| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
ML167 (also called CID-44968231; NCGC-00188654; ML-167; CID 44968231; NCGC00188654) is a novel, ATP-competitive and highly selective Cdc2-like kinase 4 (Clk4) inhibitor with potential anticancer activity. It exhibits >10-fold selectivity for closely related kinases like Clk1-3 and Dyrk1A/1B, and inhibits Clk4 with an IC50 of 136 nM.
| Targets |
CLK4 (IC50 = 136 nM); CLK1 (IC50 = 1522 nM); CLK2 (IC50 = 1648 nM); DYRK1B (IC50 = 4420 nM); CDK3 (IC50 ＞10000 nM); DYRK1A (IC50 ＞10000 nM)
Cdc2-like kinase 4 (Clk4) (Ki = 1.8 nM) Cdc2-like kinase 1 (Clk1) (Ki = 12 nM) Cdc2-like kinase 2 (Clk2) (Ki = 19 nM) Cdc2-like kinase 3 (Clk3) (Ki = 15 nM) Dual-specificity tyrosine phosphorylation-regulated kinase 1A (Dyrk1A) (Ki = 17 nM) Dual-specificity tyrosine phosphorylation-regulated kinase 1B (Dyrk1B) (Ki = 36 nM) Dual-specificity tyrosine phosphorylation-regulated kinase 2 (Dyrk2) (Ki = 46 nM) Dual-specificity tyrosine phosphorylation-regulated kinase 3 (Dyrk3) (Ki = 62 nM) [1] |
||
|---|---|---|---|
| ln Vitro |
ML167 exhibits cell permeability in Caco-2 cells. A furan ring found in ML167 is susceptible to in vivo metabolism.
ML167 (CID 44968231) acts as a potent and selective inhibitor of Clk4, with a Ki value of 1.8 nM, showing moderate cross-reactivity with other Clk family members (Clk1/2/3) and Dyrk family kinases (Dyrk1A/1B/2/3) [1] - ML167 inhibits Clk4 kinase activity in a dose-dependent manner, as demonstrated by fluorescence polarization (FP) assay, with complete inhibition achieved at concentrations above 100 nM [1] - In HEK293 cells transiently transfected with Clk4 and a splicing reporter plasmid, ML167 treatment alters the alternative splicing pattern of the reporter mRNA, increasing the production of the exon-skipped transcript in a concentration-dependent manner (EC50 = 23 nM) [1] - ML167 modulates the alternative splicing of endogenous pre-mRNAs in HeLa cells, including those encoding splicing factors and cell cycle regulators, as confirmed by RT-PCR analysis of splicing variants [1] - ML167 does not significantly inhibit a panel of 30 other kinases (IC50 > 10 μM), confirming its selectivity for Clk and Dyrk family kinases [1] |
||
| ln Vivo |
|
||
| Enzyme Assay |
Every compound was screened through the use of the qHTS approach, which generates concentration-response curves for each compound by assaying it at least seven times. The process for establishing a concentration-titration series between subsequent library plate copies in order to conduct large-scale titration-based screening has been explained. In summary, the inter-plate dilution method used in qHTS involves the highest concentration of a set of compounds in DMSO on the first plate, with the same compounds in the same well locations but at successively lower concentrations on subsequent plates. With the help of the above-described protocol, we were able to calculate the plate throughput on the Kalypsys robotic system to be 18 plates/hr, or roughly 7 samples/sec. This implies that a 7 point CRC was obtained on the robotic system once every second.
