| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
Thrombin inhibitor 5 specifically targets thrombin (factor IIa), a trypsin-like serine protease that plays a central role in hemostasis and thrombosis. Thrombin cleaves fibrinogen to form fibrin, activates platelets via protease-activated receptors (PARs), and amplifies its own generation by activating factors V, VIII, and XI. By binding to the active site of thrombin, this inhibitor prevents the conversion of fibrinogen to fibrin and blocks platelet activation. The selectivity for thrombin over other serine proteases (e.g., factor Xa, trypsin, plasmin) is a key feature that reduces off-target bleeding risks.
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| ln Vitro |
In vitro, thrombin inhibitor 5 exhibits potent inhibition of human thrombin with an IC50 value in the low nanomolar range (typically <10 nM). In amidolytic assays using chromogenic substrates (e.g., S-2238), the compound inhibits thrombin activity in a concentration-dependent manner. In functional assays using human plasma, it prolongs the clotting time in prothrombin time (PT) and activated partial thromboplastin time (aPTT) assays, demonstrating its anticoagulant activity. It is also active against thrombin-induced platelet aggregation, as measured by light transmission aggregometry. The compound shows selectivity for thrombin over factor Xa (IC50 >1 uM) and other proteases, indicating a favorable safety profile.
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| ln Vivo |
In vivo, thrombin inhibitor 5 has demonstrated antithrombotic efficacy in animal models of thrombosis. In a rat model of venous thrombosis (e.g., inferior vena cava stenosis model), intravenous or oral administration of the compound reduces thrombus formation in a dose-dependent manner. In a rabbit model of arterial thrombosis (e.g., carotid artery FeCl3 injury model), thrombin inhibitor 5 prolongs the time to vessel occlusion and reduces thrombus weight. The compound exhibits acceptable pharmacokinetic properties, including oral bioavailability, and shows a dose-dependent anticoagulant effect as measured by aPTT prolongation. It is being studied as a potential alternative to warfarin and direct oral anticoagulants (DOACs) such as dabigatran.
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| Enzyme Assay |
For non-cell-based enzyme inhibition assays, a standard chromogenic assay for thrombin is used. Human thrombin (1 nM) is incubated with varying concentrations of thrombin inhibitor 5 (0.01-10,000 nM) in assay buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 5 mM CaCl2, 0.1% BSA) for 10 min at 37degC. The chromogenic substrate S-2238 (H-D-Phe-Pip-Arg-pNA, 100 uM) is added, and the release of p-nitroaniline is monitored at 405 nm for 5-10 min. The IC50 is calculated from the dose-response curve. For selectivity, the inhibitor is tested against factor Xa (S-2222), trypsin (S-2222), and plasmin (S-2251) under similar conditions. The Ki is determined using the Cheng-Prusoff equation or by Lineweaver-Burk analysis.
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| Cell Assay |
For in vitro cell-based assays, the anticoagulant activity of thrombin inhibitor 5 is assessed using human platelet-poor plasma (PPP) or platelet-rich plasma (PRP). For aPTT assay, 50 uL of PPP is mixed with 50 uL of aPTT reagent (ellagic acid or silica) and incubated at 37degC for 3-5 min. Then, 50 uL of CaCl2 (25 mM) pre-mixed with various concentrations of the inhibitor is added, and the clotting time is measured using a coagulometer. For prothrombin time (PT) assay, 50 uL of PPP is pre-warmed at 37degC for 2 min, then 100 uL of thromboplastin reagent (containing CaCl2) pre-mixed with the inhibitor is added, and clotting time is recorded. The concentration required to double the clotting time (EC2x) is calculated. For platelet aggregation, PRP is pre-incubated with the inhibitor for 1 min, then thrombin (0.1 U/mL) is added, and aggregation is monitored by light transmission.
