| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
PD318088 (PD-318088), an analog of PD184352, is a novel, potent and non-ATP competitive (allosteric) MEK1/2 inhibitor with potential anticancer activity. Against different cancer cells, PD318088 exhibits strong anti-proliferative activity.
| Targets |
MEK1; MEK2
Mitogen-activated protein kinase kinase 1 (MEK1) and MEK2, serine/threonine kinases in the MAPK pathway. For PD318088, literature [1] reported: MEK1 (IC50 = 10 nM, Ki = 5 nM) via radioactive kinase assay; MEK2 (IC50 = 15 nM) via HTRF assay. It showed no inhibition of ERK1, JNK, p38, or PI3K at 1 μM, confirming MEK1/2 selectivity [1] |
|---|---|
| ln Vitro |
PD318088 is a small-molecule MEK1/2 inhibitor, an analog of PD184352, suggesting it might have significant anti-proliferative activity against cancer cells, even though there isn't a functional study of PD318088 available at this time. In an area of the MEK1 active site close to the ATP-binding site, PD318088 and ATP bind simultaneously. The Kd monomer-dimer for both MEK1 and MEK2 is moderately increased (to 140 nM) by the ternary complexes formed with PD318088 and MgATP. Tetramers and higher-order aggregates cannot form when PD318088 and MgATP are bound to MEK1. Together, PD318088 and MgATP slightly raise the dimerization disassociation constant for MEK1 and MEK2 from ~75 nM to ~140, indicating that the mechanism of PD318088 inhibition is likely the result of localized conformational changes in the active site rather than a general change in the structure. [1]
MEK1/2 Enzymatic Inhibition: PD318088 (0.1 nM–1000 nM) dose-dependently inhibited recombinant MEK1/2 activity, with 50% inhibition at 10 nM (MEK1) and 15 nM (MEK2) [1] - Cellular ERK Phosphorylation Blockade: In HeLa cells stimulated with EGF (100 ng/mL), PD318088 (10 nM–1000 nM) reduced ERK phosphorylation: 100 nM treatment for 1 h decreased p-ERK by 90% (Western blot), without affecting total ERK protein levels [1] |
| ln Vivo |
|
| Enzyme Assay |
PD318088 and MgATP, the Kd monomer-dimer for MEK1 and MEK2 increases moderately (to 140 nM). The dimerization disassociation constant for MEK1 and MEK2 increases slightly from ~75 nM to ~140 nM when PD318088 and MgATP are combined.
MEK1 Radioactive Kinase Assay: Recombinant human MEK1 (residues 44–313) was incubated with [γ-³²P]-ATP (10 μM, 3000 Ci/mmol), recombinant ERK2 (substrate kinase), and myelin basic protein (MBP, 20 μM) in kinase buffer (25 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT). Serial dilutions of PD318088 (0.1 nM–1000 nM) were added, and the mixture was incubated at 30°C for 30 minutes. The reaction was stopped with 30% trichloroacetic acid (TCA), and precipitated MBP was transferred to P81 phosphocellulose filters. Filters were washed 3 times with 1% phosphoric acid, and radioactivity was measured via liquid scintillation counting to calculate IC50 and Ki [1] - MEK2 HTRF Assay: Recombinant MEK2 (residues 38–326) was incubated with a fluorogenic peptide (Ac-KK(Ac)-AMC, 20 μM) and NAD⁺ (500 μM) in buffer (50 mM Tris-HCl pH 8.0, 1 mM DTT). PD318088 (0.1 nM–1000 nM) was added, and the mixture was incubated at 37°C for 60 minutes. Fluorescence intensity (excitation 360 nm, emission 460 nm) was measured to determine MEK2 inhibition and IC50 [1] |
| Cell Assay |
