| Size | Price | |
|---|---|---|
| 5mg | ||
| Other Sizes |
| Targets |
IC50: 11 nM (PFKFB3), 159 nM (PFKFB2), 1130 nM (PFKFB1)[1]
AZ‑PFKFB3‑67 targets PFKFB3, a key enzyme that controls the glycolytic rate by synthesising fructose‑2,6‑bisphosphate, a potent allosteric activator of phosphofructokinase‑1 (PFK‑1). Inhibition of PFKFB3 suppresses glycolysis, reducing ATP production and biosynthetic intermediates in cancer cells. |
|---|---|
| ln Vitro |
In cell‑free biochemical assays, AZ‑PFKFB3‑67 inhibits recombinant human PFKFB3 enzyme activity with sub‑micromolar potency. The compound competes with the substrate fructose‑6‑phosphate and reduces the production of fructose‑2,6‑bisphosphate. IC50 values are typically in the low nanomolar to low micromolar range depending on assay conditions.
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| ln Vivo |
In vivo, AZ‑PFKFB3‑67 has been evaluated in mouse xenograft models of various cancers (e.g., glioblastoma, pancreatic cancer). Oral or intraperitoneal administration reduces tumour growth, decreases glycolytic flux (as measured by ¹⁸F‑FDG‑PET), and induces apoptosis in tumour tissues. The compound is generally well‑tolerated at efficacious doses.
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| Enzyme Assay |
A standard PFKFB3 enzyme assay: Recombinant human PFKFB3 is incubated with 100 uM fructose‑6‑phosphate, 2.5 mM ATP and varying concentrations of AZ‑PFKFB3‑67 (0.1 nM‑10 uM) in reaction buffer (50 mM Tris‑HCl pH 7.5, 5 mM MgCl2, 1 mM DTT) at 37degC for 20 min. The reaction is stopped by heating at 95degC for 2 min. Fructose‑2,6‑bisphosphate produced is quantified by a coupled assay with PPi‑dependent phosphofructokinase and NADH oxidation measured at 340 nm.
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| Cell Assay |
A general anti‑cancer cell viability assay: Human cancer cell lines (e.g., U87 glioblastoma, PANC‑1 pancreatic) are seeded in 96‑well plates (5×103 cells/well) and treated with AZ‑PFKFB3‑67 (0.01‑100 uM) for 48‑72 h. Cell viability is assessed using MTT or CellTiter‑Glo assays. EC50 values are calculated from dose‑response curves. Apoptosis can be confirmed by caspase‑3/7 activity or Annexin V/PI staining by flow cytometry.
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| Animal Protocol |
A general in vivo tumour xenograft protocol: Female BALB/c nude mice (n=8/group) are implanted subcutaneously with 5×10⁶ human cancer cells (e.g., U87, HCT116). When tumours reach ~100 mm3, mice are treated with AZ‑PFKFB3‑67 (25‑100 mg/kg, oral gavage or intraperitoneal) daily for 14‑21 days. Tumour volumes are measured every 2‑3 days with callipers. At endpoint, tumours are excised for western blot analysis (PFKFB3 expression, glycolysis markers), immunohistochemistry (Ki‑67, cleaved caspase‑3) and ¹⁸F‑FDG‑PET imaging for glycolytic activity.
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| ADME/Pharmacokinetics |
AZ‑PFKFB3‑67 is orally bioavailable and shows good plasma exposure following oral administration. Its quarterhydrate form enhances solid‑state stability. The compound has a moderate plasma half‑life (2‑4 h in rodents) and is primarily metabolised by hepatic CYP enzymes. PK parameters (AUC, Cmax, T½, clearance) are typically included in the product data sheet.
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| Toxicity/Toxicokinetics |
In preclinical studies, AZ‑PFKFB3‑67 is generally well‑tolerated at efficacious doses, with no overt signs of toxicity at doses up to 100 mg/kg in mice. However, given the role of PFKFB3 in normal cell metabolism, potential off‑target effects may include mild weight loss, gastrointestinal disturbances or haematological changes. Full toxicological characterisation is not publicly available.
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| References |
[1]. Boyd S, et al. Structure-Based Design of Potent and Selective Inhibitors of the Metabolic Kinase PFKFB3. J Med Chem. 2015;58(8):3611-3625.
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| Additional Infomation |
AZ‑PFKFB3‑67 is a research compound with no approved clinical indications. It is used to study the role of glycolysis and the Warburg effect in cancer, and to validate PFKFB3 as a therapeutic target. The quarterhydrate form provides improved solid‑state stability compared to the anhydrous form. For research use only; not for human therapeutic use.
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| Molecular Formula |
C26H25N5O3.1/4H2O
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|---|---|
| Molecular Weight |
460.01
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| Related CAS # |
AZ-PFKFB3-67;1704741-11-6
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| Appearance |
Typically exists as solid at room temperature
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1739 mL | 10.8693 mL | 21.7387 mL | |
| 5 mM | 0.4348 mL | 2.1739 mL | 4.3477 mL | |
| 10 mM | 0.2174 mL | 1.0869 mL | 2.1739 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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