| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
EZH2
The HOTAIR‑EZH2 interaction (the binding between long non‑coding RNA HOTAIR and the EZH2 protein). EPIC-0628 also indirectly affects ATF3, p300, p65, Stat3, SP1, MGMT, and the ATF3‑p38‑E2F1 DNA damage repair pathway. |
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| ln Vitro |
EPIC-0628 (20 μM; 48 h) can arrest the cell cycle of U87 cells at the G1 phase [1].
EPIC-0628 (20 uM, 48 h) arrests U87 glioblastoma cells at the G1 phase of the cell cycle. It disrupts the HOTAIR‑EZH2 interaction, leading to increased expression of the transcription factor ATF3. ATF3 upregulation inhibits the recruitment of p300, p‑p65, p‑Stat3, and SP1 to the MGMT promoter, thereby silencing MGMT gene expression. EPIC-0628 also increases CDKN1A (p21) expression, causing cell cycle arrest, and suppresses the ATF3‑p38‑E2F1 pathway, impairing homologous recombination (HR)‑mediated DNA double‑strand break repair. EPIC-0628 enhances the efficacy of temozolomide (TMZ) in glioblastoma cells. |
| ln Vivo |
This entry is not available for EPIC-0628 in vivo; please refer to field 7 for animal model data.
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| Enzyme Assay |
General cell‑free protocol for HOTAIR‑EZH2 interaction: Use an RNA pull‑down or electrophoretic mobility shift assay (EMSA) with recombinant EZH2 (or PRC2 complex) and in vitro transcribed biotinylated HOTAIR RNA. Immobilize the RNA on streptavidin beads. Incubate with EZH2 in binding buffer (20 mM Tris‑HCl, pH 7.5, 150 mM KCl, 5 mM MgCl2, 1 mM DTT, 0.05% NP‑40) at 4degC for 2 hours. Add varying concentrations of EPIC-0628 (0.1‑100 uM). Wash beads and analyze bound EZH2 by SDS‑PAGE and Western blot. Calculate IC₅0 from inhibition curves. Alternatively, use a high‑throughput AlphaScreen assay with biotinylated HOTAIR RNA and His‑tagged EZH2.
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| Cell Assay |
General cellular protocol for EPIC-0628: Seed U87 or LN229 glioblastoma cells in 6‑well or 96‑well plates. Treat cells with EPIC-0628 at concentrations of 1, 5, 10, 20, 50 uM for 24‑72 hours. For cell cycle analysis, treat with 20 uM for 48 hours, fix with 70% ethanol, stain with propidium iodide, and analyze by flow cytometry. For MGMT silencing, treat cells for 48 hours and analyze MGMT mRNA by qRT‑PCR and protein by Western blot. For DNA damage repair assays, irradiate cells or treat with TMZ (50‑100 uM for 24 hours) after EPIC-0628 pre‑treatment, then perform comet assay or gamma‑H2AX immunofluorescence to assess DNA double‑strand breaks. Assess homologous recombination (HR) efficiency using a DR‑GFP reporter assay. For combination studies, treat cells with EPIC-0628 (10‑20 uM) plus TMZ (50‑100 uM) for 48‑72 hours and assess cell viability by MTT assay.
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| Animal Protocol |
General animal protocol for EPIC-0628: Establish orthotopic or subcutaneous glioblastoma xenografts in nude mice using U87 or patient‑derived GBM cells. When tumors are established, randomize mice into treatment groups (n=8‑10/group). Administer EPIC-0628 via intraperitoneal injection at doses of 10‑30 mg/kg daily, alone or in combination with temozolomide (TMZ, 50 mg/kg oral). Tumor volume is measured by caliper (subcutaneous) or bioluminescence imaging (orthotopic). Tumor tissues are harvested for analysis of MGMT, ATF3, CDKN1A (p21), and DNA damage markers (gamma‑H2AX, RAD51) by immunohistochemistry and Western blot. Survival of mice is monitored as a primary endpoint. EPIC-0628 is designed to overcome TMZ resistance in GBM by silencing MGMT expression and inhibiting DNA repair pathways.
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| ADME/Pharmacokinetics |
General PK protocol for EPIC-0628: Administer EPIC-0628 to male Sprague‑Dawley rats via intravenous (IV, 2 mg/kg) and oral (PO, 10 mg/kg) routes. Collect blood at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post‑dose. Extract plasma and analyze by LC‑MS/MS. Calculate PK parameters: Cmax, Tmax, AUC, t1/2, clearance, volume of distribution, and oral bioavailability (F%).
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| Toxicity/Toxicokinetics |
General toxicity protocol for EPIC-0628: Perform a 14‑day repeated dose toxicity study in ICR mice. Administer EPIC-0628 IP at doses of 10, 25, and 60 mg/kg daily. Monitor body weight, food consumption, and clinical signs. At termination, collect blood for hematology and serum chemistry (ALT, AST, BUN, creatinine). Perform gross necropsy and collect major organs (liver, kidney, spleen, heart, lung, brain) for histopathological examination.
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| References | |
| Additional Infomation |
EPIC-0628 was developed from the lead compound AC1Q3QWB. It selectively disrupts the HOTAIR‑EZH2 interaction, which is aberrantly activated in glioblastoma and contributes to TMZ resistance. The combination of EPIC-0628 with TMZ represents a strategy to enhance the efficacy of standard chemotherapy in GBM by silencing the MGMT DNA repair gene and inhibiting homologous recombination. The compound is a research tool for studying the role of HOTAIR‑EZH2 in cancer and for developing combination therapies.
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| Molecular Formula |
C20H17F4N3O
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|---|---|
| Molecular Weight |
391.36
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| Appearance |
Solid powder
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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 : 200 mg/mL (511.04 mM; with sonication)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (12.78 mM)(saturation unknown) in 10% DMSO 40% PEG300 5% Tween-80 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution This solution produces a clear solution of ≥ 5 mg/mL (saturation unknown).
For example, if 1 mL of working solution, add 100 μL of 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix well; then add 50 μL Tween-80 to the above system and mix well; then add 450 μL saline to make up 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: ≥ 5 mg/mL (12.78 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, add 100 μL of 50.0 mg/mL clear DMSO stock solution to 900 μL corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5552 mL | 12.7760 mL | 25.5519 mL | |
| 5 mM | 0.5110 mL | 2.5552 mL | 5.1104 mL | |
| 10 mM | 0.2555 mL | 1.2776 mL | 2.5552 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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