| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
RETV804M inhibitor (lead compound in the study) targets RET V804M mutant kinase (IC50 = 1.2 nM), wild-type RET (wt-RET, IC50 = 3.5 nM), and kinase insert domain receptor (KDR/VEGFR2, IC50 = 4.8 nM); it exhibits >200-fold selectivity over other kinases (e.g., EGFR, HER2, MET, FGFR1) [1]
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| ln Vitro |
Against Ba/F3 cells stably expressing RET V804M (Ba/F3-RET V804M), RETV804M inhibitor showed potent antiproliferative activity with an IC50 of 2.7 nM after 72 hours of treatment; for Ba/F3-wt-RET cells, the IC50 was 8.9 nM [1]
- In RET V804M-positive TT thyroid medullary carcinoma cells, the inhibitor inhibited cell proliferation with an IC50 of 3.1 nM and suppressed RET-mediated signaling activation [1] - Western blot analysis revealed that RETV804M inhibitor (5 nM, 2 hours) dose-dependently reduced phosphorylation of RET (Tyr905), ERK1/2 (Thr202/Tyr204), and AKT (Ser473) in Ba/F3-RET V804M cells, without affecting total RET, ERK1/2, or AKT protein levels [1] - Flow cytometry analysis showed that RETV804M inhibitor (10 nM, 48 hours) induced G1 cell cycle arrest in Ba/F3-RET V804M cells (G1 phase ratio increased from ~45% to ~68%) and apoptosis (apoptotic rate ~25% vs. ~3% in control) [1] - The compound did not inhibit proliferation of RET-null A549 non-small cell lung cancer cells or normal human bronchial epithelial cells (NHBE) at concentrations up to 1 μM, indicating high cancer cell selectivity [1] - Kinase selectivity panel screening (468 human kinases) showed that the inhibitor only significantly inhibited RET (V804M/wt) and KDR, with no meaningful inhibition of other kinases at 1 μM [1] |
| ln Vivo |
In the Ba/F3-RET V804M xenograft model in nude mice, oral administration of RETV804M inhibitor at 5 mg/kg, 15 mg/kg, and 45 mg/kg once daily for 14 days resulted in tumor growth inhibition (TGI) rates of 52%, 76%, and 91%, respectively [1]
- The 45 mg/kg dose reduced tumor weight from ~0.9 g (vehicle control) to ~0.08 g, with no significant body weight loss (<3% weight change) or obvious toxicity signs [1] - In the TT thyroid medullary carcinoma xenograft model, oral administration of 15 mg/kg daily for 21 days achieved a TGI rate of 73% and reduced CD31-positive microvessel density (tumor angiogenesis marker) by ~55% [1] - Immunohistochemical staining of tumor tissues demonstrated that RETV804M inhibitor (45 mg/kg) significantly decreased phosphorylation levels of RET (Tyr905) and ERK1/2, and increased the number of TUNEL-positive apoptotic cells (3.2-fold vs. control) [1] |
| Enzyme Assay |
Kinase activity assay was performed using a homogeneous time-resolved fluorescence (HTRF) method. The reaction mixture contained recombinant RET V804M, wt-RET, or KDR kinase, biotinylated peptide substrate, ATP (Km = 10 μM for RET V804M), and serial dilutions of RETV804M inhibitor. After incubation at 30°C for 60 minutes, a mixture of streptavidin-conjugated europium cryptate and XL665-labeled anti-phosphotyrosine antibody was added. HTRF signals were measured at 620 nm and 665 nm, and IC50 values were calculated by fitting dose-response curves of kinase activity inhibition [1]
- Kinase selectivity panel assay: The inhibitor was tested against 468 human kinases at 1 μM using the same HTRF method. Inhibition rates were quantified, and selectivity scores were calculated based on the ratio of inhibition of RET V804M to other kinases [1] |
| Cell Assay |
Antiproliferative assay: Ba/F3-RET V804M, Ba/F3-wt-RET, TT, A549, or NHBE cells were seeded in 96-well plates at 2×10³ cells/well and incubated overnight. Serial dilutions of RETV804M inhibitor were added, and cells were cultured for 72 hours. Cell viability was assessed using a tetrazolium salt-based colorimetric assay, and IC50 values were determined [1]
- Western blot assay: Ba/F3-RET V804M cells were seeded in 6-well plates and treated with different concentrations of RETV804M inhibitor for 2 hours. Cells were lysed in buffer containing protease and phosphatase inhibitors, and total proteins were separated by SDS-PAGE. Membranes were probed with primary antibodies against p-RET (Tyr905), RET, p-ERK1/2, ERK1/2, p-AKT, AKT, and β-actin, followed by HRP-conjugated secondary antibodies. Chemiluminescent signals were detected and quantified [1] - Cell cycle and apoptosis assay: Ba/F3-RET V804M cells were treated with RETV804M inhibitor (10 nM) for 48 hours. For cell cycle analysis, cells were fixed, stained with propidium iodide (PI), and analyzed by flow cytometry. For apoptosis analysis, cells were stained with Annexin V-FITC and PI, then detected by flow cytometry [1] |
| Animal Protocol |
Ba/F3-RET V804M xenograft model: Female nude mice (6-7 weeks old) were subcutaneously inoculated with 2×10⁶ Ba/F3-RET V804M cells into the right flank. When tumors reached an average volume of 100 mm³, mice were randomly divided into four groups (n=6 per group): vehicle control, RETV804M inhibitor 5 mg/kg, 15 mg/kg, and 45 mg/kg. The compound was formulated in a mixture of DMSO, Cremophor EL, and normal saline (volume ratio 1:1:8) and administered via oral gavage once daily for 14 consecutive days. Tumor volume (length × width² / 2) and body weight were recorded every 2 days [1]
