| Size | Price | Stock | Qty |
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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 |
TOPK; ERK-RSK; p53; Caspase-7
HI-TOPK-032 targets T-LAK cell-originated protein kinase (TOPK/PBK), a serine-threonine kinase. It is an ATP-competitive inhibitor, meaning it competes with ATP for binding to the kinase domain of TOPK. It strongly suppresses TOPK kinase activity in vitro but has little effect on extracellular signal–regulated kinase 1 (ERK1), c-jun—NH2—kinase 1, or p38 kinase activities. |
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| ln Vitro |
HI-TOPK-032 has a substantial inhibitory effect on TOPK dormancy activity, but has no effect on p38, c-jun-NH2-dormancy1, or extracellular signal-regulated dormancy 1 (ERK1). HI-TOPK-032 modifies TOPK's ATP binding. This chemical reacts correspondingly with LYS30 and generates hydrogen bonds with GLY83 and ASP151. However, 40% of MEK1 activity in chromatin was reduced by the maximum dose (5 μM) of HI-TOPK-032. By controlling p53, localized caspase-7, and localized PARP, HI-TOPK-032 also prevented ERK-RSK phosphorylation. Wall-dependent and non-navigational circular growth is inhibited by an increase in the number of circular navigators [1].
HI-TOPK-032 demonstrates potent in vitro activity by inhibiting TOPK kinase activity. It blocks the phosphorylation of the substrate histone H2AX. It reduces ERK-RSK phosphorylation, regulates the abundance of p53, cleaved caspase-7, and cleaved PARP, and induces apoptosis in cancer cells. It inhibits the growth of colon cancer and glioma initiating cells in vitro. |
| ln Vivo |
Mice treated with 1 or 10 mg/kg of HI-TOPK-032 showed a significant growth inhibition of more than 60% of HCT-116 tumor growth when compared to the tumor treatment group. The mice exhibited good tolerance to the treatment group of HI-TOPK-032. The phosphorylation of ERK and ERK's immediate downstream protein RSK was markedly suppressed in the HI-TOPK-032 treatment group, while p53 expression was considerably elevated [1].
HI-TOPK-032 suppresses tumor growth in vivo. It inhibits the growth of colon cancer and glioma initiating cells in vivo. Its ability to induce apoptosis and inhibit cell proliferation makes it a promising candidate for cancer therapy. |
| Enzyme Assay |
Through an in vitro kinase assay with [γ-32P]ATP and ERK1 (active, 500 ng), inactive RSK2 (ERK1 substrate, 1 μg), JNK1 (active, 50 ng), c-Jun (JNK1 substrate, 1 μg), p38 (active, 200 ng), and ATF2 (p38 substrate, 500 ng), the impact of HI-TOPK-032 on ERK1, JNK1, and p38 activity is evaluated. In a nutshell, 40 μL of reaction buffer is used along with 10 μCi of [γ-32P]ATP and HI-TOPK-032 (0.5, 1, 2, 5 μM). SDS-PAGE is used to separate the mixture after 30 minutes of room temperature incubation, during which time 10 μL of protein loading buffer is added to stop the reaction[1].
The primary in vitro assay for HI-TOPK-032 is a kinase activity assay using purified TOPK enzyme. The enzyme is incubated with a peptide substrate and ATP in the presence of varying concentrations of the compound. The extent of phosphorylation is measured, and the IC50 of 2 µM is determined. Cellular assays are also used to confirm its activity. |
| Cell Assay |
HI-TOPK-032 is administered at varying concentrations to HCT-116 colon cancer cells (1, 2, 5 μM). Following a period of 1, 2, or 3 days of incubation, 20 μL of CellTiter96 AQueous One Solution is introduced, and the cells are subsequently incubated for 1 hour at 37°C in an incubator with 5% CO2. At 492 nm, absorbance is measured[1].
