| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
| Targets |
Bcl-xL (IC50 = 1.1 nM)
BCL-XL (IC50 = 1.1 nM) |
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| ln Vitro |
NSC 241240-induced caspase 3/7 activity, PARP cleavage, and annexin V labeling are enhanced by WEHI-539. In Ovcar-4 (5 μM in Ovcar-4) and Ovsaho (1 μM in Ovsaho) cells, WEHI-539 alone significantly increases PARP cleavage[2].
WEHI-539 binds to BCL-XL with high affinity (IC50 = 1.1 nM) and selectivity. The compound shows selectivity for BCL-XL over other BCL-2 family members including BCL-2 and MCL-1. By neutralizing BCL-XL, WEHI-539 promotes apoptosis in cells that are dependent on BCL-XL for survival, including certain cancer cell lines. In cellular assays, the compound induces apoptosis and inhibits cell proliferation in BCL-XL-dependent cancer cells. WEHI-539 kills mouse embryonic fibroblasts that lack MCL-1. |
| ln Vivo |
In vivo, WEHI-539 demonstrates efficacy in animal models of cancers that are dependent on BCL-XL. The compound shows anti-tumor activity and induces apoptosis in tumor tissues. Pharmacodynamic markers including activation of caspases and cleavage of PARP are observed in treated tumors. The hydrochloride salt form may offer improved solubility and formulation properties. |
| Enzyme Assay |
The assay buffer (stock 50 mM HEPES and 100 mM NaCl, pH 7.5) is prepared fresh daily and adjusted to 5 mM DTT, casein (0.1 mg/mL sodium salt; aliquots stored at -20 °C) and Tween 20.
The prosurvival BCL-2 family protein BCL-X(L) is often overexpressed in solid tumors and renders malignant tumor cells resistant to anticancer therapeutics. Enhancing apoptotic responses by inhibiting BCL-X(L) will most likely have widespread utility in cancer treatment and, instead of inhibiting multiple prosurvival BCL-2 family members, a BCL-X(L)-selective inhibitor would be expected to minimize the toxicity to normal tissues. We describe the use of a high-throughput screen to discover a new series of small molecules targeting BCL-X(L) and their structure-guided development by medicinal chemistry. The optimized compound, WEHI-539 (7), has high affinity (subnanomolar) and selectivity for BCL-X(L) and potently kills cells by selectively antagonizing its prosurvival activity. WEHI-539 will be an invaluable tool for distinguishing the roles of BCL-X(L) from those of its prosurvival relatives, both in normal cells and notably in malignant tumor cells, many of which may prove to rely upon BCL-X(L) for their sustained growth.[1] Binding affinity to BCL-XL is measured using fluorescence polarization or surface plasmon resonance assays with recombinant BCL-XL protein. Selectivity profiling is conducted against other BCL-2 family members including BCL-2, MCL-1, and BFL-1/A1 to confirm specificity. Competition binding assays with fluorescently labeled BH3 peptides determine the affinity and mode of binding. |
| Cell Assay |
Ovcar-8, Ovcar-3, Ovcar-4, and Ovsaho cells are grown in the RPMI medium, Igrov-1, Cov-362 and Cov-318 cells are grown in DMEM, and Fuov-1 cells are grown in a DMEM/F-12 nutrient solution. DMSO is used to prepare a 20 mM solution of ABT-737, ABT-199, and WEHI-539. Cells are plated in 96-well plates for cell growth assays (5,000 cells/well for all cell lines, 2,500 cells/well for Ovcar-8). Cells receive drug therapy the following day. After 72 hours, the culture medium is taken out, the cells are fixed with 100 μL of cold 10% Trichloroacetic acid (TCA), incubated on ice for 30 minutes, and stained with 0.4% sulforhodamine B (SRB). The data are examined using the Graphpad Prism 4 program. A four-parameter Hill equation is fitted using non-linear regression. For drug combination studies, cells are exposed simultaneously to a range of NSC 241240 concentrations along with a fixed concentration of BH3 mimics, which is predicted from single agent studies to result in a 5% growth inhibition: ABT-737, 1 μM in Ovcar-8, Ovcar-3 and Igrov-1, 2 μM in Ovcar-4 and Ovsaho and 6 μM in Cov-362; ABT-199, 1 μM in Ovcar-4, 2 μM in Ovcar-3, Igrov-1, Cov-362 and Ovsaho and 3 μM in Ovcar-8; WEHI-539, 0.2 μM in Igrov-1, 0.3 μM in Ovcar-8, 1 μM in Ovcar-3 and Ovsaho, 3.1 μM in Cov-362 and 5 μM in Ovcar-4. SRB staining is used to determine the number of living cells. It calculates a combination index (CI)[2].
