| ln Vitro |
RN-0001 (various concentrations; reaction at room temperature for 30 minutes, reaction at 30°C for 15 minutes) showed a weak direct calcineurin inhibitory effect in cell-free experiments (IC50 = 2276 nM), and the inhibitory effect was slightly enhanced after forming a complex with cyclosporine A (IC50 = 1551 nM) [1]. RN-0001 (various concentrations) maintained the mitochondrial calcium retention capacity in cell-free mitochondrial isolation experiments, with an EC50 of 916 nM [1]. RN-0001 (various concentrations) showed reduced immunosuppressive activity and inhibited IL-2 release in cell-based immunocellular experiments, with an EC50 of 347.90 nM [1]. RN-0001 (0.01-10 μM; 24 hours) increased the survival rate of ethanol-damaged AML12 hepatocytes (IC50 = 102.7 μM) and RAW264.7 macrophages (IC50 = 60.73 μM) in a concentration-dependent manner, and was superior to Debio-025 at the same dose [1]. RN-0001 (0.01-10 μM; 24 hours) reduced LDH release, apoptosis, and abnormal expression of adipogenesis, inflammation, oxidative stress, and apoptosis-related proteins in ethanol-damaged AML12 hepatocytes in a concentration-dependent manner [1].
|
|---|---|
| ln Vivo |
RN-0001 (10 mg/kg; intraperitoneal injection; once daily for 11 days) effectively alleviated ethanol-induced liver damage, mitochondrial dysfunction, steatosis and inflammatory response in a mouse model of alcoholic liver disease [1]. RN-0001 (10 mg/kg; intraperitoneal injection; once every two days for 90 days) was well tolerated in healthy mice with no adverse reactions observed [1].
|
| Cell Assay |
Apoptosis analysis [1]
Cell Types: AML12 cells Tested Concentrations: 0.01, 0.1, 1 and 10 μM Incubation Duration: 24 hours Experimental Results: Apoptosis was reduced. |
| Animal Protocol |
Animal/Disease Models:C57BL/6J (male, 7 weeks old, NIAAA chronic plus binge drinking ethanol feeding model) [1]
Doses: 10 mg/kg Route of Administration: Intraperitoneal injection; once daily Experimental Results: Partially reversed alcohol-induced weight loss. Reduced serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), and total cholesterol (TC) levels. Reduced serum levels of pro-inflammatory cytokines TNF-α and IL-6. Restored liver weight/body weight ratio to normal. Reduced liver free fatty acid (FFA) and TG levels. Reduced hepatic fat droplets and macrophage infiltration, thereby reducing non-alcoholic fatty liver disease activity score (NAS). It reversed the abnormal expression of key proteins involved in mitochondrial function (DRP1, FIS1, MFN1, MFN2, OPA1), oxidative stress (CYP2E1, iNOS), antioxidation (p-NRF2, NRF2, SOD1, CAT), and apoptosis (cleaved caspase-3, Cyto C, Bcl2, BAX). Mitochondrial swelling was reduced after ethanol exposure. |
| References |
| Molecular Formula |
C66H120N12O13
|
|---|---|
| Molecular Weight |
1289.73
|
| CAS # |
252731-57-0
|
| Appearance |
Typically exists as solids at room temperature
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 0.7754 mL | 3.8768 mL | 7.7536 mL | |
| 5 mM | 0.1551 mL | 0.7754 mL | 1.5507 mL | |
| 10 mM | 0.0775 mL | 0.3877 mL | 0.7754 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
