| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
M2I-1 (full name: Mad2 Inhibitor-1) is a first-in-class small molecule inhibitor that targets the binding of Mad2 to Cdc20, an essential protein-protein interaction (PPI) within the mitotic SAC (spindle assembly checkpoint). In FP assays, it can also suppress the Mad2-F-Mbp1 interaction. Computational and biochemical analyses revealed that M2I-1 disrupts the conformational dynamics of Mad2, which is essential for the formation of complexes with Cdc20. Research on cells showed that M2I-1 reduces the SAC response, suggesting that the substance may be active in cells. This study thus identifies the formation of the SAC-specific complex between Mad2 and Cdc20 as a protein-protein interaction that is amenable to small molecule targeting.
| Targets |
Mad2
M2I-1 (Mad2 Inhibitor-1) targets the protein-protein interaction (PPI) between Mad2 and Cdc20 (core components of the mitotic spindle assembly checkpoint, SAC) [1] |
||
|---|---|---|---|
| ln Vitro |
M2I-1, the initial small molecule inhibitor, is crucial for preventing Mad2 from binding to Cdc20, making it a PPI within the SAC.[1]
M2I-1 was identified via a fluorescence polarization-based screen as a small molecule inhibitor that disrupts the binding of Mad2 to Cdc20, an essential PPI within the spindle assembly checkpoint (SAC) [1] - Computational and biochemical analyses indicated that M2I-1 disturbs the conformational dynamics of Mad2, which is critical for the formation of the Mad2-Cdc20 complex [1] - Cellular studies using HeLa cells revealed that M2I-1 weakens the SAC response, confirming its activity in interfering with SAC function in mammalian cells [1] |
||
| ln Vivo |
|
||
| Enzyme Assay |
M2I-1, also known as Mad2 Inhibitor-1, is a novel class of small molecule inhibitor that inhibits Mad2's binding to Cdc20, a crucial protein-protein interaction (PPI) that occurs within the mitotic spindle assembly checkpoint (SAC). The disruption of Mad2's conformational dynamics is crucial for the formation of complexes with Cdc20. In FP experiments, it can also inhibit the Mad2-F-Mbp1 interaction.
Fluorescence polarization-based binding assay for Mad2-Cdc20 interaction: Recombinant Mad2 and Cdc20 proteins were prepared; fluorescently labeled Cdc20 (or its functional fragment) was incubated with Mad2 in the presence of serial concentrations of M2I-1 (or vehicle control); fluorescence polarization signals were measured to quantify the binding affinity between Mad2 and Cdc20, and the inhibitory effect of M2I-1 on this interaction was determined [1] - Conformational dynamics analysis of Mad2: Biochemical assays (e.g., circular dichroism spectroscopy, limited proteolysis) and molecular docking simulations were performed to evaluate the effect of M2I-1 on Mad2 conformational changes; these assays assessed the structural alterations of Mad2 induced by M2I-1 that prevent its binding to Cdc20 [1] |
||
| Cell Assay |
M2I-1 was found to interfere with Mad2's conformational dynamics, which is essential for the formation of complexes with Cdc20, based on biochemical and computational analyses. M2I-1 was found to reduce the SAC response in cellular studies, suggesting that the substance may be active in cells.
SAC response assay in HeLa cells: HeLa cells were treated with M2I-1 (or DMSO as vehicle control) at different concentrations; the cells were synchronized in mitosis (e.g., via nocodazole treatment to induce spindle damage and activate SAC); the duration of mitotic arrest was monitored by time-lapse microscopy or flow cytometry (to detect mitotic markers such as phospho-histone H3); the attenuation of SAC-mediated mitotic arrest in M2I-1-treated cells was quantified to evaluate the compound’s effect on SAC function [1] |
||
| Animal Protocol |
|
||
| References | |||
| Additional Infomation |
M2I-1 is the first small molecule inhibitor targeting the Mad2-Cdc20 protein-protein interaction, a key regulatory step in the spindle assembly checkpoint (SAC)[1]. The spindle assembly checkpoint (SAC) is a cellular surveillance mechanism that ensures the correct separation of chromosomes during mitosis by blocking the process of mitosis when chromosomes are incorrectly connected to the spindle; SAC dysregulation is associated with genomic instability and cancer[1]. M2I-1 exerts its inhibitory effect by interfering with the conformational kinetics of Mad2, which is necessary for its functional interaction with Cdc20; this mechanism makes M2I-1 a valuable pharmacological tool for elucidating SAC function and a promising lead compound for the development of anticancer therapies (because SAC dysfunction is associated with tumorigenesis)[1].
|
| Molecular Formula |
C19H24N4O4S
|
|
|---|---|---|
| Molecular Weight |
404.48
|
|
| Exact Mass |
404.151
|
|
| Elemental Analysis |
C, 56.42; H, 5.98; N, 13.85; O, 15.82; S, 7.93
|
|
| CAS # |
312271-03-7
|
|
| Related CAS # |
|
|
| PubChem CID |
2883762
|
|
| Appearance |
Solid powder
|
|
| Density |
1.3±0.1 g/cm3
|
|
| Index of Refraction |
1.622
|
|
| LogP |
5.02
|
|
| Hydrogen Bond Donor Count |
2
|
|
| Hydrogen Bond Acceptor Count |
6
|
|
| Rotatable Bond Count |
6
|
|
| Heavy Atom Count |
28
|
|
| Complexity |
639
|
|
| Defined Atom Stereocenter Count |
0
|
|
| SMILES |
O=C(/C(C(N1)=O)=C\C2=CC=C(N(CC(C)C)CC(C)C)C([N+]([O-])=O)=C2)NC1=S
|
|
| InChi Key |
BWEKPQUKWLNUKX-UHFFFAOYSA-N
|
|
| InChi Code |
InChI=1S/C19H24N4O4S/c1-11(2)9-22(10-12(3)4)15-6-5-13(8-16(15)23(26)27)7-14-17(24)20-19(28)21-18(14)25/h5-8,11-12H,9-10H2,1-4H3,(H2,20,21,24,25,28)
|
|
| Chemical Name |
5-[[4-[bis(2-methylpropyl)amino]-3-nitrophenyl]methylidene]-2-sulfanylidene-1,3-diazinane-4,6-dione
|
|
| 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) |
DMSO: ~80 mg/mL ( 197.78 mM)
Water: Insoluble Ethanol: Insoluble |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.18 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 (6.18 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: 10% DMSO+40% PEG300+5% Tween-80+45% saline：≥ 2.5 mg/mL (~6.2 mM) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4723 mL | 12.3616 mL | 24.7231 mL | |
| 5 mM | 0.4945 mL | 2.4723 mL | 4.9446 mL | |
| 10 mM | 0.2472 mL | 1.2362 mL | 2.4723 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.
ACS Chem Biol.2015 Jul 17;10(7):1661-6. |
|---|
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
