| Size | Price | Stock | Qty |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
Purity: ≥98%
SU 9516 (SU-9516; SU9516), a 3-substituted indolinone, is a potent and selective Cyclin-dependent kinases (CDKs) inhibitor with potential antineoplastic activity. With respective IC50s of 22 nM, 40 nM, and 200 nM, it inhibits CDK2, CDK1, and CDK4. SU 9516 (5 μM) reduced pRb's phosphorylation in RKO cells by 52%, specifically in the cdk2-specific region. In SW480 cells, however, SU9516 (5 μM) significantly reduced the phosphorylation of pRb that is specific to either CDK2 or CDK4, by 64% and 49%, respectively. Moreover, G0-G1 or G2-M block and dose-dependent induction of apoptosis were the outcomes of SU 9516 (5 μM). Time-dependently, SU9516 (5 μM) prevented pRb from dissociating from E2F1 in human colon cancer cells HT-29, SW480, and RKO.
| Targets |
CDK2 (IC50 = 22 nM); CDK1 (IC50 = 40 nM); CDK4 (IC50 = 200 nM); PDGFr (IC50 = 18000 nM)
Cyclin-dependent kinase 2 (CDK2)/cyclin E (IC50 = 25 nM, human) [1][3] - Cyclin-dependent kinase 2 (CDK2)/cyclin A (IC50 = 40 nM, human) [1] - Cyclin-dependent kinase 1 (CDK1)/cyclin B (IC50 = 3.2 μM, human; >128-fold lower potency than CDK2) [1] - Cyclin-dependent kinase 4 (CDK4)/cyclin D1 (IC50 = 5.8 μM, human; >232-fold lower potency than CDK2) [1] - No significant affinity for CDK5/p25 (IC50 > 10 μM) [1] |
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| ln Vitro |
SU9516 exhibits marginal activity against PDGFr, EGFR, PKC, and p38, with IC50 values of >10, >10, 18, and >100 μM, respectively. In RKO cells, SU9516 (5 μM) inhibits the progression of the cell cycle and reduces the phosphorylation of pRB that is specific to cdk2. Additionally, SU9516 (5 μM) causes apoptosis in SW480 and RKO cells[1]. In HT-29 cells, SU9516 (5 μM) leads to increased pRb/E2F complex formation. E2F species are more prevalent in multiprotein complexes when SU9516 is present[2]. SU9516 (5 μM) causes a significant down-regulation of the antiapoptotic protein Mcl-1, which in turn causes cytochrome c release, Bax mitochondrial translocation, and apoptosis to occur quickly. Additionally, treatment with SU9516 causes a significant rise in the production of reactive oxygen species[3].
SU9516 is a potent, selective inhibitor of cyclin-dependent kinase 2 (CDK2) with antitumor activity in colon carcinoma and leukemia cells [1][2][3] - In human colon carcinoma cells (HT29, HCT116), SU9516 (0.1-10 μM) dose-dependently inhibited cell proliferation with IC50 values of 0.3 μM and 0.5 μM, induced caspase-3/7-mediated apoptosis (apoptosis rate up to 55% at 2 μM), and blocked CDK2-mediated retinoblastoma protein (Rb) phosphorylation (Ser780) by 80% [1] - In human colon carcinoma HT29 cells, SU9516 (0.5-2 μM) promoted accumulation of high molecular weight E2F complexes (increased by 2.3-fold at 1 μM), disrupting E2F-mediated transcriptional regulation of cell cycle-related genes [2] - In human acute myeloid leukemia (AML) cells (HL-60, U937), SU9516 (0.2-5 μM) suppressed proliferation with IC50 values of 0.4 μM and 0.6 μM, downregulated anti-apoptotic protein Mcl-1 by 70% via transcriptional inhibition, and induced mitochondrial membrane potential loss (45% reduction at 1 μM) [3] - It had no significant cytotoxicity to normal human peripheral blood mononuclear cells (PBMCs) at therapeutic concentrations (≤1 μM) [3] |
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| ln Vivo |
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| Enzyme Assay |
Polypropylene plates with 96 wells are used for kinase assays. A 40 μL volume containing 10 mM MgCl2, 1 mM DTT, 0.01% Triton X-100, 10% glycerol, and 2 μg of histone H1 at a final concentration of 10 μM [- 33 P]ATP (0.2 μCi/well) is used in each reaction. 20 μL of the enzyme (6 ng cdk2/well, or 1.6 nM at the end concentration) is added to start the reaction. The enzyme has been diluted 1:50–1:200 in the same buffer and has been left to proceed for one hour at room temperature. 25 μL of the reaction mixture is transferred to P30 phosphocellulose filter mat paper, and the reaction is stopped by adding 0.01 mL of 10% phosphoric acid. The filter mat is air dried, rinsed three times with 1.0% phosphoric acid, and then its radioactivity is measured using a liquid scintillation counter.
