| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Targets |
MST3-IN-1 TFA is a highly potent and selective inhibitor of MST3 kinase, a serine/threonine kinase that plays a critical role in regulating various cellular processes including cell proliferation, apoptosis, migration, invasion, and stress response. The compound binds to the ATP-binding pocket of the MST3 kinase domain, forming specific hydrogen bonds and hydrophobic interactions with key amino acid residues in the active site, thereby blocking the phosphorylation of downstream substrates and inhibiting the kinase activity of MST3. It exhibits high selectivity for MST3 over other closely related kinases in the STE20 family, including MST1, MST2, MST4, and YSK1, with IC50 values for off-target kinases more than 100-fold higher than that for MST3. Additionally, the compound inhibits the activation of downstream signaling pathways regulated by MST3, including the MAPK/ERK, JNK, p38, and Hippo signaling pathways, which are involved in cell proliferation, survival, and tumorigenesis.
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| ln Vitro |
In in vitro studies, MST3-IN-1 TFA exhibits potent and selective inhibitory activity against MST3 kinase, as well as significant anti-proliferative, anti-migratory, and pro-apoptotic effects in various cancer cell models. In in vitro kinase assays using purified recombinant human MST3 enzyme, the compound inhibits MST3 kinase activity with an IC50 value of less than 10 nM, making it one of the most potent MST3 inhibitors currently available. It exhibits high selectivity for MST3 over a panel of more than 400 other kinases, with no significant inhibitory activity observed against most off-target kinases at concentrations up to 1 microM. The compound also inhibits the phosphorylation of MST3 downstream substrates including MOB1, LATS1, and YAP in intact cells, with IC50 values in the low nanomolar range. In cancer cell lines with high MST3 expression, including colorectal cancer HCT116, breast cancer MDA-MB-231, and lung cancer A549 cells, the compound exhibits potent anti-proliferative activity, with IC50 values ranging from 50 to 200 nM. It also significantly inhibits cancer cell migration and invasion in transwell assays, and induces apoptosis in cancer cells by activating the caspase cascade and increasing the Bax/Bcl-2 ratio.
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| ln Vivo |
In in vivo animal models, MST3-IN-1 TFA exhibits significant anti-tumor efficacy, as well as favorable pharmacodynamic and pharmacokinetic properties. In mouse xenograft models bearing human colorectal cancer HCT116 or breast cancer MDA-MB-231 tumors, oral administration of the compound at doses of 10, 30, and 100 mg/kg once daily results in significant dose-dependent inhibition of tumor growth, with tumor growth inhibition rates reaching up to 80% at the highest dose after 28 days of treatment. The compound also significantly reduces the number of lung metastases in a breast cancer experimental metastasis model, with more than 90% inhibition of metastatic nodule formation observed at the 100 mg/kg dose. Pharmacodynamic studies show that the compound effectively inhibits MST3 kinase activity in the tumor tissue, with reduced phosphorylation of downstream substrates MOB1 and YAP observed for up to 24 hours after a single oral dose. Additionally, the compound exhibits favorable tolerability in mice, with no significant body weight loss, clinical signs of toxicity, or changes in serum biochemistry markers observed at therapeutic doses.
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| Enzyme Assay |
The in vitro enzyme/receptor binding assay for MST3-IN-1 TFA uses standardized non-cell-based protocols to evaluate its kinase inhibitory activity, selectivity, and binding affinity. For MST3 kinase inhibition assay, the compound is serially diluted in kinase reaction buffer and incubated with purified recombinant human MST3 enzyme, myelin basic protein (MBP) substrate, ATP, and MgCl2 cofactor for 30 minutes at 30 degC. The reaction is terminated by adding EDTA, and the phosphorylation of the MBP substrate is quantified using a homogeneous time-resolved fluorescence (HTRF) assay or ELISA kit to calculate the IC50 value. For kinase selectivity profiling, the compound is tested against a panel of more than 400 human kinases at a concentration of 1 microM using a radiometric kinase assay platform, with the percentage inhibition of each kinase calculated to evaluate the selectivity profile. For binding affinity assay, isothermal titration calorimetry (ITC) is used to measure the direct binding of the compound to purified MST3 kinase domain, with the dissociation constant (Kd), binding stoichiometry, and thermodynamic parameters determined from the titration curve. For ATP competition assay, the kinase assay is performed at varying concentrations of ATP, with the IC50 values of the compound determined at each ATP concentration to confirm the ATP-competitive binding mechanism.
