| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
CDK9/CycT1 (IC50 = 11 nM (in-house assay) or 5 nM (Merck Millipore panel))[1]
CDK2 (selectivity ratio CDK2/CDK9 = 98 (in-house) or CDK2/CycE IC50 = 470 nM (Merck Millipore))[1] Other CDKs: CDK1/CycB IC50 = 690 nM, CDK3/CycE IC50 = 570 nM, CDK5/p35 IC50 = 800 nM, CDK6/CycD3 IC50 = 4400 nM, CDK7/CycH/MAT1 IC50 >10000 nM (Merck Millipore panel)[1] |
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| ln Vitro |
BAY-958 is a lead compound for Atuveciclib (formerly known as BAY-1143572) which is novel, potent, oral and highly selective PTEFb/CDK9 inhibitor. It inhibits CDK9/CycT1 with an IC50 of 13 nM and is more than 100-fold more selective for CDK9 over CDK2. It also inhibits GSK3 kinase with IC50 values of 45 nM and 87 nM for GSK3α and GSK3β respectively. Atuveciclib is currently in Phase I clinical trial. Selective inhibition of exclusively transcription-regulating PTEFb/CDK9 is a promising new approach in cancer therapy. Starting from lead compound BAY-958, lead optimization efforts strictly focusing on kinase selectivity, physicochemical and DMPK properties finally led to the identification of the orally available clinical candidate atuveciclib (BAY 1143572). Structurally characterized by an unusual benzyl sulfoximine group, BAY 1143572 exhibited the best overall profile in vitro and in vivo, including high efficacy and good tolerability in xenograft models in mice and rats. BAY 1143572 is the first potent and highly selective PTEFb/CDK9 inhibitor to enter clinical trials for the treatment of cancer.
Kinase Assay: Merck Millipore CDK assays: Assays were performed according to the Merck Millipore KinaseProfilerTM standard protocols, with an ATP concentration of 10 μm.. Cell Assay: HeLa human cervical tumor cells (CCL‐2) were obtained from the American Type Culture Collection (Manassas, USA) and MOLM‐13 human acute myeloid leukemia cells (ACC 554) were obtained from the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany). Authentication of cell lines was conducted at the German Collection of Microorganisms and Cell Cultures via PCR‐based DNA profiling of polymorphic short tandem repeats. Cells were propagated under the suggested growth conditions in a humidified 37 °C incubator. Proliferation assays were conducted in 96‐well plates at densities of 3000 (HeLa) and 5000 (MOLM‐13) cells per well in the growth medium containing 10 % fetal calf serum (FCS). Cells were treated in quadruplicate with serial dilutions of test compounds for 96 h. Relative cell numbers were quantified by crystal violet staining (HeLa) or CellTitre‐Glo Luminescent Cell Viability Assay (Promega) (MOLM‐13). IC50 values (inhibitory concentration at 50 % of maximal effect) were determined by means of a four‐parameter fit on measurement data which were normalized to vehicle (DMSO) treated cells (=100 %) and measurement readings taken immediately before compound exposure (=0 %). BAY-958 potently inhibited CDK9/CycT1 with IC50 of 11 nM (in-house) and showed high selectivity vs. CDK2 (IC50 ratio 98-fold).[1] In Merck Millipore KinaseProfiler panel, BAY-958 had IC50 of 5 nM for CDK9/CycT1, 470 nM for CDK2/CycE, 690 nM for CDK1/CycB, 570 nM for CDK3/CycE, 800 nM for CDK5/p35, 4400 nM for CDK6/CycD3, and >10000 nM for CDK7/CycH/MAT1.[1] BAY-958 exhibited antiproliferative activity against HeLa cells with IC50 of 1000 nM, and against MOLM-13 cells with IC50 of 280 nM.[1] In rat hepatocytes and liver microsomes, BAY-958 showed high metabolic stability.[1] The compound had low aqueous solubility of 11 mg/L at pH 6.5, moderate Caco-2 permeability (Papp A-B 22 nm/s), and a high efflux ratio of 15.[1] Blood/plasma partitioning in rats was about 3:1.[1] |
| ln Vivo |
In an in vivo pharmacokinetic study in rats, BAY 1143572 showed low blood clearance (CLb 1.1 L/h/kg). The volumes of distribution (V ss) of BAY 1143572 is 1.0 L/kg. BAY 1143572 shows significantly improved oral bioavailability of 54 %. The blood/plasma ratios is about 1. It does not show significant inhibition of cytochrome P450 activity, with IC50 values >20 μM. The administration of BAY 1143572 in immunocompromized NOD/Shi-scid/IL-2Rγ null (NOG) mice xenografted with patient-derived ATL cells greatly reduced the infiltration of ATL cells into organs, such as liver and bone marrow. Decreased human soluble IL2R levels in serum were also observed, which indicated a reduction of ATL tumor burden.
