphocytic Leukemia|Refractory Hairy Cell Leukemia|Relapsing Chronic Myelogenous Leukemia|Small Intestine Lymphoma|Unspecified Childhood Solid Tumor, Protocol Specific National Cancer Institute (NCI) June 2009 Phase 1
NCT00918931 Terminated Leukemia|Systemic Mastocytosis M.D. Anderson Cancer Center|Gemin X June 2009 Phase 2
NCT00719901 Terminated Refractory Multiple Myeloma|Stage I Multiple Myeloma|Stage II Multiple Myeloma|Stage III Multiple Myeloma National Cancer Institute (NCI) July 2008 Phase 1|Phase 2

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Chemical Information

Download Obatoclax Mesylate (GX15-070) SDF
Molecular Weight (MW) 413.49
Formula

C20H19N3O.CH4O3S

CAS No. 803712-79-0
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms N/A
Solubility (25°C) * In vitro DMSO 83 mg/mL (200.73 mM)
Water <1 mg/mL (<1 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo 30% PEG400/0.5% Tween80/5% propylene glycol 30 mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name (Z)-2-(5-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-4-methoxy-5H-pyrrol-2-yl)-1H-indole mesylate

Customer Product Validation (9)


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Rating
Source Cancer Lett 2014 348, 20-8. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Immunohistochemical staining
Cell Lines Mice
Concentrations 3 mg/kg
Incubation Time 3 weeks
Results Tumors from 3 mice in each group were analyzed with H&E and immunohistochemical staining. Interestingly, H&E staining revealed substantially increased necrosis in the tumors for GX15-070 treated groups compared to the vehicle control group, especially for the combination (AZD2281 and GX15-070) group.

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Rating
Source Cell Death Dis 2012 19, 3:e351. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Electron microscopic morphology
Cell Lines T1682 cells
Concentrations 100, 1000 nM
Incubation Time 1, 6, 48 h
Results Gx15-070 treatment of TET cells leads to both growth inhibition and cell death. Gx15-070 induces autophagy-dependent necroptosis in acute lymphoblastic leukemia cells, it's performed electron microscopy (EM) in TET cells, which demonstrated changes in mitochondrial structure as early signs of autophagy after 1 h Gx15-070 treatment. After 6 h treatment, cytoplasmic vacuolization and mitochondrial swelling with early signs of necroptotic cell death were evident. Intriguingly, cytoplasmic vacuolization, late-stage necroptotic or autophagic cell death were the main features 48 h after Gx15-070 (1 uM) treatment.

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Rating
Source Cell Death Dis 2012 19, 3:e351. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Western blot
Cell Lines T1682 cells
Concentrations 100, 250, 500, 1000 nM
Incubation Time 6 h
Results Gx15-070 induced a progressive reduction of AKT Ser473 and Thr308 phosphorylation and a reduction of p-RPS6 phosphorylation, suggesting inhibition of mTOR pathway, in line with previous reports. Moreover, Gx15-070 induced phosphorylation of AMPKα, a Ser/Thr kinase normally activated when ATP level is reduced.AMPK phosphorylation and LC3B cleavage appeared within 1 h after Gx15-070 treatment.

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Source Cell death dis 2012 3, e299. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Viability assay
Cell Lines BM3.3 CD8 T cells
Concentrations 0.1-10uM
Incubation Time
Results T-cell activation induced resistance to Bcl-2 inhibition by ABT-737 (no binding of A1 and Mcl-1) and by Antimycin A (no binding of A1 only), but had no impact on the proapoptotic potency of the pan-Bcl-2 inhibitor obatoclax.

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Source Exp Cell Res 2014 322, 217-25. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Immunostaining
Cell Lines HUH6 cells
Concentrations 0.1, 0.3 uM
Incubation Time 24 h
Results In HUH6 cells mitochondria with an intact membrane potential were observed in the JC-1 staining in both, control and TNF-a treated cultures (C, D). The number of reddish points indicating active mitochondria decreased when the HUH6 cells were treated with obatoclax and decreased even further when the treatment was performed in combination with TNF-a, which reflects the breakdown of mitochondrial membranes decreased aggregation of JC-1. This was concentration dependent (E-H).

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Rating
Source BMC Cancer 2011 11, 24. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Cell proliferation assays
Cell Lines SET-2 cells
Concentrations 1-10000 nM
Incubation Time 72 h
Results Incubation of SET-2 cells with sub-optimal concentrations of the pan-Bcl-2 family protein inhibitor obatoclax in cell proliferation assays lowered the GI 50 of NVP-BSK805 by 3 to 4 fold.

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Source Dr. Christine Hawkins of La Trobe University. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Clonogenic survival assay
Cell Lines MEF cells
Concentrations 0-300 ng/mL
Incubation Time 24 h
Results

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Rating
Source Dr. Zhang of Tianjin Medical University. Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Hoechst staining
Cell Lines MDB-MA-231 cells
Concentrations 1 μM
Incubation Time
Results

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Rating
Source Obatoclax Mesylate (GX15-070) purchased from Selleck
Method Cell Viability/Western Blot
Cell Lines Nalm-6 ABT-R cells
Concentrations 5-10 uM
Incubation Time 18 h
Results Nalm-6 ABT-R cells were sensitive to obatoclax; however, Nalm-6 cells were equally sensitive (IC50: 10 umol/L). Reh parental cells (IC50: 5umol/L) were also more sensitive to obatoclax compared with ABT-R cells, as measured by trypan blue exclusion. Mcl-1 protein levels were downregulated following obatoclax treatment. NOXA levels also increased, however these changes did not correlate with cell death.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Tel: +1-832-582-8158 Ext:3

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