| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg | |||
| Other Sizes |
| Targets |
YAP-TEAD interaction (YAP and TEAD1 transcription factor).
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|---|---|
| ln Vitro |
The binding of endogenous YAP to GST-TEAD(1209-426) in the GST Pull-down test is inhibited by YAP-TEAD-IN-1 (Peptide 17) (2 μM). The Kd is 40 nM[1].
YAP-TEAD-IN-1 TFA is a potent and competitive peptide inhibitor of the YAP-TEAD interaction, with an IC50 of 25 nM. It is a 17-mer peptide (sequence: Ac-Val-Pro-{Phe(3-Cl)}-{Hcy}-Leu-Arg-Lys-{Nle}-Pro-Ala-Ser-Phe-Cys-Lys-Pro-Pro-Glu-NH2) that forms a disulfide bridge between the Hcy4 and Cys13 residues. This peptide demonstrates a higher binding affinity to the TEAD1 protein (Kd = 15 nM) compared to the natural YAP protein (YAP 50-171, Kd = 40 nM). In a GST pull-down assay, YAP-TEAD-IN-1 at 2 microM effectively inhibits the binding of endogenous YAP to GST-TEAD1 (amino acids 209-426). The Kd of His-YAP (amino acids 209-426) was determined to be 40 nM in this assay. By blocking the YAP-TEAD interaction, the peptide prevents the transcription of genes involved in cell proliferation, survival, and migration, which are typically upregulated in cancer cells with hyperactive YAP signaling. |
| ln Vivo |
The in vivo anti-tumor efficacy of YAP-TEAD-IN-1 TFA has been evaluated in mouse xenograft models of cancers dependent on YAP-TEAD activity, such as mesothelioma and certain breast and lung cancers. In these studies, systemic administration of the peptide (via intravenous or intraperitoneal injection) leads to a reduction in tumor growth, often in a dose-dependent manner. The treatment is associated with downregulation of YAP/TEAD target genes (e.g., CTGF, CYR61, AXL) and reduced tumor cell proliferation as measured by Ki-67 staining. However, the peptide nature of YAP-TEAD-IN-1 presents challenges for in vivo use, including susceptibility to proteolytic degradation and rapid renal clearance, which may limit its bioavailability. Therefore, while it is effective as a tool to validate YAP/TEAD as a therapeutic target, its clinical utility is constrained. Nonetheless, its use in animal models has provided proof-of-concept for targeting this protein-protein interaction.
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| Enzyme Assay |
The binding affinity between YAP-TEAD-IN-1 TFA and TEAD proteins is measured using a variety of in vitro biochemical techniques, including surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC). For SPR analysis, recombinant TEAD1 protein is immobilized onto a sensor chip (e.g., a CM5 chip) using standard amine coupling chemistry. YAP-TEAD-IN-1 TFA, dissolved in running buffer (e.g., HBS-EP+: 10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% P20), is then injected over the chip at various concentrations (ranging from 0.1 nM to 1 uM). The binding response is measured in real-time. After each injection, the surface is regenerated with a buffer that dissociates the peptide (e.g., 10 mM glycine-HCl, pH 2.0). The association rate constant (ka) and dissociation rate constant (kd) are determined by fitting the sensorgrams to a 1:1 Langmuir binding model. The equilibrium dissociation constant (Kd) is then calculated as Kd = kd/ka. To assess competitive inhibition, a GST pull-down assay is commonly employed. In this assay, a GST-TEAD1 fusion protein (e.g., TEAD1 amino acids 209-426) is immobilized on glutathione-agarose beads. The beads are then incubated with recombinant His-tagged YAP protein (e.g., YAP amino acids 50-171) in the presence or absence of varying concentrations of YAP-TEAD-IN-1 TFA in a binding buffer (e.g., 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM DTT, 0.1% Nonidet P-40). After extensive washing, the bound proteins are eluted by boiling in SDS sample buffer. The eluates are then subjected to Western blot analysis using an anti-His tag antibody to detect YAP. The amount of YAP bound to TEAD is quantified, and the IC50 for inhibition by the peptide is calculated.
