| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Pyruvate kinase-R (PKR) (activator)
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| ln Vitro |
In human red blood cells, etabopivit (20 μM, 4 h) can lower sickle point (PoS) and increase hemoglobin-oxygen affinity [1].
(Rac)-Etavopivat is an isomer of Etavopivat. Etavopivat is an orally active erythrocyte pyruvate kinase-R (PKR) activator. In vitro, Etavopivat activates PKR, leading to increased ATP levels and decreased 2,3-DPG levels in red blood cells. This shifts the oxygen dissociation curve to the left, increasing hemoglobin oxygen affinity and reducing sickling under hypoxic conditions. The racemate contains both active and inactive enantiomers; the specific enantiomer (the R-enantiomer or S-enantiomer) responsible for PKR activation is not specified for the racemate. For research purposes, the racemate may be used as a comparator to the pure active enantiomer. |
| ln Vivo |
Cynomolgus monkeys treated with etabopipate (oral, 3–22 mg/kg once daily for five days) showed increases in 2,3-DPG and ATP at 8 and 22 mg/kg, respectively [1].
No specific in vivo data for (Rac)-Etavopivat. The active enantiomer, Etavopivat (FT-4202), has been extensively studied in vivo. In preclinical models of sickle cell disease, Etavopivat reduces red blood cell sickling, improves red blood cell survival, decreases hemolysis, and reduces markers of inflammation and vaso-occlusion. In Phase 2 clinical trials, Etavopivat has shown efficacy in increasing hemoglobin levels, reducing markers of hemolysis, and improving patient-reported outcomes. (Rac)-Etavopivat would be expected to have reduced potency compared to the pure active enantiomer. |
| Enzyme Assay |
Recombinant human pyruvate kinase-R (PKR) enzyme is expressed in E. coli or insect cells and purified. The PKR activity assay measures the conversion of phosphoenolpyruvate (PEP) and ADP to pyruvate and ATP, coupled to NADH oxidation via lactate dehydrogenase (LDH). The reaction mixture contains Tris-HCl buffer (pH 7.4), KCl, MgCl2, PEP (0.5-5 mM), ADP (0.5-5 mM), NADH (0.2-0.5 mM), and LDH. The reaction is initiated by adding PKR enzyme (10-50 ng). The decrease in absorbance at 340 nm (NADH oxidation) is monitored over 5-10 minutes at 37degC. The initial velocity (V0) is calculated. For activator screening, compounds are tested at varying concentrations (0.001-100 uM), and EC50 values are determined. Etavopivat is an activator, not an inhibitor, so the assay measures increased enzymatic activity. (Rac)-Etavopivat would be expected to activate PKR with an EC50 value approximately twice that of the pure active enantiomer. No specific EC50 is publicly available for the racemate.
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| Cell Assay |
Human erythroblast cell lines (e.g., BEL-A or HUDEP-2) or primary human red blood cells are cultured in appropriate media. Cells are treated with (Rac)-Etavopivat (0.01-100 uM) for 24-72 hours. Red blood cell PKR activity is measured in cell lysates using the PKR activity assay described above. Intracellular ATP levels are measured using a bioluminescence assay (e.g., CellTiter-Glo). 2,3-DPG levels are measured by an enzymatic colorimetric assay (2,3-DPG assay kit). The ratio of ATP to 2,3-DPG is calculated. Hemoglobin oxygen affinity is measured using a Hemox Analyzer or by assessing the P50 value (partial pressure of oxygen at which hemoglobin is 50% saturated). For sickling assays, red blood cells from sickle cell disease patients are treated with (Rac)-Etavopivat (0.1-10 uM) and exposed to hypoxic conditions (e.g., 2% O2 or sodium metabisulfite). The percentage of sickled red blood cells is quantified by light microscopy. Cell viability is assessed by MTT or by counting cells with trypan blue. Apoptosis (Annexin V/PI staining) and red blood cell morphology are also assessed.
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| Animal Protocol |
For in vivo studies of the active enantiomer Etavopivat, the compound is administered orally to rodent models of sickle cell disease (e.g., Berkeley SCD mice or Townes SCD mice). Doses typically range from 5-50 mg/kg once or twice daily for 2-8 weeks. Endpoints include red blood cell PKR activity (ex vivo assay), ATP and 2,3-DPG levels in red blood cells, complete blood count (CBC, with particular focus on hemoglobin, hematocrit, red blood cell count, reticulocyte count), markers of hemolysis (bilirubin, lactate dehydrogenase, haptoglobin), red blood cell survival (biotinylation labeling method), and histopathological examination of spleen and liver for signs of extramedullary hematopoiesis and iron deposition. For the racemate, similar protocols would be used but with dose adjustments (typically 2× the active enantiomer dose) due to the presence of the inactive isomer. SCD mouse models also assess vaso-occlusion (e.g., mesenteric microcirculation studies) and inflammation (plasma IL-6, TNF-alpha). No specific in vivo data for the racemate is publicly available.
