Physicochemical Properties
| Molecular Formula | C18H25CLN6 |
| Molecular Weight | 360.884301900864 |
| Exact Mass | 360.182 |
| CAS # | 2079068-74-7 |
| PubChem CID | 126558497 |
| Appearance | Off-white to brown solid powder |
| LogP | 2.9 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 25 |
| Complexity | 439 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | ClC1=CN=C(N=C1C1C=NN(C)C=1CC1CC1)NC1CCC(CC1)N |
| InChi Key | RVZJFCNYSSUDCU-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C18H25ClN6/c1-25-16(8-11-2-3-11)14(9-22-25)17-15(19)10-21-18(24-17)23-13-6-4-12(20)5-7-13/h9-13H,2-8,20H2,1H3,(H,21,23,24) |
| Chemical Name | 4-N-[5-chloro-4-[5-(cyclopropylmethyl)-1-methylpyrazol-4-yl]pyrimidin-2-yl]cyclohexane-1,4-diamine |
| 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 | CKIα CDK7 1.3 nM (Kd) CDK9 4 nM (Kd) |
| ln Vitro | Casein Kinase inhibitor A51 (0.05-3.2 μM; 18 hours) treatment of RKO cells eliminated most of the Ser45 phosphorylation signal and the subsequent GSK3 phosphorylation cascade, which led to the stabilization of β-catenin, in a manner similar to CKIα depletion[1]. At 160 nM or less, the casein kinase inhibitor A51 is very effective at causing leukemia cells to undergo apoptosis, mostly due to its ability to stabilize p53[1]. MYC, MDM2, and the anti-apoptotic oncogene MCL1 are all eliminated by the casein kinase inhibitor A51 (0.08-2 μM; 6.5 hours). A51, a casein kinase inhibitor, significantly decreases the mRNA expression of MDM2 and MYC while increasing the expression of Wnt targets AXIN2 and CCND1 (Cyclin D1)[1]. |
| ln Vivo | When the percentage of leukemia cells in the bone marrow is greater than 1.5% of total cells, 8 days after leukemia cell injection, oral medication (Casein Kinase inhibitor A51; 5 mg/kg/day) is started. Every mouse treated with A51 exhibits normal organ morphology, histology, and blood counts[1]. Rapid oral absorption is shown in pharmacokinetic investigations of the Casein Kinase inhibitor A51 at 20 mg/kg, with Tmax values of 0.5-2 hr, Cmax values of 1060 ng/mL, T1/2 values of 2.5 hr, and area under the curve (AUC) values of 3680 (ng*hr/mL)[1]. |
| Cell Assay |
Cell Viability Assay[1] Cell Types: MV4-11 cells Tested Concentrations: 0.08 μM, 0.6 μM, 2 μM Incubation Duration: 6.5 hrs (hours) Experimental Results: Abolishes the expression of MYC, MDM2, and the anti-apoptotic oncogene MCL1. |
| References |
[1]. Small Molecules Co-targeting CKIα and the Transcriptional Kinases CDK7/9 Control AML in Preclinical Models. Cell. 2018 Sep 20;175(1):171-185.e25. |
| Additional Infomation | CK1alpha/CDK7/CDK9 Inhibitor BTX-A51 is the ditosylated salt of A51, an orally bioavailable inhibitor of casein kinase 1alpha (CK1alpha) and cyclin-dependent kinases 7 and 9 (CDK7 and CDK9), with potential antineoplastic activity. Upon administration, BTX-A51 binds to and inhibits the activity of CK1alpha, CDK7, and CDK9. Blocking the phosphorylation and kinase activity of CK1alpha prevents the enhanced binding of murine double minute X (MDMX) to p53, the formation of CK1alpha and MDM2 complex, and the resulting inhibition of p53. This induces p53-mediated cell cycle arrest, slowing tumor cell proliferation. Blocking the phosphorylation and kinase activity of CDK7 and CDK9 prevents the positive transcription elongation factor b (PTEFb)-mediated activation of RNA polymerase II (RNA Pol II) and leads to the inhibition of gene transcription of various anti-apoptotic proteins. This also induces cell cycle arrest and apoptosis, slowing tumor cell proliferation. CK1alpha, a serine/threonine kinase and a leukemic stem cell target, acts as a tumor suppressor in several cancers through the negative regulation of Wnt/beta-catenin signaling and p53. It negatively regulates p53 by phosphorylating MDMX, thus enhancing binding of MDMX to p53, as well as by forming a complex with MDM2. CDK7, a serine/threonine kinase, plays a role in controlling cell cycle progression, transcriptional regulation, and promotes the expression of key oncogenes such as c-Myc through the phosphorylation of RNA Pol II. CDK9, also a serine/threonine kinase, regulates elongation of transcription through phosphorylation of RNA Pol II at serine 2 (p-Ser2-RNAPII). It is upregulated in various tumor cell types and plays a key role in the regulation of RNA Pol II-mediated transcription of anti-apoptotic proteins. Tumor cells are dependent on anti-apoptotic proteins for their survival. |
Solubility Data
| Solubility (In Vitro) | DMSO : 100 mg/mL (277.10 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.93 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 (6.93 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 (6.93 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 | 2.7710 mL | 13.8550 mL | 27.7100 mL | |
| 5 mM | 0.5542 mL | 2.7710 mL | 5.5420 mL | |
| 10 mM | 0.2771 mL | 1.3855 mL | 2.7710 mL |