| Targets |
Menin (a scaffold protein that interacts with the mixed-lineage leukemia (MLL) protein). BMF-219 is a specific, irreversible inhibitor of the Menin-MLL interaction.
|
|---|---|
| ln Vitro |
In cell-free binding assays, BMF-219 covalently binds to Menin, irreversibly inhibiting the Menin-MLL interaction. The compound has an EC₅0 of 33 nM in human cells. By blocking the Menin-MLL interaction, BMF-219 disrupts the leukemogenic transcriptional program driven by MLL fusion proteins. The specific biochemical IC₅0 values are not provided in the available data. In cellular assays, BMF-219 inhibits the proliferation of Menin-MLL-dependent leukemia cells and induces differentiation of these cells.
|
| Enzyme Assay |
A general cell-free protocol for assessing Menin-MLL inhibition: A time-resolved fluorescence resonance energy transfer (TR-FRET) or AlphaLISA assay is used to measure the interaction between Menin and the MLL peptide. Recombinant His-tagged Menin protein (10 nM) is incubated with a biotinylated MLL peptide (10 nM, corresponding to the MLL binding site) in assay buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.01% Tween-20, 1 mM DTT, 0.1% BSA). Varying concentrations of BMF-219 (0.01 nM to 10 uM) are added. After a 30-minute pre-incubation, europium-labeled anti-His antibody (2 nM) and streptavidin-labeled XL665 (20 nM) are added. The TR-FRET signal is measured after 1 hour. The IC₅0 for inhibition of the Menin-MLL interaction is calculated. For the determination of irreversibility, a dilution or washout experiment is performed. Menin protein is pre-incubated with BMF-219 for 1 hour, then the mixture is diluted 100-fold or the compound is removed by gel filtration, and the residual Menin-MLL interaction is measured. No recovery of activity indicates irreversible inhibition.
|
| Cell Assay |
A general cellular protocol for assessing Menin inhibition and MLL-fusion protein activity: Menin-MLL-dependent leukemia cells (e.g., MLL-AF9-transformed cells or MV4-11 cells) are seeded in 12-well plates at 2×10⁵ cells/well in RPMI-1640 medium containing 10% FBS. The cells are treated with various concentrations of BMF-219 (0.1, 0.5, 1, 5, 10, 25, 50, 100 nM) for 48-72 hours. Cell viability is assessed using the MTT or CellTiter-Glo assay. For differentiation analysis, cells are treated with BMF-219 (1-10 nM) for 5-7 days, and cell surface markers of differentiation (e.g., CD11b, CD14) are analyzed by flow cytometry. For apoptosis analysis, cells are stained with Annexin V-FITC and PI and analyzed by flow cytometry. For gene expression analysis, total RNA is extracted, and the expression of Menin-MLL target genes (e.g., HOXA9, MEIS1, FLT3) is measured by qRT-PCR. GAPDH is used as a loading control. For Western blot analysis, cells are treated with BMF-219 (1-100 nM) for 24 hours, and the expression of MLL fusion protein target proteins (e.g., HOXA9, MEIS1) is measured.
|
| Animal Protocol |
A general animal protocol for evaluating BMF-219 in a Menin-MLL-driven leukemia xenograft model: Female NOD/SCID mice are injected intravenously with 1×10⁶ MLL-AF9-transformed or MV4-11 human leukemia cells. After 7 days, when the leukemia is established (monitored by bioluminescence imaging if the cells are labeled with luciferase), the mice are randomized into treatment groups (n=10 per group). BMF-219 is formulated in a suitable vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) and administered via oral gavage at doses of 10, 30, and 100 mg/kg once daily for 28 days. A vehicle control group and a positive control group (e.g., menin inhibitor MI-503 at 100 mg/kg) are included. Leukemia progression is monitored weekly by bioluminescence imaging, by measuring the percentage of human CD45+ cells in the peripheral blood by flow cytometry, and by spleen palpation. At the end of the study, the mice are euthanized. The spleen and liver are weighed. The spleen, liver, and bone marrow are collected for flow cytometry analysis (human CD45+, hCD33+, hCD34+). The bone marrow cells are also used for RNA extraction and qRT-PCR analysis of HOXA9 and MEIS1. Histopathological examination of the liver and spleen is performed (H&E staining).
