| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
WRN (Werner syndrome RecQ helicase). WRN inhibitor 8 targets the helicase domain of WRN, inhibiting its ATP‑dependent 3′→5′ DNA unwinding activity.
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| ln Vitro |
In cell‑free enzymatic assays, WRN inhibitor 8 potently inhibits the helicase activity of WRN. In cellular assays, it selectively induces DNA damage (gammaH2AX foci) and cell death in MSI‑H cancer cell lines (such as HCT116, LoVo, and DLD‑1 MSI) while having minimal effect on MSS cells (such as HT‑29 and SW480) or normal cells. The specific IC₅0 values are not provided in the available data.
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| ln Vivo |
In vivo, WRN inhibitor 8 demonstrates selective antitumor activity in MSI‑H xenograft mouse models, including colorectal and endometrial cancer PDX models. WRN inhibition precipitates replication fork collapse and double‑strand breaks, validating WRN as a promising therapeutic target for MSI‑H tumors. Dose‑dependent tumor growth inhibition and regression have been observed.
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| Enzyme Assay |
A general cell‑free protocol for assessing WRN helicase inhibition: A fluorescence‑based DNA unwinding assay is used as described for VVD‑214 (see V88650 field 5). Recombinant WRN protein (10‑50 nM) is incubated with a fluorophore‑labeled forked DNA substrate (50 nM) in helicase buffer (25 mM Tris‑HCl, pH 7.5, 2 mM MgCl2, 2 mM ATP, 50 mM KCl, 1 mM DTT, 0.1 mg/mL BSA) with varying concentrations of WRN inhibitor 8 (0.1 nM to 10 uM). The reaction is initiated by adding ATP and incubated at 37degC for 30‑60 minutes. The reaction is stopped with 20 mM EDTA and 0.5% SDS. The increase in fluorescence (excitation 485 nm, emission 528 nm) is measured. The IC₅0 for helicase inhibition is calculated. For selectivity profiling, WRN inhibitor 8 is tested against other RecQ helicases (BLM, RecQ1, RecQ5) and other ATPases.
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| Cell Assay |
A general cellular protocol for WRN inhibitor 8: MSI‑high colorectal cancer cells (e.g., HCT116, LoVo) and MSS control cells (e.g., HT‑29, SW480) are seeded in 96‑well plates at 5,000‑10,000 cells/well. Cells are treated with serial dilutions of WRN inhibitor 8 (0.01 nM to 10 uM) for 72‑120 hours. Cell viability is measured using CellTiter‑Glo assay. The selectivity index (IC₅0 MSS / IC₅0 MSI) is calculated. For DNA damage assessment, cells are treated for 24‑48 hours, fixed, and stained with an anti‑gammaH2AX (Ser139) antibody. The number of gammaH2AX foci per nucleus is quantified by immunofluorescence microscopy or high‑content imaging. Cell cycle analysis is performed by propidium iodide staining and flow cytometry. Apoptosis is assessed by Annexin V/PI staining and flow cytometry.
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| Animal Protocol |
A general animal protocol for WRN inhibitor 8: Female NOD/SCID mice are subcutaneously implanted with 5×10⁶ HCT116 MSI‑high colorectal cancer cells or other MSI‑H PDX models (e.g., endometrial cancer PDX). When tumors reach ~150‑200 mm3, mice are randomized into treatment groups (n=8‑10 per group). WRN inhibitor 8 is formulated in a suitable vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween‑80, 45% saline) and administered via oral gavage or intraperitoneal (IP) injection at doses of 10, 30, and 100 mg/kg once or twice daily for 21‑28 days. Tumor volume is measured twice weekly using a caliper. At the end of the study, mice are euthanized, and tumors are harvested for IHC (gammaH2AX, Ki‑67, cleaved caspase‑3), Western blot (gammaH2AX, cleaved PARP), and TUNEL staining.
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| ADME/Pharmacokinetics |
General PK protocol for WRN inhibitor 8: Male Sprague‑Dawley rats are administered the compound via oral gavage (PO, 10 mg/kg) and intravenous (IV, 2 mg/kg). Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours. Plasma concentrations are quantified by LC‑MS/MS. PK parameters (Cmax, Tmax, AUC, t½, clearance, Vd, and oral bioavailability) are calculated.
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| Toxicity/Toxicokinetics |
General toxicity protocol for WRN inhibitor 8: A 14‑day repeat‑dose oral toxicity study is performed in ICR mice. WRN inhibitor 8 is administered via oral gavage at doses of 10, 30, and 100 mg/kg/day for 14 days. Parameters include clinical signs, body weight, food consumption, hematology (complete blood count, platelet count, differential), serum chemistry (ALT, AST, BUN, creatinine), and histopathology of major organs (bone marrow, gastrointestinal tract, liver, kidneys, spleen, lungs, heart). Special attention is given to the bone marrow (for potential myelosuppression) and the gastrointestinal tract (for replication stress‑related toxicity).
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| References | |
| Additional Infomation |
WRN inhibitor 8 has the CAS number 3026502-95-1. The compound is part of a new class of synthetic lethal agents targeting WRN helicase in MSI‑H tumors. This synthetic lethality strategy is based on the principle that simultaneous inactivation of two genes leads to cell death, whereas the loss of a single gene allows cell survival. In MSI‑H tumors, which have defective DNA mismatch repair (dMMR), WRN is synthetically lethal. The therapeutic potential of WRN inhibition is being actively explored as a novel precision oncology strategy for MSI‑H cancers that are resistant to immune checkpoint blockade or other therapies. WRN inhibitor 8 is a chemical probe for preclinical studies. The specific molecular structure is not provided in the available data.
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| Molecular Formula |
C22H23F2N3O4S
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|---|---|
| Molecular Weight |
463.50
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| Exact Mass |
463.138
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| CAS # |
3026502-95-1
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| PubChem CID |
170718255
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
32
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| Complexity |
802
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CS(=O)(=O)/C=C/[C@H](C1CC1)NC(=O)C2=CN=C(N=C2OC3=CC=CC=C3)C(C4CC4)(F)F
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| InChi Key |
SGERSEZXZIVUEY-IENJSVCTSA-N
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| InChi Code |
InChI=1S/C22H23F2N3O4S/c1-32(29,30)12-11-18(14-7-8-14)26-19(28)17-13-25-21(22(23,24)15-9-10-15)27-20(17)31-16-5-3-2-4-6-16/h2-6,11-15,18H,7-10H2,1H3,(H,26,28)/b12-11+/t18-/m1/s1
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| Chemical Name |
2-[cyclopropyl(difluoro)methyl]-N-[(E,1S)-1-cyclopropyl-3-methylsulfonylprop-2-enyl]-4-phenoxypyrimidine-5-carboxamide
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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 | 2.1575 mL | 10.7875 mL | 21.5750 mL | |
| 5 mM | 0.4315 mL | 2.1575 mL | 4.3150 mL | |
| 10 mM | 0.2157 mL | 1.0787 mL | 2.1575 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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