Thromboxane A2 (TXA2) synthase - inhibitor (no IC50/Ki/EC50/DC50 values reported in this study). [1]
Ozagrel sodium targets thromboxane A2 (TXA2) synthase, an enzyme involved in the production of thromboxane A2 from prostaglandin H2. By inhibiting TXA2 synthase, ozagrel sodium reduces the production of TXA2, a potent vasoconstrictor and platelet aggregator. This shifts the balance toward prostacyclin (PGI2), which has vasodilatory and anti-platelet aggregation effects. The compound's mechanism of action results in antiplatelet aggregation and vasodilation. It is a selective inhibitor of human platelet aggregation.

In vitro, ozagrel sodium selectively inhibits human platelet aggregation with an IC50 of 53.12 μM. The compound inhibits TXA2 synthase activity in various cell types. Its antiplatelet effects have been characterized in platelet aggregation assays using various inducers. The compound's effects on TXA2 production and PGI2 production have been studied in cellular models. Studies have examined its effects on vasoconstriction and vascular tone in isolated blood vessel preparations. The compound's selectivity for TXA2 synthase over other enzymes has been confirmed.

Ozagrel sodium (5 mg/kg, single intravenous dose, 30 min prior to MCAO) significantly improved behavioral performance in the adhesive removal test at 7 days after stroke (73.80 ± 20.47 s vs control 129.90 ± 17.00 s, p < 0.05), although no significant difference was observed at day 1. [1]
Ozagrel sodium (5 mg/kg, single intravenous dose, 30 min prior to MCAO) significantly improved performance in the treadmill test at day 1 (42.30 ± 8.03 s vs control 29.47 ± 6.95 s, p < 0.05), day 3 (46.00 ± 10.33 s vs control 35.10 ± 6.35 s, p < 0.05), and day 7 (52.60 ± 10.60 s vs control 32.24 ± 8.44 s, p < 0.01) after MCAO. [1]
Ozagrel sodium significantly reduced infarct volume at 7 days after MCAO (255.10 ± 63.14 mm³ vs control 394.31 ± 86.85 mm³, p < 0.05). [1]
Ozagrel sodium significantly decreased the density of GFAP-positive astrocytes in the peri-infarct region at 7 days after MCAO (2027 ± 311 optical density vs control 2409 ± 300, p < 0.05). [1]
Ozagrel sodium significantly increased the number of NeuN-positive surviving neurons in the peri-infarct region at 7 days after MCAO (62 ± 13 cells/mm² vs control 34 ± 3 cells/mm², p < 0.05). [1]
Ozagrel sodium significantly reduced the number of TUNEL-positive apoptotic cells in the peri-infarct region at 7 days after MCAO (118 ± 14 cells/mm² vs control 148 ± 21 cells/mm², p < 0.05). [1]
In vivo, ozagrel sodium has been shown to reduce plasma TXB2 levels (a stable metabolite of TXA2) and increase the ratio of 6-keto-PGF12/TXB2 following intravenous administration in animal studies. It inhibits platelet aggregation induced by various agents and has a preventive effect on cerebral infarction induced by middle cerebral artery occlusion in rats. The compound has been used clinically for the treatment of cerebral vasospasm and asthma. Studies have demonstrated its ability to inhibit cerebral vasospasm and improve blood flow in ischemic conditions.

No enzyme activity assay for Ozagrel sodium was performed in this study. The compound is described as a selective TXA2 synthase inhibitor based on its known pharmacological properties. [1]
In cell-free biochemical assays, ozagrel sodium is evaluated for its inhibitory activity against thromboxane A2 synthase. Enzyme activity assays measure the compound's ability to inhibit the conversion of prostaglandin H2 to thromboxane A2. The compound's IC50 of 53.12 μM against human platelet aggregation is determined using platelet-rich plasma. Its selectivity for TXA2 synthase is confirmed by demonstrating lack of inhibition of other enzymes in the arachidonic acid cascade. These assays confirm the compound's mechanism as a selective TXA2 synthase inhibitor.

