| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Pyruvate kinase M2 (PKM2) (allosteric activator, EC50: 73 nM)
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| ln Vitro |
ML202 is a highly specific allosteric activator of the tumor-specific isoform of human pyruvate kinase M2 (hPK-M2) with an EC50 value of 73 nM. It enhances the cooperativity of phosphoenolpyruvate (PEP) binding, while having negligible effect on adenosine diphosphate (ADP) binding. ML202 is inactive against other pyruvate kinase isoforms (hPyk-M1, -R, and -L isoforms), demonstrating high selectivity for PKM2. PKM2 is a key regulator of aerobic glycolysis in cancer cells (the Warburg effect). Activation of PKM2 shifts metabolism from lactate production toward pyruvate entry into the tricarboxylic acid (TCA) cycle, potentially reducing cancer cell proliferation.
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| ln Vivo |
No specific in vivo data reported for ML202. As a PKM2 activator with high isoform selectivity, ML202 has potential for in vivo efficacy in xenograft models of cancers that express PKM2. PKM2 activation in vivo would be expected to reduce tumor glycolysis, decrease lactate production, and potentially inhibit tumor growth. In vivo studies would involve administration to tumor-bearing mice, assessment of tumor PKM2 activity, glycolytic flux (by ¹⁸F-FDG PET imaging), and tumor growth inhibition. ML202 is a research tool for studying PKM2 biology and the role of glycolysis in cancer.
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| Enzyme Assay |
Recombinant human PKM2 enzyme is expressed in E. coli and purified. The PKM2 activity assay measures the conversion of phosphoenolpyruvate (PEP) and ADP to pyruvate and ATP, coupled to NADH oxidation via lactate dehydrogenase (LDH). The reaction mixture contains Tris-HCl buffer (pH 7.4), KCl, MgCl2, PEP (0.1-10 mM), ADP (0.1-10 mM), NADH (0.2-0.5 mM), and LDH. The reaction is initiated by adding PKM2 enzyme (10-50 ng). The decrease in absorbance at 340 nm (NADH oxidation) is monitored over 5-10 minutes at 37degC. For activator screening, compounds are tested at varying concentrations (0.1-1000 nM) and the fold-increase in activity relative to control (DMSO vehicle) is calculated. EC50 values are determined from dose-response curves (EC50 = 73 nM for ML202). The selectivity against other pyruvate kinase isoforms (PKM1, PKR, PKL) is assessed using the same assay but with the respective recombinant enzyme isoforms. ML202 is inactive against PKM1, PKR, and PKL at concentrations up to 10 uM, demonstrating high isoform selectivity.
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| Cell Assay |
Human cancer cell lines that express PKM2 (e.g., HeLa, HCT116, A549, MCF-7) are cultured in DMEM or RPMI-1640 with 10% FBS and antibiotics. Cells are seeded in 96-well plates or 6-well plates and treated with ML202 (0.1-1000 nM) for 24-72 hours. Pyruvate kinase activity is measured in cell lysates using the PK activity assay described above; specific activity is normalized to total protein content (BCA assay). Lactate production is measured in the culture supernatant using a colorimetric lactate assay kit. Glucose consumption is measured by assaying glucose levels in the culture medium (glucose oxidase assay). Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) are measured using a Seahorse XF analyzer to assess glycolytic flux and mitochondrial respiration. Cell proliferation is assessed by MTT, CCK-8, or CellTiter-Glo assays. The effect of ML202 on the metabolic phenotype can be assessed by measuring the ratio of lactate production to glucose consumption. Apoptosis can be assessed by Annexin V/PI staining and caspase-3/7 activity. The EC50 for PKM2 activation is 73 nM; cellular effects are typically observed at low nanomolar to micromolar concentrations depending on the cell line and assay. ML202 is cell-permeable due to its lipophilic properties.
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| Animal Protocol |
No published in vivo animal study for ML202. As a PKM2 activator with high isoform selectivity, ML202 has potential for in vivo efficacy studies in xenograft models of PKM2-expressing cancers. A typical protocol would involve establishing subcutaneous tumor xenografts in immunodeficient mice (e.g., nude mice or NSG mice) using HCT116, HeLa, or other PKM2-expressing cancer cell lines. When tumors reach 100-200 mm3, mice are randomized and treated with ML202 formulated in a suitable vehicle (e.g., 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline, or 10% DMSO + 90% corn oil). Dosing is once daily by intraperitoneal injection (IP) or oral gavage at doses ranging from 1-50 mg/kg for 2-4 weeks. Endpoints: tumor volume measured by calipers, tumor weight at necropsy, body weight, survival. Plasma and tumor tissue are collected for pharmacokinetic analysis and pharmacodynamic evaluation: PKM2 activity in tumor lysates, lactate levels in tumor tissue and plasma (by colorimetric assay), glucose levels (by colorimetric assay), and ATP levels (by bioluminescence). Glycolytic flux can be assessed in vivo by ¹⁸F-FDG PET/CT imaging, which measures tumor glucose uptake. Histological analysis of tumors (H&E, Ki-67 for proliferation, TUNEL for cell death) can be performed. The effect on tumor metabolism can be assessed by measuring the lactate-to-glucose ratio. These protocols are generic and not validated for ML202 specifically.