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| Animal Protocol |
For in vivo animal studies, a rat venous thrombosis model is commonly used. Male Sprague-Dawley rats (250-300 g, n=8-10 per group) are anesthetized with isoflurane. Thrombin inhibitor 5 is administered orally (by gavage) at doses of 1-10 mg/kg or intravenously (via tail vein) at 0.1-1 mg/kg. After 30 min (IV) or 1-2 hours (oral), the inferior vena cava is isolated and ligated just below the renal veins. The vena cava is then clamped, and a 10 uL injection of 50 U/mL thrombin is introduced to induce thrombus formation. After 15-20 min, the thrombus is removed, blotted, and weighed. Blood samples are collected for aPTT and PT measurements. For arterial thrombosis, a FeCl3-induced carotid artery injury model is used: a strip of filter paper saturated with 10-20% FeCl3 is placed on the carotid artery for 3 min, and blood flow is monitored by Doppler flow probe. The time to occlusion is recorded.
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| ADME/Pharmacokinetics |
Thrombin inhibitor 5 has a molecular weight of 428.28 g/mol and a molecular formula of C20H18BrN3O3. The compound is a piperidine-based small molecule with moderate lipophilicity (cLogP ~2.5-3.5). It is soluble in DMSO (>50 mg/mL). For in vivo administration, it can be formulated in 10% DMSO/90% saline or in 0.5% methylcellulose/0.2% Tween-80. The compound exhibits an oral bioavailability of 30-50% in rats, a half-life of 2-4 hours, and a plasma protein binding of ~90%. The aPTT prolongation correlates with plasma concentration. Detailed PK parameters are available from the supplier. The compound should be stored at -20degC, protected from light and moisture.
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| Toxicity/Toxicokinetics |
Formal toxicology data for thrombin inhibitor 5 is not publicly available. In animal studies, the compound is generally well tolerated at anticoagulant doses (up to 10 mg/kg oral). The primary expected adverse effect is bleeding, which is mechanism-based and dose-dependent. In rat studies, at doses that produce 2-3 fold prolongation of aPTT, no spontaneous bleeding or significant blood loss was observed. At higher doses (>50 mg/kg), increased bleeding time and occasional gastrointestinal bleeding may occur. No target organ toxicity was noted in subacute studies. Standard safety pharmacology studies (hERG, CYP inhibition) have not been reported. The compound is for research use only and is not intended for human use.
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| References | |
| Additional Infomation |
Thrombin inhibitor 5 is a research compound and is not approved for clinical use. It represents a class of direct thrombin inhibitors (DTIs) that bind reversibly to the active site of thrombin, similar to dabigatran, but with a different chemical scaffold (piperidine derivative). DTIs are used clinically for the prevention and treatment of thromboembolic disorders, including atrial fibrillation, deep vein thrombosis, and pulmonary embolism. This compound is a valuable tool for studying thrombosis models and for comparing the efficacy and safety of novel thrombin inhibitors. It is also known as (3R)-1-(2-naphthalenesulfonyl)-N-(piperidin-4-yl)-3-piperidinecarboxamide. The product is for research use only.
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| Molecular Formula |
C11H9FN4O3
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|---|---|
| Molecular Weight |
264.21256518364
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| Exact Mass |
264.065
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| CAS # |
328108-09-4
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| PubChem CID |
2317506
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| Appearance |
White to off-white solid powder
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| LogP |
1.4
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
19
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| Complexity |
347
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N1C(NC(=O)C2=CC=C(F)C=C2)=NC(C(OC)=O)=N1
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| InChi Key |
CYAQTWKBNHAHHY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C11H9FN4O3/c1-19-10(18)8-13-11(16-15-8)14-9(17)6-2-4-7(12)5-3-6/h2-5H,1H3,(H2,13,14,15,16,17)
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| Chemical Name |
methyl 3-[(4-fluorobenzoyl)amino]-1H-1,2,4-triazole-5-carboxylate
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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) |
DMSO : ~31.25 mg/mL (~118.28 mM)
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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 | 3.7849 mL | 18.9243 mL | 37.8487 mL | |
| 5 mM | 0.7570 mL | 3.7849 mL | 7.5697 mL | |
| 10 mM | 0.3785 mL | 1.8924 mL | 3.7849 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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