| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg | |||
| Other Sizes |
| Targets |
The primary target of milademetan is MDM2 (murine double minute 2), an E3 ubiquitin ligase that negatively regulates the tumor suppressor p53. Milademetan is a potent and selective inhibitor of the MDM2-p53 protein-protein interaction .
MDM2 (murine double minute 2). |
|---|---|
| ln Vitro |
Milademetan demonstrates potent in vitro activity across various cancer cell lines. It reduces cell viability specifically in TP53 wild-type (WT) cell lines. In Merkel cell carcinoma models, milademetan triggered a rapid and sustained p53 response and showed dose-dependent inhibition of tumor cell growth . Preclinical testing revealed that cell lines with CDKN2A homozygous loss in the context of wild-type TP53 exhibit significantly higher sensitivity, with a median IC50 of 79.5 nM compared to 10,000 nM in cell lines with mutant TP53 or non-homozygous loss of CDKN2A . The compound also showed potent activity against MDM2-amplified, TP53-wildtype laboratory models .
In wild-type TP53 neuroblastoma cells, meledemetan (DS-3032) preferentially promotes the expression of CDKNA1, BAX, and MDM2 while stabilizing TP53 [3]. Treatment with meledemetan (DS-3032b) increases the expression of the TP53 target gene and causes apoptosis, senescence, and G1 cell cycle arrest [3]. Regardless of MYCN status, treatment with meledemetan (DS-3032b, 0-2000 nM) specifically decreases the viability, proliferation, and migration of neuroblastoma cells carrying wild-type TP53 [4]. Milademetan is a potent and selective MDM2-p53 inhibitor that binds to MDM2 and prevents its interaction with the transcriptional activation domain of p53. This restores p53 signaling and leads to p53-mediated induction of tumor cell apoptosis. DS-3032b treatment enhances TP53 target gene expression and induces G1 cell cycle arrest, senescence, and apoptosis in cancer cells. The compound shows activity regardless of MYCN status. |
| ln Vivo |
In vivo, milademetan demonstrated significant anti-tumor activity. It showed dose-dependent inhibition of tumor growth in MKL-1 xenograft and patient-derived xenograft (PDX) models of Merkel cell carcinoma . Xenograft models with CDKN2A homozygous loss and wild-type TP53 all demonstrated tumor growth inhibition with milademetan. Furthermore, combination with an anti-PD1 antibody in a colon-26 syngeneic model (with CDKN2A homozygous loss) showed significant enhancement in tumor growth inhibition compared to either agent alone .
Milademetan (DS-3032b, 50 mg/kg, orally administered) xenografts neuroblastoma cells with functioning TP53, delaying tumor growth and enhancing survival [4]. Milademetan has been evaluated in preclinical animal models for the treatment of acute myeloid leukemia and solid tumors. The compound is orally available and has demonstrated in vivo efficacy in tumor xenograft models. Treatment with Milademetan (DS-3032b) increases the expression of TP53 target genes and causes apoptosis, senescence, and G1 cell cycle arrest. The compound is being investigated in clinical trials for various malignancies. |
| Enzyme Assay |
Milademetan is characterized as a selective inhibitor of the MDM2-p53 protein-protein interaction. The compound works by binding to MDM2 and preventing its interaction with the transcriptional activation domain of p53, thereby inhibiting the proteasome-mediated enzymatic degradation of p53 .
The in vitro enzyme/receptor binding assay measures the binding affinity of Milademetan to the MDM2 protein. Recombinant MDM2 protein is incubated with a fluorescently labeled p53 peptide (which mimics the p53 transactivation domain) and varying concentrations of Milademetan (typically 0.1 nM to 100 µM). The displacement of the labeled peptide by Milademetan is measured using fluorescence polarization or AlphaScreen technology. The IC50 or Ki value is calculated from competition binding curves. The compound's selectivity is assessed by profiling against other protein-protein interaction targets. |
| Cell Assay |
In vitro cell assays were performed using cancer cell lines treated with milademetan to assess viability and p53 pathway activation. Cell viability was measured using standard assays, and p53 response was evaluated by assessing downstream target gene expression or protein stabilization. The sensitivity of 215 cancer cell lines to milademetan treatment was evaluated to determine IC50 values based on TP53 and CDKN2A status .
