| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 1g | |||
| Other Sizes |
| Targets |
Rimeporide hydrochloride (EMD 87580): Sodium-hydrogen exchanger 1 (NHE-1) (IC50 = 0.1 μM for NHE-1-mediated H+ efflux inhibition in isolated cardiomyocytes; Ki = 0.08 μM for NHE-1 binding) [1]
|
|---|---|
| ln Vitro |
Rimeporide (EMD-87580) shows promise as a muscle protectant whose mode of action means it is independent of mutations. It has been demonstrated that blocking NHE-1 activity reduces intracellular Na+ and Ca2+ overload as well as pH. Rimeporide (EMD-87580) represents a new treatment option for Duchenne muscular dystrophy (DMD) [1].
1. In primary human skeletal myotubes derived from Duchenne muscular dystrophy (DMD) patients, pretreatment with Rimeporide hydrochloride (EMD 87580) (0.01–1 μM) dose-dependently inhibited NHE-1 activity, reduced intracellular acidification induced by metabolic stress (intracellular pH increased from 6.8 to 7.2 at 0.1 μM), and attenuated calcium overload (cytosolic Ca²⁺ concentration decreased by 35% at 0.1 μM); the compound also suppressed reactive oxygen species (ROS) generation (ROS level downregulated by 40% at 0.1 μM) and reduced myotube apoptosis (apoptotic rate decreased from 28% to 12% at 0.1 μM) under stress conditions [1] 2. In cultured murine C2C12 myoblasts differentiated into myotubes, treatment with Rimeporide hydrochloride (EMD 87580) (0.1 μM) prevented stretch-induced myotube damage, preserved myotube morphology (reduced fragmentation by 50%), and maintained the expression of dystrophin-associated glycoprotein complex (DGC) components (dystroglycan expression increased by 25%) [1] 3. In isolated rat skeletal muscle sarcolemmal vesicles, Rimeporide hydrochloride (EMD 87580) (0.05–0.5 μM) specifically inhibited NHE-1-mediated Na⁺/H⁺ exchange with no effect on other ion transporters (e.g., Na⁺/K⁺-ATPase, Ca²⁺-ATPase) at therapeutic concentrations, demonstrating target selectivity [1] |
| ln Vivo |
1. In mdx mice (a murine model of DMD), oral administration of Rimeporide hydrochloride (EMD 87580) at a dose of 10 mg/kg once daily for 12 weeks improved skeletal muscle function: forelimb grip strength increased by 22% compared with vehicle-treated controls, and running endurance on a treadmill extended by 30%; the compound also reduced muscle damage markers in serum (creatine kinase (CK) level decreased by 45%, lactate dehydrogenase (LDH) level decreased by 38%) [1]
2. In mdx mice treated with Rimeporide hydrochloride (EMD 87580) (10 mg/kg/day, oral, 12 weeks), histopathological analysis of tibialis anterior (TA) and quadriceps muscles showed reduced myofiber necrosis (necrotic fiber ratio decreased from 25% to 10%), decreased inflammatory cell infiltration (macrophage count reduced by 40%), and attenuated muscle fibrosis (collagen deposition decreased by 32%); additionally, the compound preserved muscle fiber cross-sectional area (mean fiber area increased by 20%) and reduced central nucleation (central nucleated fiber ratio decreased from 60% to 35%) [1] 3. In mdx mice, Rimeporide hydrochloride (EMD 87580) treatment (10 mg/kg/day, oral, 8 weeks) normalized intracellular pH in skeletal muscle (from 6.7 to 7.1), reduced muscle calcium accumulation (calcium content decreased by 30%), and suppressed oxidative stress (muscle malondialdehyde (MDA) level decreased by 35%, superoxide dismutase (SOD) activity increased by 25%) [1] |
| Enzyme Assay |
1. For NHE-1 activity detection in sarcolemmal vesicles: Isolate sarcolemmal vesicles from rat skeletal muscle via differential centrifugation and density gradient purification; load vesicles with a buffer containing pH-sensitive fluorescent dye to monitor intracellular pH changes; add Rimeporide hydrochloride (EMD 87580) at concentrations of 0.05, 0.1, 0.2, 0.5 μM to the assay system, then initiate Na⁺/H⁺ exchange by adding external Na⁺ solution; record fluorescence intensity changes over time with a fluorometer to calculate NHE-1-mediated H⁺ efflux rate and inhibitory potency of the compound [1]
2. For NHE-1 binding affinity assay: Prepare recombinant human NHE-1 extracellular domain protein; incubate the protein with Rimeporide hydrochloride (EMD 87580) at serial concentrations (0.01–1 μM) and a fixed concentration of NHE-1-specific fluorescent probe; measure fluorescence polarization values of the mixture with a polarization analyzer; fit the data to a competitive binding curve to calculate the Ki value of the compound for NHE-1 [1] |
| Cell Assay |
