| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 100mg |
|
||
| 500mg |
|
||
| Other Sizes |
| Targets |
MYF-03-69 targets all four members of the TEAD transcription factor family, with IC₅₀ values of 385 nM for TEAD1, 143 nM for TEAD2, 558 nM for TEAD3, and 173 nM for TEAD4. Additionally, MYF-03-69 inhibits TEAD transcriptional activity with an IC₅₀ of 56 nM. The compound irreversibly modifies TEAD proteins by covalently binding to the conserved cysteine in the TEAD palmitate-binding pocket, thereby disrupting YAP-TEAD interaction.
|
|---|---|
| ln Vitro |
MYF-03-69 selectively inhibits the proliferation of Hippo signaling-defective mesothelioma cells (e.g., NCI-H226, MSTO-211H) while showing little cytotoxicity to normal mesothelial cells or Hippo signaling-intact cancer cells. At concentrations of 0.1-10 μM for 48 hours, MYF-03-69 induces G1 phase cell cycle arrest in NCI-H226 and MSTO-211H cells. At 2 μM, MYF-03-69 inhibits TEAD protein palmitoylation, downregulates YAP-TEAD target genes (such as CTGF, CYR61, and ANKRD1), and upregulates the pro-apoptotic gene BMF. Incubation of recombinant TEAD2 YBD protein with a 10-fold molar excess of MYF-03-69 at room temperature for 1 hour achieves 100% labeling efficiency. PRISM screening across 903 cancer cell lines reveals a high correlation between TEAD-YAP dependency and sensitivity to MYF-03-69.
|
| ln Vivo |
Further optimization of MYF-03-69 led to MYF-03-176, an orally bioavailable compound that demonstrates strong antitumor efficacy in a malignant pleural mesothelioma mouse xenograft model via oral administration. In cell viability screens, MYF-03-69 shows selective antiproliferative effects against Hippo signaling-defective mesothelioma cells (such as NCI-H226 and MSTO-211H), with less effect on Hippo signaling-intact mesothelioma cells (NCI-H2452) and non-cancerous mesothelial cells (MeT-5A).
|
| Enzyme Assay |
TEAD Palmitoylation Inhibition Assay: 1 μM TEADs-YBD recombinant protein is incubated with indicated concentrations of MYF-03-69 at 37°C for 2 hours, followed by the addition of palmitoyl alkyne-coenzyme A in a total volume of 50 μL. After a 30-minute reaction, 5 μL of 10% SDS and 5 μL of click reaction reagents are added to initiate the click reaction for 1 hour. After adding 4× loading buffer, samples are subjected to Western blot analysis using IRDye 800CW Streptavidin for biotin detection and anti-His-tag antibody.
Mass Spectrometry Labeling Assay: Recombinant TEAD2 YBD protein is incubated with a 10-fold molar excess of MYF-03-69 at room temperature for 1 hour, followed by mass spectrometry analysis to detect labeling efficiency. Results show 100% labeling after 1 hour incubation.
Co-crystallization Assay: 300 μM TEAD1 pre-incubated with 600 μM MYF-03-69 for 1 hour is dispensed in crystallization buffer (3 M (NH₄)₂SO₄ and 0.1 M Tris pH 9.0) and incubated against 25 μL of crystallization buffer in a 384-well hanging-drop vapor diffusion microtiter plate at 20°C for 3 days to obtain crystals.
Molecular Docking: Docking of MYF-03-69 is performed with the covalent docking protocol from Schrödinger suite software (release 2019-02) with default parameters in the TEAD2 structure (PDB code:).
Cellular Thermal Shift Assay (CETSA) : CETSA is used to detect MYF-03-69 binding to TEAD proteins in cells.
|
| Cell Assay |
Cell Proliferation Inhibition Assay: NCI-H226, MSTO-211H, NCI-H2452 mesothelioma cells, and MeT-5A mesothelial cells are seeded in culture plates and treated with 0-10 μM MYF-03-69 for 5 days. Cell viability is assessed by CellTiter-Glo or MTT assays to calculate GR₅₀ values.
Cell Cycle Analysis: NCI-H226 and MSTO-211H cells are treated with 0.1-10 μM MYF-03-69 for 48 hours, and cell cycle analysis is performed by flow cytometry to detect G1 phase cell cycle arrest.
Western Blot Analysis: Cells are treated with 2 μM MYF-03-69 for indicated times and lysed. Protein expression levels of TEAD target genes (CTGF, CYR61, ANKRD1) and the pro-apoptotic gene BMF are detected by Western blot.
TEAD Transcriptional Activity Reporter Assay: A stable cell line is established using a TEAD luciferase reporter lentivirus to monitor TEAD transcriptional activity. Cells are treated with MYF-03-69, and TEAD transcriptional inhibition is assessed by luciferase activity detection (IC₅₀ = 56 nM).
PRISM High-Throughput Screening: PRISM screening is performed on 903 barcoded cancer cell lines. Log2(AUC) values are calculated to assess sensitivity to MYF-03-69, and correlation analysis is conducted with CRISPR knockout scores from the DepMap database.
|
| Animal Protocol |
MPM Xenograft Mouse Model: A CDX mouse model is established using NCI-H226 human malignant pleural mesothelioma cells. The optimized MYF-03-69 analog MYF-03-176 is administered orally to evaluate antitumor efficacy.
|
| ADME/Pharmacokinetics |
MYF-03-69 has a molecular weight of 443.42 and the molecular formula C₂₂H₂₀F₃N₅O₂. It is soluble in DMSO at 80 mg/mL (180.42 mM). The optimized analog MYF-03-176 has good oral bioavailability.
|
| References |
[1]. Covalent Disruptor of YAP-TEAD Association Suppresses Defective Hippo Signaling. BioRxiv. doi: https://doi.org/10.1101/2022.05.10.491316. Now published in eLife doi: 10.7554/eLife.78810
[2]. Structure-Based Design of Y-Shaped Covalent TEAD Inhibitors. J Med Chem. 2023 Apr 13;66(7):4617-4632. [3]. YAP/TAZ mediates resistance to KRAS inhibitors through inhibiting proapoptosis and activating the SLC7A5/mTOR axis. JCI Insight. 2024 Dec 20;9(24):e178535. |
| Molecular Formula |
C19H19F4N4O2CL
|
|---|---|
| Molecular Weight |
446.83
|
| CAS # |
2857937-59-6
|
| Appearance |
White to off-white solid powder
|
| Synonyms |
MYF-03-176 HCl; MYF03-176 HCl
|
| 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: 100 mg/mL (243.68 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.09 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2380 mL | 11.1899 mL | 22.3799 mL | |
| 5 mM | 0.4476 mL | 2.2380 mL | 4.4760 mL | |
| 10 mM | 0.2238 mL | 1.1190 mL | 2.2380 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
