| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Menin (menin-KMT2A interaction). [1][2]
In a homogeneous time-resolved fluorescence assay, the average IC50 of JNJ-75276617 against human menin was 0.1 ± 0.05 nM across 10 independent experiments. Against mouse menin, IC50 was 0.045 ± 0.002 nM. Against dog menin, IC50 was ≤0.066 ± 0.004 nM. [2] In a fluorescence polarization binding assay, the Ki values for JNJ-75276617 against wild-type menin, M327I mutant menin, and T349M mutant menin were 0.02 nM, 0.4 nM, and 3.8 nM, respectively. [2] |
|---|---|
| ln Vitro |
JNJ-75276617 decreased menin binding to target gene promoters (MEIS1, HOXA9, HOXA10) in KMT2A-r MOLM-14 and NPM1c OCI-AML3 cells at concentrations from 0.1 to 1.0 μM, with fold decreases ranging from 1.4- to 24.7-fold compared to DMSO control. [2]
JNJ-75276617 suppressed mRNA expression of menin-KMT2A target genes (MEIS1, JMJD1C, FLT3, MEF2C, PBX3) and increased differentiation markers (ITGAM/CD11b, MNDA) in KMT2A-r and NPM1c AML cell lines. [2] JNJ-75276617 showed antiproliferative activity in AML cell lines: MOLM-14 (IC50 <0.1 μM), MOLM-13 (IC50 <0.1 μM), MV4-11 (IC50 <0.1 μM), THP-1 (IC50 >15 μM), EOL-1 (IC50 = 0.116 μM), OCI-AML3 (IC50 = 0.045 μM), and in B-ALL RS4;11 (IC50 = 0.040 μM). Normal peripheral blood mononuclear cells were not impacted up to 10 μM for 7 days. [2] JNJ-75276617 induced apoptosis in MOLM-14, OCI-AML3, and RS4;11 cells in a dose-dependent manner after 7 days of treatment. [2] JNJ-75276617 induced expression of CD11b and CD14 differentiation markers in MOLM-14 and OCI-AML3 cells after 3 and 7 days. In KMT2A-AF9-transduced mouse bone marrow cells, 200 nM JNJ-75276617 for 10 days increased neutrophil-like morphology (28.9% ± 2.6% vs DMSO 10.65% ± 1.35%). [2] JNJ-75276617 demonstrated antiproliferative activity against 6 of 13 NPM1c primary AML patient samples at days 7 and 14, and induced CD11b expression. It also showed antiproliferative effects and dose-dependent apoptosis in a primary KMT2A-r B-ALL patient sample. [2] JNJ-75276617 showed synergistic antiproliferative effects with gilteritinib in MOLM-13 cells (p<0.001). [2] JNJ-75276617 combined with venetoclax or with venetoclax plus azacitidine showed synergistic antiproliferative effects in MOLM-13 cells. [2] JNJ-75276617 retained potent antiproliferative activity in MEN1M327I and MEN1T349M mutant MV4-11 cells (IC50 = 22.6 nM and 53.59 nM, respectively). [2] JNJ-75276617 treatment (0.3 μM for 4 days) upregulated HLA-A and HLA-DR mRNA and protein expression in primary AML samples, and this was CIITA-dependent. [1] JNJ-75276617 pretreatment (0.3 μM for 4 days) enhanced T-cell-mediated cytotoxicity in allogeneic and autologous settings in primary AML samples, particularly those with NPM1c and DNMT3A mutations. [1] |
| ln Vivo |
In subcutaneous MOLM-14 AML xenograft mice, oral JNJ-75276617 (as oxalate salt) at 30, 50, and 100 mg/kg daily for 5 weeks induced dose-dependent tumor regressions of 70%, 97%, and 99%, respectively (p<0.0001 each). At 50 and 100 mg/kg, 10/10 and 9/9 tumors failed to regrow after 3 months off treatment. [2]
In a KMT2A-AF6 AML PDX model (CBAM-68552), JNJ-75276617 treatment significantly increased lifespan by >89% across all dose levels (p<0.05), reduced human CD45+ leukemic cells, and increased differentiation markers (CD11b, CD13, CD14). [2] In NPM1c AML PDX models (LEXFAM-2734 and AM7577), JNJ-75276617 at 30, 50, or 100 mg/kg resulted in median survival >133 days and >147 days, respectively (vehicle: 74 days and 53.5 days), with significant increased lifespan (>66% to >169%, p<0.0001). [2] In a KMT2A-AF4 B-ALL PDX model (CBAB-62871), JNJ-75276617 at 30 and 100 mg/kg daily increased lifespan by >48% and >160%, respectively (p=0.002), and reduced leukemia burden in bone marrow. [2] In a disseminated MOLM-13 model, JNJ-75276617 (10 mg/kg) combined with azacitidine (2 mg/kg) and venetoclax (100 mg/kg) induced a 277% increased lifespan (p<0.0001), superior to the azacitidine+venetoclax regimen (53% ILS). [2] In a disseminated OCI-AML3 NPM1c model, JNJ-75276617 combinations with azacitidine and venetoclax yielded superior survival vs azacitidine+venetoclax, though efficacy was mostly driven by JNJ-75276617. [2] |
| Enzyme Assay |
A homogeneous time-resolved fluorescence (HTRF) assay was used to quantify the ability of JNJ-75276617 to displace a fluorescein isothiocyanate (FITC)-labeled menin-binding motif 1 peptide (derived from KMT2A) from binding to terbium-labeled human, dog, and mouse menin. The compound was incubated with menin and FITC-MBM1 peptide, and the TR-FRET signal was monitored using a plate reader. Inhibitor potency was reported at 300 or 420 minutes when equilibrium was reached. Percentage inhibition and potency were calculated using Prism software. [2]
