The proliferation of HAEC cells is increased by tirofiban (0.25, 1, 3 μg/mL; 72 hours) [1]. The HUVEC migration scratch is closed in 18 hours by tirofiban (24 hours) [1]. After 30 minutes, tirofiban (0.25, 1 μg/mL; 1 hour) increases endothelial cell proliferation and causes the generation of VEGF [1].

By raising HR, LVESP, dp/dtmax, and lowering LVEDP, tirofiban (60 μg/kg; IV; once) exhibits activity in enhancing contractility, ventricular compliance, and cardiac function [2]. Tirofiban (60 μg/kg; intravenous injection; once) diminishes the no-reflow region following reperfusion following Acute Myocardial Infarction and increases eNOS activity [2]. In a squeezing model, tirofiban (50 μg/per; irrigation; single dose) showed anticoagulant effects, with a 59% 24-hour patency rate following microvascular anastomosis [3].

Cell Proliferation Assay[1]
Cell Types: HAEC cells
Tested Concentrations: 0.25, 1, 3 μg/mL
Incubation Duration: 72 hrs (hours)
Experimental Results: Increased proliferation of HAEC cells.

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Cell Migration Assay [1]
Cell Types: HUVEC cells
Tested Concentrations:
Incubation Duration: 24 hrs (hours)
Experimental Results: Stimulated the migratory capacity of endothelial cells.

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Western Blot Analysis[1]
Cell Types: HAEC cells
Tested Concentrations: 0.05, 0.12, 0.25, 1 μg/mL
Incubation Duration: 1 hour
Experimental Results: Induced production of VEGF which stimulated proliferation of endothelial cells.

Animal/Disease Models: Male SD (Sprague-Dawley) rats (10 to 15-week-age; 270-330 g)[2].
Doses: 60 μg/kg
Route of Administration: intravenous (iv) injection; once.
Experimental Results: Increased contraction force, ventricular compliance, and improved heart function. decreased the size of no-reflow and infarct.

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Animal/Disease Models: SD (Sprague-Dawley) rats (350-400 g; crush injury model)[3]
Doses: 50 µg/per (50 µg/mL, 1 mL for each)
Route of Administration: Irrigate 1 mL within the vessel lumen (before placement of the last suture); once.
Experimental Results: demonstrated anticoagulant effect with patency rates of 59%.

Absorption, Distribution and Excretion
It is cleared from the plasma largely by renal excretion, with about 65% of an administered dose appearing in urine and about 25% in feces, both largely as unchanged tirofiban.
22 to 42 L
213 - 314 mL/min [Healthy subjects]
152 - 267 mL/min [patients with coronary artery disease]
Not highly bound to plasma proteins; protein binding is concentration-independent over the range of 0.01 to 25 ug/mL. Unbound fraction in human plasma is 35%.
Tirofiban is approximately 65% bound to plasma proteins ... The steady-state volume of distribution of tirofiban ranges from 22-42 L. It is not known whether tirofiban is distributed into milk or crosses the placenta in humans; however, the drug is distributed into milk in rats and crosses the placenta in pregnant rats and rabbits.
About 65 and 25% of a single dose of tirofiban is excreted in urine and feces, respectively, principally as unchanged parent drug. Plasma clearance of tirofiban in healthy individuals ranges from 213-314 mL/minute, with renal clearance accounting for 39-69% of plasma clearance. In patients with coronary artery disease, the plasma clearance of tirofiban ranges from 152-267 mL/minute and does not appear to be influenced by gender or race; renal clearance in these patients accounts for 39% of plasma clearance. Plasma clearance is about 19-26% lower in geriatric patients (those exceeding 65 years of age) with coronary artery disease than in younger patients. Plasma clearance appears to be independent of dose in healthy individuals and is not appreciably affected by mild to moderate hepatic insufficiency. In patients with renal impairment (creatinine clearance less than 30 mL per minute), including those requiring hemodialysis, plasma clearance of tirofiban is decreased by greater than 50% compared with that in individuals with normal renal function. Tirofiban is removed by hemodialysis.

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Metabolism / Metabolites
Metabolism appears to be limited.
Metabolism appears to be limited.

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Biological Half-Life
2 hours
Elimination: Approximately 2 hours.

Protein Binding
65%

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Interactions
Concurrent use /with other platelet aggregation inhibitors (especially inhibitors of platelet receptor GP IIb/IIIa) is not recommended.
Data from a large clinical study indicate that concomitant administration of tirofiban and levothyroxine or omeprazole was associated with a higher clearance of tirofiban; the clinical importance of this effect is not known.

[1]. Tirofiban induces VEGF production and stimulates migration and proliferation of endothelial cells. Vascul Pharmacol. 2014 May-Jun;61(2-3):63-71.

[2]. Effects of tirofiban on the reperfusion-related no-reflow in rats with acute myocardial infarction. J Geriatr Cardiol. 2013 Mar;10(1):52-8.

