| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Danoprevir sodium targets the NS3/4A serine protease of the hepatitis C virus (HCV). The compound exhibits an IC50 of 0.29 nM against NS3/4A protease. It shows IC50 values of 0.2-0.4 nM against HCV genotypes 1a, 1b, 4, 5, and 6, and IC50 values of 1.6 nM and 3.5 nM against genotypes 2b and 3a, respectively. Additionally, Danoprevir is an inhibitor of the SARS-CoV 3CL protease with an IC50 of 0.05 μM.
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|---|---|
| ln Vitro |
Danoprevir sodium is a potent inhibitor of the HCV NS3/4A protease. In a continuous fluorescence resonance energy transfer (FRET) assay, the compound exhibits an IC50 of 0.29 nM against NS3/4A protease. In Huh7 cells harboring an HCV genotype 1b replicon, Danoprevir shows an EC50 of 1.8 nM. At 10 μM, Danoprevir does not inhibit a panel of 79 proteases, ion channels, transporters, and cell surface receptors, demonstrating high selectivity. The first binding of Danoprevir to NS3/4A lasts for more than 5 hours, maintaining an inhibitory state. In HCV subgenomic replicon cell lines containing individual mutations, V36M, R109K, and V170A substitutions do not confer resistance to Danoprevir, whereas the R155K substitution confers high-level resistance.
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| ln Vivo |
Danoprevir has been investigated in multiple Phase II/III clinical trials for the treatment of patients with chronic hepatitis C. In a Phase IIb trial in treatment-naïve patients with HCV-1 infection, Danoprevir plus pegylated IFNα-2a and ribavirin resulted in undetectable levels of HCV-RNA in the majority of patients without evidence of viral resistance. However, the high-dose Danoprevir arm was prematurely terminated because of grade 4 ALT elevations. Phase I trials demonstrated that ritonavir boosting improved the pharmacokinetic profile of Danoprevir. Danoprevir is also in Phase II clinical development for the treatment of chronic hepatitis B (HBV) infection (China, initiated December 2023).
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| Enzyme Assay |
The enzymatic activity of Danoprevir is measured using a continuous fluorescence resonance energy transfer (FRET) assay. The detailed protocol is as follows: The assay buffer contains 25 μM NS4A peptide, 50 mM Tris-HCl (pH 7.5), 15% (vol/vol) glycerol, 0.6 mM lauryldimethylamine oxide, 10 mM dithiothreitol, and 0.5 μM of a fluorescein/QXL520-labeled FRET substrate (Ac-DE-Dap(QXL520)-EE-Abu-ψ-[COO]-AS-Cys(5-FAMsp)-NH2). The reaction is initiated by adding 50 pM K2040 enzyme. The reaction is carried out in 96-well plates in the dark, and fluorescence data are collected. Control conditions include no inhibitor but with enzyme. The initial reaction rate is calculated by analyzing the linear phase of the reaction to obtain the IC50 value. For binding kinetics analysis of the Danoprevir-NS3/4A complex, 10 nM NS3/4A and two-fold Danoprevir are pre-incubated in 1× assay buffer for 15 minutes to form the complex, which is then rapidly diluted 200-fold into assay buffer containing the substrate. The reaction proceeds for more than 5 hours to monitor the dissociation of the complex.
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| Cell Assay |
The cellular activity of Danoprevir is evaluated using Huh7 cells harboring HCV replicons. The detailed protocol is as follows: Huh7 cells stably expressing HCV subgenomic replicons are seeded into culture plates and allowed to attach overnight. Subsequently, serial dilutions of Danoprevir are added to the culture medium, and the cells are incubated at 37°C in a 5% CO₂ incubator. After incubation, HCV RNA levels or reporter gene activity from the replicon is measured to assess antiviral activity. In this system, Danoprevir exhibits an EC50 of 1.8 nM against HCV genotype 1b replicon. In Huh7.5 cells transfected with recombinant viruses, Danoprevir shows anti-pathogen inhibitory activity against HCV genotypes 1, 4, and 6 with IC50 values of 2-3 nM, which is >100-fold lower than that against genotypes 2/3/5 (280-750 nM).
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| Animal Protocol |
In vivo studies of Danoprevir are conducted using oral gavage administration. The transmembrane transport characteristics of Danoprevir are evaluated using cell lines transfected with human transport proteins, including Lewis lung cancer porcine kidney (LLC-PK1) cells, Madin-Darby canine kidney (MDCK) cells, Chinese hamster ovary (CHO) cells, and human hepatocytes. Danoprevir transport involves organic anion transporting polypeptide (OATP) 1B1, OATP1B3, P-glycoprotein (P-gp), and multidrug resistance protein-2 (MRP2), but not breast cancer resistance protein (BCRP). Ritonavir and ciclosporin inhibit the transport of Danoprevir by human hepatocytes.
