| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg |
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| Targets |
E3 Ligase Ligand-Linker Conjugates 1 targets anti-apoptotic Bcl-2 family proteins, including Bcl-xL, Bcl-2, Bcl-W, and Mcl-1. Key EC₅₀ values are as follows:
- XZ-14455: EC₅₀ = 5.0 nM (RS4;11 cancer cells), 1.47 nM (NCI-H146 cancer cells), >2000 nM (normal WI38 cells) [1] - XZ-14424: EC₅₀ = 1.6 nM (RS4;11 cancer cells), 1.4 nM (NCI-H146 cancer cells), 1.2 nM (IR-induced senescent WI38 cells) [1] - XZ-13861: EC₅₀ = 32.8 nM (RS4;11 cancer cells), 3.12 nM (NCI-H146 cancer cells), 10.51 nM (IR-induced senescent WI38 cells) [1] |
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| ln Vitro |
CHK1-IN-3 has a low affinity for hERG (IC50 > 40 µM) and efficiently suppresses the growth of hematological malignant cell lines, particularly Z-138 (IC50: 0.013 µM) [1].
1. Selective killing of senescent cells: - IR-induced senescent WI38 cells (IR-SC WI38) and normal WI38 cells were treated with increasing concentrations of E3 Ligase Ligand-Linker Conjugates 1 (e.g., XZ-13861, XZ-14439) for 72 hours. Viable cells were quantified by flow cytometry (PI exclusion). Compounds selectively inhibited IR-SC WI38 cells in a dose-dependent manner, with minimal effect on normal WI38 cells [1] - XZ-14439 depleted Bcl-xL in IR-SC WI38 cells in both dose-dependent (increasing concentrations) and time-dependent (fixed concentration, extended incubation) manners, detected by Western blot [1] 2. Cytotoxicity against cancer cells: - RS4;11 and NCI-H146 cancer cells were treated with increasing concentrations of E3 Ligase Ligand-Linker Conjugates 1 for 72 hours. Cell viability was measured by MTS assay, and EC₅₀ values were calculated relative to vehicle control. Compounds exhibited potent anti-proliferative activity against both cell lines [1] 3. Protein degradation activity: - IR-SC WI38 cells were treated with 1 µM of compounds (e.g., XZ-15416, XZ-15405, XZ-15418, XZ-15421, PZ-15227) for 6 hours, or RS4;11 cells with 200 nM for 16 hours. Western blot analysis showed depletion of Bcl-xL in both cell types, with some compounds also affecting Bcl-2, Bcl-W, or Mcl-1 [1] |
| ln Vivo |
In a Z-138 cell-seeded xenograft model, CHK1-IN-3 as a single drug dramatically suppressed tumor growth without changing body weight [1].
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| Cell Assay |
1. Senescent cell viability assay:
- Seed normal WI38 and IR-induced senescent WI38 cells, then treat with serial concentrations of E3 Ligase Ligand-Linker Conjugates 1 for 72 hours. Digest cells with 0.25% trypsin + 1 mM EDTA, harvest in PBS containing 2% FBS, and stain with propidium iodide (100 ng/ml) for 1 minute. Centrifuge to remove unbound PI, resuspend in PBS + 2% FBS, and analyze viable (PI⁻) cells by flow cytometry at a constant flow rate. Calculate viability as a percentage of vehicle-treated control [1] 2. Cancer cell viability assay: - Seed RS4;11 and NCI-H146 cancer cells, treat with increasing concentrations of E3 Ligase Ligand-Linker Conjugates 1 for 72 hours. Measure cell viability using MTS assay, and calculate EC₅₀ values relative to vehicle-treated control [1] 3. Protein degradation Western blot assay: - Treat IR-SC WI38 cells with increasing concentrations of E3 Ligase Ligand-Linker Conjugates 1 (e.g., XZ-14439) for 18 hours, or at a fixed concentration for extended time periods. Treat RS4;11 cells with 200 nM compounds for 16 hours. Harvest cells in RIPA lysis buffer containing 1% Phosphatase Inhibitor Cocktail 3 and 1% Protease Inhibitor Cocktail. Resolve 15-30 µg of protein per lane on a 12% SDS-PAGE gel, transfer to a PVDF membrane, block with TBS-T blocking buffer (5% nonfat milk), and probe with primary antibodies (Bcl-xL, Bcl-2, Bcl-W, Mcl-1, β-actin) overnight at 4°C or 1 hour at room temperature. Incubate with peroxidase-conjugated secondary antibody for 1 hour, wash with TBS-T, and detect proteins with ECL Western Blotting Detection Reagents [1] |
| References | |
| Additional Infomation |
1. Mechanism of action: E3 ligase ligand-linker conjugates are a class of proteolytic targeting chimeras (PROTACs) that covalently link E3 ubiquitin ligands (e.g., cereblon ligands) to Bcl-2 family protein inhibitors. They recruit E3 ubiquitin ligands to anti-apoptotic Bcl-2 family proteins, induce their ubiquitination, and degrade them via the ubiquitin-proteasome system [1]. 2. Therapeutic potential: These compounds are potential senescent cell scavengers that can treat age-related diseases (e.g., Alzheimer's disease, osteoarthritis, idiopathic pulmonary fibrosis) by selectively killing senescent cells. They also exhibited potent anticancer activity against hematologic malignancies (RS4;11) and solid tumor cells (NCI-H146) [1]
3. Selectivity: These compounds showed much higher selectivity for senescent and cancer cells than for normal cells, with EC₅₀ values >2000 nM in normal WI38 cells (e.g., XZ-14455) [1] |
| Molecular Formula |
C20H23N9O
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|---|---|
| Molecular Weight |
405.46
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| Exact Mass |
405.202
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| CAS # |
2097252-39-4
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| PubChem CID |
138454800
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
1.4
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
30
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| Complexity |
598
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| Defined Atom Stereocenter Count |
1
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| SMILES |
N(C1N=C(C(=CN=1)C1C=NN(C)C=1)NC)C1C=C(C(=NC=1)C#N)O[C@H]1CNCCC1
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| InChi Key |
SLCNIAWQMQCAFR-OAHLLOKOSA-N
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| InChi Code |
InChI=1S/C20H23N9O/c1-22-19-16(13-8-26-29(2)12-13)11-25-20(28-19)27-14-6-18(17(7-21)24-9-14)30-15-4-3-5-23-10-15/h6,8-9,11-12,15,23H,3-5,10H2,1-2H3,(H2,22,25,27,28)/t15-/m1/s1
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| Chemical Name |
5-[[4-(methylamino)-5-(1-methylpyrazol-4-yl)pyrimidin-2-yl]amino]-3-[(3R)-piperidin-3-yl]oxypyridine-2-carbonitrile
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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 | 2.4663 mL | 12.3317 mL | 24.6633 mL | |
| 5 mM | 0.4933 mL | 2.4663 mL | 4.9327 mL | |
| 10 mM | 0.2466 mL | 1.2332 mL | 2.4663 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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