| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
WDR5 (WD repeat domain 5 protein) – MLL (Mixed Lineage Leukemia) protein-protein interaction. [1]
Binding affinity to WDR5: IC50 = 0.90 ± 0.20 nM, Ki < 1 nM (determined by a fluorescence polarization-based competitive binding assay). [1] IC50: 0.90 nM (WDR5), 12.7 nM (HMT)[1] Ki: <1 nM (WDR5)[1] |
|---|---|
| ln Vitro |
- MLL HMT Activity Inhibition: MM-589 potently inhibits the histone H3 lysine 4 (H3K4) methyltransferase (HMT) activity of the MLL core complex in a cell-free AlphaLISA functional assay, with an IC50 value of 12.7 ± 1.5 nM. [1]
- Enzymatic Selectivity: In a previously published HMT assay, compound MM-589 effectively inhibits MLL HMT activity (IC50 = 12.7 nM) but shows no or minimal effect (up to 100 μM) on the HMT activity of other SET1 family members, including MLL2, MLL3, MLL4, SET1a, and SET1b. [1] - Cell Growth Inhibition in MLL-translocated Leukemia Cells: MM-589 potently and selectively inhibits the cell growth of human acute leukemia cell lines harboring MLL translocations. [1] - In MOLM-13 cells (harboring MLL-AF9 fusion), the IC50 for cell growth inhibition is 0.21 ± 0.02 μM after a 4-day treatment (with 10,000 cells/well in 96-well plates). [1] - In MV4-11 cells (harboring MLL-AF4 fusion), the IC50 for cell growth inhibition is 0.25 ± 0.01 μM after a 4-day treatment (with 10,000 cells/well in 96-well plates). [1] - The compound displays >30-fold selectivity for these MLL-translocated cell lines over the HL-60 leukemia cell line (lacking MLL translocation), which has an IC50 of 8.56 ± 1.14 μM. [1] - Long-term Treatment Effect: With a 7-day treatment (starting with 50,000 cells/mL in 24-well plates, with re-dosing on day 4), MM-589 achieves >95% maximum cell growth inhibition in both MOLM-13 and MV4-11 cell lines, compared to approximately 75% maximum inhibition with a 4-day treatment. The IC50 values improve by 2-3 times with the longer treatment. [1] - Stereospecificity: The enantiomer of MM-589 (Compound 19, where the chiral center in the phenylglycine residue is changed from D- to L-configuration) has an IC50 of 14.3 μM and 35.0 μM in MOLM-13 and MV4-11 cells, respectively, making it >50-100 times less potent than MM-589. [1] In human leukemia cell lines carrying the MLL translocation, MM-589 (0.01-10 μM, 4 or 7 days) potently and specifically suppresses cell proliferation [1]. |
| ln Vivo |
MM-589 demonstrates in vivo anti-leukemic activity in animal models. MM-589 derivatives administered by intravenous injection effectively suppress tumor cell expansion and prolong survival in mouse models of MLL leukemia. Studies indicate that MLL-WDR5 interaction inhibitor derivatives with improved pharmacokinetic properties have the potential to be trialed in cancer patients. Compared to MM-401, MM-589 exhibits superior pharmacodynamic profiles in in vivo studies due to its improved cellular permeability and metabolic stability. No significant toxicity to normal hematopoietic function has been observed during treatment.
