| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 100mg |
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| Other Sizes |
| Targets |
CLK1 5 nM (IC50) CLK2 42 nM (IC50) CLK4 108 nM (IC50) DYRK1A 1521 nM (IC50)
CLK1-IN-3 targets Cdc2-like kinase 1 (CLK1) with high potency and selectivity. The compound inhibits CLK1 with an IC50 of 5 nM and shows over 300-fold selectivity for CLK1 over Dyrk1A. It also inhibits CLK2 and CLK4 with IC50 values of 42 nM and 108 nM, respectively. CLK1 is a kinase involved in the regulation of alternative splicing and has been implicated in cancer and other diseases. By inhibiting CLK1, CLK1-IN-3 modulates splicing and induces autophagy. |
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| ln Vitro |
CLK1-IN-3 (compound 10ad) has the capacity to inhibit both Clk1 and Clk2, which gives it anti-tumor potential [1]. In a dose-dependent manner, CLK1-IN-3 (10 μM-1000 μM) may bind to Clk1 protein and efficiently block its degradation [1]. In HeLa, BNLCL.2, and HCT 116 cells, CLK1-IN-3 (0–10 μM, 24 h) initiates autophagy [1]. The autolysosomal marker SQSTM1/p62 is degraded more readily when CLK1-IN-3 is present [1].
In vitro studies demonstrate that CLK1-IN-3 (10 μM-1000 μM) can effectively bind to CLK1 protein and inhibit its degradation in a dose-dependent manner. The compound (0-10 μM, 24 h) induces autophagy in HeLa, BNLCL.2, and HCT 116 cells. It stimulates the degradation of SQSTM1/p62, a marker of autolysosomes. CLK1-IN-3 shows potential in anti-tumor applications due to dual inhibition of CLK1 and CLK2. |
| ln Vivo |
In the acetaminophen (HY-66005, APAP)-induced ALI model, CLK1-IN-3 (compound 10ad) (0-40 mg/kg, intraperitoneal injection, once) can significantly inhibit ALI without obvious hepatocyte death[1]. With an acceptable pharmacokinetic profile and a relatively long half-life of 5.29 hours, CLK1-IN-3 (10 mg/kg; IV, PO, IP, once) has an oral bioavailability of 19.5%[1].
In vivo, CLK1-IN-3 (0-40 mg/kg, IP, once) significantly suppresses acute liver injury (ALI) without apparent liver cell death in the acetaminophen-induced ALI model. The compound has a relatively long half-life (T1/2 = 5.29 h) and an oral bioavailability of 19.5%. CLK1-IN-3 (10 mg/kg; IV, PO, IP, once) shows an acceptable pharmacokinetic profile. These findings support the potential of CLK1-IN-3 for treating acute liver injury. |
| Enzyme Assay |
The kinase inhibitory activity of CLK1-IN-3 can be assessed using in vitro kinase assays. In a typical assay, recombinant CLK1 kinase is incubated with a peptide substrate, ATP (including radiolabeled ³³P-ATP), and varying concentrations of CLK1-IN-3. The incorporation of phosphate into the substrate is measured, and the IC50 is calculated from dose-response curves. The IC50 for CLK1 inhibition is 5 nM. The selectivity against Dyrk1A, CLK2, and CLK4 can be assessed using similar assays.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: Hela cells, BNLCL.2 and HCT 116 cells Tested Concentrations: 0.2, 1, 5, and 10 μM Incubation Duration: 24 h Experimental Results: Elevated the expression level of LC3II protein (a marker of autophagosomes) as well as the ratio of LC3II to LC3I (a sensitive index of autophagy) in a dose-dependent and time-dependent manner. The cellular activity of CLK1-IN-3 is assessed using HeLa, BNLCL.2, and HCT 116 cells. Cells are treated with CLK1-IN-3 (0-10 μM, 24 h). Autophagy is assessed by monitoring the conversion of LC3-I to LC3-II and the degradation of SQSTM1/p62 by Western blotting. The compound's ability to bind CLK1 protein and inhibit its degradation is assessed by Western blotting or immunofluorescence. Cell viability is measured using MTT assays. |
| Animal Protocol |
Animal/Disease Models: Male C57BL/6 mice(8 weeks, injected acetaminophen (HY-66005) (500 mg/kg, ip))[1]