No cell-based assay for Ozagrel sodium was performed in this study. [1]
Cellular assays for ozagrel sodium involve evaluating its effects on platelet aggregation and TXA2 production. The compound's ability to inhibit platelet aggregation induced by various agonists (such as ADP, collagen, and arachidonic acid) is assessed using aggregometry. Its effects on TXA2 production are measured using immunoassays for TXB2. The compound's effects on vascular smooth muscle tone and vasoconstriction are studied in isolated blood vessel preparations. Studies have examined its effects on endothelial function and prostacyclin production.

Animals: Male Sprague-Dawley rats (270-300 g, 10-12 weeks old) were used. Animals were housed in temperature-humidity and light-dark controlled rooms with food and tap water ad libitum. Rats were randomly assigned to groups (n=10 per group). [1]
Drug administration: Ozagrel sodium was dissolved in 0.9% saline at a concentration of 5 mg/mL. A single dose of 5 mg/kg was intravenously administered via the tail vein at 30 minutes prior to MCAO. [1]
MCAO model: Transient MCAO was induced using the intraluminal suture method. Under inhalation anesthesia with isoflurane, the right common carotid artery (CCA) and external carotid artery (ECA) were exposed. The occipital artery and superior thyroid artery were dissected and coagulated. The right internal carotid artery (ICA) was exposed and the pterygopalatine artery was ligated. Two loose ties (5-0 silk suture) were made around the ECA stump. A small puncture opening was made in the ECA and a 3-0 monofilament with rounded tip was inserted through the opening and advanced into the ICA for 19-20 mm beyond the bifurcation to block the origin of the middle cerebral artery. Body temperature was maintained at 37°C using a heating pad. After 90 minutes of occlusion, animals were reanesthetized and reperfused by removing the suture; the ECA was then ligated. [1]
Behavioral evaluation: Adhesive removal test was performed using adhesive-backed paper squares on the distal radial region of each forelimb. Animals were given three trials with a cut-off time of 180 s; data presented as mean time to remove the left dot. Treadmill test was performed with acceleration from 20 to 80 m/min with a cut-off time of 300 s. Tests were conducted at 1, 3, and 7 days after MCAO. All animals were trained three times for one week before MCAO. [1]
Infarct volume measurement: At 7 days after MCAO, rats were sacrificed under deep anesthesia with 5% halothane. Brains were dissected and five 2-mm thick coronal slices were prepared using a rodent brain matrix. Slices were stained with 2% TTC (2-3-5-triphenyltetrazolium chloride) at 37°C for 15 minutes. Infarct area was measured using image analysis software, determined by subtracting the area of the non-infarct ipsilateral hemisphere from that of the contralateral tissue to correct for cerebral edema. [1]
Immunohistochemistry: Coronal tissue sections (12 μm thickness) from the precentral gyrus at the boundary area of cerebral infarct (located 24 mm posterior to the frontal pole) were prepared. Sections were blocked in normal goat serum for 1 hour at room temperature, then incubated with mouse anti-GFAP antibody (1:200) or mouse anti-NeuN antibody (1:100) overnight at 4°C. Sections were then incubated with Alexa 488-conjugated goat anti-mouse IgG (1:200) for 1 hour at room temperature, counterstained, and observed under fluorescence microscope and confocal scanning laser microscope. [1]
TUNEL assay: Apoptosis was detected by TUNEL assay using an in situ cell death detection kit with Cy2-conjugated streptavidin, observed by confocal scanning laser microscope. [1]
Image analysis: Optical density of GFAP, NeuN, and TUNEL-positive cells in the ischemia penumbra was quantified using image analysis software. [1]
Animal models for ozagrel sodium include models of cerebral ischemia, thrombosis, and asthma. The compound has been studied in rat models of middle cerebral artery occlusion, where it has a preventive effect on cerebral infarction. Its antiplatelet effects have been evaluated in models of thrombosis. The compound's bronchodilatory effects have been studied in models of asthma. Studies have examined its effects on cerebral vasospasm, blood flow, and platelet function.

