| Targets |
(3S,4R)-ME-344 specifically targets heme oxygenase 1 (HO-1) in the mitochondria. It binds to and alters the structure of HO-1, promoting its translocation from the rough endoplasmic reticulum to the mitochondria, although this effect is only observed in drug-sensitive cells like H460 and SHP-77. By disrupting HO-1 localization, the compound impairs mitochondrial function, reduces ATP synthesis, and triggers ROS generation. It is also reported to inhibit mitochondrial NADH dehydrogenase (Complex I) of the electron transport chain, thereby blocking oxidative phosphorylation and energy production. Its primary mechanism involves disrupting redox homeostasis in cancer cells.
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| ln Vitro |
In vitro studies have shown that (3S,4R)-ME-344 exhibits antitumor effects by disrupting mitochondrial redox balance and inhibiting HO-1 function. It reduces ATP production and induces ROS in a cell-specific manner, with effects limited to drug-sensitive cells such as H460 and SHP-77. The compound has demonstrated efficacy in AML cell lines and patient samples, especially in Mcl-1 overexpressing and Ara-C resistant models. In MTT assays, cells are incubated with the compound for 72 hours, followed by MTT addition (0.5 mg/ml, 4 h incubation) and absorbance reading at 550 nm. (3S,4R)-ME-344 is also used in actin polymerization assays at 20 microM final concentration.
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| ln Vivo |
In vivo studies have been conducted with its parent compound, ME-344. A Phase 1 clinical trial in patients with refractory solid tumors characterized its safety and PK profile. The maximum tolerated dose (MTD) and dose-limiting toxicities (DLTs) were determined; five DLTs were reported, including grade 3 neuropathy in four patients at doses of 15 and 20 mg/kg. In a Phase 1b study, ME-344 was combined with topotecan for patients with small cell lung, ovarian, and cervical cancers to evaluate safety and the recommended Phase 2 dose (RP2D). ME-344 has also shown efficacy in leukemia and lung tumor models. These studies are crucial for assessing the compound's translational potential.
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| Enzyme Assay |
(3S,4R)-ME-344 is an isoflavone small molecule that inhibits HO-1 and disrupts mitochondrial function by directly binding to the enzyme. A typical non-cellular protocol involves using purified HO-1 enzyme in a reaction buffer with hemin as a substrate. The compound is added at varying concentrations (0.1-100 microM) and incubated for 30 min at 37degC. Enzyme activity is measured by bilirubin production at 450 nm using a spectrophotometer or microplate reader. Inhibition is calculated relative to a control with 0.1% DMSO. Controls include an HO-1 inhibitor like SnPP (tin protoporphyrin IX). The IC50 is determined by nonlinear regression in GraphPad Prism.
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| Cell Assay |
For in vitro cell assays, cells (e.g., H460, SHP-77, or AML cells) are seeded in 96-well plates at 5,000-10,000 cells/well and cultured overnight. Cells are treated with (3S,4R)-ME-344 at concentrations from 0.1 to 100 microM for 24-72 h. Cell viability is measured by MTT assay: add 0.5 mg/mL MTT (4 h incubation), then 10% SDS in 0.1 M HCl to solubilize, and read absorbance at 550 nm with background subtraction at 690 nm. Apoptosis is assessed by flow cytometry using Annexin V/PI staining. ROS is measured with DCFH-DA probe. Mitochondrial membrane potential uses JC-1 staining. Each experiment is performed in triplicate.
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| Animal Protocol |
For in vivo animal studies using ME-344, tumor xenograft models (e.g., AML or lung cancer) are typically used. For example, in leukemia models, mice are treated with ME-344 as a single agent or in combination. A standard protocol for subcutaneous xenografts involves dosing mice (n=10/group) with ME-344 when tumors reach ~100 mm3. The compound is dissolved in a formulation of 10% DMSO, 40% PEG300, 5% Tween 80, and 45% saline to a working concentration of 2 mg/mL. Dosing is IV or IP at 10-20 mg/kg once daily. Tumor volume and body weight are measured every 3 days. After 2-4 weeks, tumors are excised, weighed, and analyzed.
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| ADME/Pharmacokinetics |
The parent compound ME-344 was evaluated in a Phase 1 clinical trial in patients with refractory solid tumors. The study was open-label, dose-escalation with 30 patients (median age 65, 67% female) to determine MTD and DLTs. Dose-limiting toxicities included grade 3 neuropathy reported in 4 patients at 15-20 mg/kg. The RP2D was determined as 10 mg/kg IV weekly. ME-344 has a short half-life, is cleared via hepatic metabolism, and is safe as a single agent with manageable toxicities. Other ME-344 studies obtained PK samples to characterize plasma concentration-time profiles and exposure (AUC). Detailed PK parameters (Cmax, T1/2, Vd, CL) have not been disclosed for the (3S,4R)-enantiomer specifically.
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| Toxicity/Toxicokinetics |
The parent compound ME-344 was generally well-tolerated in clinical studies; however, dose-limiting toxicities were reported. In a Phase 1 dose-escalation trial of 30 patients, dose-limiting toxicities (DLTs) included grade 3 neuropathy in four patients, with two patients each at 15 mg/kg and 20 mg/kg. The most common adverse events were gastrointestinal (nausea, diarrhea) and fatigue. No grade 4 or 5 treatment-related toxicities were reported in this study. ME-344 can also induce oxidative stress due to its mechanism of action, but this is part of its antitumor efficacy rather than a toxic side effect. Comprehensive preclinical toxicology studies for the enantiomer are not publicly available.
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| References | |
| Additional Infomation |
(3S,4R)-ME-344 is the (3S,4R)-enantiomer of ME-344 and is available from commercial suppliers. Synonyms: ME-344, (3S,4R)-ME-344 enantiomer. Molecular formula: C22H20O4. Molecular weight: 348.39. CAS: 1374524-84-1. Storage: -20degC for powder, -80degC for solutions; shipping with blue ice. For research use only-not for human use. Purity: typically ≥98% by HPLC. Replaces: DPQ, TUDCA, Picroside II. Appearance: white to off-white solid. Solubility: DMSO 70 mg/mL, ethanol <1 mg/mL, PBS <1 mg/mL.
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| Molecular Formula |
C22H20O4
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|---|---|
| Molecular Weight |
348.39
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| CAS # |
1374524-84-1
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| Related CAS # |
ME-344; 1374524-68-1
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| Appearance |
Off-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 | 2.8703 mL | 14.3517 mL | 28.7035 mL | |
| 5 mM | 0.5741 mL | 2.8703 mL | 5.7407 mL | |
| 10 mM | 0.2870 mL | 1.4352 mL | 2.8703 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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