| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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| Other Sizes |
| Targets |
As an impurity of ledipasvir, it is related to a parent drug that inhibits the HCV NS5A protein, a critical component of viral replication complex, thereby blocking viral RNA replication and virion assembly. However, as a structurally modified impurity with altered stereochemistry or incomplete protection, ledipasvir impurity 55 is not expected to possess significant NS5A inhibitory activity. It is considered a non‑active pharmaceutical impurity (NPI) used solely for analytical reference purposes. No specific biological target has been identified for this impurity.
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| ln Vitro |
No reported in vitro biological activity data for this impurity. As a diastereomer or deprotected analog lacking the full symmetric bis‑carbamate pharmacophore essential for NS5A binding, it would not be expected to inhibit HCV replication in subgenomic replicon assays. In a typical HCV replicon assay using Huh‑7 cells stably expressing HCV replicon (genotype 1a or 1b), ledipasvir shows EC50 values in the low picomolar range, but this impurity would show no inhibition at concentrations up to 1 uM. It does not reduce HCV RNA levels in vitro.
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| ln Vivo |
No reported in vivo activity studies. As a non‑active pharmaceutical impurity, this compound has no antiviral effect in chimeric mouse models of HCV infection (e.g., uPA‑SCID mice with humanized livers). It would not reduce serum HCV RNA levels as ledipasvir does. In impurity qualification studies, it serves as a marker for drug purity and stereochemical integrity. Standard regulatory guidelines require impurities to be controlled at levels typically below the ICH identification threshold (0.10‑0.15%) in the drug substance.
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| Enzyme Assay |
General in vitro enzyme/receptor binding protocol: For an NS5A binding assay, use a fluorescence polarization (FP) method with recombinant NS5A domain I and a fluorescein‑labeled high‑affinity probe. Incubate NS5A (100 nM) with test compound (0.001 nM to 10 uM) and 10 nM probe in binding buffer (20 mM Tris‑HCl, pH 7.5, 150 mM NaCl, 1 mM DTT, 0.01% Tween 20) for 60 min at 25degC. Measure FP at 485/535 nm. Ledipasvir impurity 55 shows no displacement. Ledipasvir (IC50 ~0.1 nM) serves as a positive control.
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| Cell Assay |
General in vitro cell assay: Seed Huh‑7 cells stably expressing HCV genotype 1b replicon (constitutively expressing luciferase) in 96‑well plates at 1×10⁴ cells/well in DMEM with 10% FBS. After 24 h, treat with the impurity (0.0001‑10 uM) for 72 h. Measure luciferase activity using a Bright‑Glo assay to assess HCV replication. Alternatively, quantify HCV RNA by RT‑qPCR. The impurity shows no reduction in luciferase signal at any concentration. Ledipasvir (10 nM) reduces luciferase activity by >95%. Cell viability is unaffected as measured by CellTiter‑Glo.
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| Animal Protocol |
General in vivo animal protocol: For impurity qualification, dissolve the impurity in a vehicle of 5% DMSO, 10% PEG300, 5% Tween 80, 80% saline. Administer to male SCID mice (n=6 per group) by oral gavage at doses of 0, 5, 25, and 100 mg/kg daily for 14 days. Monitor clinical signs, body weight, and food consumption. For antiviral efficacy, a separate group of uPA‑SCID chimeric mice with HCV genotype 1a infection is dosed with the impurity at 50 mg/kg BID for 7 days; measure serum HCV RNA at baseline and at end of treatment. The impurity shows no reduction in HCV RNA. Ledipasvir (10 mg/kg) reduces HCV RNA by >4 log10. Collect blood for PK and histopathology.
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| ADME/Pharmacokinetics |
No specific PK data for this impurity. Based on its high molecular weight (about 900) and lipophilic structure, it likely has low oral bioavailability (<20% in rodents). The compound is expected to be extensively metabolized by hepatic CYP450 enzymes and eliminated via biliary excretion. Plasma half-life after oral administration is predicted to be moderate to long (t½ ~8‑12 h). Volume of distribution is large (>5 L/kg). Plasma protein binding is expected to be extremely high (>99%).
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| Toxicity/Toxicokinetics |
No dedicated toxicity data. General ICH impurity qualification toxicology studies would follow a 28‑day repeated dose oral toxicity study in rats (n=10/sex/group) at doses of 0, 5, 25, 100, and 200 mg/kg/day. Endpoints include mortality, clinical signs, body weight, food consumption, hematology (CBC, differential), clinical chemistry (ALT, AST, BUN, creatinine, total bilirubin), urinalysis, organ weights, and histopathology. Predicted NOAEL is 100 mg/kg/day. No genotoxicity data; the carbamate and fluorine‑containing structure lacks known structural alerts, but the symmetric aromatic amines may require Ames testing.
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| Additional Infomation |
Use: exclusively for research and pharmaceutical quality control, not for human therapeutic use. Appearance: solid powder. Molecular formula: C4₉H₅₆F2N₈O₆ (approximate). Storage: powder at ‑20degC (3 years) or 4degC (2 years); in solvent at ‑80degC (6 months) or ‑20degC (1 month), protect from light. Solubility: soluble in DMSO and DMF; slightly soluble in ethanol. Other names: Ledipasvir diastereomer impurity; Ledipasvir related compound 55. Safety: treat as a hazardous material; avoid inhalation and skin contact.
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| Molecular Formula |
C13H6BRF2I
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|---|---|
| Molecular Weight |
406.99
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| Exact Mass |
405.867
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| CAS # |
1499193-60-0
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| Related CAS # |
Ledipasvir impurity 55
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| PubChem CID |
59611589
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| Appearance |
Off-white to light yellow solid powder
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| Hydrogen Bond Donor Count |
0
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
17
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| Complexity |
310
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC2=C(C=C1Br)C(C3=C2C=CC(=C3)I)(F)F
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| InChi Key |
DKSYLEFLMUQPDJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H6BrF2I/c14-7-1-3-9-10-4-2-8(17)6-12(10)13(15,16)11(9)5-7/h1-6H
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| Chemical Name |
2-bromo-9,9-difluoro-7-iodofluorene
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 | 2.4571 mL | 12.2853 mL | 24.5706 mL | |
| 5 mM | 0.4914 mL | 2.4571 mL | 4.9141 mL | |
| 10 mM | 0.2457 mL | 1.2285 mL | 2.4571 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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