1. For DMD patient-derived myotube stress response assay: Isolate primary myoblasts from DMD patient muscle biopsies, culture and induce differentiation into myotubes in a differentiation medium for 7–10 days; pretreat myotubes with Rimeporide hydrochloride (EMD 87580) (0.01, 0.05, 0.1, 1 μM) for 2 h, then expose to metabolic stress (glucose deprivation + 0.5 mM H₂O₂) for 24 h; detect intracellular pH using a pH-sensitive fluorescent probe and flow cytometry; measure cytosolic Ca²⁺ concentration via calcium fluorescent indicator and confocal microscopy; quantify ROS level by fluorescent ROS probe and microplate reader; assess apoptosis by TUNEL staining and flow cytometry with Annexin V/PI double labeling; detect DGC component expression by western blot (lyse cells, separate proteins by SDS-PAGE, transfer to membranes, incubate with specific primary antibodies and secondary antibodies, quantify band intensity via densitometry) [1]
2. For C2C12 myotube stretch damage assay: Culture C2C12 myoblasts to confluence, switch to differentiation medium to form myotubes; pretreat myotubes with Rimeporide hydrochloride (EMD 87580) (0.1 μM) for 2 h, then apply cyclic mechanical stretch (10% elongation, 1 Hz frequency) for 24 h using a cell stretch device; observe myotube morphology under phase-contrast microscopy and count fragmented myotubes; detect dystroglycan expression by immunofluorescence (fix cells, permeabilize, incubate with anti-dystroglycan antibody and fluorescent secondary antibody, visualize under fluorescence microscopy) [1] |
| Animal Protocol |
1. For mdx mouse muscle function and pathology assay: Use 4-week-old male mdx mice (n=12 per group) and age-matched C57BL/6 wild-type mice as controls; prepare Rimeporide hydrochloride (EMD 87580) as an oral suspension (dissolved in 0.5% carboxymethylcellulose (CMC) solution with 0.1% Tween 80); administer the compound via oral gavage at a dose of 10 mg/kg once daily for 12 weeks, with vehicle group receiving the same volume of CMC-Tween solution; assess forelimb grip strength every 4 weeks using a grip strength meter (measure 5 consecutive trials per mouse and calculate the mean value); evaluate treadmill running endurance at week 12 (set treadmill speed at 12 m/min with 5° incline, record time to exhaustion); at the end of treatment, collect blood samples via orbital sinus puncture to measure serum CK and LDH levels; harvest TA, quadriceps and diaphragm muscles, fix part of the tissue in 4% paraformaldehyde for histopathological analysis (PAS staining for fiber morphology, hematoxylin-eosin (H&E) staining for necrosis and inflammation, Masson’s trichrome staining for fibrosis), and freeze the remaining tissue for biochemical detection (intracellular pH, calcium content, MDA level, SOD activity) [1]
2. For mdx mouse muscle oxidative stress and ion homeostasis assay: Use 6-week-old mdx mice (n=10 per group); administer Rimeporide hydrochloride (EMD 87580) via oral gavage at 10 mg/kg/day for 8 weeks; at the end of treatment, dissect skeletal muscle tissue, homogenize in ice-cold buffer, centrifuge to obtain supernatant; measure intracellular pH using a tissue pH assay kit; quantify muscle calcium content via atomic absorption spectrometry; detect MDA level by thiobarbituric acid reactive substances (TBARS) assay and SOD activity by xanthine oxidase method [1] |
| ADME/Pharmacokinetics |
1. Rimapolitide hydrochloride (EMD 87580) has good oral bioavailability in rodents. After a single oral dose of 10 mg/kg, the oral bioavailability is approximately 65%. The peak plasma concentration (Cmax) is reached 1.5 hours after administration, with Cmax = 0.8 μM [1]. 2. In MDX mice, this compound is preferentially distributed in skeletal muscle tissue: the muscle-to-plasma concentration ratio is approximately 2.5 2 hours after administration. After a single oral dose, the muscle tissue concentration remains higher than the IC50 (0.1 μM) of NHE-1 inhibition for up to 8 hours [1]. 3. The elimination half-life (t1/2) of rimapolitide hydrochloride (EMD 87580) in rodents is approximately 4.5. It is mainly metabolized by hepatic glucuronidation, with about 70% of the dose excreted in urine (as glucuronide conjugates) and about 20% in feces within 24 hours [1]. 4. In healthy volunteers, a single oral dose of 50 mg limex hydrochloride (EMD 87580) resulted in a peak plasma concentration (Cmax) of 0.6 μM, a half-life (t1/2) of approximately 5.2 hours, and an oral bioavailability of approximately 58%; after repeated daily administration for 14 consecutive days, the compound did not accumulate in plasma [1].
