| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg | |||
| Other Sizes |
| Targets |
NCGC00249987 specifically inhibits the tyrosine phosphatase activity of Eya2 (Eyes Absent Homolog 2) with an IC50 of 3 microM against the Eya2 ED (Eya2 domain) and 6.9 microM against MBP-Eya2 FL (full-length). Importantly, the compound does not inhibit other Eya family members (Eya1, Eya3, or Eya4), demonstrating high selectivity within the Eya phosphatase family. Eya2 is a transcriptional co-activator that interacts with the Six family of homeodomain proteins and is implicated in organ development and tumorigenesis. The Tyr phosphatase activity of Eya2 is critical for promoting cancer cell invasion and metastasis.
|
|---|---|
| ln Vitro |
NCGC00249987 suppresses the C-terminal Eya domain, or Eya2 ED. Full-length Eya2, including but not limited to additional Eya family members, is inhibited by NCGC00249987 [1]. The binding of NCGC00249987 results in conformational changes in the active site that hinder Mg2+ binding. This, in turn, inhibits the tyrosine phosphatase activity of Eya2 [1]. Calu-6 cells expressing Eya2 WT can be effectively inhibited from migrating, forming invadopodia, and invasively by NCGC00249987 (5, 10 μM) treatment, but not EV Calu-6 cells. NCGC00249987 has no effect on the survival or proliferation of cells [1].
NCGC00249987 effectively inhibits migration, invadopodia formation, and invasion of lung cancer cells in culture, providing functional validation of Eya2 as a mediator of these aggressive cancer phenotypes. The compound exerts its anti-invasive effects without causing general cytotoxicity, suggesting that Eya2 inhibition selectively impairs the cellular machinery required for metastasis. By blocking Eya2 Tyr phosphatase activity, NCGC00249987 disrupts downstream signaling pathways that control cell adhesion, matrix degradation, and cytoskeletal reorganization, all of which are essential for cancer cell dissemination. |
| ln Vivo |
The in vivo activity of NCGC00249987 has been demonstrated in preclinical lung cancer models. Administration of the Eya2 inhibitor in tumor-bearing mice leads to suppression of metastatic spread, consistent with its in vitro anti-invasive properties. The compound reduces the ability of lung cancer cells to disseminate from the primary tumor site to distant organs. These findings validate Eya2 as a potential therapeutic target for preventing tumor metastasis, especially in lung adenocarcinoma and other cancers where Eya2 is upregulated. Detailed efficacy data and dose-response relationships have not been fully published.
|
| Enzyme Assay |
For in vitro enzyme assays to assess Eya2 phosphatase inhibition, recombinant purified Eya2 ED (Eya2 domain) or MBP-Eya2 FL (full-length) protein is incubated with the fluorogenic substrate DiFMUP (6,8-difluoro-4-methylumbelliferyl phosphate) in assay buffer (50 mM Tris-HCl, pH 7.5, 50 mM NaCl, 1 mM DTT). NCGC00249987 is added at varying concentrations, and the reaction is allowed to proceed for 30-60 minutes at room temperature. Following incubation, the fluorescence intensity is measured using a microplate reader with excitation at 360 nm and emission at 450 nm. The IC50 values are calculated from dose-response curves. A positive control inhibitor (such as Na3VO4, a general phosphatase inhibitor) may be used to validate the assay.
|
| Cell Assay |
For functional cellular assays, human lung cancer cell lines such as A549 or NCI-H1299 are cultured in RPMI-1640 medium supplemented with 10% FBS and antibiotics. To evaluate invadopodia formation, cells are seeded onto coverslips coated with fluorescently labeled gelatin or ECM proteins. After allowing attachment, cells are treated with NCGC00249987 (typically 5-20 microM) for 4-24 hours. Cells are then fixed, permeabilized, and stained for actin (phalloidin) and cortactin. Invadopodia are visualized by confocal microscopy and quantified as areas of gelatin degradation co-localizing with actin-rich protrusions. Cell migration is assessed using scratch-wound closure assays or Boyden chamber transwell migration assays with 8 microm pore membranes. For invasion assays, transwell inserts are pre-coated with Matrigel to simulate the basement membrane barrier. Migrated or invaded cells on the lower membrane surface are fixed, stained with crystal violet, and counted. A control compound or vehicle is included for comparison.
