| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
ALK2-IN-2 targets activin receptor-like kinase 2 (ALK2) (IC50 = 0.04 μM for human ALK2 kinase activity; Ki = 0.02 μM, competitive inhibition mode against ATP) [1]
ALK2-IN-2 shows high selectivity over other ALK family members and kinases (ALK1: IC50 = 3.2 μM; ALK3: IC50 = 2.8 μM; ALK4: IC50 = 4.5 μM; TGF-βR1: IC50 > 10 μM; selectivity index > 80 vs. ALK2) [1] |
|---|---|
| ln Vitro |
- ALK2 kinase inhibitory activity: ALK2-IN-2 potently and selectively inhibited recombinant human ALK2 kinase activity in a dose-dependent manner, with IC50 = 0.04 μM and Ki = 0.02 μM. Kinetic analysis confirmed it competes with ATP for binding to the ALK2 ATP-binding pocket [1]
- Inhibition of ALK2-dependent Smad signaling: ALK2-IN-2 (0.01-1 μM) dose-dependently suppressed BMP6-induced Smad1/5/8 phosphorylation (p-Smad1/5/8) in C2C12 myoblasts. At 0.5 μM, p-Smad1/5/8 levels were reduced by 78% compared to BMP6-stimulated control. It did not affect TGF-β1-induced Smad2/3 phosphorylation [1] - Antiproliferative activity: The compound inhibited proliferation of ALK2-overexpressing cancer cells. IC50 values were 0.3 μM (H226, non-small cell lung cancer), 0.5 μM (A549, lung cancer), and 0.7 μM (MCF-7, breast cancer). It had minimal cytotoxicity to normal human foreskin fibroblasts (NHF, IC50 > 20 μM) [1] - High kinase selectivity: ALK2-IN-2 (10 μM) showed no significant inhibition against 45 other kinases (e.g., EGFR, VEGFR2, CDK2, ERK1) tested, confirming high target selectivity [1] |
| Enzyme Assay |
- ALK2 kinase activity assay: Recombinant human ALK2 kinase domain was mixed with ATP (10 μM), fluorescently labeled peptide substrate (derived from Smad1), and ALK2-IN-2 at gradient concentrations (0.001-1 μM) in kinase buffer (pH 7.5). The mixture was incubated at 37°C for 1 hour, and phosphorylated substrate was detected by homogeneous time-resolved fluorescence (HTRF) assay. IC50 was calculated by plotting inhibition rate against drug concentration. Kinetic analysis with varying ATP concentrations confirmed competitive inhibition [1]
- Kinase selectivity assay: Recombinant ALK1, ALK3, ALK4, TGF-βR1, and 45 other kinases were separately mixed with their corresponding substrates, ATP, and ALK2-IN-2 (10 μM) in kinase buffer. After 37°C incubation for 1 hour, enzyme activity was detected by HTRF assay to evaluate selectivity [1] |
| Cell Assay |
- Smad signaling inhibition assay: C2C12 myoblasts were seeded into 6-well plates (5×10⁵ cells/well) and incubated overnight. Cells were pre-treated with ALK2-IN-2 (0.01-1 μM) for 1 hour, then stimulated with BMP6 (50 ng/mL) or TGF-β1 (10 ng/mL) for 15 minutes. Cells were lysed, and p-Smad1/5/8, Smad1/5/8, p-Smad2/3, Smad2/3, and GAPDH proteins were detected by western blot. Band intensities were quantified by densitometry [1]
- Cell viability assay: Cancer cells (H226, A549, MCF-7) and NHF cells were seeded into 96-well plates (5×10³ cells/well) and treated with ALK2-IN-2 (0.01-20 μM) for 72 hours. Cell viability was measured by tetrazolium salt-based assay, and IC50 values were calculated [1] |
| References | |
| Additional Infomation |
Chemical Classification: ALK2-IN-2 is a small molecule ALK2 inhibitor belonging to the 3-(4-aminosulfonylnaphthyl)pyrazolo[1,5-a]pyrimidine derivative class [1] - Mechanism of Action: This compound binds to the ATP-binding pocket of ALK2, competitively inhibiting its serine/threonine kinase activity. This blocks BMP6-mediated Smad1/5/8 phosphorylation and its downstream signaling, thereby inhibiting the proliferation of ALK2-overexpressing cancer cells without affecting TGF-β1/Smad2/3 signaling [1] - Target Background: ALK2 is a type I receptor of the TGF-β/BMP superfamily, involved in regulating cell proliferation, differentiation and tissue homeostasis. Abnormal activation of ALK2 is closely associated with the development and progression of various cancers (e.g., lung cancer, breast cancer) and fibrotic diseases [1]
- Therapeutic potential: ALK2-IN-2 is a highly effective, selective, and cell-penetrating ALK2 inhibitor that has shown good efficacy against ALK2-driven cancers. It provides a lead compound for the development of therapies targeting ALK2-dependent malignancies and fibrotic diseases [1] |
| Molecular Formula |
C28H27N5O2S
|
|---|---|
| Molecular Weight |
497.6112844944
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| Exact Mass |
497.188
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| CAS # |
2254409-25-9
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| PubChem CID |
138454752
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
4.1
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
36
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| Complexity |
851
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| Defined Atom Stereocenter Count |
0
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| SMILES |
S(C1C=CC(=C2C=CC=CC2=1)C1C=NN2C=C(C=NC2=1)C1C=CC(=CC=1)C(C)N1CCCC1)(N)(=O)=O
|
| InChi Key |
IPESYYFOXATSFJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C28H27N5O2S/c1-19(32-14-4-5-15-32)20-8-10-21(11-9-20)22-16-30-28-26(17-31-33(28)18-22)24-12-13-27(36(29,34)35)25-7-3-2-6-23(24)25/h2-3,6-13,16-19H,4-5,14-15H2,1H3,(H2,29,34,35)
|
| Chemical Name |
4-[6-[4-(1-pyrrolidin-1-ylethyl)phenyl]pyrazolo[1,5-a]pyrimidin-3-yl]naphthalene-1-sulfonamide
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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 : ~125 mg/mL (~251.20 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.18 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume 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: ≥ 2.08 mg/mL (4.18 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.18 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0096 mL | 10.0480 mL | 20.0961 mL | |
| 5 mM | 0.4019 mL | 2.0096 mL | 4.0192 mL | |
| 10 mM | 0.2010 mL | 1.0048 mL | 2.0096 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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