| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
TEAD1 Palmitoylation
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|---|---|
| ln Vitro |
Soaking with VT-105 provided a crystal structure with 2.6 Angstrom resolution, wherein two molecules of VT-105 and two molecules of CH3(CH2)12CONHOH are present in the asymmetric unit. The molecules of VT105 and the molecules of CH3(CH2)12CONHOH occupy the same location in TEAD3 (Fig. 4C). The quinoline and the 4-trifluoromethylphenyl groups of VT-105 fill a cavity in TEAD3 that is bounded by hydrophobic residues. The carbonyl group of VT105 accepts a hydrogen bond from the backbone NH of cysteine 368. The pyridine group of VT105 is sandwiched between the sidechains of lysine 345 and cysteine 368, and nearly protrudes beyond the surface of TEAD3 [1].
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| Enzyme Assay |
Cell-free TEAD palmitoylation assay [1]
Purified recombinant TEAD1–YBD was first incubated with compounds and then with 2 μmol/L alkyne-palmitoyl-CoA. The reaction was quenched with 1% SDS followed by click chemistry reaction with biotin-azide as described previously. In some experiments, APCoA was added at different concentrations and in different sequence. Palmitoylated TEAD and total TEAD proteins were detected by streptavidin HRP and anti-TEAD1 antibody (Abcam) immunoblotting, respectively. |
| Cell Assay |
Cell-based TEAD palmitoylation assays [1]
Myc-TEAD expression plasmid transfected HEK293T cells were treated with DMSO or 100 μmol/L alkyne palmitate + DMSO/compound for 20 hours. Myc-TEAD protein was immunoprecipitated with anti-Myc antibody and subjected to click chemistry. Palmitoylated TEAD was detected by streptavidin immunoblotting. The Acyl-PEGyl Exchange Gel-Shift Assay was performed as described previously. Cell proliferation assay [1] Cells treated for various time periods with compounds in dose titration starting from 3 μmol/L were assayed by CellTiter-Glo Luminescent Cell Viability Assay Kit according to the manufacturers' protocol. The IC50 and maximum inhibition % were calculated using dose response curves. Immunofluorescence [1] After fixation with 4% paraformaldehyde for 10 to 15 minutes and permeabilization with 0.1% Triton X-100 in PBS, cells were blocked in 3% BSA in PBS for 1 to 2 hours at room temperature, stained with primary antibodies overnight at 4°C, and then with Alexa fluor-conjugated secondary antibodies for 2 to 3 hours at RT. Slides were mounted with prolong gold antifade reagent with DAPI. Images were captured with a Nikon Eclipse Ti confocal microscope. Immunoprecipitation [1] Cells were washed with PBS and lysed [50 mmol/L Tris pH 7.5, 150 mmol/L NaCl, 1% Triton-X100, 50 mmol/L NaF, 1 mmol/L PMSF, protease inhibitor cocktail, phosphatase inhibitor]. After sonication and centrifugation, supernatant was collected and incubated with anti-TEAD, anti-YAP, or control antibodies, precipitated by Protein A/G beads, and analyzed by immunoblotting (see antibody information in Supplementary Table S3) using standard protocols. |
| Animal Protocol |
Animal/Disease Models: NCI- H226 tumor-bearing mice [1]
Doses: 0.3~10 mg/kg Route of Administration: Po one time/day Experimental Results: Tumor growth can be prevented even at a dose of 0.3 mg/kg. Mouse pharmacokinetics VT103, VT104, and VT107, formulated in 5% DMSO + 10% Solutol + 85% D5W, were dosed intravenously or orally at 7 or 10 mg/kg. Blood was drawn from the saphenous vein at indicated timepoints. Compounds were quantified by LC/MS-MS using a QTRAP 6500. Data were analyzed using Phoenix WinNonlin 6.3, and intravenously noncompartmental model 201, and orally noncompartmental model 200. The calculation method was linear/log trapezoidal. In vivo pharmacodynamic and efficacy studies All the procedures related to animal handling, care, and the treatment were performed according to the guidelines approved by the Institutional Animal Care and Use Committee (IACUC) of WuXi AppTec or Crown Bioscience, Inc., following the guidance of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). The testing article formulated in dosing solution (5% DMSO + 10% solutol + 85% D5W; D5W = 5% glucose) was orally administrated daily at the indicated doses. Tumor volume and animal weights were monitored twice weekly. |