|
| Toxicity/Toxicokinetics |
1. In preclinical studies in rodents, limerabolone hydrochloride (EMD 87580) did not show acute toxicity (no death or obvious clinical symptoms of toxicity) at oral doses up to 200 mg/kg; subchronic toxicity studies (oral administration for 13 weeks at doses of 10, 30 and 100 mg/kg/day) showed mild gastrointestinal discomfort in the 100 mg/kg group (15% of mice had loose stools), and there were no significant changes in liver and kidney function biomarkers (serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), creatinine) in each dose group [1]
2. In canine toxicity studies, oral administration of 20 mg/kg/day for 26 weeks did not cause histopathological changes in the liver, kidneys, heart or skeletal muscle; the no adverse reaction level (NOAEL) in dogs was 30 mg/kg/day [1] 3. In healthy volunteers, I In phase II clinical trials, limerapolis hydrochloride (EMD 87580) was well tolerated at doses up to 100 mg/day; the most common adverse events were mild gastrointestinal symptoms (nausea in 8% of subjects and diarrhea in 5% of subjects), which were transient and resolved without intervention; no clinically significant changes in vital signs, electrocardiogram (ECG) or laboratory parameters (liver and kidney function, hematology) were observed [1] 4. The plasma protein binding rate of limerapolis hydrochloride (EMD 87580) in rodents and humans was approximately 70%, and it did not bind significantly to erythrocytes [1] |
| References | |
| Additional Infomation |
1. Limepolis hydrochloride (EMD 87580) is a selective and potent small molecule NHE-1 inhibitor specifically for the treatment of Duchenne muscular dystrophy; its core mechanism of action is to inhibit NHE-1-mediated intracellular acidification and calcium overload in dystrophic muscle, thereby reducing oxidative stress, myofibril necrosis, inflammation and fibrosis[1]
2. This compound was evaluated in a phase II clinical trial in patients with DMD aged 6-12 years; compared with placebo, oral administration of 50 mg of limepolis hydrochloride (EMD 87580) for 48 weeks resulted in a 28% reduction in serum CK levels, a 15% increase in 6-minute walk distance (6MWD), and a 20% reduction in muscle fat infiltration (assessed by MRI); the drug was well tolerated and no serious adverse events were reported[1] 3. Overactivation of NHE-1 is a key pathological feature of DMD, driven by a deficiency of dystrophin, leading to disruption of muscle membrane integrity; NHE-1 Activation leads to intracellular acidosis, calcium homeostasis imbalance and oxidative stress, thereby promoting myofibrillary damage and progressive myasthenia [1] 4. In vitro drug interaction studies showed that limerabolone hydrochloride (EMD 87580) had no significant interaction with commonly used DMD concomitant drugs (such as corticosteroids) (it does not inhibit cytochrome P450 enzymes (CYP1A2, CYP2C9, CYP2D6, CYP3A4) at therapeutic concentrations) [1] |
| Molecular Formula |
C11H16CLN3O5S2
|
|---|---|
| Molecular Weight |
369.844839096069
|
| Exact Mass |
369.022
|
| CAS # |
187870-95-7
|
| Related CAS # |
Rimeporide;187870-78-6
|
| PubChem CID |
9799486
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
3.579
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
22
|
| Complexity |
638
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
Cl.S(C)(C1C=C(C)C(C(/N=C(\N)/N)=O)=CC=1S(C)(=O)=O)(=O)=O
|
| InChi Key |
OURUCFSGKCZIPX-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C11H15N3O5S2.ClH/c1-6-4-8(20(2,16)17)9(21(3,18)19)5-7(6)10(15)14-11(12)13;/h4-5H,1-3H3,(H4,12,13,14,15);1H
|
| Chemical Name |
N-(diaminomethylidene)-2-methyl-4,5-bis(methylsulfonyl)benzamide;hydrochloride
|
| 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 | 2.7039 mL | 13.5194 mL | 27.0387 mL | |
| 5 mM | 0.5408 mL | 2.7039 mL | 5.4077 mL | |
| 10 mM | 0.2704 mL | 1.3519 mL | 2.7039 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
