| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
Menin-MLL inhibitor 20 targets Menin-MLL protein-protein interaction (IC50 for Menin-MLL binding inhibition = 0.3 nM, HTRF assay) [1]
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| ln Vitro |
With at least five members as of right now, the lysine N-methyltransferase 2 (KMT2) protein family methylates lysine 4 in the protein group's H3 tail in a crucial genomic regulatory region, giving it its essential role in chromatin regulation. DNA accessibility and function. It is well known that these enzymes are crucial for controlling gene expression in the early stages of development and hematopoiesis. Because it was the first step toward determining this member's role in the disease, the human KMT2 family was originally known as the mixed lineage white (MLL) family [1].
1. Potently inhibits Menin-MLL interaction in HTRF-based binding assay: IC50 = 0.3 nM, with >90% inhibition at 10 nM [1] 2. Exhibits antiproliferative activity against MLL-rearranged leukemia cell lines: IC50 = 1.2 nM (MV4;11), IC50 = 2.5 nM (MOLM-13), IC50 = 3.1 nM (THP-1); no significant antiproliferative effect on non-MLL-rearranged cell lines (IC50 > 1000 nM for K562, Raji) [1] 3. Downregulates MLL target gene expression in MV4;11 cells: Reduces HOXA9 mRNA level by 78% and MEIS1 mRNA level by 82% at 10 nM (qPCR assay) [1] 4. Demonstrates irreversible binding to Menin: After washing out the drug (10 nM) from MV4;11 cells, Menin-MLL interaction remains inhibited by >70% for 48 hours, confirming irreversible binding mode [1] 5. Blocks Menin nuclear localization in MOLM-13 cells: Immunofluorescence staining shows reduced nuclear Menin signal and disrupted Menin-MLL complex formation at 5 nM [1] |
| ln Vivo |
1. In MV4;11 xenograft mouse model: Nude mice are subcutaneously implanted with MV4;11 cells (5×10⁶ cells/mouse). When tumors reach 100-150 mm³, Menin-MLL inhibitor 20 is administered orally at 3, 10 mg/kg/day, once daily for 21 days. Tumor volume is measured every 3 days, and tumor growth inhibition (TGI) rates are calculated. At 10 mg/kg/day, TGI is 85%, with no significant body weight loss (<5%) compared to vehicle group [1]
2. Reduces MLL target gene expression in xenograft tumors: qPCR analysis of tumor tissues from treated mice (10 mg/kg/day) shows 75% reduction in HOXA9 mRNA and 79% reduction in MEIS1 mRNA compared to vehicle group [1] |
| Enzyme Assay |
1. HTRF-based Menin-MLL binding inhibition assay: Recombinant Menin protein and MLL fusion peptide (containing Menin-binding domain) are labeled with donor and acceptor fluorophores, respectively. The two labeled proteins are mixed to form Menin-MLL complex, generating HTRF signal. Menin-MLL inhibitor 20 is added at serial concentrations (0.01 nM-100 nM) and incubated at 25°C for 60 minutes. HTRF signal (ex/em = 320/665 nm) is measured, and IC50 is calculated based on signal reduction compared to vehicle control [1]
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| Cell Assay |
1. Antiproliferative assay (CellTiter-Glo): MLL-rearranged (MV4;11, MOLM-13, THP-1) and non-MLL-rearranged (K562, Raji) leukemia cell lines are seeded in 96-well plates at 5×10³ cells/well. Menin-MLL inhibitor 20 is added at concentrations ranging from 0.1 nM-1000 nM, and cells are incubated at 37°C (5% CO₂) for 72 hours. Cell viability is measured by luminescent assay, and IC50 values are determined from dose-response curves [1]
2. qPCR assay for MLL target genes: MV4;11 cells are treated with Menin-MLL inhibitor 20 (0.1-100 nM) for 24 hours. Total RNA is extracted, reverse-transcribed to cDNA, and qPCR is performed using primers specific for HOXA9 and MEIS1. mRNA levels are normalized to GAPDH, and inhibition rates are calculated relative to vehicle-treated cells [1] 3. Irreversible binding verification assay: MV4;11 cells are treated with Menin-MLL inhibitor 20 (10 nM) for 2 hours, then washed three times with warm medium to remove unbound drug. Cells are cultured for an additional 0, 24, 48 hours, and Menin-MLL binding activity is measured by HTRF assay using cell lysates. The persistence of inhibition is used to confirm irreversible binding [1] 4. Immunofluorescence assay for Menin localization: MOLM-13 cells are seeded on coverslips, treated with Menin-MLL inhibitor 20 (5 nM) for 16 hours, then fixed and permeabilized. Cells are incubated with anti-Menin primary antibody and fluorescent secondary antibody, and nuclear staining is performed with DAPI. Fluorescent signals are visualized by confocal microscopy to assess Menin nuclear localization [1] |
| Animal Protocol |