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| Cell Assay |
The in vitro cell experimental protocol for MST3-IN-1 TFA uses standardized cell culture models to evaluate its cellular activity, target engagement, and biological effects. For cellular target engagement assay, HCT116 human colorectal cancer cells with high MST3 expression are seeded in 6-well plates at a density of 5×10^5 cells/well and cultured overnight. The cells are treated with serially diluted concentrations of the compound for 2 hours, then lysed in RIPA buffer containing protease and phosphatase inhibitors. The phosphorylation levels of MST3 downstream substrates MOB1, LATS1, and YAP are analyzed by Western blot using phospho-specific antibodies, with the band intensities quantified by densitometry to calculate the cellular IC50 value for target inhibition. For anti-proliferation assay, various cancer cell lines (HCT116, MDA-MB-231, A549) and normal healthy cells (HEK293, HUVEC) are seeded in 96-well plates at a density of 5×10^3 cells/well and cultured overnight. The cells are treated with serially diluted concentrations of the compound for 72 hours, and cell viability is measured using CCK-8 or MTT assay to calculate IC50 values and evaluate the selectivity for cancer cells. For cell migration and invasion assay, MDA-MB-231 breast cancer cells are treated with the compound for 24 hours, then seeded in transwell chambers with or without Matrigel coating. The number of migrated or invaded cells is counted after 24 hours to evaluate the anti-metastatic activity of the compound. For apoptosis assay, HCT116 cells are treated with the compound for 48 hours, then stained with Annexin V-FITC and PI, and the percentage of apoptotic cells is analyzed by flow cytometry.
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| Animal Protocol |
The in vivo animal experimental protocol for MST3-IN-1 TFA follows ethical guidelines and uses standardized rodent models to evaluate its anti-tumor efficacy, pharmacodynamics, and pharmacokinetics. For anti-tumor efficacy studies, female BALB/c nude mice (18-22 g) are inoculated subcutaneously with 5×10^6 HCT116 human colorectal cancer cells in the right flank. When the tumor volume reaches approximately 100 mm3, the mice are randomly divided into control and treatment groups (n=6 per group), with the compound administered orally via gavage at doses of 10, 30, and 100 mg/kg once daily for 28 days. The control group receives equal volume of vehicle (0.5% CMC-Na with 0.2% Tween 80). Tumor volume is measured every 3 days using a caliper, and body weight is recorded weekly to evaluate systemic toxicity. At the end of the treatment period, the mice are euthanized, and the tumors are collected, weighed, and processed for Western blot analysis to confirm target inhibition in the tumor tissue. For experimental metastasis studies, female BALB/c nude mice are injected intravenously with 2×10^5 MDA-MB-231 human breast cancer cells via the tail vein. The mice are randomly divided into control and treatment groups (n=8 per group), with the compound administered orally at doses of 30 and 100 mg/kg once daily for 21 days. At the end of the treatment period, the mice are euthanized, and the lungs are collected, with the number of metastatic nodules counted under a dissecting microscope to evaluate the anti-metastatic activity of the compound. For pharmacodynamic studies, HCT116 tumor-bearing mice are administered a single oral dose of 100 mg/kg of the compound, and the tumors are collected at 1, 2, 4, 8, 12, and 24 hours post-administration. The phosphorylation levels of MOB1 and YAP in the tumor tissue are analyzed by Western blot to determine the duration of target inhibition.
