| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
EGFR (WT) 0.24 nM (IC50) EGFRL858R/T790M 7.25 nM (IC50) EGFRL858R 0.46 nM (IC50) EGFRT790M 1.79 nM (IC50)
Neptinib targets the epidermal growth factor receptor (EGFR) kinase family, including wild-type and mutant forms (L858R, L858R/T790M, T790M, and likely exon 19 deletions). It binds irreversibly to the ATP-binding pocket via a covalent bond (Michael addition) to Cys797, blocking downstream signaling pathways (PI3K/AKT, RAS/MAPK). |
|---|---|
| ln Vitro |
In vitro, Neptinib is a potent inhibitor of the EGFR kinase family, with IC₅0 values of 0.24 nM (EGFRwt), 7.25 nM (EGFRL858R/T790M), 0.46 nM (EGFRL858R), and 1.79 nM (EGFRT790M). It is therefore a potential fourth-generation EGFR-TKI with activity against the T790M and L858R/T790M resistance mutations, which cause resistance to first- and second-generation TKIs.
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| ln Vivo |
In vivo, Neptinib has a significant inhibitory effect on tumor growth in mouse non-small cell lung cancer (NSCLC) transplant models with different EGFR mutations. Its improved pharmacokinetics (PK) compared to Afatinib leads to stronger antitumor activity. It is currently in preclinical research and has not yet entered human clinical trials.
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| Enzyme Assay |
For EGFR kinase assays, recombinant EGFR enzymes (WT, L858R, T790M, L858R/T790M) are incubated with Neptinib (0.01-1000 nM) in kinase buffer (50 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM DTT, 0.01% Triton X-100) with 10 uM ATP and a biotinylated peptide substrate for 30-60 min at 30degC. The reaction is stopped, and phosphorylated peptide is detected by TR-FRET. For off-target selectivity, the compound is tested against a panel of kinases (e.g., HER2, HER4, BLK, BMX, MKNK2, MAP4K5, SGK) at 100 nM.
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| Cell Assay |
For cellular assays, NSCLC cell lines (PC9, H1975, HCC827, H3255) are seeded in 96-well plates (5,000-10,000 cells/well) and treated with Neptinib (0.1-1000 nM) for 72 h. Viability is assessed by CellTiter-Glo. IC₅0 is calculated. For apoptosis, cells are treated with Neptinib (10-500 nM) for 48 h and stained with Annexin V/PI. For Western blot, cells are lysed after 2-8 h of treatment and probed with anti-p-EGFR (Tyr1068), total EGFR, p-ERK, and total ERK.
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| Animal Protocol |
For in vivo efficacy, female BALB/c nude mice (6-8 weeks) are implanted subcutaneously with EGFR-mutant NSCLC cells (e.g., H1975, L858R/T790M; PC9, exon 19 deletion). When tumors reach 150-200 mm3, mice are randomized (n=8-10). Neptinib is formulated in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline, and administered orally at 10-50 mg/kg once daily for 14-28 days. Tumor volume is measured with calipers. A control group receives vehicle; a positive control receives Afatinib (10-20 mg/kg, oral). For PK/PD, blood and tumors are collected at 2, 4, 8, 12, 24 h post dose for compound concentration (LC-MS/MS) and p-EGFR analysis.
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| ADME/Pharmacokinetics |
Neptinib is soluble in DMSO (100 mg/mL). It is orally bioavailable and has improved PK over Afatinib (higher Cmax, longer t½). The elimination half-life (t½) is 4-8 hours in mice. Its improved PK properties are attributed to its structural modifications, which reduce first-pass metabolism and increase systemic exposure.
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| Toxicity/Toxicokinetics |
Preclinical toxicology: Neptinib is well tolerated in mice at doses up to 50 mg/kg. No significant body weight loss or hepatotoxicity is reported. Skin rash and diarrhea (class effects of EGFR inhibitors) are observed at high doses (>100 mg/kg) but are less severe than with Afatinib. No genotoxicity.
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| References | |
| Additional Infomation |
Neptinib (NEP010) is a research compound in preclinical development for EGFR-mutant NSCLC, including T790M-resistant mutations. It is not an FDA-approved drug. Its improved PK and strong antitumor activity make it a promising candidate for further development. For research use only.
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| Molecular Formula |
C22H23CLFN5O2
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|---|---|
| Molecular Weight |
443.90
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| Exact Mass |
443.152
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| CAS # |
2085843-51-0
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| PubChem CID |
132164916
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| Appearance |
Typically exists as solids at room temperature
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
31
|
| Complexity |
610
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC1=C(C=CC(=C1)NC1=C2C=C(C(=CC2=NC=N1)OCC)NC(/C=C/CN(C)C)=O)F
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| InChi Key |
RTAAZCCTCLCXHX-AATRIKPKSA-N
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| InChi Code |
InChI=1S/C22H23ClFN5O2/c1-4-31-20-12-18-15(11-19(20)28-21(30)6-5-9-29(2)3)22(26-13-25-18)27-14-7-8-17(24)16(23)10-14/h5-8,10-13H,4,9H2,1-3H3,(H,28,30)(H,25,26,27)/b6-5+
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| Chemical Name |
(E)-N-[4-(3-chloro-4-fluoroanilino)-7-ethoxyquinazolin-6-yl]-4-(dimethylamino)but-2-enamide
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| Synonyms |
NEP010
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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 | 2.2528 mL | 11.2638 mL | 22.5276 mL | |
| 5 mM | 0.4506 mL | 2.2528 mL | 4.5055 mL | |
| 10 mM | 0.2253 mL | 1.1264 mL | 2.2528 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
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