Physicochemical Properties
| Molecular Formula | C24H33CLN6O2S |
| Molecular Weight | 505.0758 |
| Exact Mass | 504.21 |
| Elemental Analysis | C, 57.07; H, 6.59; Cl, 7.02; N, 16.64; O, 6.34; S, 6.35 |
| CAS # | 2247481-21-4 |
| Related CAS # | 2247481-21-4 |
| PubChem CID | 134814484 |
| Appearance | Light yellow to yellow solid powder |
| LogP | 4 |
| Hydrogen Bond Donor Count | 3 |
| Hydrogen Bond Acceptor Count | 9 |
| Rotatable Bond Count | 10 |
| Heavy Atom Count | 34 |
| Complexity | 667 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | XSDZPPRQLIZLDG-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C24H33ClN6O2S/c1-32-11-8-27-17-2-4-18(5-3-17)30-22-12-19(20(25)13-28-22)21-14-34-23(31-21)29-16-24(15-26)6-9-33-10-7-24/h12-14,17-18,27H,2-11,16H2,1H3,(H,28,30)(H,29,31) |
| Chemical Name | 4-[[[4-[5-chloro-2-[[4-(2-methoxyethylamino)cyclohexyl]amino]pyridin-4-yl]-1,3-thiazol-2-yl]amino]methyl]oxane-4-carbonitrile |
| Synonyms | JSH-150; JSH150; JSH 150 |
| 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) |
Biological Activity
| Targets | CDK9/cyclinT1 (IC50 = 1 nM); CDK16/Cyclin Y (IC50 = 292 nM); Cdk1/cyclin B (IC50 = 1.34 μM); CDK14/Cyclin Y (IC50 = 1.68 μM); CDK7/Cyclin H/MNAT1(IC50 = 1.72 μM); cdk2/cyclin A (IC50 = 2.86 μM); Cdk5/p25 (IC50 = 4.64 μM) |
| ln Vitro | JSH-150 demonstrates an IC50 of 1 nM in the biochemical assay against CDK9 kinase and attains selectivity over other members of the CDK kinase family that is approximately 300–10,000 times higher. High selectivity over other 468 kinases/mutants is another feature of it. With regard to melanoma, neuroblastoma, hepatoma, colon cancer, lung cancer, and leukemia cell lines, JSH-150 exhibits strong antiproliferative properties. It can suppress MCL-1 and C-Myc expression, halt the cell cycle, and cause apoptosis in leukemia cells in a dose-dependent manner. It can also inhibit the phosphorylation of RNA Pol II.[1] |
| ln Vivo | JSH-150 at a dosage of 10 mg/kg can virtually completely stop the growth of tumors in the xenograft mouse model inoculated with MV4-11 cells. JSH-150 is a promising drug candidate for leukemia and other cancers, and its good in vivo PK/PD profile and high selectivity make it an excellent pharmacological tool to study CDK9-mediated physiology and pathology.[1] |
| Enzyme Assay | The kinase reaction system comprises 10 μM ATP, 0.5 μL of serially diluted compounds 11–43 (including JSH–150), and 5 μL of CDK9 substrate PDKtide (0.2 μg/μL) with CDK9/CyclinK kinase (3 ng/μL). Each tube's reaction is initiated instantly by adding ATP, and it continues at 37 °C for an hour. 5 μL solvent reactions are conducted in a 384-well plate after the tube has been cooled for five minutes at room temperature. Subsequently, 5 microliters of ADP-Glo reagent are introduced into every well to halt the reaction and utilise the residual ADP in 40 minutes. Lastly, a luminescence signal is generated by adding 10 μL of kinase detection reagent into the well and incubating it for 30 minutes. The dose-response curve is fitted with Prism 7.0, and the luminosity signal is measured using an automated plate reader. |
| Cell Assay | Investigation of Signaling Pathways. For two hours, DMSO, serially diluted JSH-150, and 1 μM compound 7 are applied to MV4-11, HL-60, and MEC-1 cells. Following that, the cells are lysed in cell lysis buffer and rinsed in 1 × PBS. For immunoblotting, the following antibodies are used: phospho-CDK9 (Thr186), CDK9, phospho-RNA Pol II (Ser2), phospho-RNA Pol II (Ser5), RNA Pol II, XIAP, MCL-1, C-MYC, BCL-2, and GAPDH. |
| Animal Protocol | Mice: Female nu/nu mice aged five weeks are utilized. Cells are taken during exponential growth before being implanted. Five million MV4-11 cells in PBS are combined 1:1 with Matrigel and injected into the right flank subcutaneous space of nu/nu mice. When MV4-11 tumors are 200–400 mm3 in size, daily oral administration is started. Then, for efficacy research, the animals are randomly assigned to treatment groups consisting of five mice apiece. Every day, JSH-150 is given orally via gavage in an HKI solution (0.5% methocellulose/0.4% Tween80 in ddH2O). JSH-150 or its vehicle as a control are given in a range of dosages. JSH-150 is administered orally to female nu/nu mice with established MV4-11 tumor xenografts at doses of 10, 20, and 30 mg/kg/d, or as a vehicle. Following JSH-150 treatment, daily measurements of body weight and tumor growth are made. Volume of tumor is computed. |
| References |
[1]. Eur J Med Chem . 2018 Oct 5:158:896-916. |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~100 mg/mL (~198 mM) Ethanol: ~100 mg/mL (~198 mM) Water: ~100 mg/mL (~198 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.95 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 25.0 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: ≥ 2.5 mg/mL (4.95 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 25.0 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (4.95 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 25.0 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 | 1.9799 mL | 9.8994 mL | 19.7988 mL | |
| 5 mM | 0.3960 mL | 1.9799 mL | 3.9598 mL | |
| 10 mM | 0.1980 mL | 0.9899 mL | 1.9799 mL |