| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
The primary target of Cloricromene hydrochloride is phospholipase A2 (PLA2), an enzyme that releases arachidonic acid from membrane phospholipids. By inhibiting PLA2 activation, Cloricromene blocks the liberation of arachidonic acid, thereby suppressing the synthesis of downstream pro-inflammatory and pro-thrombotic eicosanoids, such as thromboxane A2 (TXA2), prostaglandins, and leukotrienes. This results in reduced platelet aggregation, decreased leukocyte activation and adhesion, and vasodilation. The compound also may have direct effects on platelet and leukocyte membranes, altering their fluidity and responsiveness to agonists. By inhibiting PLA2, Cloricromene also reduces the formation of platelet-activating factor (PAF), a potent phospholipid mediator of inflammation and thrombosis.
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| ln Vitro |
In vitro studies demonstrate that Cloricromene hydrochloride is a potent inhibitor of platelet aggregation induced by various agonists, including collagen, ADP, arachidonic acid, and thrombin. It inhibits the release of arachidonic acid from platelet membranes by interfering with PLA2 activation, as shown by reduced production of thromboxane B2 (TXB2, a stable metabolite of TXA2) in stimulated platelets. The compound also suppresses leukocyte function, including chemotaxis, adhesion to endothelium, and release of reactive oxygen species (ROS) and proteolytic enzymes. In whole blood or platelet-rich plasma (PRP), Cloricromene (1-100 uM) concentration-dependently inhibits aggregation, with IC50 values typically in the low micromolar range. It also inhibits the expression of adhesion molecules (e.g., P-selectin, CD11b/CD18) on activated platelets and leukocytes. In addition, Cloricromene has been shown to improve erythrocyte deformability and reduce blood viscosity, contributing to its hemorheological effects. The compound is not cytotoxic to vascular endothelial cells or smooth muscle cells at therapeutic concentrations.
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| ln Vivo |
In vivo studies using animal models of thrombosis and ischemia have demonstrated the antithrombotic and protective effects of Cloricromene hydrochloride. In rat models of arterial and venous thrombosis, intravenous administration of Cloricromene (1-10 mg/kg) significantly reduces thrombus formation and improves vessel patency. In models of ischemia-reperfusion injury (e.g., in the heart, brain, or intestine), pretreatment with Cloricromene reduces infarct size, limits leukocyte infiltration, and preserves tissue function. The compound also improves microcirculatory blood flow in models of peripheral arterial disease and reduces the severity of endotoxin-induced shock. In a rabbit model of experimental thrombosis, Cloricromene hydrochloride inhibits platelet aggregation and reduces thrombus weight. It also has been shown to reduce neointimal hyperplasia after vascular injury, suggesting a potential role in preventing restenosis. These in vivo effects are attributed to its combined antiplatelet, anti-leukocyte, and vasodilatory actions.
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| Enzyme Assay |
Non-cell-based assays for studying Cloricromene hydrochloride often focus on its ability to inhibit PLA2 activity. A common protocol uses a commercial PLA2 assay kit (e.g., from Cayman Chemical or Abcam). Purified secretory PLA2 (sPLA2) or cytosolic PLA2 (cPLA2) is incubated with a fluorescent or radiolabeled phospholipid substrate in assay buffer (e.g., 100 mM Tris-HCl pH 7.5, 5 mM CaCl2, 0.1% BSA). Cloricromene is added at varying concentrations (0.1-100 uM) and pre-incubated with the enzyme for 10-15 minutes. The reaction is initiated by adding the substrate. After incubation at 37degC for 30-60 minutes, the reaction is stopped, and the product (e.g., free arachidonic acid or lysophospholipid) is extracted and quantified. For the fluorescent assay, the increase in fluorescence (e.g., λex 485 nm, λem 535 nm) is measured. IC50 values are calculated from the dose-response curve. To study the compound's effect on arachidonic acid release from membranes, a cell-free system using isolated platelet or leukocyte membranes can be used. Membranes are incubated with 3H-arachidonic acid-labeled phospholipid substrate in the presence of CaCl2 and varying concentrations of Cloricromene. The reaction is terminated, and the released radiolabeled arachidonic acid is separated by thin-layer chromatography (TLC) and quantified by liquid scintillation counting.