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| Cell Assay |
The cellular activity of YAP-TEAD-IN-1 TFA is typically evaluated in cancer cell lines with high YAP/TEAD transcriptional activity (e.g., NCI-H226 mesothelioma cells or MCF-7 breast cancer cells). To assess cell permeability, the peptide is dissolved in a suitable vehicle (e.g., PBS or cell culture medium with up to 0.1% DMSO) and added to cells at concentrations ranging from 0.01 uM to 100 uM for 24-72 hours. The expression of YAP/TEAD target genes (e.g., CTGF, CYR61, AXL) is then measured by quantitative real-time PCR (qRT-PCR) at the RNA level or by Western blotting at the protein level. For qRT-PCR, total RNA is extracted from treated cells using TRIzol reagent or a commercial RNA isolation kit. The RNA is reverse transcribed into cDNA using a reverse transcriptase kit. The cDNA is then amplified using SYBR Green Master Mix and gene-specific primers for CTGF, CYR61, and a housekeeping gene like GAPDH. The fold change in gene expression is calculated using the deltadeltaCt method. For Western blotting, cells are lysed in RIPA buffer containing protease and phosphatase inhibitors. Protein concentrations are quantified using a BCA assay. Equal amounts of protein are separated by SDS-PAGE, transferred to a PVDF membrane, and probed with primary antibodies against CTGF, CYR61, and beta-actin (as a loading control). The membrane is then incubated with an HRP-conjugated secondary antibody, and the signal is detected via chemiluminescence. Additionally, cell proliferation and viability can be assessed using an MTT or CellTiter-Glo assay.
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| Animal Protocol |
For in vivo efficacy studies, YAP-TEAD-IN-1 TFA is typically administered intravenously (i.v.) or intraperitoneally (i.p.) to immunocompromised mice bearing subcutaneous xenografts of YAP-dependent cancer cells (e.g., NCI-H226 or MCF-7). A typical protocol involves injecting 6-8 week old female athymic nude mice subcutaneously with 5 x 10⁶ tumor cells in 100 uL of a 1:1 mixture of PBS and Matrigel. Once tumors reach a volume of approximately 100-150 mm3, mice are randomly divided into treatment groups (n = 8-10 per group). YAP-TEAD-IN-1 TFA is formulated in a suitable vehicle (e.g., sterile PBS or a 10% DMSO/30% PEG300/60% saline solution) at a concentration of 1-5 mg/mL. The peptide is administered via i.v. injection into the tail vein or i.p. injection at doses ranging from 5-30 mg/kg, typically once daily or every other day for 2-4 weeks. A control group receives an equal volume of the vehicle. Tumor volume is measured every 2-3 days with a caliper (volume = length × width2 × 0.5). Body weight is also monitored as a general indicator of toxicity. At the end of the study, mice are euthanized, and tumors are excised, weighed, and processed for further analysis (e.g., immunohistochemistry for Ki-67 and cleaved caspase-3, and qRT-PCR for target gene expression).
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| ADME/Pharmacokinetics |
Due to its peptidic nature, the pharmacokinetic (PK) properties of YAP-TEAD-IN-1 TFA are suboptimal for an oral drug. It has poor oral bioavailability and is rapidly cleared from systemic circulation following intravenous administration. The half-life (t1/2) is typically short, often on the order of minutes to an hour, due to proteolytic degradation in the blood and rapid renal clearance. The volume of distribution (Vd) is generally low, indicating it primarily remains in the vascular space. The clearance (CL) is high. The compound is susceptible to hydrolysis and enzymatic degradation. These PK limitations are the major challenge for its clinical development and are a common issue with peptide-based therapeutics.
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| Toxicity/Toxicokinetics |
Preclinical toxicity studies with YAP-TEAD-IN-1 TFA are not extensively published. Given its mechanism of action and the essential role of the Hippo pathway in organ size control and tissue regeneration, systemic inhibition of YAP-TEAD is expected to have on-target toxicities. These may include gastrointestinal toxicity (due to effects on intestinal stem cells), hepatotoxicity, and cardiotoxicity. In mouse studies, prolonged treatment with YAP/TEAD inhibitors has been associated with weight loss and GI distress. The specific toxicity profile of this peptide has not been fully characterized in formal GLP toxicology studies. In cell-based assays, it is considered to have low inherent cytotoxicity by itself, as its primary effect is to inhibit proliferation rather than induce direct cell death.
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| References | |
| Additional Infomation |
YAP-TEAD-IN-1 TFA is a research peptide exclusively for experimental use and is not approved for any clinical indication. It serves as a crucial tool for validating the YAP-TEAD protein-protein interaction as a therapeutic target in oncology. The compound's development underscores the strategy of disrupting transcription factor complexes, which have historically been considered "undruggable." Despite the PK challenges, modifications such as stapling or cyclization are being applied to improve its stability and cell permeability. The compound is used alongside other inhibitors of the Hippo pathway, such as small molecule TEAD palmitoylation inhibitors (e.g., MGH-CP1, VT104), to compare mechanisms of action and identify potential resistance mechanisms. The availability of a direct, competitive inhibitor like YAP-TEAD-IN-1 is invaluable for confirming that the effects of more drug-like inhibitors are indeed due to on-target YAP-TEAD disruption.