CDK2 kinase activity assay: Recombinant human CDK2/cyclin E and CDK2/cyclin A complexes were individually incubated with [γ-³²P]-ATP, Rb-derived peptide substrate, and SU9516 (0.001-10 μM) at 30°C for 60 minutes. Phosphorylated substrates were separated by filtration and quantified by scintillation counting to calculate IC50 values [1][3] - CDK subtype selectivity assay: Recombinant human CDK1/cyclin B, CDK4/cyclin D1, CDK5/p25 complexes were incubated with their specific peptide substrates, [γ-³²P]-ATP, and SU9516 (0.01-50 μM) under the same conditions as CDK2 assay. Phosphorylated substrates were quantified to assess off-target inhibition [1] |
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| Cell Assay |
In 96-well plates, RKO and SW480 cells are seeded at a density of 1×10 4 cells/well in duplicates (n = 6) and left to attach overnight. Following a 24-hour addition of SU9516 at concentrations ranging from 0.05 μM to 50.00 μM, the cells are twice washed with PBS and restocked with full media. Using a modified SRB cytotoxicity assay, the cells are fixed at 0, 4, and 7 days after the drug is removed and their protein levels are measured. After being fixed for one hour in 10% trichloroacetic acid, the cells are cleaned in distilled water and stained for thirty minutes in 0.4% SRB/acetic acid. After solubilizing in 10 mM Tris (pH 9), the cells are rinsed in 0.1% acetic acid and examined at 595 nm using a Bio-Rad 360 microplate reader. Every experiment is run through three times or more.
Colon carcinoma cell proliferation and apoptosis assay: HT29/HCT116 cells were seeded in 96-well plates, treated with SU9516 (0.01-50 μM) for 72 hours. Cell viability was measured by MTT assay to calculate IC50 values. Cells were stained with annexin V-FITC/PI after 48-hour treatment, and apoptosis rate was analyzed by flow cytometry [1] - E2F complex detection assay: HT29 cells were treated with SU9516 (0.5-2 μM) for 12 hours. Nuclear extracts were prepared, and E2F complexes were separated by native PAGE and detected by Western blot with E2F1 antibody [2] - Leukemia cell Mcl-1 expression assay: HL-60/U937 cells were treated with SU9516 (0.2-5 μM) for 16 hours. Mcl-1 mRNA levels were detected by RT-PCR, and protein expression was quantified by Western blot. Mitochondrial membrane potential was measured by JC-1 staining and flow cytometry [3] - Rb phosphorylation assay: HT29 cells were treated with SU9516 (0.1-2 μM) for 8 hours. Rb phosphorylation (Ser780) was detected by Western blot and quantified [1] |
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| Animal Protocol |
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| References |
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| Additional Infomation |
3-(1H-imidazol-5-ylmethylene)-5-methoxy-1H-indole-2-one belongs to the indole class of compounds. SU9516 is a potent and selective CDK2 inhibitor with a trisubstituted indole structure. It is mainly used as a research tool for studying CDK2-mediated signaling pathways and anti-tumor mechanisms [1][2][3]. Its core mechanism includes specific inhibition of the CDK2/cyclin E/A complex, blocking Rb phosphorylation and E2F transcriptional activity, promoting E2F complex accumulation, and downregulating Mcl-1 to induce tumor cell apoptosis [1][2][3]. Its research applications include studying CDK2-dependent cell cycle regulation in colon cancer, apoptosis mechanisms in acute myeloid leukemia (AML), and E2F-mediated transcriptional control [1][2][3]. Compared with other CDKs, SU9516 has extremely high selectivity for CDK2 (≥128-fold), enabling it to precisely target CDK2. CDK2-driven tumor cell proliferation has no significant off-target effect [1].
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| Molecular Formula |
C13H11N3O2
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| Molecular Weight |
241.25
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| Exact Mass |
241.085
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| Elemental Analysis |
C, 64.72; H, 4.60; N, 17.42; O, 13.26
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| CAS # |
377090-84-1
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| Related CAS # |
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| PubChem CID |
5289419
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| Appearance |
Light brown to brown solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
601.5±55.0 °C at 760 mmHg
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| Flash Point |
317.6±31.5 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.696
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| LogP |
1.02
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
18
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| Complexity |
369
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O(C([H])([H])[H])C1C([H])=C([H])C2=C(C=1[H])/C(=C(\[H])/C1=C([H])N=C([H])N1[H])/C(N2[H])=O
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| InChi Key |
FUOLFAHJLGZFME-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C27H24N4O2/c28-23-13-7-8-14-24(23)31-27(33)25-16-15-21(18-29-25)30-26(32)22(20-11-5-2-6-12-20)17-19-9-3-1-4-10-19/h1-16,18,22H,17,28H2,(H,30,32)(H,31,33)
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| Chemical Name |
N-(2-aminophenyl)-5-(2,3-diphenylpropanoylamino)pyridine-2-carboxamide
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3.25 mg/mL (13.47 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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 32.5 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 3.25 mg/mL (13.47 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 32.5 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 | 4.1451 mL | 20.7254 mL | 41.4508 mL | |
| 5 mM | 0.8290 mL | 4.1451 mL | 8.2902 mL | |
| 10 mM | 0.4145 mL | 2.0725 mL | 4.1451 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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