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| Cell Assay |
For cell-based studies, human platelets or leukocytes are isolated from fresh whole blood. For platelet aggregation studies, platelet-rich plasma (PRP) is prepared by centrifugation of whole blood (e.g., 200g for 10 min). PRP is diluted to a standard platelet count (e.g., 2×10^8 platelets/mL). Cloricromene hydrochloride is dissolved in DMSO or saline and added to PRP at varying concentrations (0.1-100 uM) for 5-10 min at 37degC. Aggregation is induced by the addition of an agonist (e.g., collagen 1-5 ug/mL, ADP 5-20 uM, arachidonic acid 0.5-1 mM, or thrombin 0.1-1 U/mL) in an aggregometer. The aggregation trace is recorded for 5-10 minutes, and the maximum aggregation percentage and aggregation slope are calculated. For leukocyte adhesion studies, human umbilical vein endothelial cells (HUVECs) are cultured in 96-well plates until confluent. HUVECs are activated with TNF-alpha (10 ng/mL) for 4-6 hours to upregulate adhesion molecules. Leukocytes (e.g., isolated human neutrophils) are pre-incubated with Calcein-AM (a fluorescent dye) for 30 min, then washed. The labeled leukocytes are added to the HUVEC monolayer in the presence or absence of Cloricromene (1-50 uM). After 30 min of co-incubation, non-adherent cells are washed off, and the fluorescence of adherent cells is measured (λex 485 nm, λem 535 nm). The percentage of adhesion is calculated relative to control wells. For ROS production, neutrophils are isolated and loaded with dihydrorhodamine 123 (DHR 123) or DCFH-DA. Cells are stimulated with phorbol 12-myristate 13-acetate (PMA, 100 nM) or fMLP (1 uM) with or without Cloricromene (1-50 uM). The fluorescence intensity is measured over time in a fluorescence plate reader (λex 488 nm, λem 520 nm). Flow cytometry can also be used to measure ROS production and adhesion molecule expression (e.g., CD11b, CD18) on the surface of activated neutrophils or platelets.
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| Animal Protocol |
For in vivo studies, animal models of thrombosis and ischemia are used. In a rat model of ferric chloride-induced arterial thrombosis, male Sprague-Dawley rats (250-300 g) are anesthetized, and the carotid artery is exposed. A piece of filter paper soaked in 25-50% FeCl3 is placed on the artery for 1-3 minutes to induce thrombosis. Cloricromene hydrochloride is administered intravenously (i.v.) as a bolus (e.g., 1, 3, 10 mg/kg) or by continuous infusion 10-30 minutes before FeCl3 application. Blood flow is monitored using a Doppler flow probe distal to the injury site. The time to occlusion and the duration of flow are recorded. At the end of the experiment, the thrombus is excised and weighed. In a mouse model of pulmonary thromboembolism, mice are injected i.v. with a mixture of collagen and epinephrine to induce fatal thromboembolism. Cloricromene (5-20 mg/kg, i.v.) is administered 5 min before the pro-thrombotic mixture. Survival is monitored for 15 minutes. For ischemia-reperfusion studies, a rat model of cerebral ischemia (middle cerebral artery occlusion, MCAO) or myocardial ischemia (left anterior descending coronary artery ligation) is used. Cloricromene is administered i.v. (1-10 mg/kg) at the onset of reperfusion. Infarct size is measured by TTC staining, and neurological scores (for MCAO) or left ventricular function (for myocardial ischemia) are assessed. Blood samples can be collected for measurement of platelet aggregation ex vivo and for biochemical markers of inflammation (e.g., TXB2, LTB4, IL-6, TNF-alpha).
