| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg | |||
| Other Sizes |
| Targets |
- TEAD1, TEAD2, TEAD3, TEAD4 (pan-TEAD inhibitor)[1]
- IC50 values for inhibiting palmitoylation of TEAD1-4 are in the submicromolar range in vitro[1]
- Cellular YAP-TEAD transcriptional activity IC50: 56 nM (NCI-H226 mCherry reporter assay)[1]
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|---|---|
| ln Vitro |
- Inhibition of TEAD Palmitoylation: In an in vitro assay using recombinant TEAD2 YBD protein, MYF-03-69 potently inhibits palmitoylation. Mass spectrometry analysis shows that a 10-fold molar excess of MYF-03-69 leads to 100% labeling of TEAD2 protein after 1 hour at room temperature, modifying cysteine C380.[1]
- Inhibition of TEAD Transcriptional Activity: In Hippo/YAP reporter NCI-H226 (NF2-deficient MPM) cells, a 72-hour treatment with MYF-03-69 decreases YAP-TEAD transcriptional activity in a dose-dependent manner with an IC50 of 56 nM, while the non-covalent control MYF-03-69-NC has minimal effect.[1] - Regulation of Downstream Gene Expression: A 24-hour treatment with MYF-03-69 in NCI-H226 and MSTO-211H cells leads to significant downregulation of canonical YAP target genes (CTGF, CYR61, IGFBP3, KRT34, NPPB) and upregulation of the pro-apoptotic gene BMF. Protein levels of CYR61 and AXL are also decreased.[1] - Anti-proliferative Activity: In 5-day cell growth assays, MYF-03-69 potently inhibits the growth of Hippo signaling-defective MPM cells (NCI-H226, MSTO-211H) but shows no effect on Hippo signaling-intact (NCI-H2452) or non-cancerous (MeT-5A) mesothelial cells. This selective anti-proliferative effect is also observed in 3D spheroid cultures.[1] - Cell Cycle Arrest: Treatment with MYF-03-69 for 48 hours induces G1 phase cell cycle arrest in NCI-H226 and MSTO-211H cells.[1] - High-throughput Screening in Cancer Cell Lines: PRISM screening across 903 cancer cell lines reveals that sensitivity to MYF-03-69 highly correlates with TEAD1 and YAP1 dependency scores. Sensitive cell lines identified include YAP/TEAD co-dependent, YAP-dependent, or TEAD-dependent cells such as liposarcoma (94T778), liver cancer (SKHEP-1), and cholangiocarcinoma (HuCCT1), with nanomolar IC50 values.[1] |
| Enzyme Assay |
- In Vitro Palmitoylation Assay: 1 μM of recombinant TEADs-YBD protein is pre-incubated with indicated concentrations of MYF-03-69 at 37°C for 2 hours. Palmitoyl alkyne-coenzyme A is then added as a lipidation substrate in a 50 μL reaction volume. After 30 minutes, SDS and click reagents are added to initiate the click reaction. After another hour, 4x loading buffer is added, and samples are analyzed by Western blot. Streptavidin is used for biotin detection, and an anti-His tag antibody is used for protein detection.[1]
- Mass Spectrometry: TEAD2 protein is incubated with a 10-fold molar excess of MYF-03-69 for 1 hour at room temperature. The reaction is analyzed by LC-MS. Proteins are desalted on-column and eluted with a gradient. Full-scan mass spectra are acquired. Deconvolution is performed using software. To identify the modification site, the protein is digested with trypsin, and the resulting peptides are analyzed by CE-MS/MS. Data is searched against a database using Mascot.[1] |
| Cell Assay |
- Cell Proliferation Assay: For 2D assays, cells are seeded at 200 cells/well in 384-well plates. MYF-03-69 is added using a workstation the next day. After 5 days, cell viability is measured using a CellTiter-Glo kit. For 3D spheroid assays, NCI-H226 and MSTO-211H cells are seeded at 200 cells/well in ultra-low attachment plates, and viability is measured using a 3D CellTiter-Glo kit.[1]