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| ADME/Pharmacokinetics |
No specific pharmacokinetic data for (Rac)-Etavopivat. For the active enantiomer Etavopivat (FT-4202), the compound is orally bioavailable with good absorption. In humans, Phase 1/2 studies have reported pharmacokinetic parameters: Tmax ~1-2 hours, terminal half-life suitable for once-daily dosing (approximately 6-12 hours), dose-proportional AUC and Cmax. Metabolism is primarily via CYP3A4, and drug-drug interaction potential exists. Excretion is primarily in feces and urine. The racemate would be expected to have similar PK parameters to the pure active enantiomer, but with differences in enantiomer-specific clearance. Molecular weight: 457.5 (for the racemate). Solubility: soluble in DMSO. Storage: powder at -20degC for 3 years; in solvent at -80degC for 6 months.
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| Toxicity/Toxicokinetics |
No specific toxicity data for (Rac)-Etavopivat. For the active enantiomer Etavopivat (FT-4202), preclinical toxicity studies have been conducted to support clinical development. In clinical trials, Etavopivat has been generally well tolerated. Common adverse events include mild gastrointestinal disturbances (nausea, diarrhea) and headache. No significant drug-induced liver injury, cardiotoxicity (QT prolongation), or genotoxicity has been reported. As a PKR activator, the primary expected on-target effect is modulation of red blood cell metabolism, which is the intended therapeutic mechanism. Long-term safety data from ongoing clinical trials will further characterize the risk-benefit profile. The racemate is not used clinically; only the pure active enantiomer has been advanced to clinical trials.
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| References | |
| Additional Infomation |
(Rac)-Etavopivat ((Rac)-FT-4202, CAS: 2622070-93-1) is the racemic isomer of Etavopivat (FT-4202), an orally active erythrocyte pyruvate kinase-R (PKR) activator. PKR is a key enzyme in the glycolytic pathway in red blood cells. Activation of PKR increases ATP production and reduces 2,3-DPG levels, shifting the hemoglobin oxygen dissociation curve to the left and increasing oxygen affinity. This reduces red blood cell sickling under hypoxic conditions in sickle cell disease (SCD). Etavopivat (the active enantiomer) has been investigated in Phase 2/3 clinical trials for SCD and other hemoglobinopathies. (Rac)-Etavopivat is a research reagent used for comparative studies. Molecular formula: C22H23N3O6S, molecular weight: 457.5. Appearance: white to off-white solid powder. Solubility: soluble in DMSO. Storage: powder at -20degC. Not approved for clinical use. For research use only. References: Etavopivat is an orally active erythrocyte pyruvate kinase-R (PKR) activator that can be used in studies of sickle cell disease and other haemoglobinopathies.
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| Molecular Formula |
C22H23N3O6S
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| Molecular Weight |
457.499524354935
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| Exact Mass |
457.13
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| CAS # |
2622070-93-1
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| Related CAS # |
Etavopivat;2245053-57-8
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| PubChem CID |
135338361
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| Appearance |
White to off-white solid powder
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| LogP |
-0.8
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
32
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| Complexity |
832
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| Defined Atom Stereocenter Count |
0
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| SMILES |
S(C1=CN=C2C(=C1)OCCO2)(N1CC2CN(C(C(C3C=CC=CC=3)CO)=O)CC=2C1)(=O)=O
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| InChi Key |
KZFFYEPYCVDOGE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H23N3O6S/c26-14-19(15-4-2-1-3-5-15)22(27)24-10-16-12-25(13-17(16)11-24)32(28,29)18-8-20-21(23-9-18)31-7-6-30-20/h1-5,8-9,19,26H,6-7,10-14H2
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| Chemical Name |
1-[5-(2,3-dihydro-[1,4]dioxino[2,3-b]pyridin-7-ylsulfonyl)-1,3,4,6-tetrahydropyrrolo[3,4-c]pyrrol-2-yl]-3-hydroxy-2-phenylpropan-1-one
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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 | 2.1858 mL | 10.9290 mL | 21.8579 mL | |
| 5 mM | 0.4372 mL | 2.1858 mL | 4.3716 mL | |
| 10 mM | 0.2186 mL | 1.0929 mL | 2.1858 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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