In an MOLM-13 human acute myeloid leukemia xenograft model in mice, daily oral administration of BAY-958 hydrochloride at 30 or 40 mg/kg resulted in marked inhibition of tumor growth with treatment-to-control (T/C) ratios of 0.16 and 0.12, respectively, at the end of the experiment. Treatment was well tolerated with body weight change less than 10% and no fatal toxicities.[1] |
| Enzyme Assay |
CDK9/CycT1 kinase assay: Recombinant full-length His-tagged human CDK9 and CycT1 were used. The biotinylated peptide biotin-Ttds-YISPLKSPYKISEG (C-terminus amide) served as substrate. 50 nL of 100-fold concentrated test compound in DMSO was pipetted into a 384-well plate. 2 μL of CDK9/CycT1 in assay buffer (50 mM Tris-HCl pH 8.0, 10 mM MgCl2, 1.0 mM dithiothreitol, 0.1 mM sodium orthovanadate, 0.01% Nonidet P-40) was added and pre-incubated for 15 min at 22°C. Kinase reaction started by adding 3 μL of ATP (final 10 μM) and substrate (final 1 μM) in assay buffer, incubated 25 min at 22°C. Reaction stopped with TR-FRET detection reagents (streptavidin-XL665, anti-RB(pSer807/pSer811) antibody, LANCE EU-labeled anti-mouse IgG) in EDTA/BSA/HEPES buffer. After 1 h incubation, fluorescence emissions at 620 nm and 665 nm (excitation 350 nm) were measured. The ratio 665/620 nm indicated phosphorylated substrate. IC50 calculated by four-parameter fit.[1]
CDK2/CycE assay: Similar protocol using GST-tagged human CDK2 and CycE, with same biotinylated peptide substrate. ATP final concentration 10 μM, substrate 0.75 μM. CDK2/CycE concentration ~130 ng/mL. Otherwise identical conditions.[1] Merck Millipore CDK assays: Performed according to KinaseProfiler standard protocols with ATP concentration of 10 μM.[1] |
| Cell Assay |
Proliferation assay: HeLa human cervical tumor cells (3000 cells/well) or MOLM-13 human acute myeloid leukemia cells (5000 cells/well) were seeded in 96-well plates in growth medium containing 10% fetal calf serum. Cells were treated in quadruplicate with serial dilutions of BAY-958 for 96 hours. Relative cell numbers were quantified by crystal violet staining for HeLa or CellTitre-Glo Luminescent Cell Viability Assay for MOLM-13. IC50 values were determined by four-parameter fit on data normalized to vehicle (DMSO)-treated cells (100%) and measurement readings taken immediately before compound exposure (0%).[1]
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| Animal Protocol |
Immunocompromized NOD/Shi-scid/IL-2Rγ null (NOG) mice xenografted with patient-derived ATL cells and in vivo pharmacokinetic in rats For in vivo efficacy study in MOLM-13 human AML model: 2×10^6 MOLM-13 cells suspended in 100% Matrigel were inoculated subcutaneously into left flank of female NMRI nu/nu mice (18-21 g, 5-6 weeks). Treatment started 3 days after tumor cell inoculation. BAY-958 hydrochloride was administered orally once daily at 30 or 40 mg/kg. PEG400/water 80:10 was used as vehicle control. Body weight and tumor areas (longest diameter × perpendicular) measured at least twice weekly. T/C ratios calculated by dividing mean tumor area of treatment group by mean tumor area of vehicle group at the time vehicle group was sacrificed.[1] |
| ADME/Pharmacokinetics |
Metabolic stability in rat hepatocytes: Liver cells distributed in Williams' Medium E containing 5% FCS at density 1.0×10^6 viable cells/mL. Test compound added at 1 μM final concentration. Suspensions shaken at 580 rpm, aliquots removed at 2,8,16,30,45,90 min, mixed with equal volume cold acetonitrile, frozen, centrifuged, supernatants analyzed by LC-MS/MS. Half-life determined from concentration-time plot, intrinsic clearance calculated. Hepatic in vivo blood clearance (CLb) and maximal oral bioavailability (Fmax) calculated using well-stirred liver model.[1]