- TEAD Reporter Assay: NCI-H226 cells are transduced with a TBS-mCherry lentivirus and sorted by GFP. Selected cells are plated at 1,000 cells/well in black 384-well plates. MYF-03-69 is added the next day for 72 hours. Cells are then stained with Hoechst 33342, and mCherry and Hoechst signals are read using a high-content imager.[1] - Western Blot: After treatment, cells are lysed. Proteins are separated by SDS-PAGE and transferred to nitrocellulose membranes. Membranes are incubated with specific primary antibodies (e.g., anti-CYR61, anti-AXL, anti-TEAD, anti-YAP), followed by secondary antibodies, and visualized using an imager.[1] - RT-PCR: Total RNA is extracted from cells treated with MYF-03-69. cDNA is synthesized from 500 ng of purified RNA using a reverse transcription system. TaqMan probes (for genes like CTGF, CYR61, BMF) are used for RT-PCR reactions.[1] - Co-immunoprecipitation (Co-IP): NCI-H226 cells are treated with indicated doses of MYF-03-69 for 24 hours. Co-IP experiments are then performed using a magnetic bead-based kit according to the manufacturer's instructions to monitor endogenous YAP-TEAD interaction.[1] - Cell Cycle Analysis: Cells are seeded in 6-well plates and treated with MYF-03-69 for 48 hours. Cells are then harvested and fixed in cold 70% ethanol overnight. Subsequently, samples are treated with RNase A and stained with propidium iodide (PI) solution before analysis using a flow cytometer. Data is analyzed with FlowJo software.[1] |
| ADME/Pharmacokinetics |
The paper notes that the optimized analog MYF-03-176 shows decent PK properties including low clearance and high oral bioavailability.[1]
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| Toxicity/Toxicokinetics |
SLC-ABPP proteomic analysis in NCI-H226 cells treated with up to 25 μM MYF-03-69 for 3 hours shows that only 7 out of 12,498 detected cysteines are significantly labeled (competition ratio CR>2), suggesting that MYF-03-69 exhibits quite low reactivity towards the cysteine proteome (high selectivity).[1]
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| References | |
| Additional Infomation |
- Mechanism of Action: MYF-03-69 is a "Y-shaped" molecule that acts by forming a covalent bond with a conserved cysteine in the palmitate-binding pocket (PBP) of TEAD proteins (C359 in TEAD1, C380 in TEAD2). The co-crystal structure confirms that it occupies both the hydrophobic lipid tunnel and a hydrophilic side pocket. This covalent binding inhibits TEAD palmitoylation, disrupts the YAP-TEAD protein-protein interaction, and ultimately suppresses downstream gene transcription in the Hippo pathway.[1]
- Indications: MYF-03-69 shows therapeutic potential in cancers with dysregulated Hippo signaling, particularly mesothelioma. PRISM screening results also suggest potential efficacy in other YAP- or TEAD-dependent cancers, including liver cancer, liposarcoma, and lung cancer.[1] |
| Molecular Formula |
C22H20F3N5O2
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|---|---|
| Molecular Weight |
443.421714782715
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| Exact Mass |
443.156
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| CAS # |
2416418-11-4
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| PubChem CID |
162623428
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
2.4
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
32
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| Complexity |
656
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C(N1C[C@@H](OCC2=CC=C(C(F)(F)F)C=C2)[C@H](N2C=C(C3=CC=CN=C3)N=N2)C1)(=O)C=C
|
| InChi Key |
AZGCPBIUNCLWPF-WOJBJXKFSA-N
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| InChi Code |
InChI=1S/C22H20F3N5O2/c1-2-21(31)29-12-19(30-11-18(27-28-30)16-4-3-9-26-10-16)20(13-29)32-14-15-5-7-17(8-6-15)22(23,24)25/h2-11,19-20H,1,12-14H2/t19-,20-/m1/s1
|
| Chemical Name |
1-[(3R,4R)-3-(4-pyridin-3-yltriazol-1-yl)-4-[[4-(trifluoromethyl)phenyl]methoxy]pyrrolidin-1-yl]prop-2-en-1-one
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| Synonyms |
MYF-03-69; TEAD-IN-3; MYF03-69; 2416418-11-4; MYF-0369; orb1687446; CHEMBL5417564;
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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 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)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~225.52 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.64 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 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: 2.5 mg/mL (5.64 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 25.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2552 mL | 11.2760 mL | 22.5520 mL | |
| 5 mM | 0.4510 mL | 2.2552 mL | 4.5104 mL | |
| 10 mM | 0.2255 mL | 1.1276 mL | 2.2552 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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