Pharmacokinetic data for ozagrel sodium show that the compound is a sodium salt with a molecular weight of 232.21 g/mol and a molecular formula of C13H11N2NaO2. The CAS number is 130952-46-4. The compound is soluble in water (44 mg/mL) and DMSO (10 mM). It should be stored as a powder at -20°C for up to 3 years or at 4°C for up to 2 years. Following intravenous administration of 1 μg/kg/min, plasma concentrations of ozagrel reach 1657.3 ng/mL. The compound has been used clinically.

Minor side effects associated with Ozagrel sodium include headache, nausea, and elevated liver enzyme levels. [1]
Ozagrel sodium is generally well-tolerated. Common side effects include gastrointestinal reactions and allergic reactions such as nausea, vomiting, urticaria, and rash. The compound is a potent inhibitor of TXA2 synthase and has antiplatelet and vasodilatory effects. It has been used clinically for the treatment of cerebral vasospasm and asthma. Standard safety precautions for handling pharmaceutical compounds apply. The compound is for research use only and not for human administration except in approved clinical settings.

Preischemic neuroprotective effect of minocycline and sodium ozagrel on transient cerebral ischemic rat model. PMID: 25555371 DOI: 10.1016/j.brainres.2014.12.051

Ozagrel belongs to the cinnamic acid class of drugs. Ozagrel has been used in clinical trials for the treatment of dry eye syndrome.

---

Ozagrel sodium is a selective thromboxane A2 synthase inhibitor that modulates the arachidonic acid cascade, reducing TXA2 and increasing PGI2. TXA2 is a potent platelet aggregator and vasoconstrictor, while PGI2 inhibits platelet aggregation and is a vasodilator. Ozagrel sodium improves the PGI2/TXA2 balance during acute cerebral ischemia. [1]
Ozagrel sodium has been widely used for the treatment of thrombotic or lacunar stroke. [1]
This study demonstrated that Ozagrel sodium administered as a single dose prior to MCAO exerted direct neuroprotection against ischemic stroke, resulting in smaller infarct volume, greater neuronal survival, fewer activated astrocytes, reduced apoptosis, and improved behavioral outcomes. The mechanism involves suppression of platelet activity, inflammatory enzymes, edema, and vasoconstriction in the ischemic penumbra. [1]
Ozagrel sodium may be useful for stroke prevention in cerebrovascular procedures such as carotid endarterectomy, bypass procedures, endovascular angioplasty, thromboembolectomy, thrombolysis, cerebral angiography, and aneurysm clipping where temporary cerebral blood flow occlusion carries a risk of ischemic stroke. [1]
The difference in effectiveness between minocycline and Ozagrel sodium may relate to differences in diversity of effects; Ozagrel sodium suppresses decrease of cerebral blood flow but does not have an effect on free radicals, whereas minocycline suppresses free radicals in the ischemic period and has more multiple effects. [1]
Ozagrel sodium (OKY-046 sodium) is a potent and selective thromboxane A2 (TXA2) synthase inhibitor. It is an antiplatelet agent that selectively inhibits human platelet aggregation with an IC50 of 53.12 μM. By inhibiting TXA2 synthase, ozagrel sodium reduces TXA2 production and promotes PGI2 production, leading to vasodilation and inhibition of platelet aggregation. The compound belongs to the cinnamic acid class of drugs. It has been used clinically for the treatment of cerebral vasospasm and asthma. The CAS number is 130952-46-4.

C13H11N2NAO2

250.23

250.072

130952-46-4

5282440

White to off-white solid powder

468ºC at 760 mmHg

236.8ºC

0.694

1

3

4

17

283

0

C1=CC(=CC=C1CN2C=CN=C2)/C=C/C(=O)O

SHZKQBHERIJWAO-AATRIKPKSA-N

InChI=1S/C13H12N2O2/c16-13(17)6-5-11-1-3-12(4-2-11)9-15-8-7-14-10-15/h1-8,10H,9H2,(H,16,17)/b6-5+

(E)-3-[4-(imidazol-1-ylmethyl)phenyl]prop-2-enoic acid

Ozagrel sodiumOKY-046 sodiumOKY046 sodiumOKY 046 sodium

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

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

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 Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

---

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
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

---

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

---

 (Please use freshly prepared in vivo formulations for optimal results.)