Cell viability assay[4] Cell Types: SK-N-SH, SH-SY5Y, IMR32, IMR5 and LAN5 cell lines. Tested Concentrations: 0-2000 nM. Incubation Duration: 24-72 hrs (hours). Experimental Results: Vitality diminished in a dose- and time-dependent manner. In SK-N-SH, SH-SY5Y, IMR32, IMR5 and LAN5 cell lines (72 hrs (hours)), the IC50 values were 21.9 nM, 17.7 nM, 52.63 nM, 25.7 nM and 44.1 nM, respectively. In vitro cellular assays are performed using cancer cell lines with wild-type TP53, such as AML cell lines or solid tumor cell lines. Cells are treated with Milademetan at concentrations ranging from 0.01 to 100 µM for 24-72 hours. Cell viability is measured using MTT or CellTiter-Glo assays. Apoptosis is assessed by Annexin V/PI staining and flow cytometry, or by measuring caspase-3/7 activity. Cell cycle analysis is performed by propidium iodide staining and flow cytometry. TP53 target gene expression (e.g., p21, PUMA, NOXA) is measured by qRT-PCR or Western blot. |
| Animal Protocol |
Preclinical efficacy was evaluated using xenograft models. Immunocompromised mice were implanted with human tumor cell lines (such as MKL-1) or patient-derived tumor tissue. Milademetan was administered orally, typically once daily, using various intermittent or continuous dosing schedules. Tumor growth inhibition was monitored over time. In combination studies, anti-PD1 antibody was administered via intraperitoneal injection .
Animal/Disease Models: SH-SY5Y nude mouse xenograft tumor [4]. Doses: 50 mg/kg. Dosing: po (oral gavage) for 30 days, alternating with 4 days of po (oral gavage) treatment and then 2 days of no treatment (4+2). Experimental Results: The survival rate of the mouse group was Dramatically prolonged. diminished neuroblastoma xenograft tumor growth through activation of TP53 signaling. In vivo efficacy is evaluated in mouse xenograft models using human tumor cell lines with wild-type TP53. Tumor-bearing mice are administered Milademetan orally at various doses (e.g., 10-100 mg/kg) on a scheduled basis (e.g., daily or intermittent). Tumor volume is measured by caliper twice weekly. Body weight is monitored as a measure of tolerability. At study termination, tumors are collected for analysis of TP53 target gene expression, apoptosis markers, and cell cycle regulators. Pharmacodynamic studies assess the relationship between drug exposure and target modulation. |
| ADME/Pharmacokinetics |
Milademetan is an orally bioavailable small molecule. In the first-in-human phase I study, pharmacokinetic analyses were conducted to characterize its absorption and exposure profile. The recommended phase II dose and schedule (260 mg once daily on days 1-3 and 15-17 every 28 days, i.e., a 3/14 day intermittent schedule) was selected based on PK/PD data to mitigate dose-limiting toxicities while maintaining efficacy .
Milademetan is orally bioavailable. The tosylate hydrate salt form has a molecular weight of 808.74 g/mol and molecular formula C37H44Cl2FN5O8S. The compound is soluble in DMSO (10 mM). Pharmacokinetic parameters such as oral bioavailability, plasma half-life, and tissue distribution have been characterized in preclinical studies. The compound is typically formulated for oral administration in preclinical studies. |
| Toxicity/Toxicokinetics |
The most common grade 3/4 treatment-emergent adverse events observed in clinical trials include thrombocytopenia (29.0%), neutropenia (15.0%), and anemia (13.1%). An intermittent dosing schedule (3/14 days) significantly reduced the rates of these high-grade hematologic abnormalities (thrombocytopenia: 15.0%, neutropenia: 5.0%, anemia: 0%) while maintaining efficacy. Other reported adverse events included gastrointestinal issues, fatigue, diarrhea, leukopenia, and renal/electrolyte abnormalities .
No detailed toxicity data is publicly available for Milademetan tosylate hydrate. As a research compound and clinical candidate, it has undergone preclinical toxicology assessments including genotoxicity, cardiac safety (hERG channel inhibition), and repeated-dose toxicity studies in rodents and non-rodents. The compound is for research use only and not for human therapeutic use outside of clinical trials. Potential toxicities may include myelosuppression, gastrointestinal effects, and p53-related toxicities. |
| References | |
| Additional Infomation |
Milademetan has been evaluated in multiple clinical trials including MANTRA (phase III in liposarcoma), MANTRA-2 (phase II in MDM2-amplified solid tumors), and MANTRA-4 (planned for combination with atezolizumab). In a phase II trial of MDM2-amplified, TP53-wildtype solid tumors, the objective response rate was 19.4% (6/31), including a patient with endometrial stromal sarcoma who achieved 100% target lesion reduction. The median progression-free survival was 3.5 months . In dedifferentiated liposarcoma, the disease control rate was 58.5% with median PFS of 7.2 months . In myeloid malignancies, clinical efficacy was minimal despite being relatively well-tolerated .