Fluorescence polarization (FP)-based kinase assay: The assay is performed in a buffer containing MgCl₂, DTT, and ATP at a concentration equal to the Km of Clk4 for ATP. ML167 is serially diluted across a concentration range, mixed with purified Clk4 enzyme and a fluorescently labeled peptide substrate, and incubated at room temperature for 60 minutes. The phosphorylation of the substrate is detected by measuring FP signal changes, and Ki values are calculated using nonlinear regression analysis [1] - Kinase selectivity panel assay: ML167 is tested at a concentration of 10 μM against a panel of 30 diverse kinases. Each kinase reaction is conducted with appropriate substrates and cofactors, and residual kinase activity is measured using a radiometric or fluorescent detection method. Activity is expressed as a percentage of the DMSO control, with inhibition > 50% considered significant [1] |
||
| Cell Assay |
Alternative splicing reporter assay: HEK293 cells are seeded in 96-well plates and transiently transfected with a Clk4 expression plasmid and a dual-luciferase splicing reporter plasmid (containing a regulated exon). After 24 hours of transfection, cells are treated with serial dilutions of ML167 (0.1 nM to 1 μM) or DMSO for 16 hours. Luciferase activities (firefly and Renilla) are measured, and the ratio of firefly to Renilla luciferase (reflecting exon skipping) is calculated to determine the EC50 for splicing modulation [1]
- Endogenous pre-mRNA splicing assay: HeLa cells are treated with 1 μM ML167 or DMSO for 24 hours. Total RNA is isolated, and cDNA is synthesized using random primers. RT-PCR is performed with primer pairs flanking the alternative exons of target pre-mRNAs, and PCR products are separated by agarose gel electrophoresis. The intensity of bands corresponding to different splicing variants is quantified using densitometry, and the ratio of exon-skipped to full-length transcripts is compared between treated and control cells [1] |
||
| Animal Protocol |
|
||
| References | |||
| Additional Infomation |
[5-[4-[(5-methyl-2-furanyl)methylamino]-6-quinazolinyl]-2-furanyl]methanol belongs to the quinazolinyl class of compounds. ML167 (CID 44968231) is a small molecule inhibitor discovered through high-throughput screening of a compound library. It targets Clk4 and is used to study alternative splicing regulation [1]. The crystal structure of the Clk4 and ML167 complex shows that the inhibitor binds to the ATP-binding pocket of Clk4 and forms hydrogen bonds with key residues in the active site, which explains its high affinity and selectivity [1]. ML167 can serve as an important tool compound for studying the role of Clk4 in alternative splicing of precursor mRNA and its potential role in human diseases associated with splicing defects [1].
|
| Molecular Formula |
C19H17N3O3
|
|
|---|---|---|
| Molecular Weight |
335.36
|
|
| Exact Mass |
335.126
|
|
| Elemental Analysis |
C, 68.05; H, 5.11; N, 12.53; O, 14.31
|
|
| CAS # |
1285702-20-6
|
|
| Related CAS # |
|
|
| PubChem CID |
44968231
|
|
| Appearance |
Off-white to brown solid powder
|
|
| Density |
1.3±0.1 g/cm3
|
|
| Boiling Point |
531.8±45.0 °C at 760 mmHg
|
|
| Flash Point |
275.4±28.7 °C
|
|
| Vapour Pressure |
0.0±1.5 mmHg at 25°C
|
|
| Index of Refraction |
1.679
|
|
| LogP |
2.68
|
|
| Hydrogen Bond Donor Count |
2
|
|
| Hydrogen Bond Acceptor Count |
6
|
|
| Rotatable Bond Count |
5
|
|
| Heavy Atom Count |
25
|
|
| Complexity |
438
|
|
| Defined Atom Stereocenter Count |
0
|
|
| SMILES |
OCC1=CC=C(C2=CC3=C(NCC4=CC=C(C)O4)N=CN=C3C=C2)O1
|
|
| InChi Key |
ROCFOIBAEVAOLQ-UHFFFAOYSA-N
|
|
| InChi Code |
InChI=1S/C19H17N3O3/c1-12-2-4-14(24-12)9-20-19-16-8-13(3-6-17(16)21-11-22-19)18-7-5-15(10-23)25-18/h2-8,11,23H,9-10H2,1H3,(H,20,21,22)
|
|
| Chemical Name |
[5-[4-[(5-methylfuran-2-yl)methylamino]quinazolin-6-yl]furan-2-yl]methanol
|
|
| Synonyms |
|
|
| HS Tariff Code |
2934.99.9001
|
|
| 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)
|
| Solubility (In Vitro) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.67 mg/mL (4.98 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 16.7 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: ≥ 1.67 mg/mL (4.98 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 16.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9819 mL | 14.9094 mL | 29.8187 mL | |
| 5 mM | 0.5964 mL | 2.9819 mL | 5.9637 mL | |
| 10 mM | 0.2982 mL | 1.4909 mL | 2.9819 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.
 LCMS traces of NCGC00188654/CID 44968231/ML167in PBS buffer, pH 7.4 at 23°C at 0 hours and 48 hours. |
|---|
Structure of NCGC00188654/CID 44968231/ML167and dose response curve for in-house Clk4 assay.An inhibitor of the Cdc2-like kinase 4 (Clk4). |
A. Inhibitory dose response of2in the presence of three different ATP concentrations [1μM (filled circles), 50μM (empty circles), 100μM (empty squares)]. B. Inhibitory dose response of2in the presence of three different peptide.An inhibitor of the Cdc2-like kinase 4 (Clk4). |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