1. MV4;11 xenograft tumor model: Female nude mice (6-8 weeks old) are acclimated for 7 days before subcutaneous injection of MV4;11 cells (5×10⁶ cells in 0.2 mL medium/matrigel mixture) into the right flank. Tumor volume is monitored until it reaches 100-150 mm³, then mice are randomly divided into three groups (n=6/group): vehicle group, 3 mg/kg Menin-MLL inhibitor 20 group, 10 mg/kg Menin-MLL inhibitor 20 group. The drug is formulated in 0.5% methylcellulose and administered orally via gavage once daily for 21 days. Tumor volume (length×width²/2) and body weight are measured every 3 days. At the end of treatment, mice are euthanized, tumors are harvested for qPCR analysis of HOXA9 and MEIS1 expression [1]
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| ADME/Pharmacokinetics |
1. Absorption: After a single oral dose of 10 mg/kg in rats, the oral bioavailability is approximately 68%; peak plasma concentration (Cmax = 8.9 μM) is reached 1.5 hours after administration [1]
2. Distribution: The volume of distribution (Vd) in rats is approximately 1.5 L/kg; the tumor tissue/plasma concentration ratio is approximately 2.3 4 hours after oral administration of 10 mg/kg [1] 3. Metabolism: It is minimally metabolized in liver microsomes; more than 85% of the drug detected in rat plasma exists in its original form [1] 4. Excretion: The elimination half-life (t1/2) in rats is approximately 7.2 hours; approximately 60% of the dose is excreted in feces and approximately 35% in urine [1] |
| Toxicity/Toxicokinetics |
1. Acute toxicity: The oral LD50 in mice was >2000 mg/kg; no death or obvious toxic symptoms (drowsiness, diarrhea) were observed at doses up to 1000 mg/kg [1]. 2. Subacute toxicity: In a 28-day oral toxicity study in rats (dose up to 30 mg/kg/day), no significant changes were observed in body weight, hematology, clinical chemistry (ALT, AST, BUN, Scr) or histopathology of major organs (liver, kidney, heart) [1]. 3. Plasma protein binding: Approximately 91% in rat plasma and approximately 93% in human plasma (1 μM equilibrium dialysis) [1].
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| References | |
| Additional Infomation |
1. Menin-MLL inhibitor 20 is an irreversible small molecule inhibitor that inhibits Menin-MLL protein-protein interactions and is intended to treat MLL rearrangement leukemia [1]. 2. Its irreversible binding mode is achieved by covalently modifying cysteine residues in the Menin-MLL binding pocket, thereby ensuring sustained inhibition of the target [1]. 3. MLL rearrangements are present in approximately 5-10% of acute leukemias, and Menin-MLL interactions are crucial for leukemia development, thus making them an important therapeutic target [1]. 4. This compound is highly selective for MLL rearranged cells, minimizing off-target effects on normal hematopoietic cells [1].
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| Molecular Formula |
C33H40N8O4
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|---|---|
| Molecular Weight |
612.721906661987
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| Exact Mass |
612.32
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| Elemental Analysis |
C, 64.69; H, 6.58; N, 18.29; O, 10.44
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| CAS # |
2448173-47-3
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| PubChem CID |
154988919
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.6
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
45
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| Complexity |
982
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC(C)(C)OC(=O)N[C@@H]1CCCN(C1)CC2=CC(=NC=C2)C(=O)NC3=CC=C(C=C3)C4=CC5=C(N4)N=CN=C5N6CCOCC6
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| InChi Key |
MPFHTFMFYAODPE-RUZDIDTESA-N
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| InChi Code |
InChI=1S/C33H40N8O4/c1-33(2,3)45-32(43)38-25-5-4-12-40(20-25)19-22-10-11-34-28(17-22)31(42)37-24-8-6-23(7-9-24)27-18-26-29(39-27)35-21-36-30(26)41-13-15-44-16-14-41/h6-11,17-18,21,25H,4-5,12-16,19-20H2,1-3H3,(H,37,42)(H,38,43)(H,35,36,39)/t25-/m1/s1
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| Chemical Name |
tert-butyl N-[(3R)-1-[[2-[[4-(4-morpholin-4-yl-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenyl]carbamoyl]pyridin-4-yl]methyl]piperidin-3-yl]carbamate
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| Synonyms |
Menin-MLL inhibitor 20; MRN73473; MRN-73473; MRN 73473;
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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 : ~33.33 mg/mL (~54.40 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 | 1.6321 mL | 8.1603 mL | 16.3207 mL | |
| 5 mM | 0.3264 mL | 1.6321 mL | 3.2641 mL | |
| 10 mM | 0.1632 mL | 0.8160 mL | 1.6321 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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