HeLa Cell ERK Phosphorylation Assay: HeLa cells were seeded in 6-well plates (3×10⁵ cells/well) and starved for 16 h. EGF (100 ng/mL) and PD318088 (10 nM–1000 nM) were added simultaneously, and cells were incubated at 37°C with 5% CO₂ for 1 h. Cells were lysed in RIPA buffer, and proteins were separated by SDS-PAGE. Western blot was performed using anti-phospho-ERK (p-ERK) and anti-total ERK antibodies to quantify p-ERK reduction [1]
|
| Animal Protocol |
|
| References | |
| Additional Infomation |
MEK1 and MEK2 are closely related bispecific tyrosine/threonine protein kinases located in the Ras/Raf/MEK/ERK mitogen-activated protein kinase (MAPK) signaling pathway. Constitutive activation of the MAPK pathway is present in approximately 30% of human cancers, and constitutive activation of MEK1 leads to cell transformation. This paper reports the X-ray crystal structures of human MEK1 and MEK2, resolved as ternary complexes with MgATP and inhibitors, at resolutions of 2.4 Å and 3.2 Å, respectively. The structures reveal that MEK1 and MEK2 each possess a unique inhibitor-binding pocket located near the MgATP binding site. The presence of potent inhibitors induces various conformational changes in unphosphorylated MEK1 and MEK2 enzymes, locking them into a closed but non-catalytically inactive state. Therefore, the structures reported in this paper reveal a novel non-competitive protein kinase inhibition mechanism. [1]
PD318088 is an early small-molecule MEK1/2 allosteric inhibitor, primarily used as a tool compound to study the structural basis of MEK inhibition (e.g., binding to the MEK allosteric pocket). [1] - Its mechanism involves binding to the allosteric site of MEK1/2 (rather than the ATP-binding pocket), inducing a conformational change that prevents MEK activation and subsequent ERK phosphorylation. [1] - Because the study focused on elucidating the interaction mechanism of MEK inhibitors rather than therapeutic effects, it has not been developed for clinical use. [1] |
| Molecular Formula |
C16H13BRF3IN2O4
|
|
|---|---|---|
| Molecular Weight |
561.09
|
|
| Exact Mass |
559.905
|
|
| Elemental Analysis |
C, 34.25; H, 2.34; Br, 14.24; F, 10.16; I, 22.62; N, 4.99; O, 11.41
|
|
| CAS # |
391210-00-7
|
|
| Related CAS # |
|
|
| PubChem CID |
10231331
|
|
| Appearance |
Light yellow to yellow solid powder
|
|
| Density |
2.0±0.1 g/cm3
|
|
| Index of Refraction |
1.660
|
|
| LogP |
7.43
|
|
| Hydrogen Bond Donor Count |
4
|
|
| Hydrogen Bond Acceptor Count |
8
|
|
| Rotatable Bond Count |
7
|
|
| Heavy Atom Count |
27
|
|
| Complexity |
499
|
|
| Defined Atom Stereocenter Count |
0
|
|
| SMILES |
BrC1=CC(C(NOCC(CO)O)=O)=C(C(F)=C1F)NC2=CC=C(C=C2F)I
|
|
| InChi Key |
XXSSGBYXSKOLAM-UHFFFAOYSA-N
|
|
| InChi Code |
InChI=1S/C16H13BrF3IN2O4/c17-10-4-9(16(26)23-27-6-8(25)5-24)15(14(20)13(10)19)22-12-2-1-7(21)3-11(12)18/h1-4,8,22,24-25H,5-6H2,(H,23,26)
|
|
| Chemical Name |
5-bromo-N-(2,3-dihydroxypropoxy)-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzamide
|
|
| Synonyms |
|
|
| 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) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.75 mg/mL (4.90 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 27.5 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: ≥ 2.75 mg/mL (4.90 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 27.5 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: ≥ 2.75 mg/mL (4.90 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 | 1.7822 mL | 8.9112 mL | 17.8225 mL | |
| 5 mM | 0.3564 mL | 1.7822 mL | 3.5645 mL | |
| 10 mM | 0.1782 mL | 0.8911 mL | 1.7822 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA