| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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Purity: ≥98%
TBB (NSC 231634; Casein Kinase II Inhibitor I) is a novel, potent, cell-permeable and ATP-competitive inhibitor of protein kinase CK2 with an IC50 of 0.15 μM for rat liver CK2. TBB induces apoptosis and caspase-dependent degradation of haematopoietic lineage cell-specific protein 1 (HS1) in Jurkat cells. Inhibitors of CK2 kinase inhibit cell proliferation and induce apoptosis in numerous cancer cell lines. Due to these properties, they are considered potentially useful in anticancer therapy. I
| Targets |
TBB is a highly specific inhibitor of protein kinase CK2 (formerly casein kinase II).
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| ln Vitro |
Casein kinase 2, or CK2, has the highest inhibitory capacity of TBB, according to studies employing a panel of 33 protein kinases (human CK2: IC50=1.6 μM at 100 μM ATP). Additionally, three less powerful kinases are inhibited by TBB: glycogen synthase kinase 3β (GSK3β) (IC50=11.2 μM), phosphorylase kinase (IC50=8.7 μM), and CDK2 (IC50=15.6 μM). Compared to CK2, the IC50 values of all other examined kinases were 50 times higher [1]. When administered according to the recommended dosage schedule, TBB (60 μM TBB) either by itself or in conjunction with the anticancer drugs CPT or TRAIL may decrease the viability of androgen-insensitive PC-3 cells. TBB's effect on PC-3 cell apoptosis is not the cause of its time-dependent activity [2]. Tested against 33 protein kinases (Ser/Thr-specific or Tyr-specific), TBB is an ATP/GTP competitive inhibitor of the protein kinase casein kinase 2 (CK2). While three kinases (phosphorylase kinase, glycogen synthase kinase 3L, and cyclin-dependent kinase 2/ Cyclin A) were moderately inhibited, with an IC50 value one to two orders of magnitude higher than CK2 (IC50=0.9 μM), only CK2 was significantly inhibited (>85%) in the presence of 10 μM TBB (and 100 μM ATP). Additionally, in cultivated Jurkat cells, TBB suppresses endogenous CK2 [3].
In assays using retinal endothelial cells (RECs), TBB was among the CK2 inhibitors (like emodin and DRB) that mimicked the effects of broad-spectrum kinase inhibitors (H7, H89). It significantly decreased REC proliferation, migration, tube formation, tube collapse, and secondary sprouting, as summarized in Table 2 of the literature. |
| ln Vivo |
In the mouse OIR model, the degree of retinal neovascularization was reduced by about 60% following treatment with TBB (60 mg/kg per day for 6 days) [4].
In a mouse model of oxygen-induced retinopathy (OIR), intraperitoneal treatment with TBB at a dose of 60 mg/kg per day resulted in an approximately 60% reduction in retinal neovascularization compared to untreated or vehicle-treated control groups. The inhibitory effect primarily targeted the neovascular tufts, while the main vascular tree showed minimal changes. Retinal structure in treated animals showed no adverse changes compared to normoxic controls. |
| Cell Assay |
Cell Viability and Proliferation Assay: Bovine retinal endothelial cells (RECs) were plated in 96-well plates in complete medium with 10% FCS (proliferation) or 0.5% FCS (viability). Inhibitors, including TBB, were added at the time of cell seeding. The number of viable cells was determined on days 4-7 using an MTS cell proliferation assay.[4]
Tube Formation Assay on Reconstituted Basement Membrane Matrix: RECs were seeded on a reconstituted basement membrane matrix (Matrigel) in 96-well plates. For testing effects on tube formation, inhibitors were added at the time of cell seeding. To assess effects on tube stabilization/collapse, inhibitors were added 24 hours after seeding. Capillary-like tube structures were photographed and analyzed over time.[4] Secondary Sprouting Assay on Matrigel: RECs were seeded on basement membrane matrix and allowed to form and collapse tubes over 2-3 days. Subsequently, a synergistic combination of four angiogenic growth factors (VEGF, IGF-I, FGF-2, PlGF) was added to stimulate secondary sprout formation. Inhibitors were added along with the growth factors. The extent of sprouting was assessed after 5-6 days by measuring the number of living cells in colonies using the MTS assay.[4] Wound Migration Assay: Confluent monolayers of RECs on plastic were wounded and then incubated in low-serum medium with or without the combination of four growth factors. Inhibitors were added at the time of wounding. Cell migration into the wound area was quantified after 7 days. |
| Animal Protocol |
Oxygen-Induced Retinopathy (OIR) Mouse Model: Seven-day-old C57BL/6J mouse pups were exposed to 75% oxygen for 5 days and then returned to room air (relative hypoxia) for an additional 5 days to induce retinal neovascularization. Starting on day 11 (the first day back in room air), mice were treated with TBB intraperitoneally.
Drug Formulation and Dosing: TBB was dissolved in a vehicle consisting of PBS (pH 7.2) with 20% polyethylene glycol 400 (PEG 400) and 2% Tween-80. It was administered intraperitoneally at a dose of 60 mg/kg body weight per day. The total injection volume was 50 µL or less per mouse. Control mice received the PEG-Tween vehicle alone. Tissue Analysis: At the end of the experiment, mice were euthanized and perfused with fluorescein-dextran. Retinas were dissected, flat-mounted, and photographed under a fluorescence microscope to visualize vasculature. Paraffin-embedded eyes were sectioned and stained with hematoxylin and eosin. Nuclei of vascular cells on the vitreal side of the inner limiting membrane (preretinal nuclei) were counted on multiple sections per eye as a quantitative measure of neovascularization. |
| References |
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| Additional Infomation |
TBB was considered the most specific CK2 inhibitor known at the time. This study provided the first evidence linking the protein kinase CK2 to the angiogenesis process. The anti-angiogenic effects of TBB observed in vitro and in vivo suggest that CK2 is a key signaling molecule in pathological neovascularization, such as neovascularization in proliferative diabetic retinopathy. Inhibiting CK2 offers a potential therapeutic strategy for treating diseases involving abnormal blood vessel growth.
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| Molecular Formula |
C6HBR4N3
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|---|---|
| Molecular Weight |
434.7082
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| Exact Mass |
430.69
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| CAS # |
17374-26-4
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| PubChem CID |
1694
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| Appearance |
Off-white to yellow solid powder
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| Density |
2.8±0.1 g/cm3
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| Boiling Point |
552.5±45.0 °C at 760 mmHg
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| Melting Point |
262-266ºC
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| Flash Point |
288.0±28.7 °C
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| Vapour Pressure |
0.0±1.5 mmHg at 25°C
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| Index of Refraction |
1.801
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| LogP |
3.96
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
13
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| Complexity |
184
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
OMZYUVOATZSGJY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C6HBr4N3/c7-1-2(8)4(10)6-5(3(1)9)11-13-12-6/h(H,11,12,13)
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| Chemical Name |
4,5,6,7-tetrabromo-1H-benzo[d][1,2,3]triazole
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| Synonyms |
Casein Kinase II Inhibitor I, Tetrabromobenzotriazole, NSC 231634, NSC231634, NSC-231634
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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 |
| 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) |
DMSO : ≥ 100 mg/mL (~230.04 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.75 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 (5.75 mM) 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; with ultrasonication. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.75 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3004 mL | 11.5019 mL | 23.0038 mL | |
| 5 mM | 0.4601 mL | 2.3004 mL | 4.6008 mL | |
| 10 mM | 0.2300 mL | 1.1502 mL | 2.3004 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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