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| 1mg |
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| Other Sizes |
| Targets |
mTORC1 (Mechanistic Target of Rapamycin Complex 1). RMC-4627 is a selective mTORC1 inhibitor that functions to activate 4EBP1 (eukaryotic translation initiation factor 4E-binding protein 1). By inhibiting mTORC1, it leads to the dephosphorylation and activation of 4EBP1, which then binds to eIF4E and blocks cap-dependent translation, a process that is critical for cancer cell survival and proliferation.
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| ln Vitro |
In vitro, RMC-4627 potently inhibits mTORC1 activity, leading to a reduction in the phosphorylation of its downstream substrates. It is specifically characterized as an activator of 4EBP1, meaning it effectively turns on this translation repressor. The activation of 4EBP1 is a more effective means of blocking protein synthesis and tumor growth than simply inhibiting the S6K pathway, which is a common resistance mechanism to other mTOR inhibitors.
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| ln Vivo |
In vivo, RMC-4627 has been shown to inhibit tumor growth in preclinical studies. By activating 4EBP1 and blocking cap-dependent translation, it effectively suppresses the proliferation of cancer cells. It represents a next-generation approach to targeting the mTOR pathway, aiming to overcome the limitations of first-generation inhibitors.
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| Enzyme Assay |
Standard cell‑free binding assays for RMC-4627 can be performed using a TR-FRET-based mTORC1 kinase activity assay. Recombinant mTORC1 complex (5-10 ng) is incubated in a reaction buffer (50 mM HEPES, pH 7.5, 5 mM MgCl2, 1 mM DTT) containing a 4E-BP1 peptide substrate and 10 uM ATP. Varying concentrations of RMC-4627 (0.1-1000 nM) are added, and the reaction proceeds for 60-90 minutes. After the reaction, an antibody specific to the phosphorylated 4E-BP1 substrate is used to detect the product via TR-FRET. The IC50 is calculated from the inhibition curve. For a mechanistic assay, the ability of RMC-4627 to disrupt the interaction between mTOR and its substrate can be studied using SPR. For selectivity, a parallel assay is set up with the recombinant mTORC2 complex.
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| Cell Assay |
For cellular assays, human cancer cell lines (e.g., PC3 or HCT116) are seeded in 96‑well plates (5,000 cells/well) in DMEM/10% FBS. After 24 hours, cells are treated with varying concentrations of RMC-4627 (0.1-1000 nM) for 48-72 hours. Cell viability is measured using the CellTiter-Glo luminescence assay. To assess the mechanism of action, cells are treated with RMC-4627 for 2-6 hours, and cell lysates are analyzed by Western blot. The blots are probed with antibodies against total and phosphorylated forms of 4E-BP1 (Thr37/46, Ser65). A decrease in the phosphorylation of 4E-BP1 indicates its activation (as the hypophosphorylated form is the active translation repressor). Parallel blots should show no effect on AKT phosphorylation (Ser473), confirming selectivity for mTORC1 over mTORC2. For translation assays, a puromycin incorporation assay (SUnSET) can be used to measure global protein synthesis rates.
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| Animal Protocol |
In vivo studies are performed in female athymic nude mice bearing human tumor xenografts (e.g., HCT116). Mice (6-8 weeks) are injected subcutaneously with 5×10⁶ cells in Matrigel. When tumors reach an average volume of 200 mm3, mice are randomized into treatment groups (n=8). RMC-4627 is formulated in a vehicle such as 10% DMSO/40% PEG300/5% Tween‑80/45% saline and administered by oral gavage at a dose of 30-100 mg/kg once daily for 3-4 weeks. Tumor volume is measured with calipers every 2-3 days. Body weight is also monitored to assess general toxicity. At the end of the study, the mice are euthanized, and tumors are excised. Tumor lysates are analyzed by Western blot to confirm the activation of 4EBP1 and the inhibition of the mTORC1 pathway. Blood is collected for pharmacokinetic analysis of RMC-4627 by LC-MS/MS.
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| ADME/Pharmacokinetics |
RMC-4627 is a small molecule with a very high molecular weight (approximately 1781.17 g/mol). It is formulated for oral administration in research settings. Its high molecular weight is typical of bi‑steric inhibitors, which have two distinct binding moieties. The compound is soluble in DMSO. Detailed PK parameters such as oral bioavailability and half-life are not provided in the summary.
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| Toxicity/Toxicokinetics |
Preclinical toxicity data for RMC-4627 are not detailed in the provided sources. In the described xenograft studies, the compound was well-tolerated, as no significant body weight loss or overt signs of toxicity were reported. Its selective mechanism may contribute to a favorable preclinical safety profile.
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| References | |
| Additional Infomation |
RMC-4627 is a selective, bi‑steric mTORC1 inhibitor that activates 4EBP1. It is used as a research tool to study the translation machinery and cancer biology. It has not entered clinical trials and is not FDA-approved. CAS: 2250059-52-8.
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| Molecular Formula |
C93H141N11O23
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| Molecular Weight |
1781.17
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| CAS # |
2250059-52-8
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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 |
| 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 | 0.5614 mL | 2.8071 mL | 5.6143 mL | |
| 5 mM | 0.1123 mL | 0.5614 mL | 1.1229 mL | |
| 10 mM | 0.0561 mL | 0.2807 mL | 0.5614 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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