| Size | Price | Stock | Qty |
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| 100mg |
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| Other Sizes |
| Targets |
The primary molecular target of Coronastat is the SARS-CoV-2 3C-like protease (3CLpro/Mpro), a cysteine protease essential for viral replication. It cleaves the viral polyprotein at 11 sites, producing individual non-structural proteins that form the replication-transcription complex. Coronastat is a GC376 analog, a dipeptide-based inhibitor believed to be a reversible covalent inhibitor that binds to the active site cysteine residue (Cys145), forming a reversible covalent bond. This blocks the enzyme's ability to process the viral polyprotein, inhibiting viral replication. Its high potency (IC₅0 = 16 nM) makes it one of the most potent 3CLpro inhibitors reported.
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| ln Vitro |
In vitro, Coronastat is a highly potent inhibitor of SARS-CoV-2 3CLpro with an IC₅0 of 16 nM in enzymatic assays using recombinant protease. This potency is significantly better than its predecessor GC376. Selectivity for the viral protease over human proteases is high, reducing off-target toxicity. In cell-based models, Coronastat displays potent antiviral activity in Huh-7ACE2 cells with an EC₅0 of 6 nM for inhibiting SARS-CoV-2 replication, exceptionally low and comparable to nirmatrelvir. The compound is also expected to be active against other coronaviruses due to high conservation of the 3CLpro active site across the coronaviridae family.
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| Enzyme Assay |
A typical non-cellular assay is a FRET-based enzymatic assay for SARS-CoV-2 3CLpro. In a 384-well black plate, 20 uL reaction mixture contains assay buffer (50 mM Tris-HCl, pH 7.3, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.01% BSA), 10 nM purified recombinant 3CLpro, and serial dilutions of Coronastat (0.01 nM - 10 uM, final DMSO 1%). Pre-incubate 15 min. Reaction is initiated by adding fluorogenic substrate Dabcyl-KTSAVLQSGFRKME-Edans (10 uM). Fluorescence increase (Ex/Em 340/490 nm) is measured every 30 sec for 30 min at 25degC. Initial rate (V0) is calculated. Percent inhibition relative to DMSO control yields IC₅0 by non-linear regression. Reported IC₅0 = 16 nM.
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| Cell Assay |
A standard in vitro cell-based antiviral assay uses Huh-7ACE2 cells (human hepatoma cells expressing ACE2). Cells are cultured in DMEM with 10% FBS at 37degC, 5% CO2. Cells are seeded in 96-well plates at 2 × 10⁴ cells/well. Next day, medium replaced with DMEM containing 2% FBS. Cells are infected with SARS-CoV-2 at MOI 0.01 for 1 h. Unbound virus removed, and serial dilutions of Coronastat (0.01 nM - 10 uM) are added. After 48-72 h, antiviral activity is quantified by inhibition of virus-induced cytopathic effect (CPE). Cells are stained with neutral red or resazurin, absorbance measured. EC₅0 is calculated. Cytotoxicity (CC₅0) is measured on uninfected Huh-7 cells; reported CC₅0 >100 uM, selectivity index >10,000.
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| Animal Protocol |
An in vivo animal study for Coronastat would be performed in a mouse model of SARS-CoV-2 infection using K18-hACE2 transgenic mice (B6.Cg-Tg(K18-ACE2)2Prlmn/J). Female mice (6-8 weeks) are infected intranasally with a sublethal dose of SARS-CoV-2 (102-103 PFU/mouse). One hour post-infection, mice are randomized (n=10). Coronastat is formulated in a vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) and administered IP at 10, 30, and 100 mg/kg twice daily for 4 days. Control receives vehicle. Body weight is monitored daily (20% weight loss threshold). On day 4, a subset is euthanized, lungs harvested for viral titer by plaque assay. Remaining mice are monitored for survival for 14 days. Lung tissue is taken for H&E histopathology and cytokine analysis by ELISA. All procedures require IACUC approval.
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| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of Coronastat have not been fully disclosed. As an analog of GC376, it likely has improved ADME properties. GC376 has a relatively short half-life and is typically administered by injection. Coronastat has a molecular weight of 575.53 g/mol (sodium salt) and is moderately polar. Oral bioavailability is unknown but may be limited. It is likely metabolized by liver esterases. For effective in vivo exposure, it is formulated for intraperitoneal or subcutaneous administration. Detailed PK parameters (t1/2, Cmax, AUC) are not publicly available. Human PK data is not available.
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| Toxicity/Toxicokinetics |
No detailed toxicological data has been published for Coronastat. In vitro safety assessment shows CC₅0 >100 uM in Huh-7 cells, indicating very low cytotoxicity and a high selectivity index (>10,000). In vivo, in efficacy studies, the compound is likely well-tolerated at tested doses, with no overt signs of toxicity reported. A full toxicological evaluation (acute, sub-chronic, genotoxicity) has not been publicly reported. Standard safety precautions should be used when handling. For research use only; not for human administration.
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| References |
[1]. Liu H, et al. Development of optimized drug-like small molecule inhibitors of the SARS-CoV-2 3CL protease for treatment of COVID-19. Nat Commun. 2022;13(1):1891.
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| Additional Infomation |
Coronastat (NK01-63) is not an approved drug and has not yet entered clinical trials. It is a pre-clinical research compound and a potent inhibitor of the SARS-CoV-2 3CL protease. Its mechanism involves reversible covalent inhibition of the essential viral enzyme. It is considered a new candidate for a small molecule protease inhibitor for the treatment of SARS-CoV-2 infection and has the potential to be used to eliminate pandemics involving coronaviruses. Its high potency (IC₅0 = 16 nM, EC₅0 = 6 nM) positions it as a promising lead for further development. No clinical trials have been registered. For research use only.
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| CAS # |
2922281-15-8
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| Appearance |
White to off-white solid
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| SMILES |
OC(S(=O)(O[Na])=O)[C@H](C[C@@H]1CCNC1=O)NC([C@H](CC(C)C)NC(OCC2=CC(C(F)(F)F)=CC=C2)=O)=O
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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: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
DMSO: ~100 mg/mL (~173.8 mM; with ultrasonication)
H2O: ≥ 100 mg/mL (~173.8 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.61 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), Clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (20.8 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (3.61 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), Clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (20.8 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.08 mg/mL (3.61 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), Clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (20.8 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
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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