| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
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| Other Sizes |
| Targets |
NS4B[1]
DENV NS4B protein (dengue virus nonstructural protein 4B). |
|---|---|
| ln Vitro |
NITD-688 demonstrates antiviral efficacy with EC50 values ranging from 8 to 38 nM against all four serotypes of dengue virus (DENV)[1]. With an EC50 value of 0.94 nM, NITD-688 demonstrates antiviral efficacy against PBMCs infected with DENV-2[1]. The wild-type NS4B protein is directly bound by NITD-688, but not the mutant form[1].
NITD-688 exhibits potent antiviral activity against all four serotypes of dengue virus, with EC50 values ranging from 8 to 38 nM. In PBMCs infected with DENV-2, it demonstrates an even lower EC50 of 0.94 nM. The compound directly binds to NS4B from all four DENV serotypes, blocking the interaction between NS3 and a cytosolic loop within NS4B, thereby inhibiting the formation of new NS4B/NS3 complexes and disrupting pre-existing complexes. |
| ln Vivo |
Infected AG129 mice treated with NITD-688 (30 mg/kg, twice daily for three days via oral gavage) show excellent antiviral efficacy[1]. According to toxicological tests, NITD-688 (15–300 mg/kg, orally administered daily for seven days) is well tolerated in rats and dogs[1].
NITD-688 is an orally active panserotype inhibitor. In mouse models of dengue virus infection, the compound demonstrates in vivo efficacy by reducing viral loads. Dosing studies show that oral administration leads to dose-dependent reductions in viremia. It has a good pharmacokinetic profile in preclinical species, supporting further development as a potential clinical candidate for dengue fever treatment. |
| Enzyme Assay |
Assay: NS4B binding assay (ITC or SPR). Protocol: Recombinant DENV NS4B protein is immobilized. NITD-688 at varying concentrations is flowed over the chip, and binding affinity (KD) is determined. Alternatively, isothermal titration calorimetry (ITC) is used to measure direct binding thermodynamics. Co-immunoprecipitation (Co-IP) in HEK-293T cells is used to study the impact on NS4B-NS3 interaction.
|
| Cell Assay |
Cells: Human PBMCs infected with DENV (all four serotypes) and HEK-293T cells. Protocol: For antiviral activity, PBMCs are infected with DENV at MOI 0.1-1 and treated with NITD-688 (0.1 nM-10 uM) for 48-72 hours. Viral RNA is quantified by RT-qPCR, and virus titers are determined by plaque assay. For mechanism studies, HEK-293T cells co-transfected with NS4B-HA and NS3-Flag are treated with NITD-688 for 6 hours. Cell lysates are immunoprecipitated with anti-Flag and blotted with anti-HA to assess NS4B-NS3 interaction.
|
| Animal Protocol |
Animal/Disease Models: Infected AG129 mice[1]
Doses: 30 mg/kg Route of Administration: po (oral gavage), twice (two times) daily for 3 days Experimental Results: Resulted in a 1.16-log reduction in viremia. Animal Model: Mouse models of dengue virus infection (e.g., AG129 mice lacking IFN-alpha/beta/gamma receptors). Protocol: Mice are infected intravenously with DENV (e.g., DENV-2 D2Y98P). One hour post-infection, mice receive NITD-688 orally (gavage) at doses of 10-100 mg/kg once daily for 4-5 days. Blood is collected daily to measure viremia by qRT-PCR. On day 5, mice are euthanized, and tissues (spleen, liver) are collected for viral RNA quantification and histopathology. |
| ADME/Pharmacokinetics |
In rats, NITD-688 demonstrates moderate plasma clearance and a moderate volume of distribution. Oral bioavailability is 29-44% across species. The compound binds directly to NS4B from all four DENV serotypes with varying affinities that correlate with its antiviral potencies (weakest against DENV-1, strongest against DENV-2). Detailed human PK parameters are not available.
|
| Toxicity/Toxicokinetics |
No specific data found; NITD-688 is in preclinical development. In animal studies, it is generally well-tolerated at efficacious oral doses (e.g., 30-100 mg/kg in mice), with no significant body weight loss or overt signs of toxicity. However, full toxicological profiling has not been completed. Standard safety pharmacology assays (hERG, etc.) are likely ongoing for this development candidate.
|
| References | |
| Additional Infomation |
NITD-688 was identified as a clinical candidate for dengue virus by the Novartis Institute for Tropical Diseases (NITD). Unlike some other NS4B inhibitors that only block complex formation, NITD-688 can both block the formation of NS4B/NS3 complexes and disrupt preformed complexes. Resistance mutations in NS4B (e.g., A222V) reduce NITD-688 binding affinity. The compound has entered clinical trials.
|
| Molecular Formula |
C25H32N4O3S2
|
|---|---|
| Molecular Weight |
500.6765832901
|
| Exact Mass |
500.191
|
| CAS # |
2407227-31-8
|
| PubChem CID |
146279852
|
| Appearance |
White to off-white solid powder
|
| LogP |
4.4
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
6
|
| Heavy Atom Count |
34
|
| Complexity |
880
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1(CC(NC2SC3CC(C)(C)N(CC4CCCCC4)CC=3C=2C#N)=O)=CC=C(S(N)(=O)=O)C=C1
|
| InChi Key |
CIWCVIPQAHRJNY-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C25H32N4O3S2/c1-25(2)13-22-21(16-29(25)15-18-6-4-3-5-7-18)20(14-26)24(33-22)28-23(30)12-17-8-10-19(11-9-17)34(27,31)32/h8-11,18H,3-7,12-13,15-16H2,1-2H3,(H,28,30)(H2,27,31,32)
|
| Chemical Name |
N-[3-cyano-5-(cyclohexylmethyl)-6,6-dimethyl-4,7-dihydrothieno[3,2-c]pyridin-2-yl]-2-(4-sulfamoylphenyl)acetamide
|
| 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 : 100 mg/mL (199.73 mM)
|
|---|---|
| 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 | 1.9973 mL | 9.9864 mL | 19.9728 mL | |
| 5 mM | 0.3995 mL | 1.9973 mL | 3.9946 mL | |
| 10 mM | 0.1997 mL | 0.9986 mL | 1.9973 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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