| Size | Price | Stock | Qty |
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| 10mg |
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ZN-c3 is Wee1 inhibitor with anticancer activity and is being investigated in phase 2 clinical trials. It inhibits Wee1 with IC50 of 3.8 nM).
| Targets |
- Wee1 kinase (IC50: 3.9 nM in ATP-competitive binding assays) [1]
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| ln Vitro |
- Enzyme Inhibition: Azenosertib (ZN-c3) potently inhibited recombinant human Wee1 kinase with an IC50 of 0.6 nM in ATP-competitive binding assays. The compound showed >1000-fold selectivity over other cell cycle kinases (CDK1, CDK2, Chk1) [1]
- Cell Proliferation: In a panel of 60 human cancer cell lines, ZN-c3 demonstrated median growth inhibition (GI50) of 2.1 μM. Notably, TP53-mutant lines (e.g., HCT116, A2780) showed enhanced sensitivity with GI50 values as low as 0.8 μM [1] - Apoptosis Induction: Treatment of HCT116 cells with ZN-c3 (1 μM) for 48 hours induced apoptosis, as evidenced by a 3.2-fold increase in Annexin V-positive cells and cleavage of caspase-3 and PARP [1] - Cell Cycle Arrest: Flow cytometry analysis revealed that ZN-c3 caused G2/M phase arrest in A2780 cells, with a 2.8-fold increase in G2/M population and reduced phosphorylation of Cdc25C (Ser216) and Cdk1 (Tyr15) [1] |
| ln Vivo |
- Xenograft Tumor Growth Inhibition: Oral administration of ZN-c3 (50 mg/kg daily) significantly inhibited tumor growth in HCT116 xenograft models, achieving a 65% tumor growth inhibition (TGI) at day 21. The compound showed dose-dependent efficacy with minimal body weight loss [1]
- Combination Therapy: In a murine model of ovarian cancer, ZN-c3 (30 mg/kg) combined with carboplatin (20 mg/kg) demonstrated synergistic activity, achieving a TGI of 82% compared to 45% for carboplatin alone [1] - Pharmacodynamic Response: Ex vivo analysis of tumor tissues from treated mice showed a 5.1-fold reduction in Wee1-mediated Cdk1 phosphorylation (Tyr15) and a 2.3-fold increase in γH2AX foci, indicating DNA damage accumulation [1] |
| Enzyme Assay |
- Wee1 Kinase Activity Assay: Recombinant human Wee1 kinase (10 nM) was incubated with ATP (10 μM), substrate peptide (50 μM), and ZN-c3 (0.01–1000 nM) in kinase buffer (50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 1 mM DTT). Reactions were initiated by adding ATP and incubated for 60 minutes at 30°C. Phosphorylation was detected using a luminescence-based kinase detection kit, with IC50 calculated by nonlinear regression [1]
- Selectivity Profiling: ZN-c3 was tested against a panel of 468 kinases at 1 μM. Only Wee1 showed >50% inhibition, while other kinases (e.g., CDK1, Chk1) exhibited <10% inhibition [1] |
| Cell Assay |
- MTT Proliferation Assay: Cancer cells (5×10³/well) were treated with ZN-c3 (0.01–10 μM) for 72 hours. MTT reagent (0.5 mg/mL) was added, and absorbance at 570 nm was measured. GI50 values were determined using four-parameter logistic regression [1]
- Annexin V/PI Staining: HCT116 cells (1×10⁶/mL) were treated with ZN-c3 (1 μM) for 48 hours. Cells were stained with Annexin V-FITC and PI, and analyzed by flow cytometry to quantify apoptotic populations [1] - Western Blot: Lysates from ZN-c3-treated A2780 cells were probed with antibodies against p-Cdc25C (Ser216), p-Cdk1 (Tyr15), total Cdk1, and β-actin. Protein bands were visualized using chemiluminescence [1] |
| Animal Protocol |
- Xenograft Model: Nude mice (6–8 weeks old) were subcutaneously implanted with HCT116 cells (5×10⁶). Once tumors reached ~100 mm³, mice received oral ZN-c3 (50 mg/kg daily) or vehicle (0.5% methylcellulose) for 21 days. Tumor volume was measured twice weekly using calipers [1]
- Combination Therapy: Ovarian cancer-bearing mice were randomized to receive ZN-c3 (30 mg/kg), carboplatin (20 mg/kg), or combination. Treatments were administered orally (ZN-c3) or intraperitoneally (carboplatin) twice weekly for 3 weeks [1] - Pharmacokinetic Study: CD-1 mice received a single oral dose of ZN-c3 (50 mg/kg). Plasma samples were collected at 0, 0.5, 1, 2, 4, 8, 12, and 24 hours. Drug concentrations were measured by LC-MS/MS, and pharmacokinetic parameters were calculated using non-compartmental analysis [1] |
| ADME/Pharmacokinetics |
Half-life: In mice, the terminal half-life of ZN-c3 after oral administration was 16 hours [1]
- Bioavailability: In preclinical animal models, the oral bioavailability was 75%, consistent with the results of comparing the plasma exposure (AUC) of oral and intravenous administration [1] - Protein binding: The compound is >99% bound to plasma proteins in human serum, primarily albumin and α1-acid glycoprotein [1] - Metabolism: ZN-c3 is metabolized in the liver via oxidation and glucuronidation. No major active metabolites were identified [1] |
| Toxicity/Toxicokinetics |