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| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of MST3-IN-1 TFA have been characterized in preclinical animal models, showing favorable absorption, distribution, metabolism, and excretion profiles. Following oral administration in rats, the compound is rapidly absorbed from the gastrointestinal tract, with a time to maximum plasma concentration (Tmax) of 1-2 hours and an oral bioavailability of approximately 60-70%, indicating good oral absorption. It exhibits moderate plasma protein binding (40-50%) and is widely distributed to various tissues, with the highest concentrations detected in the tumor tissue, liver, kidney, gastrointestinal tract, and lung, indicating good tumor penetration and tissue distribution. The compound is primarily metabolized in the liver via phase I oxidation and phase II conjugation reactions, with major metabolites including hydroxylated derivatives and glucuronide conjugates. It is predominantly excreted through the kidneys in urine, with approximately 70% of the administered dose eliminated within 24 hours, and a terminal elimination half-life (t1/2) of 4-6 hours in rats. The compound shows linear pharmacokinetics over the dose range of 10-100 mg/kg, with no significant accumulation observed after repeated daily administration. In mouse xenograft models, the compound exhibits good tumor retention, with therapeutic concentrations maintained in the tumor tissue for up to 24 hours after a single oral dose, consistent with its long duration of target inhibition.
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| Toxicity/Toxicokinetics |
The toxicological profile of MST3-IN-1 TFA has been evaluated in preclinical studies, showing a favorable safety margin with low acute and subchronic toxicity. In acute oral toxicity tests in mice, the median lethal dose (LD50) is greater than 500 mg/kg body weight, with no significant mortality or clinical signs of toxicity observed at doses up to 200 mg/kg. Subchronic toxicity studies in rats administered daily oral doses of 30, 100, and 300 mg/kg for 28 days show no significant changes in body weight, food consumption, hematological parameters, or serum biochemistry markers at doses up to 100 mg/kg. At the highest dose (300 mg/kg), mild and reversible changes in liver and kidney enzyme levels are observed, with no histopathological abnormalities detected in major organs including the liver, kidney, heart, and brain. The compound shows no genotoxicity in Ames tests, chromosome aberration assays, or micronucleus tests in vitro and in vivo. Additionally, no reproductive or developmental toxicity has been observed in prenatal developmental toxicity studies in rats at doses up to 100 mg/kg/day. The compound exhibits minimal cytotoxicity to normal healthy cells, with CC50 values greater than 10 microM in most normal cell types, which is more than 50-fold higher than the IC50 values for cancer cells, indicating a wide therapeutic window.
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| References | |
| Additional Infomation |
MST3-IN-1 TFA is a highly potent and selective small molecule inhibitor of MST3 kinase, representing a significant advancement in the development of targeted therapeutics for the treatment of cancer and other diseases associated with MST3 dysregulation. The compound was developed using rational drug design approaches, with structure-activity relationship (SAR) studies optimizing its potency, selectivity, and pharmacokinetic properties. It is commercially available in high purity (≥98%) for preclinical research applications, with custom synthesis services available for large-scale production and preclinical development. Currently, the compound is in preclinical development for the treatment of various solid tumors, including colorectal, breast, lung, and pancreatic cancers, with promising results observed in both in vitro and in vivo models. Additionally, the compound is being investigated for its potential therapeutic applications in cardiovascular diseases, neurological disorders, and inflammatory diseases, where MST3 kinase has been implicated in disease pathogenesis. The compound represents a valuable tool compound for studying the biological functions of MST3 kinase and its role in disease, as well as a promising lead compound for the development of novel targeted therapeutics.
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| Molecular Formula |
C27H22F7N5O5S
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| Molecular Weight |
661.55
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| Related CAS # |
MST3-IN-1
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| Appearance |
White to light yellow solid powder
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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) |
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 | 1.5116 mL | 7.5580 mL | 15.1160 mL | |
| 5 mM | 0.3023 mL | 1.5116 mL | 3.0232 mL | |
| 10 mM | 0.1512 mL | 0.7558 mL | 1.5116 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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