A fluorescence polarization binding assay was performed to assess binding of JNJ-75276617 to wild-type, M327I mutant, and T349M mutant menin. Dose-dependent displacement of a KMT2A peptide was measured. [2] |
| Cell Assay |
For the menin-KMT2A inhibitor screen, cryopreserved mononuclear cells from AML patients were thawed, resuspended in medium with DNase I, MgSO4, and heparin, then cultured in liquid medium with G-CSF, TPO, and IL-3 (all 20 ng/mL) or co-cultured on MS5 stromal cells. Cells were treated with DMSO or 0.03, 0.30, and 3.00 μM JNJ-75276617 for 14 days, with fresh medium and inhibitor added at day 7. On days 7 and 14, cells were stained with CD45-PECy7 and CD117, and fluorescence was measured using a flow cytometer. [1]
For apoptosis detection, cells were stained with annexin V and live/dead dye after treatment with JNJ-75276617 for up to 7 days, then analyzed by flow cytometry. [2] For differentiation assessment, cells were stained with CD11b and CD14 antibodies and analyzed by flow cytometry. May-Grünwald Giemsa staining was used to assess morphological changes in KMT2A-AF9-transduced mouse bone marrow cells after 10 days of treatment with 200 nM JNJ-75276617. [2] For T-cell cytotoxicity assays, primary AML samples were pretreated with 0.3 μM JNJ-75276617 for 4 days, then washed and co-cultured with anti-CD3/CD28 activated T cells from healthy donors at different effector:target ratios for 3 days. Viable cell counts were determined by annexin-V and Zombie NIR staining. [1] For autologous T-cell cytotoxicity, CD45+ blast and CD3+ T-cell populations were sorted from NPM1c AML patient samples, and AML blasts were cultured with or without their own T cells at a fixed ratio in the presence or absence of 0.3 μM JNJ-75276617. [1] For chromatin immunoprecipitation (ChIP)-qPCR, cells were cross-linked, lysed, and sonicated. Menin antibody was used for immunoprecipitation, and binding to target gene promoters (MEIS1, HOXA9, HOXA10) was quantified. [2] For the Quantigene multiplex assay, cells were treated with JNJ-75276617 or DMSO for 48-72 hours, then lysed. Target-specific probes were mixed with lysates, and signals were measured on a FLEXMAP 3D system. Absolute IC50 values were calculated by dose-response modeling. [2] |
| Animal Protocol |
Female 6- to 8-week-old immunocompromised mice were used for xenograft studies. JNJ-75276617 (as oxalate salt) was orally dosed daily at various concentrations (10, 30, 50, 100 mg/kg) for up to 4 weeks. Venetoclax (100 mg/kg) was orally dosed daily. Azacitidine (2 mg/kg) was dosed by intraperitoneal injection daily for the first week only. All experiments were performed in accordance with institutional guidelines. [2]
In the subcutaneous MOLM-14 model, tumor volume was measured, and tumor regression was assessed. In disseminated models, survival was monitored, and leukemic burden was assessed by flow cytometry for human CD45+ cells in bone marrow. [2] |
| ADME/Pharmacokinetics |
Following oral dosing in mice, JNJ-75276617 (as oxalate salt) showed adequate oral bioavailability of 34% and a plasma half-life of approximately 9 hours. [2]
|
| Toxicity/Toxicokinetics |
The IC50 for the inhibitory effect of JNJ-75276617 on hERG potassium current was >30 μM. In human induced pluripotent stem cell-derived cardiomyocytes, JNJ-75276617 did not cause any physiologically relevant effects on calcium transient duration 90%, beat rate, Ca2+ amplitude, incidence of cessation of beating, early afterdepolarization-like, or fibrillation-like events at concentrations up to 5 μM. [2]
In a kinase panel (n=109) at 1.0 μM, JNJ-75276617 showed no significant activity. In a receptor panel at 10 μM against 52 receptors, no significant interaction (>50% inhibition of control-specific binding) was observed. [2] |
| References |
|
| Additional Infomation |
Bleximenib is an orally bioavailable protein-protein interaction (PPI) inhibitor that inhibits menin-mixed lineage leukemia (MLL; mixed lineage leukemia 1; MLL1; myeloid/lymphoid leukemia; histone lysine N-methyltransferase 2A; KMT2A) proteins, exhibiting potential antitumor activity. After oral administration, Bleximenib inhibits the interaction between menin and MLL proteins and the formation of the menin-MLL complex. This reduces the expression of downstream target genes, thereby inhibiting the proliferation of leukemia cells with KMT2A gene alterations (such as gene rearrangement (KMT2A-r), duplication, and amplification) or nucleolar phosphatidylcholine 1 (NPM1) gene alterations. The menin-MLL complex plays a crucial role in the survival, growth, transformation, and proliferation of certain types of leukemia cells.