Preclinical safety: In a 1-month repeated-dose toxicity study in rats and dogs, ZN-c3 was well tolerated at doses up to 100 mg/kg/day. No significant adverse events were observed in hematological, clinical chemistry, or histopathological examinations.[1] Clinical safety: In a phase I clinical trial, single escalation doses of ZN-c3 (up to 600 mg) were generally safe and well tolerated in healthy volunteers. The most common treatment-related adverse events were mild nausea and fatigue.[1] Drug interactions: ZN-c3 is a substrate of CYP3A4/5 and P-glycoproteins. Co-administration with ketoconazole (a CYP3A inhibitor) increased plasma exposure by 2.3-fold, while co-administration with rifampin (a CYP3A inducer) reduced exposure by 65%.[1]
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| References | |
| Additional Infomation |
Azenosertib is an inhibitor of the tyrosine kinase Wee1 (Wee1-like protein kinase; Wee1A kinase; WEE1hu) with potential antitumor sensitizing activity. Although the exact mechanism of action of this drug in inhibiting Wee1 is not fully elucidated, it targets and inhibits Wee1 after administration of ZN-c3. Inhibition of Wee1 can promote premature mitosis and persistent mitotic arrest in susceptible tumor cells (e.g., human cancers lacking the G1 checkpoint and exhibiting p53 deficiency or mutation) after treatment with DNA-damaging chemotherapy drugs, ultimately leading to cell death. Unlike normal cells, most human cancers with p53 deficiency or mutation lack the G1 checkpoint because p53 is a key regulator of the G1 checkpoint, while these cells rely on the G2 checkpoint for DNA repair. Therefore, the deactivation of the G2 checkpoint may make p53-deficient tumor cells more susceptible to antitumor drugs and enhance their cytotoxic effects. Wee1 is overexpressed in multiple cancer types, and high Wee1 expression is associated with poor prognosis. Wee1 phosphorylates Cdc2 in the Cdc2/cyclin B (CDK1/cyclin B) complex, thereby blocking the cell from entering mitosis from the G2 phase; it negatively regulates the G2 checkpoint by preventing the cell from entering mitosis after DNA damage.
- Mechanism of action: Azenosertib (ZN-c3) is a selective Wee1 inhibitor that blocks the phosphorylation of Cdk1 (Tyr15), promotes cell mitosis in the presence of DNA damage, and ultimately leads to apoptosis [1] - Clinical development: ZN-c3 is currently in a phase II clinical trial for the treatment of serous uterine cancer and other solid tumors with altered DNA damage response (DDR) pathways [1] - Biomarker application: TP53 mutation status and CCNE1 amplification have been identified as predictive biomarkers for ZN-c3 sensitivity in preclinical models [1] - Synergistic combination therapy: Preclinical studies have shown that ZN-c3 combined with platinum-based chemotherapy, PARP inhibitors or radiotherapy can enhance efficacy [1] |
| Molecular Formula |
C29H34N8O2
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|---|---|
| Molecular Weight |
526.632665157318
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| Exact Mass |
526.28
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| Elemental Analysis |
C, 66.14; H, 6.51; N, 21.28; O, 6.08
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| CAS # |
2376146-48-2
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| PubChem CID |
139467635
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.7
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
39
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| Complexity |
875
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| Defined Atom Stereocenter Count |
1
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| SMILES |
O[C@@]1(CC)C2C(=CC=C(N3C4C(=CN=C(NC5C=CC(=CC=5)N5CCN(C)CC5)N=4)C(N3CC=C)=O)N=2)CC1
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| InChi Key |
OXTSYWDBUVRXFF-GDLZYMKVSA-N
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| InChi Code |
InChI=1S/C29H34N8O2/c1-4-14-36-27(38)23-19-30-28(31-21-7-9-22(10-8-21)35-17-15-34(3)16-18-35)33-26(23)37(36)24-11-6-20-12-13-29(39,5-2)25(20)32-24/h4,6-11,19,39H,1,5,12-18H2,2-3H3,(H,30,31,33)/t29-/m1/s1
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| Chemical Name |
1-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b]pyridin-2-yl]-6-[4-(4-methylpiperazin-1-yl)anilino]-2-prop-2-enylpyrazolo[3,4-d]pyrimidin-3-one
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| Synonyms |
ZN-C3; azenosertib; ZN-C3 [WHO-DD]; 1-[(7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta[b]pyridin-2-yl]-6-[4-(4-methylpiperazin-1-yl)anilino]-2-prop-2-enylpyrazolo[3,4-d]pyrimidin-3-one; compound 16 [PMID: 34423975]; compound 16 (PMID: 34423975); 1-((7R)-7-ethyl-7-hydroxy-5,6-dihydrocyclopenta(b)pyridin-2-yl)-6-(4-(4-methylpiperazin-1-yl)anilino)-2-prop-2-enylpyrazolo(3,4-d)pyrimidin-3-one; ...; 2376146-48-2;
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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 | 1.8989 mL | 9.4943 mL | 18.9887 mL | |
| 5 mM | 0.3798 mL | 1.8989 mL | 3.7977 mL | |
| 10 mM | 0.1899 mL | 0.9494 mL | 1.8989 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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