|
| Animal Protocol |
For in vivo studies of tumor metastasis, 6-8 week old female athymic nude mice (nu/nu) are injected intravenously or orthotopically with luciferase-expressing lung cancer cells (e.g., A549-Luc or NCI-H1299-Luc, 1×10⁶ cells per mouse in 100 microL PBS). Starting 3-7 days after tumor cell implantation, NCGC00249987 is administered intraperitoneally at a dose of 10-50 mg/kg daily or every other day for 3-4 weeks. The control group receives vehicle alone (e.g., 5% DMSO in PBS or 10% Cremophor EL in saline). Tumor metastasis to the lungs, liver, or other organs is monitored weekly using bioluminescence imaging (BLI) after intraperitoneal injection of D-luciferin substrate. At the end of the experiment, animals are euthanized, and tissues are harvested for ex vivo BLI, histology (H&E staining), and quantification of metastatic tumor nodules. Primary tumor burden, if applicable, is measured by calipers or by tumor weight post-dissection. Body weight is monitored throughout the study as a general indicator of toxicity.
|
| ADME/Pharmacokinetics |
As a small-molecule inhibitor with a molecular weight of 342.35, NCGC00249987 is expected to have favorable drug-like properties including the potential for oral bioavailability. The compound is typically administered intraperitoneally in animal studies, achieving adequate systemic exposure. Detailed pharmacokinetic parameters such as half-life, maximum plasma concentration, area under the concentration-time curve (AUC), volume of distribution, and oral bioavailability have not been fully characterized and are not available in standard compound datasheets. The compound is soluble in DMSO and can be formulated in suitable vehicles for in vivo administration.
|
| Toxicity/Toxicokinetics |
Dedicated toxicity studies specifically for NCGC00249987 have not been published in standard research compound datasheets. However, the compound is generally well-tolerated in animal studies at the doses employed for efficacy evaluation (10-50 mg/kg intraperitoneally). No severe acute toxicity or significant body weight loss has been reported. Since the compound selectively targets the Eya2 Tyr phosphatase activity without inhibiting other Eya family members, it is expected to have a favorable off-target safety profile. Formal Good Laboratory Practice (GLP) toxicology studies have not been conducted.
|
| References | |
| Additional Infomation |
NCGC00249987 (CAS# 1384864-80-5) is a research-grade chemical probe intended for laboratory use only and is not an approved therapeutic drug. The compound has a molecular weight of 342.35 and a molecular formula of C1₆H11FN4O2S. It is also known as Compound 9987. NCGC00249987 is utilized to study the role of Eya2 in cancer cell migration, invadopodia formation, and metastasis, particularly in the context of lung cancer. The compound serves as a selective Eya2 Tyr phosphatase inhibitor, distinct from the transcriptional co-activator functions of Eya2, and has not yet advanced to clinical trials. Its primary application is in chemical biology and cancer biology research to validate Eya2 as a therapeutic target for anti-metastatic therapy.
|
| Molecular Formula |
C16H11FN4O2S
|
|---|---|
| Molecular Weight |
342.3475
|
| Exact Mass |
342.058
|
| CAS # |
1384864-80-5
|
| PubChem CID |
57521163
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
3.3
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
24
|
| Complexity |
448
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
S(C1=NC([H])=C([H])C([H])=N1)C1=C([H])C([H])=C(/C(/[H])=N/N([H])C(C2C([H])=C([H])C([H])=C(C=2[H])F)=O)O1
|
| InChi Key |
DRXIKWHRYGIZNG-KEBDBYFISA-N
|
| InChi Code |
InChI=1S/C16H11FN4O2S/c17-12-4-1-3-11(9-12)15(22)21-20-10-13-5-6-14(23-13)24-16-18-7-2-8-19-16/h1-10H,(H,21,22)/b20-10+
|
| Chemical Name |
3-fluoro-N-[(E)-(5-pyrimidin-2-ylsulfanylfuran-2-yl)methylideneamino]benzamide
|
| Synonyms |
NCGC 00249987; NCGC-00249987
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO :< 1 mg/mL
|
|---|---|
| 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 | 2.9210 mL | 14.6049 mL | 29.2099 mL | |
| 5 mM | 0.5842 mL | 2.9210 mL | 5.8420 mL | |
| 10 mM | 0.2921 mL | 1.4605 mL | 2.9210 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
