| 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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| Other Sizes |
Purity: ≥98%
| Targets |
ERK5 (IC50 = 8 nM)
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| ln Vitro |
AX-15836 demonstrates ERK5 selectivity of more than 1,000-fold over a panel of more than 200 kinases. With a Kd of 3,600 nM, it also displays selectivity over BRD4. Peripheral blood mononuclear cells (PBMCs), endothelial cells, and oncogenic cell lines all exhibit a similar intracellular potency (4–9 nM) for AX15836. AX15836 was completely ineffective (EC50>10 M) at inhibiting the inflammatory cytokine response, indicating that the compounds' BRD inhibition component was what actually caused the cytokine reduction. Only a small number of genes exhibit differential expression in samples treated with AX15836 in HUVEC and HeLa cell types. In HeLa cells, AX15836 was able to demonstrably inhibit ERK5 that had been phosphorylated in response to EGF[1].
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| Cell Assay |
For studies on cell proliferation, cells are exposed to DMSO vehicle (0.25% final volume) or an eight-point serial dilution series of AX-15836 (starting concentration of 15 μM). For MM.1S cells, compound was added 1 h before adding recombinant human IL-6 at 5 nM.. Utilizing the CellTiter-Glo 2.0 reagent, the relative number of viable cells was calculated 3 days later. The Synergy 2 multimode reader was used to read the Luminescence[1].
Preparation of cell samples for RNA-Seq. [1] Cells were preincubated with 0.1% DMSO vehicle, 1 μM AX-15836 (ERK5 inhibitor), 5 μM AX15839 (dual ERK5/BRD inhibitor), or 1 μM I-BET762 (BRD inhibitor) for 1 h. HUVECs and HeLa cells were then stimulated with their respective agonists (10 μg/mL Pam3CSK4 or 50 ng/mL EGF) for 5 h at 37 °C. Cells were processed to total RNA using the RNeasy kit. RNA was sent for RNA-Seq. One microgram total RNA from each sample was ribodepleted using the Ribo-Zero-rRNA Removal Kit. Sequencing libraries were then prepared from ribodepleted RNA using NEBNext Ultra RNA Library Prep Kit for Illumina following the manufacturer’s recommended protocol and barcoded using standard Illumina TruSeq barcoded adapter sequences. Final libraries were size-selected using Agencourt AMPure XP beads. Purified libraries were pooled and loaded onto an Illumina HiSeq2000 sequencer for 100-base single end sequencing with 7-base index reads. |
| Animal Protocol |
Male CD-1 mice (∼12 wk old; Envigo) were used in this study. Animals were allowed to acclimate for a minimum of 48 h before the conduction of experiments. All animals were group housed in ventilated cages in the animal facility with regulated temperature from 20 °C to 26 °C and relative humidity at 30–70% with a 12/12-h day/night cycle. Pharmacokinetic studies were performed using fasted mice orally dosed at 50 mg/kg with AX-15836 formulated in 2.5% (vol/vol) dimethylacetamide: in 0.3% (wt/vol) carboxymethylcellulose. Blood was sampled for eight time points over 24 h. [1]
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| References | |
| Additional Infomation |
Unlike other members of the MAPK family, ERK5 contains a large C-terminal domain with transcriptional activation capabilities, in addition to its N-terminal classical kinase domain. ERK5 gene deletion leads to embryonic lethality, while tissue-specific deletion significantly affects erythroid development, cardiac function, and neurogenesis. Furthermore, ERK5 deletion also exhibits anti-inflammatory and anti-tumor effects. Small molecule ERK5 inhibitors have shown good activity in cellular and animal models of inflammation and tumors. This article reports the synthesis and biological characteristics of a highly efficient and selective ERK5 inhibitor. Unlike ERK5 gene deletion/deficiency and previously reported inhibitory effects, inhibition of this kinase using the novel inhibitor with the highest selectivity did not show anti-inflammatory or anti-proliferative activity. The therapeutic efficacy of previously reported ERK5 inhibitors has been confirmed to originate from non-targeting activity of the bromine domain, a conserved protein module involved in the recognition of acetyl-lysine residues during transcription. The phenotypes reported by ERK5 gene deletion or depletion are likely due to the loss of ERK5's non-catalytic function. The newly reported inhibitors will help identify which of the many reported phenotypes are caused by kinase activity and clarify which phenotypes can serve as drug targets. [1]
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| Molecular Formula |
C32H40N8O5S
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|---|---|
| Molecular Weight |
648.7756
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| Exact Mass |
648.28
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| Elemental Analysis |
C, 59.24; H, 6.21; N, 17.27; O, 12.33; S, 4.94
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| CAS # |
2035509-96-5
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| Related CAS # |
2035509-96-5
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| PubChem CID |
122705989
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| Appearance |
Off-white to yellow solid powder
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| LogP |
2.4
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
46
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| Complexity |
1170
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| Defined Atom Stereocenter Count |
0
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| SMILES |
S(C([H])([H])[H])(N1C2=C([H])C([H])=C([H])C([H])=C2C(N(C([H])([H])[H])C2=C([H])N=C(N=C12)N([H])C1C([H])=C([H])C(=C([H])C=1OC([H])([H])C([H])([H])[H])C(N1C([H])([H])C([H])([H])C([H])(C([H])([H])C1([H])[H])N1C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])C1([H])[H])=O)=O)(=O)=O
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| InChi Key |
HTFNVAVTYILUCF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C32H40N8O5S/c1-5-45-28-20-22(30(41)39-14-12-23(13-15-39)38-18-16-36(2)17-19-38)10-11-25(28)34-32-33-21-27-29(35-32)40(46(4,43)44)26-9-7-6-8-24(26)31(42)37(27)3/h6-11,20-21,23H,5,12-19H2,1-4H3,(H,33,34,35)
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| Chemical Name |
2-[2-ethoxy-4-[4-(4-methylpiperazin-1-yl)piperidine-1-carbonyl]anilino]-5-methyl-11-methylsulfonylpyrimido[4,5-b][1,4]benzodiazepin-6-one
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| Synonyms |
AX15836; AX-15836; 2035509-96-5; AX15,836; AX-15,836; AX 15,836; 2-[2-ethoxy-4-[4-(4-methylpiperazin-1-yl)piperidine-1-carbonyl]anilino]-5-methyl-11-methylsulfonylpyrimido[4,5-b][1,4]benzodiazepin-6-one; CHEMBL4541479; 2-((2-ethoxy-4-(4-(4-methylpiperazin-1-yl)piperidine-1-carbonyl)phenyl)amino)-5-methyl-11-(methylsulfonyl)-5,11-dihydro-6H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6-one; 2-(2-Ethoxy-4-(4-(4-methylpiperazin-1-yl)piperidine-1-carbonyl)phenylamino)-5-methyl-11-(methylsulfonyl)-5H-benzo[e]pyrimido[5,4-b][1,4]diazepin-6(11H)-one; AX 15836
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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 |
| 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) |
DMSO: ~100 mg/mL (~154.1 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.85 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 (3.85 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (3.85 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 | 1.5414 mL | 7.7068 mL | 15.4135 mL | |
| 5 mM | 0.3083 mL | 1.5414 mL | 3.0827 mL | |
| 10 mM | 0.1541 mL | 0.7707 mL | 1.5414 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.
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