Doses: 10, 20, and 40 mg/kg Route of Administration: IP, once Experimental Results: diminished serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels Dramatically in dose-dependent. Animal/Disease Models: Male balb/c (Bagg ALBino) mouse: (aged 8 weeks)[1] Doses: 10 mg/kg Route of Administration: IV, PO, IP, once (pharmacokinetic/PK Analysis) Experimental Results: pharmacokinetic/PK Parameters of CLK1-IN-3 in male balb/c (Bagg ALBino) mouse:[1]. IV (10 mg/kg) PO ( 10 mg/kg) IP (10 mg/kg) Cmax (ng/mL) 13166.5±875.9 1457.4±177.3 4654.3±435.3 T1/2 (h) 2.96±1.2 5.29±2.1 3.27 ±1.1 AUC0-t (ng/mL* h) 9520.5±1011.3 1860.2±411.0 5010.4±987.2 CL (L/h/kg) 1.05±0.10 5.51±1.00 3.58±0.82 F (%) 19.5% CLK1-IN-3 has been evaluated in an acetaminophen-induced acute liver injury (ALI) model in mice. The compound is administered intraperitoneally at doses of 0-40 mg/kg. Liver injury is assessed by measuring serum ALT and AST levels, and by histopathological analysis of liver tissue. The compound significantly suppresses ALI without apparent liver cell death. These results demonstrate the in vivo efficacy of CLK1-IN-3 in protecting against acute liver injury. |
| ADME/Pharmacokinetics |
CLK1-IN-3 has been characterized with an acceptable pharmacokinetic profile. When administered at 10 mg/kg (IV, PO, IP), the compound shows a relatively long half-life of 5.29 hours and an oral bioavailability of 19.5%. The compound has a molecular weight of 430.48 g/mol and is soluble in DMSO at 100 mg/mL (232.30 mM). It is typically stored at 4°C, sealed and away from moisture. Stock solutions can be stored at -20°C for several months.
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| Toxicity/Toxicokinetics |
No detailed toxicity data has been published for CLK1-IN-3. In the acute liver injury model, the compound did not cause apparent liver cell death at the tested doses. As a CLK inhibitor, it may have on-target effects on splicing regulation in normal tissues. Comprehensive toxicology studies would be required to evaluate its safety profile for potential therapeutic development. The compound is intended for research use only.
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| References | |
| Additional Infomation |
CLK1-IN-3 (CAS# 2922550-28-3) is a potent and selective inhibitor of CLK1 with an IC50 of 5 nM and over 300-fold selectivity over Dyrk1A. It also inhibits CLK2 and CLK4 with IC50 values of 42 nM and 108 nM, respectively. The compound induces autophagy in vitro and significantly suppresses acetaminophen-induced acute liver injury in vivo. It has a half-life of 5.29 h and oral bioavailability of 19.5%. The molecular formula is C24H23FN6O and molecular weight is 430.48 g/mol.
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| Molecular Formula |
C24H23FN6O
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|---|---|
| Molecular Weight |
430.477427721024
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| Exact Mass |
430.191
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| CAS # |
2922550-28-3
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| PubChem CID |
168355546
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
2.7
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
32
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| Complexity |
636
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1CCN(CC1)C2=CC=C(C=C2)C(=O)NC3=CC4=C(NC=C4C5=CC(=CN=C5)F)N=C3
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| InChi Key |
LQKHTRXKHPHEPP-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H23FN6O/c1-30-6-8-31(9-7-30)20-4-2-16(3-5-20)24(32)29-19-11-21-22(15-28-23(21)27-14-19)17-10-18(25)13-26-12-17/h2-5,10-15H,6-9H2,1H3,(H,27,28)(H,29,32)
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| Chemical Name |
N-[3-(5-fluoropyridin-3-yl)-1H-pyrrolo[2,3-b]pyridin-5-yl]-4-(4-methylpiperazin-1-yl)benzamide
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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 (~232.3 mM; with ultrasonication)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (5.81 mM) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and 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: 2.5 mg/mL (5.81 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3230 mL | 11.6149 mL | 23.2299 mL | |
| 5 mM | 0.4646 mL | 2.3230 mL | 4.6460 mL | |
| 10 mM | 0.2323 mL | 1.1615 mL | 2.3230 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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