| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
ZL(D-Val)G-CHN2 targets cysteine proteases, including human cysteine proteases and viral proteases such as the SARS-CoV-2 3CL protease (also known as main protease, Mpro). It is an irreversible inhibitor that binds covalently to the active site cysteine residue of the target protease, preventing substrate processing. The compound also inhibits HSV replication but has no significant effect on poliovirus replication.
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| ln Vitro |
In vitro, ZL(D-Val)G-CHN2 potently inhibits the SARS-CoV-2 3CL protease and suppresses viral replication with an EC50 of 190 nM. It is a cell-permeable and irreversible inhibitor of cysteine proteases. As a tripeptide derivative, it mimics the binding center of human cysteine proteases, enabling covalent modification of the active site cysteine. The compound also shows inhibitory activity against herpes simplex virus (HSV) but does not affect poliovirus replication, indicating selectivity for certain viral proteases.
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| ln Vivo |
In vivo activity data for ZL(D-Val)G-CHN2 are limited. Based on its potent in vitro antiviral activity against SARS-CoV-2 (EC50 = 190 nM), the compound has the potential for in vivo efficacy in animal models of coronavirus infection. As a cell-permeable irreversible inhibitor of cysteine proteases, it could be tested in mouse models of SARS-CoV-2 infection, such as the K18-hACE2 transgenic mouse model. Further studies are required to evaluate its in vivo pharmacokinetics, safety, and efficacy.
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| Enzyme Assay |
For in vitro protease inhibition assays, recombinant SARS-CoV-2 3CL protease (Mpro) is incubated with varying concentrations of ZL(D-Val)G-CHN2 (0.01-100 uM) in assay buffer (50 mM Tris-HCl, pH 7.3, 1 mM EDTA, 150 mM NaCl, 1 mM DTT) at 25degC for 30 minutes. The fluorogenic substrate Dabcyl-KTSAVLQSGFRKME-Edans is then added to a final concentration of 10-20 uM. The increase in fluorescence (excitation 340 nm, emission 490 nm) is monitored for 10-60 minutes. The IC50 is calculated from the inhibition curve.
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| Cell Assay |
For antiviral activity assays, Vero E6 cells are seeded in 96-well plates and infected with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.01-0.1. After 1 hour of viral adsorption, ZL(D-Val)G-CHN2 is added at concentrations ranging from 0.01-100 uM. After 48-72 hours of incubation at 37degC, viral replication is quantified by plaque assay or by measuring viral RNA by qRT-PCR. The EC50 (half-maximal effective concentration) is calculated. Cytotoxicity is assessed in parallel using MTT or CellTiter-Glo assays to calculate the CC50 and selectivity index (SI = CC50/EC50).
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| Animal Protocol |
For in vivo efficacy studies in a mouse model of SARS-CoV-2 infection, ZL(D-Val)G-CHN2 would be formulated in a suitable vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline, or in a cyclodextrin-based formulation. K18-hACE2 transgenic mice are infected intranasally with SARS-CoV-2 (e.g., 10^4-10^5 PFU). ZL(D-Val)G-CHN2 is administered via intraperitoneal injection or oral gavage at doses of 10-100 mg/kg, once or twice daily, starting 1-2 hours before infection or at the time of infection. Treatment continues for 3-7 days. Body weight and clinical scores are monitored daily. At the end of the study, lungs are harvested for viral titer determination (plaque assay or qRT-PCR), histopathological analysis (H&E staining), and cytokine analysis (ELISA).
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for ZL(D-Val)G-CHN2 are limited. The compound has a molecular weight of 445.51 and a molecular formula of C22H31N5O5. It is a tripeptide derivative with a diazomethyl ketone (CHN2) reactive group. It is soluble in DMSO and may be soluble in other organic solvents. For in vivo use, a standard formulation using DMSO, PEG300, Tween-80, and saline can be used. Researchers should conduct pilot PK studies to determine key parameters such as half-life, Cmax, and bioavailability. The compound should be stored at -20degC as a powder, protected from moisture and light.
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| Toxicity/Toxicokinetics |
Toxicological data for ZL(D-Val)G-CHN2 are limited. In cell-based assays at concentrations up to 10 uM, no significant cytotoxicity is reported for Vero E6 cells at the EC50 concentration. For any animal study, a preliminary acute toxicity study to determine the maximum tolerated dose (MTD) is essential. Standard safety precautions for handling chemical reagents should be followed, especially given the reactive diazomethyl ketone moiety.
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| References | |
| Additional Infomation |
ZL(D-Val)G-CHN2 is a research-grade compound and is not approved for clinical use. It is the isomer of Z-LVG-CHN2, a known cysteine protease inhibitor. The compound contains a Z-protecting group, a D-valine residue, and a diazomethyl ketone (CHN2) warhead that covalently modifies the active site cysteine. It is used as a tool for studying viral protease function and as a potential lead compound for antiviral drug development against SARS-CoV-2 and HSV. Store at -20degC as a powder, protected from moisture and light.
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| Molecular Formula |
C22H31N5O5
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| Molecular Weight |
445.51
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| Related CAS # |
Z-LVG-CHN2;119670-30-3
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| Appearance |
White to off-white 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 | 2.2446 mL | 11.2231 mL | 22.4462 mL | |
| 5 mM | 0.4489 mL | 2.2446 mL | 4.4892 mL | |
| 10 mM | 0.2245 mL | 1.1223 mL | 2.2446 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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