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| ADME/Pharmacokinetics |
Pharmacokinetic data for Cloricromene hydrochloride is limited. As a small molecule (MW 432.34 Da) with a coumarin structure, it is likely rapidly absorbed and metabolized. In animal studies, the compound is typically administered intravenously due to potential low oral bioavailability. Following i.v. administration in rats (e.g., 5 mg/kg), the plasma half-life (t1/2) is likely short (e.g., 0.5-2 hours), as is common for many coumarins. The volume of distribution (Vd) is moderate, suggesting distribution into tissues. Clearance (CL) is likely via hepatic metabolism, primarily by CYP450 enzymes (e.g., CYP2C9, CYP3A4), followed by biliary excretion. The compound may undergo phase II glucuronidation. Its major metabolite(s) and their activity are not well-characterized. For in vitro assays, stock solutions are prepared in DMSO (e.g., 100 mM) and diluted in assay buffer, with the final DMSO concentration kept below 0.1-0.5% to avoid solvent effects on cells or enzymes. The compound is stable in powder form at -20degC and in solution for short periods when protected from light.
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| Toxicity/Toxicokinetics |
Preclinical toxicity data for Cloricromene hydrochloride is not extensively reported. In animal models, the compound was well-tolerated at doses up to 10-20 mg/kg (i.v.) without significant adverse effects on body weight, behavior, or major organ histopathology. The acute LD50 in rodents is likely >100 mg/kg (i.v.). The primary safety concern for PLA2 inhibitors could be off-target effects on lipid metabolism and cell membrane integrity, but no significant toxicity has been reported in short-term studies. In cell-based assays, Cloricromene shows low cytotoxicity at concentrations up to 100 uM in most cell types (e.g., endothelial cells, fibroblasts). Standard safety precautions for handling include using personal protective equipment (gloves, lab coat, goggles) and working in a chemical fume hood. The compound is for research use only and is not for human use. No genotoxicity, carcinogenicity, or reproductive toxicity data is publicly available.
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| References |
[1]. G Orefice, et al. No effect of cloricromen on some coagulation parameters in patients with ischaemic cerebrovascular disease. J Int Med Res. 1994 Sep-Oct;22(5):287-91.
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| Additional Infomation |
See also: Cloricromen (note moved to).
Cloricromene hydrochloride (also known as Cloricromen hydrochloride) is a research-grade chemical compound with antithrombotic and vasodilatory properties. It is chemically described as ethyl 2-((8-chloro-3-(2-(diethylamino)ethyl)-4-methyl-2-oxo-2H-chromen-7-yl)oxy)acetate hydrochloride. It is a coumarin derivative that has been investigated for the treatment of peripheral arterial disease, ischemic stroke, and myocardial infarction, but it has not been approved for clinical use by the FDA or EMA. It is soluble in DMSO (e.g., 17 mg/mL) and has limited aqueous solubility. The product should be stored as a powder at -20degC, protected from light and moisture, where it is stable for 3 years. In solution, it should be stored at -80degC in aliquots and used within 6 months. It is a valuable tool for studying platelet and leukocyte function, thrombosis, inflammation, and ischemia-reperfusion injury in vitro and in vivo. |
| Molecular Formula |
C20H27CL2NO5
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|---|---|
| Molecular Weight |
432.34
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| Exact Mass |
431.127
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| CAS # |
74697-28-2
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| Related CAS # |
Cloricromen;68206-94-0
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| PubChem CID |
16219127
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| Appearance |
Typically exists as solid at room temperature
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| Boiling Point |
538ºC at 760 mmHg
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| Melting Point |
221-222.3ºC
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| Flash Point |
279.2ºC
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| LogP |
4.383
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
28
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| Complexity |
561
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCN(CC)CCC1=C(C2=C(C(=C(C=C2)OCC(=O)OCC)Cl)OC1=O)C.Cl
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| InChi Key |
CCCZJRFQJNGCCU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H26ClNO5.ClH/c1-5-22(6-2)11-10-15-13(4)14-8-9-16(26-12-17(23)25-7-3)18(21)19(14)27-20(15)24;/h8-9H,5-7,10-12H2,1-4H3;1H
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| Chemical Name |
ethyl 2-[8-chloro-3-[2-(diethylamino)ethyl]-4-methyl-2-oxochromen-7-yl]oxyacetate;hydrochloride
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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 | 2.3130 mL | 11.5650 mL | 23.1299 mL | |
| 5 mM | 0.4626 mL | 2.3130 mL | 4.6260 mL | |
| 10 mM | 0.2313 mL | 1.1565 mL | 2.3130 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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