| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| Other Sizes |
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| Targets |
WDR5 (Ki < 1 nM); WDR5-MLL1 interaction (IC50 = 0.9 nM); MLL1 (IC50 = 0.32 µM)
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|---|---|
| ln Vitro |
MM-401 inhibits the WDR5-MLL1 interaction with an IC50 value of 0.9 nM while maintaining a strong binding affinity to WDR5 with a Ki value of < 1 nM[1]. By preventing the MLL1-WDR5 interaction and consequently the complex assembly, MM-401 can specifically inhibit MLL1 activity (IC50 value of 0.32µM)[1]. MLL1-dependent H3K4 methylation in cells is particularly inhibited by MM-401 (20 μM; 48 h)[1]. Similar alterations to the MLL-AF9 transcriptome are induced by MM-401 as by the MLL1 deletion[1]. MM-401 (10, 20, 40 μM; 48 h) specifically prevents MLL leukemia cells from growing by causing apoptosis and cell cycle arrest[1].
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| Enzyme Assay |
Histone Methytransferase Assays[1]
The HMT assay was performed as described previously (Dou et al., 2005). For inhibitor studies, compounds at various concentrations were incubated first with the pre-assembled complex and reactions were initiated by addition of substrates. For kinetic analyses, the reaction progression curve was established to determine the linear range of the reaction at room temperature. For Lineweaver-Burk curve, reactions (0.5µM enzyme complex and 50µM substrates) were initiated and quenched after 4 minutes by addition of β-mercaptoethanol at a final concentration of 178µM. Crystal Structures[1] WDR5//MM-401 or WDR5/MM-NC-401 binary complex was obtained by mixing WDR5 and compounds at molar ratio 1: 2. The complex was crystallized by hanging-drop-vapor-diffusion at 22°C. Details see supplemental information. |
| Cell Assay |
Apoptosis Analysis[1]
Cell Types: Murine MLL-AF9 and Hoxa9/Meis1 cells Tested Concentrations: 10, 20, 40 μM Incubation Duration: 48 h Experimental Results: Specifically induced apoptosis of MLL-AF9 cells. Cell Cycle Analysis[1] Cell Types: Murine MLL-AF9 and Hoxa9/Meis1 cells Tested Concentrations: 10, 20, 40 μM Incubation Duration: 48 h Experimental Results: Induced prominent G1/S arrest in MLL-AF9 cells in a concentration dependent manner. RT-PCR[1] Cell Types: MLL-AF9 cells Tested Concentrations: 20 μM Incubation Duration: 48 h Experimental Results: Dramatically diminished H3K4me, expression of 5 Hox A genes, especially Hoxa9 and Hoxa10. Assays for Cell Viability, Wright-Giemsa staining, apoptosis, cell cycle and cell differentiation [1] Inhibitors were diluted from stock to culture media containing 0.1% DMSO final concentration. For viability assays, cells were cultured at 1×105/ml and passaged every 2 days. Viability was determined using the CellTitreGlo® Kit according to the manufacturer’s directions. Luminescence was monitored on a Molecular Dynamics plate reader. For staining, cells treated with 10, 20 and 40µM /MM-401, or 40µM MM-NC-401 or DMSO vehicle for 4 days were diluted to 2.5×105/ml in 1× PBS and fixed to glass slides by cytospin followed by Wright-Giemsa staining. Cell images were taken at 40× magnification by light microscopy. Apoptosis, cell cycle and cell differentiation analyses were performed using standard protocols (see supplemental information). Real Time-PCR, RNA-seq and CHIP assays [1] MLL1-AF9 cells were cultured for 2 days in the presence of /MM-401 or MM-NC-401. At the end of treatment, cells were harvested by centrifugation at 300×g and washed with 1xPBS. RNAs for duplicated biological samples were extracted by a standard protocol. 10ng RNAs were used for Illumina sequencing library. Four RNA seq samples were multiplexed and loaded into one lane in Hi-seq sequencer. RNA-seq analyses were described in supplemental information. CHIP assays were performed as previously described. |
| References | |
| Additional Infomation |
Here we report a comprehensive characterization of our recently developed inhibitor MM-401 that targets the MLL1 H3K4 methyltransferase activity. MM-401 is able to specifically inhibit MLL1 activity by blocking MLL1-WDR5 interaction and thus the complex assembly. This targeting strategy does not affect other mixed-lineage leukemia (MLL) family histone methyltransferases (HMTs), revealing a unique regulatory feature for the MLL1 complex. Using MM-401 and its enantiomer control MM-NC-401, we show that inhibiting MLL1 methyltransferase activity specifically blocks proliferation of MLL cells by inducing cell-cycle arrest, apoptosis, and myeloid differentiation without general toxicity to normal bone marrow cells or non-MLL cells. More importantly, transcriptome analyses show that MM-401 induces changes in gene expression similar to those of MLL1 deletion, supporting a predominant role of MLL1 activity in regulating MLL1-dependent leukemia transcription program. We envision broad applications for MM-401 in basic and translational research. [1]
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| Molecular Formula |
C31H47F3N8O7
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|---|---|
| Molecular Weight |
700.749497652054
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| Exact Mass |
700.351
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| CAS # |
1442106-11-7
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| Related CAS # |
MM-401;1442106-10-6
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| PubChem CID |
137662115
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| Appearance |
White to light yellow solid powder
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| Hydrogen Bond Donor Count |
8
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
49
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| Complexity |
1060
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| Defined Atom Stereocenter Count |
4
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| SMILES |
C(F)(F)(F)C(=O)O.O=C1NCCCC[C@@](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC)C(=O)N[C@]1([H])C1C=CC=CC=1)(C)NC(=O)C(C)C
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| InChi Key |
UCPNHHHBFDAYBP-HXXCMCGZSA-N
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| InChi Code |
InChI=1S/C29H46N8O5.C2HF3O2/c1-5-20-24(39)36-22(19-12-7-6-8-13-19)26(41)32-16-10-9-15-29(4,37-23(38)18(2)3)27(42)35-21(25(40)34-20)14-11-17-33-28(30)31;3-2(4,5)1(6)7/h6-8,12-13,18,20-22H,5,9-11,14-17H2,1-4H3,(H,32,41)(H,34,40)(H,35,42)(H,36,39)(H,37,38)(H4,30,31,33);(H,6,7)/t20-,21-,22+,29+;/m0./s1
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| Chemical Name |
N-[(3R,6S,9S,12R)-9-[3-(diaminomethylideneamino)propyl]-6-ethyl-12-methyl-2,5,8,11-tetraoxo-3-phenyl-1,4,7,10-tetrazacyclohexadec-12-yl]-2-methylpropanamide;2,2,2-trifluoroacetic acid
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| Synonyms |
MM-401 TFA; 1442106-11-7; MM-401 (TFA); CHEMBL4099772;
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 (142.70 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.57 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 (3.57 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 (3.57 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 | 1.4270 mL | 7.1352 mL | 14.2704 mL | |
| 5 mM | 0.2854 mL | 1.4270 mL | 2.8541 mL | |
| 10 mM | 0.1427 mL | 0.7135 mL | 1.4270 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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