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| ADME/Pharmacokinetics |
Danoprevir sodium exhibits low absolute bioavailability. In healthy subjects, the absolute bioavailability of 100 mg Danoprevir is 1.15%, which increases more than threefold (to 3.86%) when co-administered with ritonavir. The pharmacokinetics of intravenous 6 mg Danoprevir are not altered by oral 100 mg ritonavir; in contrast, exposure to oral 100 mg Danoprevir increases two- to threefold when co-administered with ritonavir. Oral 100 mg ciclosporin increases exposure to intravenous 2 mg Danoprevir. Danoprevir transport involves OATP1B1, OATP1B3, P-gp, and MRP2.
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| Toxicity/Toxicokinetics |
According to the Safety Data Sheet (SDS), Danoprevir (free form, CAS 850876-88-9) is classified as not a hazardous substance or mixture, with no significant acute toxicity, skin corrosion, or serious eye damage risks identified. However, in clinical trials, the high-dose Danoprevir arm was prematurely terminated due to grade 4 ALT (alanine aminotransferase) elevations, indicating a need to monitor hepatotoxicity risk. This compound is strictly for research use only and is not intended for human therapeutic use.
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| References | |
| Additional Infomation |
Danoprevir sodium (development codes: ITMN-191 sodium, R7227 sodium, RO5190591 sodium) is the sodium salt form of Danoprevir, with the molecular formula C₃₅H₄₆FN₅NaO₉S and molecular weight of 754.82. This compound is an orally active HCV NS3/4A serine protease inhibitor belonging to the 15-membered macrocyclic peptidomimetic class. On June 8, 2018, Danoprevir (Ganovo®) received approval from the National Medical Products Administration (NMPA) of China for the treatment of treatment-naïve adult patients with chronic hepatitis C virus genotype 1b infection, in combination with ritonavir, peginterferon alfa-2a, and ribavirin. This compound is for research use only and is not intended for human therapeutic use.
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| Molecular Formula |
C35H46FN5NAO9S
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|---|---|
| Molecular Weight |
754.82
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| Exact Mass |
753.282
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| Elemental Analysis |
C, 55.77; H, 6.02; F, 2.52; N, 9.29; Na, 3.05; O, 19.10; S, 4.25
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| CAS # |
916826-48-7
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| Related CAS # |
850876-88-9; 916826-48-7
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| PubChem CID |
23718928
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| Appearance |
Typically exists as solids at room temperature
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| LogP |
0
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
52
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| Complexity |
1530
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| Defined Atom Stereocenter Count |
5
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| SMILES |
CC(C)(OC(N[C@H]1CCCCCC=C[C@@H]2C[C@]2(NC([C@@H]3C[C@@H](OC(N4CC5=C(C(F)=CC=C5)C4)=O)CN3C1=O)=O)C(NS(=O)(C6CC6)=O)=O)=O)C.[Na].[H]
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| Synonyms |
ITMN-191 sodium; R7227 sodium; RO5190591 sodium
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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 | 1.3248 mL | 6.6241 mL | 13.2482 mL | |
| 5 mM | 0.2650 mL | 1.3248 mL | 2.6496 mL | |
| 10 mM | 0.1325 mL | 0.6624 mL | 1.3248 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.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT03019991 | Completed | Drug: Danoprevir Drug: Ritonavir |
Healthy | Ascletis Pharmaceuticals Co., Ltd. |
October 2015 | Phase 1 |
| NCT03020082 | Completed | Drug: Danoprevir Drug: Ritonavir |
Chronic Hepatitis C | Ascletis Pharmaceuticals Co., Ltd. |
June 2016 | Phase 3 |
| NCT03020004 | Completed | Drug: Danoprevir Drug: Ritonavir |
Chronic Hepatitis C | Ascletis Pharmaceuticals Co., Ltd. |
January 2016 | Phase 2 |
| NCT03020095 | Completed | Drug: Danoprevir Drug: Ritonavir |
Chronic Hepatitis C | Ascletis Pharmaceuticals Co., Ltd. |
November 28, 2011 | August 2015 |
| NCT01531647 | Completed | Drug: danoprevir Drug: raltegravir |
Healthy Volunteer | Hoffmann-La Roche | January 2012 | Phase 1 |
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