|
| ADME/Pharmacokinetics |
General pharmacokinetic protocol for BMF-219: Male Sprague-Dawley rats are administered BMF-219 via oral gavage (PO, 10 mg/kg) and intravenous (IV, 2 mg/kg) injection. The compound is formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. Plasma concentrations of BMF-219 are quantified by LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t½, clearance, Vd, and oral bioavailability) are calculated using non-compartmental analysis. BMF-219 is reported to be orally active, so it is expected to have good bioavailability. The molecular formula of BMF-219 is C31H34N₈O3, and the molecular weight is 566.65.
|
| Toxicity/Toxicokinetics |
General toxicity protocol for BMF-219: A 14-day repeated-dose oral toxicity study is performed in Sprague-Dawley rats. BMF-219 is administered via oral gavage at doses of 10, 30, and 100 mg/kg/day for 14 consecutive days. Clinical signs, body weight, and food consumption are monitored daily. At the end of the study, blood samples are collected for hematology (complete blood count with differential) and serum chemistry (ALT, AST, ALP, BUN, creatinine, total protein, albumin, glucose, electrolytes). Gross necropsy is performed, and the weights of major organs (liver, kidney, spleen, heart, lung, brain, testes) are recorded. Histopathological examination of these organs is conducted. Bone marrow smears are prepared to assess myelosuppression. Special attention is given to the gastrointestinal tract and bone marrow, as these are rapidly proliferating tissues that may be affected by inhibition of the Menin-MLL pathway.
|
| References | |
| Additional Infomation |
Icovamenib is an oral, highly bioavailable, irreversible menin inhibitor. Menin is an important cofactor of the menin-mixed lineage leukemia (MLL; myeloid/lymphoid leukemia; KMT2A) fusion protein, possessing potential antitumor activity. After oral administration, icovamenib specifically targets and binds to menin, thereby preventing the interaction between menin and MLL proteins and the formation of the menin-MLL complex. This reduces the expression of downstream target genes (such as MYC and Bcl2) and inhibits the proliferation of MLL-rearranged tumor cells. Menin is an important transcriptional regulator that plays a crucial role in oncogenic signaling pathways in cancers driven by oncogenic MLL fusion genes.
BMF-219 (Icovamenib) is a selective, orally active, irreversible Menin inhibitor. The molecular formula is C31H34N₈O3, and the molecular weight is 566.65. The IUPAC name is (R)-4-((3-acrylamidopiperidin-1-yl)methyl)-N-(4-(4-morpholino-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenyl)picolinamide. BMF-219 forms a stable and irreversible covalent bond with Menin, leading to sustained inhibition of the Menin-MLL interaction. It is being developed for the treatment of Menin-MLL-dependent cancers, including acute leukemias, multiple myeloma, diffuse large B-cell lymphoma, and solid tumors. The compound has also been proposed for the treatment of Type II diabetes. BMF-219 is a research compound with potential clinical applications. |
| Exact Mass |
566.275
|
|---|---|
| CAS # |
2448172-22-1
|
| PubChem CID |
154988914
|
| Appearance |
Solid powder
|
| LogP |
2.8
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
42
|
| Complexity |
924
|
| Defined Atom Stereocenter Count |
1
|
| InChi Key |
CPRLHPSXWZTPMC-XMMPIXPASA-N
|
| InChi Code |
InChI=1S/C31H34N8O3/c1-2-28(40)35-24-4-3-11-38(19-24)18-21-9-10-32-27(16-21)31(41)36-23-7-5-22(6-8-23)26-17-25-29(37-26)33-20-34-30(25)39-12-14-42-15-13-39/h2,5-10,16-17,20,24H,1,3-4,11-15,18-19H2,(H,35,40)(H,36,41)(H,33,34,37)/t24-/m1/s1
|
| Chemical Name |
N-[4-(4-morpholin-4-yl-7H-pyrrolo[2,3-d]pyrimidin-6-yl)phenyl]-4-[[(3R)-3-(prop-2-enoylamino)piperidin-1-yl]methyl]pyridine-2-carboxamide
|
| Synonyms |
Menin-MLL inhibitor 21
|
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 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.) |
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.
Link: https://clinicaltrials.gov/ct2/show/NCT07502495
Conditions:Type 2 DiabetesLink: https://clinicaltrials.gov/ct2/show/NCT07502508
Conditions:Type 2 DiabetesLink: https://clinicaltrials.gov/ct2/show/NCT07254286
Conditions:Healthy VolunteerProducts are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
