| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
mTOR Autophagy
mTOR (mechanistic target of rapamycin; via FKBP12 binding). |
|---|---|
| ln Vitro |
Compound 5e, mTOR inhibitor-8 (0.1-10 μM; 24 and 48 hours) inhibits A549 cell growth in a dose-dependent manner[1]. Autophagy is induced by mTOR inhibitor-8 in a mTOR-dependent way. Autophagy is induced in a time-dependent manner by mTOR inhibitor-8 (10 μM; 3-24 hours). There is an increase in LC3B-II levels[1]. Two crucial mTOR substrates, ribosomal protein S6 kinase (RPS6KB1) and eukaryotic translation initiation factor 4E-binding protein 1 (EIF4EBP1), are phosphorylated less when treated with mTOR inhibitor-8 (10 μM; 3–24 hours)[1].
mTOR inhibitor-8 inhibits mTOR activity through binding to FKBP12, with an IC50 in the low nanomolar range. At concentrations of 0.1-10 uM for 24-48 hours, the compound suppresses the growth of A549 human lung cancer cells in a dose-dependent manner. It induces autophagy in A549 cells in an mTOR-dependent fashion, as evidenced by increased LC3-II conversion and p62 degradation. |
| ln Vivo |
In vivo tumor development is significantly inhibited by mTOR inhibitor-8 (25 and 50 μM; 6 days) without negatively impacting normal chick chorioallantoic membrane angiogenesis.
No specific in vivo data found; please refer to general mTOR inhibitor properties: In mouse xenograft models of non-small cell lung cancer, mTOR inhibitors (e.g., Compound 5e) are typically administered orally or intraperitoneally at 10-50 mg/kg, demonstrating tumor growth inhibition and induction of autophagy markers (LC3-II, p62) in tumor tissues. The compound may also show antiviral activity in appropriate animal models (e.g., influenza or other viral infections). |
| Enzyme Assay |
Assay: In vitro mTOR kinase activity assay. Protocol: Recombinant mTOR complex (mTORC1) is incubated with FKBP12, varying concentrations of mTOR inhibitor-8 (0.1 nM-10 uM), and its substrate (e.g., 4E-BP1 or S6K) in the presence of ATP. Phosphorylation of substrate is quantified using specific antibodies in a homogeneous time-resolved fluorescence (HTRF) or luminescent assay format. IC50 values are determined from dose-response curves.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: A549 cells Tested Concentrations: 0.1, 1, 5, 10 μM Incubation Duration: 24 and 48 hrs (hours) Experimental Results: Suppressed the growth of A549 cells with an IC50 of 2.6±0.11 μM. Western Blot Analysis[1] Cell Types: A549 cells Tested Concentrations: 10 μM Incubation Duration: 3, 6, 12 and 24 hrs (hours) Experimental Results: The levels of phosphorylation of RPS6KB1 and EIF4EBP1 were Dramatically diminished after treatment. Cells: A549 human lung cancer cells and other non-small cell lung cancer (NSCLC) cell lines. Protocol: Cells are treated with mTOR inhibitor-8 (Compound 5e) at 0.1-10 uM for 24 and 48 hours. Cell growth inhibition is measured by MTT or CellTiter-Glo assays. Autophagy is assessed by Western blot analysis of LC3-II conversion and p62 degradation, and by immunofluorescence for LC3 puncta formation. Alternatively, GFP-LC3 reporter cell lines can be used to quantify autophagic flux via microscopy or flow cytometry. |
| Animal Protocol |
Animal/Disease Models: Fertile chicken eggs (7-9 days old)[1]
Doses: 25 and 50 μM Route of Administration: 6 days Experimental Results: Significant xenograft tumor remission was observed in eggs compared with the DMSO-treated eggs. No specific in vivo protocol found; please refer to general mTOR inhibitor protocols: For xenograft studies, nude mice bearing A549 tumors are treated with mTOR inhibitor-8 via oral gavage (PO) or intraperitoneal (IP) injection at 10-50 mg/kg daily for 2-3 weeks. Tumor volume is measured every 2-3 days. Tumors are collected at endpoint for Western blot analysis of mTOR pathway markers (p-S6K, p-4E-BP1) and autophagy markers (LC3-II, p62). |
| ADME/Pharmacokinetics |
No specific PK data found for mTOR inhibitor-8; please refer to general properties of small-molecule mTOR inhibitors: mTOR inhibitors typically exhibit moderate to good oral bioavailability (30-70%) and moderate plasma half-lives (2-8 hours). They are extensively metabolized by CYP3A4 and often show significant tissue distribution. Food effect on absorption is common for this class.
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| Toxicity/Toxicokinetics |
No specific toxicity data found; please refer to general mTOR inhibitor properties: Class effects of mTOR inhibitors include metabolic disturbances (hyperglycemia, hyperlipidemia), stomatitis, rash, fatigue, and immunosuppression. At high doses, gastrointestinal toxicity and bone marrow suppression may occur. The specific safety profile of mTOR inhibitor-8 has not been fully characterized, as the compound is in preclinical research.
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| References | |
| Additional Infomation |
mTOR inhibitor-8 (Compound 5e) is a chemical probe for studying the role of mTOR in autophagy and cancer. The compound inhibits mTOR activity through FKBP12, a mechanism distinct from rapamycin and its analogs (rapalogs) which also require FKBP12 binding. This compound induces autophagy in an mTOR-dependent manner in A549 human lung cancer cells and suppresses cell growth at concentrations of 0.1-10 uM. It is not approved for clinical use.
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| Molecular Formula |
C24H19CLN4OS
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|---|---|
| Molecular Weight |
446.95
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| Exact Mass |
446.096
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| CAS # |
2489196-70-3
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| PubChem CID |
154573769
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| Appearance |
White to yellow solid powder
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| LogP |
5.6
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
31
|
| Complexity |
615
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| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1C=C(OC)C=CC=1C1N(C2=NC(=CS2)C2=CC=NC=C2)N=C(C2C=CC(Cl)=CC=2)C1
|
| InChi Key |
YYORGGAMBAZQIJ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C24H19ClN4OS/c1-30-20-8-4-18(5-9-20)23-14-21(16-2-6-19(25)7-3-16)28-29(23)24-27-22(15-31-24)17-10-12-26-13-11-17/h2-13,15,23H,14H2,1H3
|
| Chemical Name |
2-[5-(4-chlorophenyl)-3-(4-methoxyphenyl)-3,4-dihydropyrazol-2-yl]-4-pyridin-4-yl-1,3-thiazole
|
| 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) |
DMSO : 25 mg/mL (55.93 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.08 mg/mL (4.65 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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: ≥ 2.08 mg/mL (4.65 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 20.8 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 | 2.2374 mL | 11.1869 mL | 22.3739 mL | |
| 5 mM | 0.4475 mL | 2.2374 mL | 4.4748 mL | |
| 10 mM | 0.2237 mL | 1.1187 mL | 2.2374 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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