| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| Other Sizes |
| Targets |
The primary molecular target of NS2B/NS3-IN-3 is the NS2B/NS3 protease complex of the Zika virus. This protease is a serine protease (catalytic triad His, Asp, Ser) essential for processing the viral polyprotein. The viral RNA genome is translated as a single polyprotein that must be cleaved into individual viral proteins by host and viral proteases. The NS2B/NS3 protease complex is responsible for cleaving at several sites within the non-structural region. Inhibition of this protease blocks viral maturation and replication. NS2B/NS3-IN-3 is a competitive inhibitor that binds to the active site, preventing substrate cleavage. The compound shows selectivity for the viral protease over host proteases and is expected to have activity against other flaviviruses due to active site conservation.
|
|---|---|
| ln Vitro |
In cells with EC68 values of 1-3 μM, NS2B/NS3-IN-3 (Compd 66) demonstrates potent antiviral action against the cellular reproduction of the Zika virus[1].
In vitro, NS2B/NS3-IN-3 (Compound 66) exhibits strong antiviral activity against Zika virus. In cell-based assays, it demonstrates potent inhibition of Zika virus replication, with EC₆₈ values (concentration for 68% maximal effect) of 1-3 uM. While the EC₅0 is not explicitly stated, this EC₆₈ range indicates activity in the low micromolar range. The compound is expected to be nontoxic to mammalian cells at effective concentrations. The exact mechanism has been confirmed through enzymatic assays using purified NS2B/NS3 protease, where the compound inhibits proteolytic activity. It is also predicted to have activity against other flaviviruses such as dengue virus (DENV). The compound is likely used as a positive control for developing new flavivirus inhibitors. |
| Enzyme Assay |
A typical non-cellular protease inhibition assay uses a FRET-based method with purified ZIKV NS2B/NS3 protease. The assay is performed in 96-well black plates. The reaction mixture (100 uL) contains 50 mM Tris-HCl (pH 9.0), 20% glycerol, 1 mM CHAPS, and 200 nM of purified ZIKV NS2B/NS3 protease. Varying concentrations of NS2B/NS3-IN-3 (0.01-100 uM) are added and pre-incubated for 15 min at room temperature. The reaction is initiated by adding a fluorogenic substrate, such as Boc-GRR-AMC (20 uM). Fluorescence increase (Ex/Em 380/460 nm) is measured every 30 sec for 30 min at 37degC. Initial linear rate (V0) is calculated. Percent inhibition relative to DMSO control is used to determine IC₅0 by non-linear regression.
|
| Cell Assay |
A standard in vitro cell-based antiviral assay uses Vero cells infected with Zika virus. Vero cells are cultured in DMEM with 10% FBS at 37degC, 5% CO2. Cells are seeded in 96-well plates at 1 × 10⁴ cells/well. Next day, medium replaced with DMEM containing 2% FBS. Cells are infected with Zika virus (e.g., MR766 strain) at MOI 0.1 for 1 h. Unbound virus removed, and fresh medium containing various concentrations of the test compound (0.1-100 uM) is added. After 48-72 h, antiviral activity is assessed by quantifying viral RNA copy number in supernatant by RT-qPCR. EC₅0 is calculated; reported EC₆₈ = 1-3 uM. Cell viability (CC₅0) is measured by MTT assay on uninfected cells.
|
| Animal Protocol |
An in vivo animal study for NS2B/NS3-IN-3 could be performed in the AG129 mouse model of Zika virus infection. Female AG129 mice (6-8 weeks) are infected intraperitoneally (IP) with 102-103 PFU of a mouse-adapted Zika virus strain (e.g., ZIKV-Dakar). Two hours post-infection, mice are randomized (n=10). NS2B/NS3-IN-3 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 5 days. Control receives vehicle. Body weight is monitored daily. On day 6, a subset is euthanized; blood, brain, spinal cord, and testes are collected for viral titer by plaque assay and RNA quantification by RT-qPCR. Remaining mice are monitored for survival for 21 days. All procedures require IACUC approval.
|
| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of NS2B/NS3-IN-3 have not been reported. As a small molecule with molecular weight 323.39 g/mol (C1₉H21N3O2), it has a molecular weight within range for good oral absorption. It has a predicted logP that is moderately lipophilic, facilitating cell membrane permeability. No experimental PK data is available. Human PK data is not available.
|
| Toxicity/Toxicokinetics |
No toxicological data is available for NS2B/NS3-IN-3. In cell-based assays, the compound is expected to be well-tolerated at concentrations effective against the virus (EC₆₈ 1-3 uM), suggesting a reasonable therapeutic window. However, formal cytotoxicity data (CC₅0) has not been disclosed. Standard safety precautions for handling research chemicals should be followed.
|
| References | |
| Additional Infomation |
NS2B/NS3-IN-3 is not an approved drug and has not entered clinical trials. It is a research tool for studying the NS2B/NS3 protease of Zika virus and other flaviviruses. Its mechanism of action is inhibition of the viral protease, an essential enzyme for viral replication. The compound is used to validate the protease as a drug target and as a lead for developing new antiviral agents against Zika and related viruses. No clinical trials have been registered. For research use only; not for diagnostic or therapeutic applications.
|
| Molecular Formula |
C19H21N3O2
|
|---|---|
| Molecular Weight |
323.388944387436
|
| Exact Mass |
323.163
|
| CAS # |
2832876-90-9
|
| Related CAS # |
NS2B/NS3-IN-3 hydrochloride;2832876-91-0
|
| PubChem CID |
163409029
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
2.7
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
24
|
| Complexity |
438
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1CNCCC1CNC(=O)C2=CC3=C(N2)C=C(C=C3)C4=COC=C4
|
| InChi Key |
WTTOXJPIYYZQSP-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C19H21N3O2/c23-19(21-11-13-3-6-20-7-4-13)18-10-15-2-1-14(9-17(15)22-18)16-5-8-24-12-16/h1-2,5,8-10,12-13,20,22H,3-4,6-7,11H2,(H,21,23)
|
| Chemical Name |
6-(furan-3-yl)-N-(piperidin-4-ylmethyl)-1H-indole-2-carboxamide
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO: 65 mg/mL (201.00 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3 mg/mL (9.28 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 30.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.0922 mL | 15.4612 mL | 30.9224 mL | |
| 5 mM | 0.6184 mL | 3.0922 mL | 6.1845 mL | |
| 10 mM | 0.3092 mL | 1.5461 mL | 3.0922 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
