| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 2mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| Other Sizes |
|
Purity: ≥98%
THZ531 (THZ-531) is a novel, potent and irreversible / covalent inhibitor of Cyclin-dependent kinases CDK12 and CDK13 (IC50 =158 nM and 69 nM for CDK12/13, respectively) with anticancer effects. CDKs, or cyclin-dependent kinases, are essential for controlling the transcription of genes. THZ531 targets a cysteine outside the kinase domain in an irreversible manner. THZ531 results in the loss of hyperphosphorylated and elongating RNA polymerase II along with a loss of gene expression. DNA damage response genes and important transcription factor genes linked to super-enhancers are significantly downregulated by THZ531. THZ531 caused a significant increase in apoptotic cell death. Thus, small molecules with the ability to specifically target CDK12 and CDK13 may aid in the identification of cancer subtypes that depend more heavily on their kinase activities.
| Targets |
CDK12 (IC50 = 158 nM); CDK13 (IC50 = 69 nM); CDK7 (IC50 = 8.5 μM); CDK9 (IC50 = 10.5 μM)
|
|---|---|
| ln Vitro |
THZ531 potently inhibits CDK12 and CDK13, with IC50s of 158 nM and 69 nM, respectively, according to the results of kinase assays; in contrast, the inhibition of CDK7 and CDK9 is more than 50-fold weaker, with IC50s of 8.5 and 10.5 μM, respectively. With an IC50 of 50 nM, THZ531 treatment causes a significant and permanent reduction in Jurkat cell proliferation. The number of cells exhibiting sub-G1 content increases in a dose- and time-dependent manner after treatment with increasing doses of THZ531, according to FACS cell cycle analysis. Over the course of the experiment, there is no increase in the percentage of apoptotic cells at 50 nM THZ531 compared to the DMSO control. With 30 to 40% of cells exhibiting Annexin V positivity after 72 hours, higher dosages of THZ531 produce a noticeable Annexin V signal. THZ531 treatment is also associated with a marked decrease in elongating Pol II[1].
|
| Enzyme Assay |
THZ531 or DMSO are applied to the cells for six hours. Cells are treated, then twice-washed in cold PBS before being lysed in the lysis buffer provided below: 50 mM Hepes pH 7.4, 150 mM NaCl, 1% Nonidet P40 substitute, 5 mM EDTA, 1 mM DTT, and protease/phosphatase cocktails. After clearing, lysates are subjected to an overnight pulldown at 4°C using either bio-THZ1 or bio-TH531. For three hours, lysates are further incubated at room temperature in order to maximize the formation of covalent bonds. After that, lysates are incubated for a further two to three hours at 4°C with streptavidin agarose for pulldown[1].
|
| Cell Assay |
THZ531 or DMSO at the recommended concentrations are applied to Jurkat cells for 72 hours after they are plated at 20,000 cells per well in fresh media in 96-well plates. Following their 24-hour inoculation with fresh media at the indicated concentrations for 72 hours, HAP1 cells are seeded in 96-well plates at a density of 12,000 cells/well. Evaluation is done on THZ531's anti-proliferative impact. Cells treated with THZ531 or DMSO for 6 hours are used to measure the impact of inhibitor washout on the anti-proliferation of Jurkat cells. The medium containing the inhibitor is then removed, and THZ531 or not is added during a 66-hour incubation. Evaluation is done on THZ531's anti-proliferative impact. There are three biological duplicates used for every proliferation assay. Fitting a non-linear regression curve is used to determine IC50s[1].
|
| References |
| Molecular Formula |
C30H32CLN7O2
|
|---|---|
| Molecular Weight |
558.0738
|
| Exact Mass |
557.23
|
| Elemental Analysis |
C, 64.57; H, 5.78; Cl, 6.35; N, 17.57; O, 5.73
|
| CAS # |
1702809-17-3
|
| Related CAS # |
1702809-17-3
|
| Appearance |
Solid powder
|
| SMILES |
CN(C)C/C=C/C(=O)NC1=CC=C(C=C1)C(=O)N2CCC[C@H](C2)NC3=NC=C(C(=N3)C4=CNC5=CC=CC=C54)Cl
|
| InChi Key |
RUBYHLPRZRMTJO-MOVYNIQHSA-N
|
| InChi Code |
InChI=1S/C30H32ClN7O2/c1-37(2)15-6-10-27(39)34-21-13-11-20(12-14-21)29(40)38-16-5-7-22(19-38)35-30-33-18-25(31)28(36-30)24-17-32-26-9-4-3-8-23(24)26/h3-4,6,8-14,17-18,22,32H,5,7,15-16,19H2,1-2H3,(H,34,39)(H,33,35,36)/b10-6+/t22-/m1/s1
|
| Chemical Name |
(E)-N-[4-[(3R)-3-[[5-chloro-4-(1H-indol-3-yl)pyrimidin-2-yl]amino]piperidine-1-carbonyl]phenyl]-4-(dimethylamino)but-2-enamide
|
| Synonyms |
THZ-531; THZ531; THZ 53
|
| 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) |
DMSO: 100~250 mg/mL (179.2~448 mM)
Ethanol: ~4 mg/mL (~7.2 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.43 mg/mL (2.56 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 14.3 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: 1.43 mg/mL (2.56 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 14.3 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: ≥ 1.43 mg/mL (2.56 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 | 1.7919 mL | 8.9594 mL | 17.9189 mL | |
| 5 mM | 0.3584 mL | 1.7919 mL | 3.5838 mL | |
| 10 mM | 0.1792 mL | 0.8959 mL | 1.7919 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.
 THZ531 covalently inhibits CDK12 and 13.Nat Chem Biol.2016Oct;12(10):876-84. |
|---|
 THZ531 co-crystal structure with CDK12-cyclin K.Nat Chem Biol.2016Oct;12(10):876-84. |
 THZ531 induces apoptosis in Jurkat cells.Nat Chem Biol.2016Oct;12(10):876-84. |
 CDK12 binds regulatory and coding regions of active genes.Nat Chem Biol.2016Oct;12(10):876-84. |
|---|
 THZ531 inhibits transcription elongation and gene expression.Nat Chem Biol.2016Oct;12(10):876-84. |
 THZ531 inhibits DDR and transcription factor gene expression.Nat Chem Biol.2016Oct;12(10):876-84. |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA