| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
Ki: 40 nM (PI5P4Kα) and 30 nM (PI5P4Kβ)[1]
PI5P4Kalpha and PI5P4Kbeta (phosphatidylinositol 5-phosphate 4-kinase alpha and beta). These enzymes phosphorylate phosphatidylinositol 5-phosphate (PI5P) at the 4th position of the inositol ring to produce phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2). CC260 is a selective inhibitor of these two isoforms, with weak or no inhibition of PI5P4Kgamma or other protein kinases. |
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| ln Vitro |
CC260 (20 μM) reduces S6K phosphorylation (Thr-389) by mTORC1 but increases Insulin-induced Akt phosphorylation at both Thr-308 and Ser-473 in cultured C2C12 myotubes[1]. Acetyl-CoA carboxylase (ACC) phosphorylation is greatly increased by CC260 (2.5 μM, 5 μM, 10 μM, and 20 μM) in a dose-dependent manner[1]. Treatment with CC260 decreases the ability of BT474 cells to withstand serum starvation; however, this ability can be restored by expressing the refractory mutant PI5P4Kβ[1]. Treatment with CC260 increases the generation of glycolytic ATP in BT474 cells[1].
In cell-free kinase assays, CC260 selectively inhibits PI5P4Kalpha and PI5P4Kbeta with Ki values of 40 nM and 30 nM, respectively. It demonstrates minimal to no inhibition against a panel of other protein kinases, including Plk1 and RSK2, indicating its high selectivity. A crystal structure of PI5P4Kbeta in complex with CC260 confirmed its binding mode, where it occupies the active site of the kinase. |
| ln Vivo |
In BT474 breast cancer cells, treatment with CC260 reduced the ability of the cells to survive under serum-starved conditions, which could be rescued by expressing a CC260-refractory mutant of PI5P4Kbeta. Inhibition of PI5P4Kalpha and PI5P4Kbeta by CC260 increases AMPK activity, and in cells where both isoforms are knocked down by siRNA, no further increase in AMPK activity is observed, confirming on-target activity. CC260 did not inhibit mTOR/FRAP1.
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| Enzyme Assay |
A standard in vitro kinase activity assay for PI5P4Ks is the thin-layer chromatography (TLC) method. The assay is performed by incubating purified recombinant PI5P4Kalpha or PI5P4Kbeta with the substrate, phosphatidylinositol 5-phosphate (PI5P), and 32P-ATP. The reaction mixture is then separated by TLC, which resolves the product (radiolabeled PI(4,5)P2) from the substrate (PI5P). The TLC plate is exposed to a phosphor screen, and the radioactivity is quantified to calculate the Ki or IC50.
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| Cell Assay |
To assess target engagement in a cellular context, a western blot assay can be used. Cells (e.g., BT474) are treated with CC260 or a negative control, and protein lysates are prepared. The lysates are then subjected to gel electrophoresis and probed with antibodies specific for phosphorylated AMPK (p-AMPK) and total AMPK. CC260 treatment is expected to increase p-AMPK levels compared to control, indicating inhibition of PI5P4K and the downstream metabolic stress response.
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| Animal Protocol |
To demonstrate in vivo target engagement, a xenograft mouse model can be used. In one such study, BT474 tumor-bearing mice were treated with CC260 or vehicle control. The study design included groups for assessing survival under metabolic stress. Tumor tissues were collected and analyzed to measure markers of the PI5P4K/AMPK signaling pathway, such as p-AMPK, TIGAR, and G6PD levels, to confirm on-target activity of the compound in vivo.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of CC260 are not extensively detailed in standard research product information. As a potent, small molecule inhibitor, it is expected to be cell-permeable and have reasonable bioavailability for in vivo studies, often being formulated in common vehicles like DMSO:PEG300:Tween80 for systemic administration. Its primary use is as a chemical probe for in vitro and in vivo studies to delineate the function of PI5P4K isoforms in various diseases.
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| Toxicity/Toxicokinetics |
Specific toxicity data for CC260 is not reported in the referenced literature. However, as a research compound, its safety profile is expected to be manageable for experimental uses. In the described studies, treatment of mice with CC260 did not result in reported deaths or severe adverse events, suggesting it is tolerated at the described dosing regimens. Long-term and off-target toxicity studies would be necessary for any potential therapeutic development.
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| References | |
| Additional Infomation |
CC260 has been used as a chemical probe to demonstrate that pharmacological inhibition of PI5P4Kalpha and PI5P4Kbeta selectively kills p53-null tumor cells by disrupting cell energy metabolism. This finding identifies PI5P4Kalpha/beta as potential therapeutic targets for cancers with TP53 mutations. The compound is also studied in diabetes research. A crystal structure of PI5P4Kbeta with CC260 is available (PDB ID: 7N81), which aids in further drug discovery for these targets. It is a research tool not intended for therapeutic use.
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| Molecular Formula |
C24H29CL2N5O2
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|---|---|
| Molecular Weight |
490.43
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| Exact Mass |
489.169
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| CAS # |
2411088-26-9
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| PubChem CID |
146347307
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| Appearance |
Off-white to gray solid powder
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| LogP |
6.5
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
33
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| Complexity |
678
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| Defined Atom Stereocenter Count |
1
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| SMILES |
N1=C2C(N(C)C(=O)[C@@H](CC3CCCC3)N2C2CCCC2)=CN=C1NC1=CC(Cl)=C(O)C(Cl)=C1
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| InChi Key |
AEZJZTFWCDAUDF-LJQANCHMSA-N
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| InChi Code |
InChI=1S/C24H29Cl2N5O2/c1-30-20-13-27-24(28-15-11-17(25)21(32)18(26)12-15)29-22(20)31(16-8-4-5-9-16)19(23(30)33)10-14-6-2-3-7-14/h11-14,16,19,32H,2-10H2,1H3,(H,27,28,29)/t19-/m1/s1
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| Chemical Name |
(7R)-8-cyclopentyl-7-(cyclopentylmethyl)-2-(3,5-dichloro-4-hydroxyanilino)-5-methyl-7H-pteridin-6-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: 160 mg/mL (326.24 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 4 mg/mL (8.16 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 sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 40.0 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: ≥ 4 mg/mL (8.16 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 40.0 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 | 2.0390 mL | 10.1951 mL | 20.3903 mL | |
| 5 mM | 0.4078 mL | 2.0390 mL | 4.0781 mL | |
| 10 mM | 0.2039 mL | 1.0195 mL | 2.0390 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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