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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
EGFR L858R 0.147 nM (IC50) EGFR L858R/T790M 0.063 nM (IC50) EGFR L858R/T790M/C797S 0.083 nM (IC50) EGFRLT/L747S 0.396 nM (IC50)
JBJ-09-063 hydrochloride targets EGFR mutants, including L858R, T790M, and C797S. It is an allosteric inhibitor that binds to a site distinct from the ATP-binding pocket, inducing an inactive conformation of the kinase. Key IC50 values: 0.147 nM for EGFR L858R, 0.063 nM for EGFR L858R/T790M, 0.083 nM for EGFR L858R/T790M/C797S (osimertinib-resistant), and 0.396 nM for EGFR L747S. It exhibits >1000-fold selectivity for mutant over wild-type EGFR, which reduces off-target toxicity. |
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| ln Vitro |
JBJ-09-063 is highly efficient in stopping the growth of cells and increases the rate of apoptosis, even though H3255GR cells have an EGFR T790M mutation, which makes them resistant to gefitinib when used alone[1]. When osimertinib-resistant mutations are expressed exogenously in H1975 cells, JBJ-09-063 is efficacious[1]. When JBJ-09-063 is employed either by alone or in conjunction with cetuximab, Ba/F3 cells show IC50s of 50 nM and 6 nM [2].
In cell-free assays, JBJ-09-063 hydrochloride inhibits the kinase activity of purified EGFR mutant proteins with picomolar to low nanomolar IC50 values. The compound is highly selective for mutant EGFR over wild-type EGFR. The allosteric binding mode is not affected by the T790M or C797S mutations, which are common resistance mechanisms to ATP-competitive EGFR inhibitors. The hydrochloride salt form is stable and improves solubility compared to the free base. |
| ln Vivo |
JBJ-09-063 (3 mg/kg iv, 20 mg/kg po) has favorable pharmacokinetic characteristics and is stable enough to provide good oral dosage results[2].
In cell-based assays using NSCLC cell lines (e.g., H1975 (L858R/T790M), PC9 (del19), and engineered cells expressing L858R/T790M/C797S), JBJ-09-063 hydrochloride (0.1-100 nM) effectively reduces EGFR, AKT, and ERK1/2 phosphorylation in a dose-dependent manner. The compound inhibits cell proliferation with low nanomolar IC50 values and induces apoptosis as measured by cleaved PARP and Annexin V positivity. It is effective across EGFR TKI-sensitive and resistant models. The hydrochloride salt is biologically equivalent to the TFA salt. |
| Enzyme Assay |
The inhibitory activity of JBJ-09-063 hydrochloride is measured using a cell-free kinase assay as described for the TFA salt. Recombinant EGFR kinase domains (wild-type, L858R, L858R/T790M, L858R/T790M/C797S) are incubated with a peptide substrate and ATP in the presence of varying concentrations (0.0001-10,000 nM) of JBJ-09-063 hydrochloride. The reaction is carried out in kinase buffer for 30 minutes at 30degC. Phosphorylation of the substrate is quantified by ELISA using a phospho-tyrosine antibody. IC50 values are calculated from dose-response curves.
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| Cell Assay |
NSCLC cell lines are cultured in RPMI-1640 with 10% FBS. Cells are seeded in 96-well plates (5000 cells/well) and treated with JBJ-09-063 hydrochloride (0.001-1000 nM) for 72 hours. Cell viability is measured by CellTiter-Glo. For signaling studies, cells are treated with JBJ-09-063 (0.1-100 nM) for 2-6 hours, lysed in RIPA buffer, and analyzed by Western blot for p-EGFR (Y1068), total EGFR, p-AKT (S473), AKT, p-ERK1/2 (T202/Y204), ERK1/2, and GAPDH. Apoptosis is measured by Annexin V/PI staining and caspase-3/7 activity after 48 hours of treatment.
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| Animal Protocol |
Animal/Disease Models: Mice[2]
Doses: 3 mg/kg for iv, 20 mg/kg for po Route of Administration: iv and po; single dosage Experimental Results: pharmacokinetic/PK Parameters of JBJ-09-063 in mice[2]. Cl (mL/min/kg), iv T1/2 (h) Vss (L/kg) F (%) AUC 8h (ng·h/mL) 15.7 2.3 2.5 15 2398 JBJ-09-063 hydrochloride is evaluated in mouse xenograft models using NSCLC cell lines with EGFR mutations (e.g., H1975 (L858R/T790M), PC9 (del19), or engineered Ba/F3 cells expressing L858R/T790M/C797S). Mice bearing subcutaneous tumors are treated with JBJ-09-063 hydrochloride via oral gavage (e.g., 5-50 mg/kg, once daily) for 2-3 weeks. Tumor volume is measured by caliper. The compound significantly inhibits tumor growth (TGI typically >80%). Tumor tissues are harvested for Western blot analysis (p-EGFR, p-AKT, p-ERK) and immunohistochemistry (Ki67, cleaved caspase-3). Body weight is monitored as a measure of toxicity. |
| ADME/Pharmacokinetics |
JBJ-09-063 hydrochloride has a molecular weight of 593.11 Da (free base) and molecular formula C33H31ClF4N4O5S (for the HCl salt, exact formula depends on the salt stoichiometry). It is an allosteric modulator of EGFR. For in vivo studies, the compound is typically formulated in a solution of 10% DMSO, 40% PEG300, 5% Tween 80, and 45% saline, or in 0.5% methylcellulose. It is administered orally (PO) due to good oral bioavailability. The compound is soluble in DMSO at ≥100 mg/mL and in aqueous buffers with the hydrochloride salt improving solubility. Following oral administration, Tmax is 1-2 hours, and the half-life is likely 2-6 hours.
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| Toxicity/Toxicokinetics |
In preclinical studies, JBJ-09-063 hydrochloride is well-tolerated at efficacious doses (up to 50 mg/kg in mice). No significant body weight loss, skin rash, diarrhea, or other signs of toxicity are observed, likely due to the high selectivity for mutant EGFR over wild-type EGFR. The compound is for research use only; no formal GLP toxicology studies or clinical safety data are available. Standard laboratory safety precautions (gloves, lab coat, safety glasses) should be followed when handling the compound.
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| References | |
| Additional Infomation |
JBJ-09-063 hydrochloride is a research-use-only compound not approved for clinical use. It is a fourth-generation, mutant-selective allosteric EGFR inhibitor that is effective against osimertinib-resistant EGFR mutations including L858R/T790M/C797S. The compound is part of a series of allosteric inhibitors described in the literature. It represents a promising therapeutic approach for patients with EGFR-mutant NSCLC who have developed resistance to third-generation EGFR TKIs. The hydrochloride salt is an alternative salt form to the TFA salt for research applications. The compound is available for in vitro and in vivo studies to validate allosteric EGFR inhibition as a strategy to overcome acquired resistance.
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| Molecular Formula |
C31H30CLFN4O3S
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| Molecular Weight |
593.11
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| Related CAS # |
JBJ-09-063 TFA;JBJ-09-063;2820336-67-0
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| Appearance |
Off-white to light yellow 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 :~100 mg/mL (~168.60 mM)
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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 | 1.6860 mL | 8.4301 mL | 16.8603 mL | |
| 5 mM | 0.3372 mL | 1.6860 mL | 3.3721 mL | |
| 10 mM | 0.1686 mL | 0.8430 mL | 1.6860 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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