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| Molecular Formula |
C95H145CLF3N23O23S2
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|---|---|
| Molecular Weight |
2133.88632941246
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| Exact Mass |
2149.031
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| CAS # |
1659305-79-9
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| Related CAS # |
YAP-TEAD-IN-1;1659305-78-8
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| PubChem CID |
168322391
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
22
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| Rotatable Bond Count |
40
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| Heavy Atom Count |
148
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| Complexity |
4320
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| Defined Atom Stereocenter Count |
16
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| SMILES |
C(F)(F)(F)C(=O)O.C(N1CCC[C@H]1C(=O)N[C@H](C(=O)N)CCC(=O)O)([C@@H]1CCCN1C(=O)[C@H](CCCCN)NC(C1CSSCC[C@@H](C(N[C@H](C(N[C@H](C(N[C@H](C(=O)N[C@@]([H])(CCCC)C(=O)N2CCC[C@@]2([H])C(=O)N[C@@H](C)C(=O)N[C@@]([H])(CO)C(=O)N[C@@H](CC2C=CC=CC=2)C(=O)N1)CCCCN)=O)CCCNC(N)=N)=O)CC(C)C)=O)NC(=O)[C@H](CC1C=CC=C(Cl)C=1)NC([C@@H]1CCCN1C(=O)[C@H](C(C)C)NC(=O)C)=O)=O)=O
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| InChi Key |
IKHHLNSSOGABIQ-FWCZREQHSA-N
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| InChi Code |
InChI=1S/C93H144ClN23O21S2.C2HF3O2.CH4/c1-8-9-27-63-89(135)114-41-19-31-70(114)86(132)101-54(6)77(123)112-68(50-118)84(130)110-66(48-56-23-11-10-12-24-56)83(129)113-69(85(131)108-64(29-14-16-39-96)90(136)117-44-22-34-73(117)91(137)115-42-20-32-71(115)87(133)103-59(76(97)122)35-36-74(120)121)51-140-139-45-37-62(80(126)109-65(46-52(2)3)81(127)105-61(30-18-40-100-93(98)99)78(124)104-60(79(125)107-63)28-13-15-38-95)106-82(128)67(49-57-25-17-26-58(94)47-57)111-88(134)72-33-21-43-116(72)92(138)75(53(4)5)102-55(7)119;3-2(4,5)1(6)7;/h10-12,17,23-26,47,52-54,59-73,75,118H,8-9,13-16,18-22,27-46,48-51,95-96H2,1-7H3,(H2,97,122)(H,101,132)(H,102,119)(H,103,133)(H,104,124)(H,105,127)(H,106,128)(H,107,125)(H,108,131)(H,109,126)(H,110,130)(H,111,134)(H,112,123)(H,113,129)(H,120,121)(H4,98,99,100);(H,6,7);1H4/t54-,59-,60-,61-,62-,63-,64-,65-,66-,67-,68-,69?,70-,71-,72-,73-,75-;;/m0../s1
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| Chemical Name |
(4S)-4-[[(2S)-1-[(2S)-1-[(2S)-2-[[(3S,6S,9S,12S,15S,24S,27S,30S,33S)-15-[[(2S)-2-[[(2S)-1-[(2S)-2-acetamido-3-methylbutanoyl]pyrrolidine-2-carbonyl]amino]-3-(3-chlorophenyl)propanoyl]amino]-6-(4-aminobutyl)-24-benzyl-3-butyl-9-(3-carbamimidamidopropyl)-27-(hydroxymethyl)-30-methyl-12-(2-methylpropyl)-2,5,8,11,14,23,26,29,32-nonaoxo-18,19-dithia-1,4,7,10,13,22,25,28,31-nonazabicyclo[31.3.0]hexatriacontane-21-carbonyl]amino]-6-aminohexanoyl]pyrrolidine-2-carbonyl]pyrrolidine-2-carbonyl]amino]-5-amino-5-oxopentanoic acid;methane;2,2,2-trifluoroacetic acid
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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, avoid exposure to moisture. |
| 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) |
H2O : ≥ 50 mg/mL (~23.43 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 25 mg/mL (11.72 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.4686 mL | 2.3431 mL | 4.6863 mL | |
| 5 mM | 0.0937 mL | 0.4686 mL | 0.9373 mL | |
| 10 mM | 0.0469 mL | 0.2343 mL | 0.4686 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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