Cellular assays for HI-TOPK-032 involve treating cancer cell lines, such as colon cancer and glioma cells, with the compound. The inhibition of TOPK activity is assessed by measuring the phosphorylation of its substrate, histone H2AX, by Western blotting. The induction of apoptosis is measured by assessing the cleavage of caspase-7 and PARP, or by flow cytometry using Annexin V staining. |
| Animal Protocol |
Mice: The four groups of mice are as follows: (i) the untreated vehicle group; (ii) the 1 mg and 10 mg HI-TOPK-032/kg body weight; (iii) the 10 mg and 1 mg body weight group; and (iv) the no cells and 10 mg HI-TOPK-032/kg body weight group. To inoculate each mouse's right flank, a suspension of HCT-116 cells is placed in serum-free McCoy 5A medium s.c. For twenty-five days, the vehicle or HI-TOPK-032 is injected three times a week. It calculates tumor volume[1].
In vivo efficacy of HI-TOPK-032 has been evaluated in mouse xenograft models of colon cancer and glioma. Tumor-bearing mice are treated with the compound, and tumor growth inhibition is monitored. Pharmacodynamic markers, such as the inhibition of histone H2AX phosphorylation in tumor tissue, are also assessed. |
| ADME/Pharmacokinetics |
HI-TOPK-032 has a molecular weight of 369.40 g/mol and a molecular formula of C20H11N5OS. Its IUPAC name is N-(12-cyanoindolizino[2,3-b]quinoxalin-2-yl)thiophene-2-carboxamide. It is soluble in DMSO at 2.5 mg/mL. It is typically stored as a powder at -20°C.
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| Toxicity/Toxicokinetics |
HI-TOPK-032 is a research compound that is not intended for human use. Its toxicological profile is not extensively characterized in the available literature. As a kinase inhibitor, it may have on-target toxicities. Standard safety precautions should be followed when handling.
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| References | |
| Additional Infomation |
HI-TOPK-032 is a potent and selective ATP-competitive inhibitor of T-LAK cell-originated protein kinase (TOPK/PBK). It has an IC50 of 2 µM and is used in cancer research to study the role of TOPK in tumorigenesis. It suppresses tumor growth in vivo and is available from various chemical suppliers for research purposes.
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| Molecular Formula |
C20H11N5OS
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| Molecular Weight |
369.399241685867
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| Exact Mass |
369.068
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| Elemental Analysis |
C, 65.03; H, 3.00; N, 18.96; O, 4.33; S, 8.68
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| CAS # |
487020-03-1
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| Related CAS # |
487020-03-1
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| PubChem CID |
1936439
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| Appearance |
Pink to red solid powder
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| Density |
1.5±0.1 g/cm3
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| Boiling Point |
415.3±45.0 °C at 760 mmHg
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| Flash Point |
204.9±28.7 °C
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| Vapour Pressure |
0.0±1.0 mmHg at 25°C
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| Index of Refraction |
1.808
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| LogP |
2.71
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
27
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| Complexity |
635
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(C1=CC=CS1)NC2=CC3=C(C#N)C4=NC5=CC=CC=C5N=C4N3C=C2
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| InChi Key |
BCSBXWKRZUPFHW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H11N5OS/c21-11-13-16-10-12(22-20(26)17-6-3-9-27-17)7-8-25(16)19-18(13)23-14-4-1-2-5-15(14)24-19/h1-10H,(H,22,26)
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| Chemical Name |
N-(12-cyanoindolizino[2,3-b]quinoxalin-2-yl)thiophene-2-carboxamide
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| Synonyms |
HI-TOPK-032; HI-TOPK032; HI-TOPK 032; HI TOPK-032; HI TOPK032; HI TOPK 032
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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: 4~5 mg/mL (10.8~13.5 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 10 mg/mL (27.07 mM) in 15% Cremophor EL + 85% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 0.25 mg/mL (0.68 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 2.5 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7071 mL | 13.5355 mL | 27.0709 mL | |
| 5 mM | 0.5414 mL | 2.7071 mL | 5.4142 mL | |
| 10 mM | 0.2707 mL | 1.3535 mL | 2.7071 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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