- TT thyroid medullary carcinoma xenograft model: Female nude mice were subcutaneously inoculated with 5×10⁶ TT cells. When tumors reached 120 mm³, mice were divided into vehicle control and RETV804M inhibitor 15 mg/kg groups (n=6 per group). Oral administration was performed once daily for 21 days, with tumor volume and body weight monitored regularly. At the end of the study, tumors were excised for immunohistochemical staining (p-RET, p-ERK1/2, CD31) and TUNEL assay [1] |
| ADME/Pharmacokinetics |
In mice, after oral administration of 15 mg/kg of the RETV804M inhibitor, the peak plasma concentration (Cmax) was 3.2 μg/mL, the area under the plasma concentration-time curve (AUC₀₋₂₄h) was 22.7 μg·h/mL, and the oral bioavailability was 65% [1]. - The terminal half-life (t₁/₂) of the compound in mice after oral administration was 5.4 hours [1]. - In vitro metabolic stability studies using human liver microsomes showed that the half-life of the compound was 135 minutes, indicating good metabolic stability [1]. - The plasma protein binding rate of the RETV804M inhibitor in mouse plasma was 93%, and the plasma protein binding rate in human plasma was 95% [1]. - The apparent volume of distribution (Vdss) in mice was 2.1 L/kg, indicating good tissue penetration [1].
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| Toxicity/Toxicokinetics |
In a 28-day repeated oral toxicity study in rats, the RETV804M inhibitor at doses up to 60 mg/kg did not cause significant weight loss, death, or histopathological abnormalities in major organs (liver, kidney, heart, lung, spleen) [1] - No significant changes were observed in hematological parameters (white blood cell count, red blood cell count, platelet count) or liver and kidney function biochemical indicators (ALT, AST, creatinine, blood urea nitrogen) [1] - At concentrations up to 10 μM, the compound did not inhibit cytochrome P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) in human liver microsomes [1]
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| References | |
| Additional Infomation |
The RETV804M inhibitor is a potent, orally active, and selective RET V804M mutant kinase inhibitor for the treatment of RET V804M-driven cancers [1]. Its mechanism of action involves selectively binding to the ATP-binding pocket of the RET V804M kinase, inhibiting its catalytic activity and blocking downstream MAPK/ERK and PI3K/AKT signaling pathways, thereby inducing cell cycle arrest and apoptosis in RET V804M-positive cancer cells [1]. RET V804M is the major acquired resistance mutation to first-generation RET inhibitors (e.g., vandetanib, cabozantinib) in RET fusion-positive cancer patients; this inhibitor aims to address the unmet medical need to overcome such resistance [1]. The compound's favorable pharmacokinetic properties (high oral bioavailability, long half-life, and good tissue penetration) and high kinase selectivity support its development as a targeted anticancer drug for RET V804M-mutant tumors [1].
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| Molecular Formula |
C19H16N6OEXACTMASS
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|---|---|
| Molecular Weight |
344.3699
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| Exact Mass |
344.138
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| Elemental Analysis |
C, 66.27; H, 4.68; N, 24.40; O, 4.65
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| CAS # |
2414373-42-3
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| PubChem CID |
154729143
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| Appearance |
Off-white to light yellow solid powder
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| Density |
1.4±0.1 g/cm3
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| Index of Refraction |
1.729
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| LogP |
-0.72
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
26
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| Complexity |
475
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(C1C=CC(=CC=1)C1C=NN2C=CC(NCC3C=CN=CC=3)=NC2=1)N
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| InChi Key |
DICBAYNNPUCOOQ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H16N6O/c20-18(26)15-3-1-14(2-4-15)16-12-23-25-10-7-17(24-19(16)25)22-11-13-5-8-21-9-6-13/h1-10,12H,11H2,(H2,20,26)(H,22,24)
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| Chemical Name |
4-(5-((pyridin-4-ylmethyl)amino)pyrazolo[1,5-a]pyrimidin-3-yl)benzamide
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| Synonyms |
DUN73423; DUN-73423; DUN 73423;
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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 | 2.9039 mL | 14.5193 mL | 29.0385 mL | |
| 5 mM | 0.5808 mL | 2.9039 mL | 5.8077 mL | |
| 10 mM | 0.2904 mL | 1.4519 mL | 2.9039 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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