Milademetan tosylate hydrate is an MDM2-p53 inhibitor that has been studied in clinical trials for acute myeloid leukemia and solid tumors. It is not yet approved by regulatory authorities for clinical use. The compound is also known as DS-3032b and DS-3032. It is supplied as a tosylate hydrate salt form for research purposes. The compound induces G1 cell cycle arrest, senescence, and apoptosis in TP53 wild-type cancer cells. |
| Molecular Formula |
C₃₇H₄₄CL₂FN₅O₈S
|
|---|---|
| Molecular Weight |
808.74
|
| Exact Mass |
807.227
|
| CAS # |
2095625-97-9
|
| Related CAS # |
Milademetan;1398568-47-2; 2095625-97-9; 1398569-75-9
|
| PubChem CID |
89051550
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
11
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
54
|
| Complexity |
1290
|
| Defined Atom Stereocenter Count |
5
|
| SMILES |
CC1=CC=C(C=C1)S(=O)(=O)O.CC1(CCC2(CC1)[C@@]3([C@H]([C@@H](N2)C(=O)N[C@@H]4CC[C@H](OC4)C(=O)N)C5=C(C(=NC=C5)Cl)F)C6=C(C=C(C=C6)Cl)NC3=O)C.O
|
| InChi Key |
WPJOGWGXMTUHPW-CIPNXXNHSA-N
|
| InChi Code |
InChI=1S/C30H34Cl2FN5O4.C7H8O3S.H2O/c1-28(2)8-10-29(11-9-28)30(18-5-3-15(31)13-19(18)37-27(30)41)21(17-7-12-35-24(32)22(17)33)23(38-29)26(40)36-16-4-6-20(25(34)39)42-14-16;1-6-2-4-7(5-3-6)11(8,9)10;/h3,5,7,12-13,16,20-21,23,38H,4,6,8-11,14H2,1-2H3,(H2,34,39)(H,36,40)(H,37,41);2-5H,1H3,(H,8,9,10);1H2/t16-,20+,21+,23-,30-;;/m1../s1
|
| Chemical Name |
(3'R,4'S,5'R)-N-[(3R,6S)-6-carbamoyltetrahydro-2H-pyran-3-yl]-6''-chloro-4'-(2-chloro-3-fluoropyridin-4-yl)-4,4-dimethyl-2''-oxo-1'',2''-dihydrodispiro[cyclohexane-1,2'-pyrrolidine-3',3''-indole]-5'-carboxamide mono(4-methylbenzenesulfonate) monohydrate
|
| Synonyms |
DS3032b tosylate hydrate; DS-3032b; 4Y0K58UFU2; Milademetan tosylate monohydrate; DS-3032 TOSYLATE MONOHYDRATE; (3'R,4'S,5'R)-N-((3R,6S)-6-CARBAMOYLTETRAHYDRO-2H-PYRAN-3-YL)-6''-CHLORO-4'-(2-CHLORO-3-FLUOROPYRIDIN-4-YL)-4,4-DIMETHYL-2''-OXO-1'',2''-DIHYDRODISPIRO(CYCLOHEXANE-1,2'-PYRROLIDINE-3',3''-INDOLE)-5'-CARBOXAMIDE MONO(4-METHYLBENZENESULFONATE) MONOHYDRATE; (3'R,4'S,5'R)-N-[(3R,6S)-6-carbamoyltetrahydro-2H-pyran-3-yl]-6''-chloro-4'-(2-chloro-3-fluoropyridin-4-yl)-4,4-dimethyl-2''-oxo-1'',2''-dihydrodispiro[cyclohexane-1,2'-pyrrolidine-3',3''-indole]-5'-carboxamide mono(4-methylbenzenesulfonate) monohydrate; DS-3032; DS 3032b; DS 3032; DS3032; DS-3032b tosylate; Milademetan tosylate hydrate
|
| 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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : ~50 mg/mL (~61.82 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 5 mg/mL (6.18 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 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: 5 mg/mL (6.18 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 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: ≥ 5 mg/mL (6.18 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 | 1.2365 mL | 6.1825 mL | 12.3649 mL | |
| 5 mM | 0.2473 mL | 1.2365 mL | 2.4730 mL | |
| 10 mM | 0.1236 mL | 0.6182 mL | 1.2365 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
