| Size | Price | Stock | Qty |
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| 5mg |
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| 50mg | |||
| Other Sizes |
| Targets |
Merbarone targets DNA topoisomerase II, an enzyme involved in DNA replication and repair. Topoisomerase II catalyzes the transient cleavage and rejoining of DNA strands, allowing the passage of another DNA helix through the break. This activity is essential for chromosome condensation, segregation, and DNA replication. Merbarone is a catalytic inhibitor of topoisomerase II, meaning it inhibits the catalytic activity of the enzyme without stabilizing the DNA-topo II cleavable complex. This distinguishes it from classic topoisomerase II poisons like etoposide.
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| ln Vitro |
Merbarone (1-100 μM) has an IC50 of 10 μM and suppresses the growth of L1210 cells in a concentration-dependent manner [3]. Human topoisomerase IIα-catalyzed DNA relaxing is inhibited by merbarone (10-200 μM; 10 minutes) with an IC50 of around 40 μM [1]. Using an IC50 of around 50 μM, merbarone (25-200 μM; 6 min) inhibits DNA breakage mediated by tyrosine isomerase II [1]. Global inhibition of topoisomerase II-mediated DNA cleavage is achieved by merbarone (100 μM; 6 min) [1]. The binding of topoisomerase II to DNA is not affected by merbarone (100 μM; 6 min) [1]. Topoisomerase II-catalyzed ATP hydrolysis is not inhibited by merbarone (200 μM; 4–16 min) [1].
In vitro, merbarone is a catalytic inhibitor of topoisomerase II with an IC50 of 120 μM. It has anti-proliferative activity against human cancer cells. Merbarone blocks topoisomerase II-mediated DNA cleavage without stabilizing the DNA-topo II-cleavable complexes. It induces apoptosis in CEM cells via a caspase-3-dependent mechanism. The compound's unique mechanism of action makes it a valuable tool for studying the role of topoisomerase II in DNA replication and cell cycle regulation. |
| ln Vivo |
In P388 mouse leukemia, merbarone (50 mg/kg; daily intraperitoneal injection for 5 days) increases longevity (ILS) by 101% [2]. Mice treated orally with merbarone (124 mg/kg) for nine days showed antitumor activity [2].
In vivo, merbarone has mild antitumor action but is also found to be nephrotoxic. It is orally bioactive. By inhibiting topoisomerase II, merbarone interferes with DNA replication and induces DNA damage, leading to cell death in rapidly dividing cancer cells. However, its nephrotoxic effects limit its therapeutic utility. The compound has been studied in preclinical models for its antitumor activity, but specific in vivo efficacy data are limited in the available literature. |
| Enzyme Assay |
Non-cellular in vitro assays for merbarone involve measuring its inhibition of topoisomerase II activity. A typical protocol uses purified recombinant human topoisomerase II enzyme and a DNA substrate (e.g., kinetoplast DNA or a plasmid DNA). The enzyme is incubated with ATP, the DNA substrate, and varying concentrations of merbarone in a reaction buffer. The reaction products are resolved by agarose gel electrophoresis to separate the decatenated or relaxed DNA from the supercoiled substrate. The IC50 for inhibition of topoisomerase II activity is determined from the concentration-response curve.
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| Cell Assay |
Cellular assays for merbarone are performed using cancer cell lines such as CEM (human T-cell leukemia) cells. Cells are treated with merbarone at various concentrations for 24-72 hours. Cell viability is measured using an MTT assay. Apoptosis is assessed by measuring caspase-3 activity using a fluorogenic substrate or by flow cytometry using annexin V staining. DNA damage can be assessed by measuring γ-H2AX foci formation. The IC50 for anti-proliferative activity is determined.
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| Animal Protocol |
In vivo animal studies for merbarone are conducted in mouse models of cancer, such as xenograft models. Mice bearing tumors are administered merbarone orally or intraperitoneally at doses such as 10-50 mg/kg. Tumor volume is measured over time to assess antitumor efficacy. Body weight and clinical signs are monitored to assess toxicity. At the end of the study, tumor tissues may be collected for analysis of topoisomerase II activity and apoptosis markers. However, specific published protocols for merbarone are limited in the available literature.
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| ADME/Pharmacokinetics |
Merbarone has a molecular weight of 263.27. It is soluble in DMSO at 53 mg/mL (201.31 mM), but is insoluble in water and ethanol. For in vivo oral administration, it can be formulated as a homogeneous suspension in CMC-Na at ≥5 mg/mL. For injection, it can be formulated in 5% DMSO, 40% PEG300, 5% Tween 80, and 50% ddH2O (0.850 mg/mL) or in 5% DMSO and 95% corn oil (0.300 mg/mL). Detailed pharmacokinetic parameters have not been fully reported.
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| Toxicity/Toxicokinetics |
Merbarone has a significant toxicity profile. It is found to be nephrotoxic. As a research chemical, it is not intended for human use and is strictly for preclinical research purposes. Standard safety precautions should be followed when handling merbarone, including the use of personal protective equipment. No specific LD50 values are available in the provided literature.
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| References |
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| Additional Infomation |
Mebalon, a non-sedated derivative of thiobarbituric acid, is a novel topoisomerase II catalytic inhibitor with antitumor activity. Mebalon interferes with DNA replication by directly binding to topoisomerase II, and its binding site may be shared with other topoisomerase II cleavage enhancers, such as etoposide. Because mebalon is neither intercalated into DNA nor stable in the DNA-topoisomerase II cleavable complex, its mechanism of action appears to be entirely novel.
Merbarone (NSC 336628) is a catalytic inhibitor of topoisomerase II with an IC50 of 120 μM. It has anti-proliferative activity against human cancer cells. The compound acts primarily by blocking topoisomerase II-mediated DNA cleavage without stabilizing the topo II-DNA covalent complex. It induces apoptosis in CEM cells via a caspase-3-dependent mechanism. Merbarone has mild antitumor action but is nephrotoxic. It is orally bioactive. Merbarone is not a clinically approved drug and has not entered clinical trials. Its primary application is in research on topoisomerase II biology and anticancer drug development. |
| Molecular Formula |
C11H9N3O3S
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|---|---|
| Molecular Weight |
263.271
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| Exact Mass |
263.036
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| CAS # |
97534-21-9
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| PubChem CID |
4990817
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| Appearance |
Pink to purple solid powder
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| Density |
1.51g/cm3
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| Index of Refraction |
1.688
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| LogP |
0.502
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
18
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| Complexity |
384
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
JARCFMKMOFFIGZ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H9N3O3S/c15-8(12-6-4-2-1-3-5-6)7-9(16)13-11(18)14-10(7)17/h1-5,7H,(H,12,15)(H2,13,14,16,17,18)
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| Chemical Name |
4,6-dioxo-N-phenyl-2-sulfanylidene-1,3-diazinane-5-carboxamide
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| Synonyms |
Merbarone RLBN-1001 NSC-336628
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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 : ~12.5 mg/mL (~47.48 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (9.50 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 sonication.
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 (9.50 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.7984 mL | 18.9919 mL | 37.9838 mL | |
| 5 mM | 0.7597 mL | 3.7984 mL | 7.5968 mL | |
| 10 mM | 0.3798 mL | 1.8992 mL | 3.7984 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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