Metabolic stability in rat liver microsomes: Similar procedure using rat liver microsomes (details not fully described but CLb values provided).[1] Caco-2 permeability: Caco-2 cells seeded at 4.5×10^4 cells/well on 24-well inserts (0.4 μm pore size), grown for 15 days in DMEM with 10% FCS, 1% GlutaMAX, penicillin/streptomycin, nonessential amino acids. Medium changed every 2-3 days. Before assay, medium replaced with FCS-free HEPES/carbonate transport buffer pH 7.2. Test compounds at 2 μM added to apical or basolateral compartment. After 2h incubation at 37°C, samples taken from both compartments, precipitated with methanol, analyzed by LC-MS/MS. Papp calculated for A-B and B-A directions; efflux ratio = Papp B-A / Papp A-B.[1] Rat pharmacokinetics: Male Wistar rats received BAY-958 intravenously at 0.3-0.5 mg/kg or intragastrically at 0.6-1 mg/kg, formulated as solutions with solubilizers (e.g., PEG400). Catheterized at jugular vein; plasma samples collected at 2 min (i.v.), 8,15,30,45 min, 1,2,4,6,8,24 h post-dose. Samples precipitated with ice-cold acetonitrile (1:5), supernatants analyzed by LC-MS/MS. PK parameters calculated using linear-log trapezoidal rule for AUC.[1] Equilibrium shake flask solubility: 2 mg solid compound in 1 mL phosphate buffer pH 6.5, stirred 24 h at RT, centrifuged. Supernatant analyzed by HPLC with UV detection against calibration curve. Solubility value in mg/L determined.[1] |
| Toxicity/Toxicokinetics |
BAY-958 did not show significant inhibition of cytochrome P450 activity, with IC50 values >20 μM. However, it had a CYP1A2 induction liability (NOEL ≤5 μg/L) which was later removed by structural modification.[1]
In the in vivo efficacy study, BAY-958 hydrochloride at 30 or 40 mg/kg daily was well tolerated with less than 10% mean body weight reduction and no fatal toxicities.[1] |
| References | |
| Additional Infomation |
BAY-958 is a triazine-based PTEFb/CDK9 inhibitor with a benzyl sulfonamide group. It was identified from Bayer's compound library and used as a lead for optimization. The binding mode of BAY-958 with CDK9 was investigated by docking experiments using the Glide docking program and a published X-ray complex of CDK9/CycT1 (PDB ID: 3MY1). The triazine core and aniline NH mediate binding to the hinge region; the benzyl sulfonamide moiety points toward the exit of the ATP pocket; the methoxyphenyl substituent points toward the ribose pocket; two hydrogen bonds to the hinge region, two hydrogen bonds from the amino part of the sulfonamide group, and a π-stacking interaction of the methoxyphenyl moiety with Phe103 gatekeeper residue were observed. A weak hydrogen bond between the para-fluoro substituent and Lys48 may also exist.[1]
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| Molecular Formula |
C17H16FN5O3S
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| Molecular Weight |
389.40404510498
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| Exact Mass |
389.095
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| CAS # |
1335490-39-5
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| Related CAS # |
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| PubChem CID |
53491924
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
2.1
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
27
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| Complexity |
575
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| Defined Atom Stereocenter Count |
0
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| SMILES |
COC1=C(C=CC(=C1)F)C2=NC(=NC=N2)NC3=CC=CC(=C3)CS(=O)(=O)N
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| InChi Key |
MGCPJSMHKDVTGW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H16FN5O3S/c1-26-15-8-12(18)5-6-14(15)16-20-10-21-17(23-16)22-13-4-2-3-11(7-13)9-27(19,24)25/h2-8,10H,9H2,1H3,(H2,19,24,25)(H,20,21,22,23)
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| Chemical Name |
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5681 mL | 12.8403 mL | 25.6805 mL | |
| 5 mM | 0.5136 mL | 2.5681 mL | 5.1361 mL | |
| 10 mM | 0.2568 mL | 1.2840 mL | 2.5681 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.
Antitumor efficacy in two AML models in mice and rats. A, B: Antitumor efficacy of BAY 1143572 in an MOLM‐13 human AML model in mice.ChemMedChem. 2017 Nov 8; 12(21): 1776–1793. |
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Antitumor efficacy of BAY‐958 hydrochloride in an MOLM‐13 human AML model in mice.ChemMedChem. 2017 Nov 8; 12(21): 1776–1793. |
Docking mode of BAY‐958 in complex with CDK9.
X‐ray structure ofN‐acetyl derivative33.ChemMedChem. 2017 Nov 8; 12(21): 1776–1793. |
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