Cellular potency is evaluated in BCL-XL-dependent cancer cell lines by measuring apoptosis induction using flow cytometry with annexin V/PI staining or by caspase activity assays. Cell viability is assessed using MTT or CellTiter-Glo assays. BCL-XL target engagement is confirmed by assessing displacement of BCL-XL from pro-apoptotic binding partners using immunoprecipitation or proximity ligation assays. |
| Animal Protocol |
In vivo efficacy is evaluated in mouse xenograft models of BCL-XL-dependent cancers. WEHI-539 hydrochloride is administered via appropriate routes at various doses. Tumor growth inhibition, apoptosis induction, and pharmacodynamic biomarkers are assessed in tumor tissues. Pharmacokinetic sampling is performed to correlate exposure with efficacy. |
| ADME/Pharmacokinetics |
Pharmacokinetic data for WEHI-539 hydrochloride are available from preclinical studies. The compound demonstrates adequate exposure and bioavailability for in vivo efficacy studies. The hydrochloride salt form may provide improved solubility and stability compared to the free base. Detailed PK parameters may vary depending on the formulation and route of administration.
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| Toxicity/Toxicokinetics |
Toxicology data for WEHI-539 hydrochloride are limited as the compound is primarily used as a research tool. Preclinical studies indicate an acceptable safety profile at efficacious doses, though comprehensive toxicology assessments for human use have not been conducted. Standard laboratory safety precautions should be observed during handling.
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| References | |
| Additional Infomation |
Background: BH3 mimics are a class of drugs that antagonize Bcl-2 family apoptosis inhibitors. We have previously demonstrated that these compounds enhance the activity of carboplatin against various ovarian cancer cell lines. However, recent clinical studies have shown that BH3 mimics antagonizing Bcl-XL are associated with significant thrombocytopenia. Therefore, we developed ABT-199, which specifically inhibits Bcl-2. Unfortunately, aberrant expression of Bcl-XL appears to be more common than Bcl-2 in ovarian cancer. Therefore, we compared the ability of ABT-199 and the Bcl-XL selective compound WEHI-539 to enhance the activity of carboplatin against ovarian cancer cell lines. Methods: We tested the efficacy of WEHI-539, ABT-737, and ABT-199 in combination with carboplatin using six ovarian cancer cell lines. We evaluated drug activity using cell growth assays, trypan blue staining, and apoptosis assays (by detecting caspase 3/7 activity, PARP lysis, and Annexin-V/propidium iodide staining). Results: We found that WEHI-539 and ABT-737 (but not ABT-199) synergized with carboplatin in cell growth assays and enhanced cell death in trypan blue staining assessment. In addition, WEHI-539 and ABT-737 enhanced carboplatin-induced caspase 3/7 activity, PARP cleavage and Annexin-V labeling, while ABT-199 did not have this effect. Conclusion: These observations suggest that compounds targeting Bcl-XL must be used for successful treatment of ovarian cancer patients with BH3 mimics, highlighting the need for development strategies to minimize thrombocytopenia caused by such compounds. [2]
WEHI-539 hydrochloride is a valuable research tool for studying the role of BCL-XL in apoptosis and cancer biology. It has been used to demonstrate the dependency of certain cancer types on BCL-XL for survival and to validate BCL-XL as a therapeutic target. The compound has not entered clinical trials and remains a tool for preclinical research. |
| Molecular Formula |
C31H30CLN5O3S2
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| Molecular Weight |
620.184603214264
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| Exact Mass |
619.15
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| Elemental Analysis |
C, 60.04; H, 4.88; Cl, 5.72; N, 11.29; O, 7.74; S, 10.34
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| CAS # |
2070018-33-4
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| Related CAS # |
WEHI-539;1431866-33-9
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| PubChem CID |
78357795
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| Appearance |
Light yellow to yellow solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
42
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| Complexity |
903
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1CC2=C(C=C(C=C2)C3=NC(=C(S3)CCCOC4=CC=C(C=C4)CN)C(=O)O)/C(=N/NC5=NC6=CC=CC=C6S5)/C1.Cl
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| InChi Key |
GJBYTVIQTMIXGA-DYICZVFTSA-N
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| InChi Code |
InChI=1S/C31H29N5O3S2.ClH/c32-18-19-10-14-22(15-11-19)39-16-4-9-27-28(30(37)38)34-29(40-27)21-13-12-20-5-3-7-24(23(20)17-21)35-36-31-33-25-6-1-2-8-26(25)41-31;/h1-2,6,8,10-15,17H,3-5,7,9,16,18,32H2,(H,33,36)(H,37,38);1H/b35-24+;
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| Chemical Name |
5-[3-[4-(aminomethyl)phenoxy]propyl]-2-[(8E)-8-(1,3-benzothiazol-2-ylhydrazinylidene)-6,7-dihydro-5H-naphthalen-2-yl]-1,3-thiazole-4-carboxylic acid;hydrochloride
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.03 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.03 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6124 mL | 8.0622 mL | 16.1244 mL | |
| 5 mM | 0.3225 mL | 1.6124 mL | 3.2249 mL | |
| 10 mM | 0.1612 mL | 0.8062 mL | 1.6124 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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