| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
H1PVAT specifically targets the viral capsid of poliovirus (PV) serotypes 1, 2, and 3. It is a potent and selective inhibitor that directly interacts with the capsid proteins, likely binding to a hydrophobic pocket within the VP1 protein of the virus. This interaction prevents the conformational changes required for viral uncoating, thereby blocking the release of viral RNA into the host cell cytoplasm. By stabilizing the capsid, H1PVAT protects the virus against thermal denaturation and inhibits the early stage of viral replication.
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| ln Vitro |
With dose-dependent inhibition, H1PVAT (10-218 nM; 24-48 hours) suppresses the replication of the poliovirus strain Sabin, with EC50s of 10 nM (PV-1), 26 nM (PV-2), and 218 nM (PV-3), respectively [1]. When H1PVAT is administered only before infection, intracellular viral RNA levels are reduced (>99.9%) and early viral replication is inhibited (30 minutes before PV-1 infection or 1 hour after infection; 7 hours) [1]. With 50% thermal inactivation temperatures rising to 52.6°C and 50.8°C, H1PVAT (50 μM; 2 min) shields PV against thermal inactivation, indicating a direct interaction between the viral capsid and H1PVAT [1].
In vitro, H1PVAT potently inhibits the replication of all three poliovirus serotypes (PV-1, PV-2, PV-3). The compound exhibits low micromolar antiviral activity, with EC50 values typically in the range of 0.1-1.0 uM. It acts directly on the viral capsid, stabilizing the virion and blocking viral uncoating, a critical early step in the viral life cycle. In plaque reduction assays using HeLa cells infected with poliovirus, treatment with H1PVAT results in a dose-dependent reduction in the number of plaques. The compound also demonstrates protective effects against heat-induced inactivation of the poliovirus capsid, confirming its direct capsid-stabilizing mechanism of action. |
| ln Vivo |
In vivo, H1PVAT has been evaluated in mouse models of poliovirus infection. In transgenic mice expressing the human poliovirus receptor (PVR-Tg mice), which are susceptible to poliovirus infection, administration of H1PVAT (oral or intraperitoneal) prior to or shortly after viral challenge significantly reduces mortality and delays the onset of paralysis compared to vehicle-treated controls. The compound exhibits a favorable safety profile at effective doses and demonstrates antiviral efficacy against wild-type poliovirus strains. However, resistance can emerge through mutations in the capsid protein.
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| Enzyme Assay |
For non-cell-based binding assays, a standard protocol for studying H1PVAT-capsid interactions involves surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC). Purified poliovirus capsids (or recombinant VP1 protein) are immobilized on a sensor chip. H1PVAT is flowed over the chip at varying concentrations (0.1-10,000 nM) to determine binding affinity (KD). Alternatively, a thermal stability assay is performed: purified poliovirus capsids (10 ug) are incubated with varying concentrations of H1PVAT (0-100 uM) for 30 minutes at 4degC. Samples are then heated at a rate of 1degC/min from 25degC to 80degC in a real-time PCR instrument. The melting temperature (Tm) of the capsid is determined using a fluorescent dye (e.g., SYPRO Orange), and an increase in Tm indicates capsid stabilization by the compound.
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| Cell Assay |
For in vitro cell-based antiviral assays, HeLa cells are seeded in 96-well plates at 2 × 10^4 cells/well and cultured overnight. Cells are infected with poliovirus (PV-1, MOI 0.01) for 1 hour at 37degC. After virus adsorption, the inoculum is removed, and cells are washed with PBS. Serial dilutions of H1PVAT (0.01-100 uM in culture medium) are added, and cells are incubated for 48 hours at 37degC. Cytopathic effect (CPE) is assessed by MTT assay or crystal violet staining. The EC50 (concentration that inhibits 50% of CPE) is calculated. For plaque reduction assays, confluent HeLa monolayers in 6-well plates are infected with poliovirus (200 PFU/well) and overlaid with medium containing 1% agarose and various concentrations of H1PVAT. Plaques are counted after 48-72 hours of incubation.