|
| Enzyme Assay |
- Fluorescence Polarization (FP)-based Competitive Binding Assay for WDR5: To determine binding affinities of compounds to WDR5, an FP-based competitive binding assay was used. 5 μL of test compound solution in DMSO was added to 120 μL of a preincubated complex solution containing N-terminal His-tagged WDR5 protein (residues 24-334) and a S-FAM labeled tracer in assay buffer (0.1 M phosphate, 25 mM KCl, 0.01% Triton, pH 6.5). The final concentrations of WDR5 and tracer were 4 nM and 0.6 nM, respectively. The plate was incubated at room temperature on a shaker for 3 hours, after which the mP values were measured using a plate reader. Ki values were calculated using a previously described equation. [1]
- Cell-free AlphaLISA MLL HMT Functional Assay: The inhibition of MLL HMT activity was evaluated using a homogeneous AlphaLISA assay. The recombinant MLL core complex (containing MLL, WDR5, ASH2L, RbBP5, DPY30) and recombinant nucleosomes were used. Serial dilutions of the compound (2.5 μL) in assay buffer with 4% DMSO and 5 μL of MLL complex solution were added to a 384-well plate and incubated for 30 minutes with gentle shaking. Then, 2.5 μL of a SAM/Nucleosome mixture was added, with final concentrations of MLL complex, SAM, and nucleosomes being 5 nM, 200 nM, and 3 nM, respectively, in 1% DMSO. The methylation reaction proceeded for 120 minutes in the dark. The reaction was stopped by adding 5 μL of a high-salt stopping solution for 15 minutes. Subsequently, 5 μL of a mixture containing anti-H3K4me1/2 AlphaLISA acceptor beads and biotinylated anti-H3 (C-terminus) antibody in detection buffer was added and incubated for 1 hour. Finally, 5 μL of streptavidin-labeled donor beads were added and incubated for 30 minutes. The plate was read using a microplate reader with excitation at 680 nm and emission at 615 nm. IC50 values were determined by fitting fluorescence intensities versus compound concentrations using a sigmoidal dose-response curve in Graphpad Prism software. [1] |
| Cell Assay |
- Cell Viability Assay (4-day treatment): To assess cell growth inhibition, MOLM-13, MV4-11, or HL-60 leukemia cells were seeded at 1 x 10⁴ cells per well in 96-well plates. Cells were treated with various concentrations of the test compound (MM-589 or others) for 4 days. The final concentration of DMSO in the culture media was 0.2%. Cell viability was determined using the WST-8 cell proliferation assay kit, following the manufacturer's instructions. The IC50 values (50% cell growth inhibition) were calculated relative to the DMSO control using Prism software. Experiments were performed three times in triplicate. [1]
- Long-term Cell Viability Assay (7-day treatment): MOLM-13 and MV4-11 cell lines were plated at a density of 5 x 10⁴ cells/mL in 24-well plates (2 mL/well) and treated with relevant concentrations of MM-589. On day 4, cell viability was measured using the WST-8 assay. Then, 10% of viable cells from each well were transferred to freshly prepared medium containing corresponding concentrations of the compound and cultured for an additional 3 days. On day 7, cell viability for each treatment was determined. [1] Cell Viability Assay[1] Cell Types: MV4-11 and MOLM-13 cells Tested Concentrations: 0.01, 0.1, 1, 10 μM Incubation Duration: 4 days or 7 days Experimental Results: Potently inhibited MV4-11 and MOLM-13 cell growth with IC50s of 0.25 and 0.21 μM, respectively. Had much weaker activity in the inhibition of cell growth of the HL-60 cell line with an IC50 of 8.6 μM. |
| Animal Protocol |
Animal Models: Establish disseminated leukemia models by intravenous injection of MLL-rearranged leukemia cells (e.g., MLL-AF9 cells) into immunodeficient mice (NSG or NOD/SCID mice) via the tail vein.
Dosing Regimen: Administer MM-589 by intravenous injection, typically once daily for 2-3 consecutive weeks. Due to the improved solubility of the TFA salt form of MM-589, it should be formulated in appropriate vehicle buffers.
Efficacy Assessment: Monitor tumor burden by bioluminescent imaging, record survival (Kaplan-Meier analysis), and detect the percentage of leukemia cells (hCD45⁺) in peripheral blood and bone marrow by flow cytometry.
Toxicity Assessment: Monitor animal body weight changes, behavioral performance, and peripheral blood cell counts to evaluate effects on normal hematopoietic function.
Pharmacodynamic Analysis: Collect bone marrow or spleen samples to detect H3K4me3 levels and target gene expression such as HOXA9.
Data Analysis: Compare survival differences and tumor burden changes between treatment and control groups.
|
| ADME/Pharmacokinetics |
- Microsomal Stability: The metabolic stability of MM-589 was evaluated in liver microsomes from different species. It demonstrated excellent stability, with a half-life (T₁/₂) greater than 60 minutes in human, mouse, and rat liver microsomes. [1]
As a macrocyclic peptidomimetic, MM-589 exhibits significantly improved metabolic stability and cellular permeability through a cyclization strategy compared to the linear peptide MM-101. This compound has good solubility in DMSO. The TFA salt form has improved solubility in aqueous buffers, making it more suitable for in vivo administration. MM-589 powder is stable for 2-3 years when stored at -20°C or 4°C protected from light; solutions are stable for 6 months at -80°C protected from light. MM-589 represents the most potent inhibitor of the WDR5-MLL interaction reported to date, and further optimization may yield a new therapy for acute leukemia. |
| Toxicity/Toxicokinetics |
According to the Safety Data Sheet, MM-589 is not classified as a hazardous substance or mixture, and GHS label elements do not apply. Toxicological testing: Not listed as a carcinogen by NTP; not listed by IARC; not regulated by OSHA; not listed by ACGIH. At the cellular level, MM-589 shows no significant toxicity to normal bone marrow cells, selectively inhibiting the proliferation of MLL-rearranged leukemia cells. The toxicological effects of this product have not been thoroughly studied. Standard laboratory safety practices should be followed when handling, and release to the environment should be avoided. MM-589 has a purity of ≥98% and is for research use only, not for human or veterinary applications.