JNJ-75276617 is being clinically investigated for acute leukemias harboring KMT2A or NPM1 alterations, as monotherapy for relapsed/refractory acute leukemia (NCT04811560), or in combination with AML-directed therapies (NCT05453903). [2] JNJ-75276617 has a unique binding mode to menin, including a hydrogen bond (salt bridge) with Asp290, which distinguishes it from other menin-KMT2A inhibitors like revumenib. [2] JNJ-75276617 demonstrated potent activity against MEN1M327I and MEN1T349M mutant cells that confer resistance to other menin-KMT2A inhibitors. [2] |
| Molecular Formula |
C32H50FN7O3
|
|---|---|
| Molecular Weight |
599.78
|
| Exact Mass |
599.395
|
| Elemental Analysis |
C, 64.08; H, 8.40; F, 3.17; N, 16.35; O, 8.00
|
| CAS # |
2654081-35-1
|
| Related CAS # |
Bleximenib oxalate;2866179-95-3;(S)-Bleximenib oxalate; 2866139-02-6; 2866179-96-4
|
| PubChem CID |
156498110
|
| Appearance |
Colorless to light yellow oil
|
| LogP |
4.3
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
10
|
| Rotatable Bond Count |
15
|
| Heavy Atom Count |
43
|
| Complexity |
861
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
CCN(C(C)C)C(=O)C1=C(C=CC(=C1)F)OC2=C(N=CN=N2)N3CCC4(C3)CN(C4)[C@H](CCCN(C)CCOC)C(C)C
|
| InChi Key |
PDUGAXSIWNMIBQ-HHHXNRCGSA-N
|
| InChi Code |
InChI=1S/C32H50FN7O3/c1-8-40(24(4)5)31(41)26-18-25(33)11-12-28(26)43-30-29(34-22-35-36-30)38-15-13-32(19-38)20-39(21-32)27(23(2)3)10-9-14-37(6)16-17-42-7/h11-12,18,22-24,27H,8-10,13-17,19-21H2,1-7H3/t27-/m1/s1
|
| Chemical Name |
N-ethyl-5-fluoro-2-[[5-[2-[(3R)-6-[2-methoxyethyl(methyl)amino]-2-methylhexan-3-yl]-2,7-diazaspiro[3.4]octan-7-yl]-1,2,4-triazin-6-yl]oxy]-N-propan-2-ylbenzamide
|
| Synonyms |
bleximenib; JNJ75276617; JNJ 75276617; 2654081-35-1; JNJ-75276617; DCN5WEN78T; N-ethyl-5-fluoro-2-[[5-[2-[(3R)-6-[2-methoxyethyl(methyl)amino]-2-methylhexan-3-yl]-2,7-diazaspiro[3.4]octan-7-yl]-1,2,4-triazin-6-yl]oxy]-N-propan-2-ylbenzamide; N-ethyl-5-fluoro-2-((5-(2-((3R)-6-(2-methoxyethyl(methyl)amino)-2-methylhexan-3-yl)-2,7-diazaspiro(3.4)octan-7-yl)-1,2,4-triazin-6-yl)oxy)-N-propan-2-ylbenzamide;
|
| 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6673 mL | 8.3364 mL | 16.6728 mL | |
| 5 mM | 0.3335 mL | 1.6673 mL | 3.3346 mL | |
| 10 mM | 0.1667 mL | 0.8336 mL | 1.6673 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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