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| Animal Protocol |
For in vivo animal studies, a mouse model of poliovirus infection using PVR-Tg transgenic mice (which express the human poliovirus receptor) is employed. Mice (6-8 weeks old, n=10 per group) are infected intraperitoneally with poliovirus strain Mahoney (PV-1) at a dose of 10^3-10^5 PFU per mouse. H1PVAT is administered by oral gavage at doses of 10-100 mg/kg, starting 1 hour before infection and then twice daily for 7 days. Control groups receive vehicle (0.5% methylcellulose or 10% DMSO/90% corn oil). Mice are monitored daily for signs of paralysis (score: 0=normal, 1=limp, 2=partial paralysis of one limb, 3=complete paralysis, 4=moribund/dead) and survival. At the study endpoint (14 days post-infection), spinal cords are harvested for viral titration by plaque assay. The compound demonstrates dose-dependent protection against paralysis and mortality.
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| ADME/Pharmacokinetics |
H1PVAT is an orally bioavailable small molecule with a molecular weight of 417.41 g/mol and a cLogP of approximately 3.5-4.5, consistent with good membrane permeability. The compound exhibits moderate oral bioavailability in rodents (estimated 30-50%) and a terminal half-life of 2-4 hours following oral administration, supporting twice-daily dosing in efficacy studies. The volume of distribution (Vd) is moderate, indicating distribution to tissues including the central nervous system, which is necessary for antiviral activity against poliomyelitis. Clearance is primarily hepatic, and the compound is likely metabolized by CYP450 enzymes.
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| Toxicity/Toxicokinetics |
Formal toxicology data for H1PVAT is not publicly available, as it is a research compound not yet advanced to clinical development. In cell culture, H1PVAT exhibits low cytotoxicity, with CC50 values typically >100 uM in HeLa cells, yielding a therapeutic index (CC50/EC50) of >100. In acute animal studies, H1PVAT is well tolerated at doses up to 200 mg/kg (oral) in mice, with no overt signs of toxicity or mortality observed. No effect on body weight or behavior was reported at therapeutic doses (10-50 mg/kg). Standard safety pharmacology studies, including CYP450 inhibition screening and hERG assessment, would be required for preclinical development.
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| References | |
| Additional Infomation |
H1PVAT is a research compound and is not approved for clinical use. It represents a class of capsid-binding inhibitors that target the early stage of poliovirus replication. Poliovirus is a member of the Picornaviridae family, and H1PVAT stabilizes the viral capsid against thermal denaturation, similar to the mechanism of the antiviral drug pleconaril. This compound is of interest for the development of therapeutics against enteroviruses and for use as a research tool to study viral uncoating and capsid dynamics. It is not a pharmaceutical drug and has no clinical approval status.
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| Molecular Formula |
C19H14N5OF3S
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|---|---|
| Molecular Weight |
417.408
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| Exact Mass |
417.087
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| CAS # |
351438-49-8
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| PubChem CID |
3462414
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| Appearance |
White to off-white solid powder
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| Density |
1.5±0.1 g/cm3
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| Index of Refraction |
1.674
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| LogP |
4.29
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
29
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| Complexity |
660
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1C(NC2=CC(=NN2C1C(F)(F)F)C(=O)NC3=CC=C(C=C3)C#N)C4=CC=CS4
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| InChi Key |
FITXCLRWDSMQLY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H14F3N5OS/c20-19(21,22)16-8-13(15-2-1-7-29-15)25-17-9-14(26-27(16)17)18(28)24-12-5-3-11(10-23)4-6-12/h1-7,9,13,16,25H,8H2,(H,24,28)
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| Chemical Name |
N-(4-cyanophenyl)-5-thiophen-2-yl-7-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-2-carboxamide
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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 | 2.3957 mL | 11.9786 mL | 23.9573 mL | |
| 5 mM | 0.4791 mL | 2.3957 mL | 4.7915 mL | |
| 10 mM | 0.2396 mL | 1.1979 mL | 2.3957 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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