|
| References | |
| Additional Infomation |
- MM-589 corresponds to Compound 18 in the study. It was synthesized via a method involving solid-phase peptide synthesis, ring-closing metathesis (RCM), catalytic hydrogenation, and removal of protecting groups. The overall synthetic route is detailed in Schemes 4 and 5 of the paper. [1]
- A cocrystal structure of MM-589 (18) in complex with WDR5 was determined at a resolution of 1.64 Å (PDB code 5VFC). The structure shows that MM-589 binds to the central channel of the WD40 propeller in WDR5 through a conserved network of hydrogen bonds and hydrophobic interactions. The smaller ring size of MM-589 (n=2) compared to earlier analogs (n=4 for compound 2, n=6 for compound 16) forms a more compact structure, facilitating optimal intramolecular hydrogen bonds and reducing conformational flexibility, which contributes to its high binding affinity. [1] - The study found a good correlation between the potencies of compounds in inhibiting MLL HMT activity (IC50) and inhibiting cell growth (IC50) in MV4-11 and MOLM-13 cell lines. [1] - The compound is stable in cell culture media for up to 7 days. [1] |
| Molecular Formula |
C30H45F3N8O7
|
|---|---|
| Molecular Weight |
686.722917318344
|
| Exact Mass |
686.336
|
| Elemental Analysis |
C, 52.47; H, 6.61; F, 8.30; N, 16.32; O, 16.31
|
| CAS # |
2253167-09-6
|
| Related CAS # |
MM-589 (racemic mixture ) (TFA);MM-589;2097887-20-0
|
| PubChem CID |
132585206
|
| Sequence |
isobutyryl-D-aMeLys(1)-Arg-Abu-D-Phg-(1).TFA
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
8
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
48
|
| Complexity |
1050
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
C(F)(F)(F)C(=O)O.O=C1NCC[C@@](C(N[C@@H](CCCNC(=N)NC)C(=O)N[C@@H](CC)C(=O)N[C@@H]1C1C=CC=CC=1)=O)(C)NC(=O)C(C)C
|
| InChi Key |
NARXNZHWAWPJIY-HTCBAMNXSA-N
|
| InChi Code |
InChI=1S/C28H44N8O5.C2HF3O2/c1-6-19-23(38)35-21(18-11-8-7-9-12-18)25(40)31-16-14-28(4,36-22(37)17(2)3)26(41)34-20(24(39)33-19)13-10-15-32-27(29)30-5;3-2(4,5)1(6)7/h7-9,11-12,17,19-21H,6,10,13-16H2,1-5H3,(H,31,40)(H,33,39)(H,34,41)(H,35,38)(H,36,37)(H3,29,30,32);(H,6,7)/t19-,20-,21+,28+;/m0./s1
|
| Chemical Name |
N-[(3R,6S,9S,12R)-6-ethyl-12-methyl-9-[3-[(N'-methylcarbamimidoyl)amino]propyl]-2,5,8,11-tetraoxo-3-phenyl-1,4,7,10-tetrazacyclotetradec-12-yl]-2-methylpropanamide;2,2,2-trifluoroacetic acid
|
| Synonyms |
MM-589 (TFA); MM589 TFA; MM 589 TFA; 2253167-09-6; 2097887-21-1; N-[(3R,6S,9S,12R)-6-ethyl-12-methyl-9-[3-[(N'-methylcarbamimidoyl)amino]propyl]-2,5,8,11-tetraoxo-3-phenyl-1,4,7,10-tetrazacyclotetradec-12-yl]-2-methylpropanamide;2,2,2-trifluoroacetic acid;
|
| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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: ~100 mg/mL (145.6 mM)
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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 | 1.4562 mL | 7.2810 mL | 14.5620 mL | |
| 5 mM | 0.2912 mL | 1.4562 mL | 2.9124 mL | |
| 10 mM | 0.1456 mL | 0.7281 mL | 1.4562 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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