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| ADME/Pharmacokinetics |
No specific pharmacokinetic data for ML202. Based on its molecular properties (MW 387.48, LogP 3.609), the compound is moderately lipophilic and is expected to have moderate to good oral bioavailability. Plasma half-life, Cmax, AUC, clearance, volume of distribution, and protein binding have not been publicly reported. Solubility: DMSO 50 mg/mL (129.04 mM). For in vivo formulation: ≥2.5 mg/mL (6.45 mM) in 10% DMSO + 90% (20% SBE-beta-CD in Saline) (clear solution). Storage: powder at -20degC for 3 years; in solvent at -80degC for 6 months. The compound is cell-permeable and can be used in vivo for metabolic studies.
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| Toxicity/Toxicokinetics |
No specific toxicity data for ML202. As a PKM2 activator, the primary expected on-target effect is modulation of glucose metabolism in PKM2-expressing cells. PKM2 is highly expressed in cancer cells but also expressed in some normal proliferative tissues (e.g., bone marrow, intestinal epithelium, immune cells). Therefore, PKM2 activation could potentially affect normal cell proliferation and metabolism. In cell-based studies, no significant cytotoxicity has been reported at relevant concentrations (low nanomolar to low micromolar). In vivo toxicity studies would assess maximum tolerated dose (MTD), body weight loss, organ toxicity (liver, kidney), hematological parameters, and blood glucose/lactate levels. No data are publicly available. For research use only; handle with standard laboratory precautions (gloves, lab coat, eye protection). ML202 is not for human use.
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| References | |
| Additional Infomation |
6-[(3-methoxyphenyl)methyl]-4-methyl-2-methylsulfinyl-5-thieno[3,4]pyrrolo[1,3-d]pyridazinone is an organosulfur heterocyclic compound, an organonitrogen heterocyclic compound, and an organoheterobicyclic compound.
ML202 (CAS: 1221186-52-2) is a highly specific allosteric activator of human pyruvate kinase M2 (PKM2), the tumor-specific isoform of pyruvate kinase. PKM2 is a key regulator of aerobic glycolysis (the Warburg effect) in cancer cells. ML202 enhances the cooperativity of phosphoenolpyruvate (PEP) binding (EC50 = 73 nM) and is inactive against other pyruvate kinase isoforms (PKM1, PKR, PKL). ML202 is a valuable research tool for studying PKM2 biology, the Warburg effect, and the metabolic reprogramming of cancer cells. Molecular formula: C18H17N3O3S2, molecular weight: 387.48. Appearance: white to off-white solid powder. Purity: ≥98% (HPLC). Solubility: DMSO 50 mg/mL (129.04 mM). Storage: store at +4degC or -20degC. Not approved for clinical use. For research use only. Reference: ML202 is a highly specific allosteric activator of human pyruvate kinase M2 (hPK-M2), which can affect the cooperativity of phosphoenolpyruvate (PEP) binding, while adenosine diphosphate (ADP) binding almost no effect. |
| Molecular Formula |
C18H17N3O3S2
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|---|---|
| Molecular Weight |
387.48
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| Exact Mass |
387.071
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| CAS # |
1221186-52-2
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| PubChem CID |
44246498
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| Appearance |
White to off-white solid powder
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| LogP |
3.609
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
26
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| Complexity |
617
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1C2=C(C3=C1C(=O)N(N=C3)CC4=CC(=CC=C4)OC)SC(=C2)S(=O)C
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| InChi Key |
MORBXZMIXGYQDB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H17N3O3S2/c1-20-14-8-15(26(3)23)25-17(14)13-9-19-21(18(22)16(13)20)10-11-5-4-6-12(7-11)24-2/h4-9H,10H2,1-3H3
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| Chemical Name |
10-[(3-methoxyphenyl)methyl]-7-methyl-4-methylsulfinyl-3-thia-7,10,11-triazatricyclo[6.4.0.02,6]dodeca-1(8),2(6),4,11-tetraen-9-one
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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: 50 mg/mL (129.04 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.45 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), 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 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.45 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5808 mL | 12.9039 mL | 25.8078 mL | |
| 5 mM | 0.5162 mL | 2.5808 mL | 5.1616 mL | |
| 10 mM | 0.2581 mL | 1.2904 mL | 2.5808 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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