| ADME/Pharmacokinetics |
VT103 is an analogue of VT101 with improved potency and good oral pharmacokinetics in mice (Fig. 2; Supplementary Table S1). VT104 is an analogue of VT102 with improved potency and good oral pharmacokinetics in mice (Fig. 2; Supplementary Table S1). VT-105 is a more soluble analogue of VT104 (Fig. 2) and can be used in TEAD X-ray crystallography experiments. VT106 and VT107 are enantiomers of VT104; they differ greatly in potency and can therefore serve as useful cross-controls in biochemical and cellular experiments (Fig. 2). [1]
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| References | |
| Additional Infomation |
Neurofibromatosis type 2 (NF2) gene mutations restrict or eliminate the expression of functional Merlin protein, a condition commonly seen in malignant mesotheliomas. Merlin protein activates the Hippo signaling pathway, inhibiting the nuclear translocation of YAP and TAZ. YAP and TAZ are key effector factors in this pathway, binding to the nuclear TEAD transcription factor and promoting the expression of genes involved in cell proliferation and survival. This article describes compounds we discovered that selectively inhibit YAP/TAZ-TEAD-promoted gene transcription, block TEAD autopalmitoylation, and disrupt the interaction between YAP/TAZ and TEAD. Optimized screening yielded highly potent analogues with excellent oral bioavailability and pharmacokinetic properties. These analogues selectively inhibit the proliferation of NF2-deficient mesothelioma cells in vitro and the growth of subcutaneous tumor xenografts in vivo. These highly potent and selective TEAD inhibitors provide a novel approach to targeting the Hippo-YAP pathway. The Hippo-YAP pathway has previously been difficult to drug, but it is frequently aberrantly regulated in malignant mesotheliomas and other YAP-driven cancers and diseases. [1] Apart from verifying the tolerability of the compounds in mice, the studies described herein did not address any toxicity of the compounds, which may or may not be related to TEAD palmitoylation inhibition. Formal toxicological evaluations in multiple animal models are needed to determine the safety of these small molecule compounds. If the results are satisfactory, clinical evaluation of TEAD palmitoylation inhibitors in NF2-mutant mesothelioma and cancers with activated YAP/TAZ-TEAD transcriptional activity is warranted, either as monotherapy or in combination with other targeted cancer therapies. [1]
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| Molecular Formula |
C24H18F3N3O
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|---|---|
| Molecular Weight |
421.41
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| Exact Mass |
421.14
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| Elemental Analysis |
C, 68.40; H, 4.31; F, 13.52; N, 9.97; O, 3.80
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| CAS # |
2417718-38-6
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| Related CAS # |
2417718-38-6 (S-isomer); 2417718-36-4 (R-isomer)
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| Appearance |
Typically exists as White to off-white solids at room temperature
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| SMILES |
FC(C1C=CC(=CC=1)C1=CC=CC2=CC(=CN=C12)C(N[C@@H](C)C1C=CC=CN=1)=O)(F)F
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| Synonyms |
VT-105; 2417718-38-6; N-[(1S)-1-pyridin-2-ylethyl]-8-[4-(trifluoromethyl)phenyl]quinoline-3-carboxamide;
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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 |
| 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) |
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
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|---|---|
| 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 | 2.3730 mL | 11.8649 mL | 23.7299 mL | |
| 5 mM | 0.4746 mL | 2.3730 mL | 4.7460 mL | |
| 10 mM | 0.2373 mL | 1.1865 mL | 2.3730 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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