[3]. The effect of tirofiban on microvascular thrombosis: crush model. Plast Reconstr Surg. 2005 Jul;116(1):205-8.

Tirofiban is a member of the class of piperidines that is L-tyrosine in which a hydrogen attached to the amino group is replaced by a butylsulfonyl group and in which the hydrogen attached to the phenolic hydroxy group is replaced by a 4-(piperidin-4-yl)butyl group. It has a role as a fibrin modulating drug, a platelet glycoprotein-IIb/IIIa receptor antagonist and an anticoagulant. It is a member of piperidines, a sulfonamide and a L-tyrosine derivative.
Tirofiban prevents the blood from clotting during episodes of chest pain or a heart attack, or while the patient is undergoing a procedure to treat a blocked coronary artery. It is a non-peptide reversible antagonist of the platelet glycoprotein (GP) IIb/IIIa receptor, and inhibits platelet aggregation.
Tirofiban is a Platelet Aggregation Inhibitor. The physiologic effect of tirofiban is by means of Decreased Platelet Aggregation.
Tirofiban is a non-peptide tyrosine derivative, with anticoagulant activity. Upon administration, tirofiban antagonizes fibrinogen binding to the platelet cell surface receptor, glycoprotein (GP) IIb/IIIA complex, one of the two purinergic receptors activated by ADP. This prevents the GP IIb/IIIa receptor complex-mediated activation of adenylyl cyclase. This results in decreased levels of cAMP, interferes with the platelet membrane function and subsequent platelet-platelet interaction, prevents the release of platelet granule constituents and prolongs bleeding time.
Tyrosine analog and PLATELET GLYCOPROTEIN GPIIB-IIIA COMPLEX antagonist that inhibits PLATELET AGGREGATION and is used in the treatment of ACUTE CORONARY SYNDROME.
See also: Tirofiban Hydrochloride (has salt form).

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Drug Indication
For treatment, in combination with heparin, of acute coronary syndrome, including patients who are to be managed medically and those undergoing PTCA or atherectomy.
FDA Label

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Mechanism of Action
Tirofiban is a reversible antagonist of fibrinogen binding to the GP IIb/IIIa receptor, the major platelet surface receptor involved in platelet aggregation. Platelet aggregation inhibition is reversible following cessation of the infusion of tirofiban.
Tirofiban inhibits platelet aggregation by reversibly binding to the platelet receptor glycoprotein (GP) IIb/IIIa of human platelets, thus preventing the binding of fibrinogen. Inhibition of platelet aggregation occurs in a dose- and concentration-dependent manner.

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Therapeutic Uses
Antithrombotic; in treatment of unstable angina.
Tirofiban is indicated, in combination with heparin, for the prevention of acute cardiac ischemic complications in patients with acute coronary syndrome (unstable angina or non-Q-wave myocardial infarction). these patients are at high risk for myocardial infarction and sudden death due to progression of total coronary artery occlusion, whether managed medically or with percutaneous coronary intervention (PCI). Note: Acute coronary syndrome is defined as prolonged (>/= 10 minutes) or repetitive symptoms of cardiac ischemia occurring at rest or with minimal exertion, associated with either ST-T wave changes on electrocardiogram or elevated cardiac enzymes. This definition includes unstable angina and non-Q-wave myocardial infarction but excludes myocardial infarction that is associated with Q waves or nontransient ST-segment elevation. /Included in US product labeling/

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Drug Warnings
Adverse cardiovascular effects that occurred in greater than 1% of patients receiving tirofiban plus heparin in clinical trials were bradycardia (4%) or dissection of the coronary artery (5%). Edema/swelling or vasovagal reactions were reported in 2% of patients receiving tirofiban and heparin in these trials.
Sweating was reported in 2% of patients receiving tirofiban and heparin in controlled clinical trials. Anaphylaxis and/or urticaria requiring discontinuance of therapy was not reported in clinical trials of tirofiban, but anaphylaxis and other severe allergic reactions have been reported during postmarketing experience. Such reactions have occurred on the first day of tirofiban infusion, during initial treatment, and during readministration of the drug. Some severe allergic reactions have been associated with severe thrombocytopenia (platelet counts less than 10,000/cu mm).
Pelvic pain occurred in 6%, leg pain in 3%, and dizziness in 3% of patients receiving tirofiban plus heparin in clinical trials. Other adverse effects reported in greater than 1% of patients receiving tirofiban plus heparin in clinical trials include headache, nausea, and fever.
The administration of tirofiban in patients with acute unstable angina or non-ST-segment elevation myocardial infarction has been associated with a small increase in the frequency of major bleeding compared with heparin and aspirin therapy alone.
For more Drug Warnings (Complete) data for TIROFIBAN (11 total), please visit the HSDB record page.

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Pharmacodynamics
Tirofiban prevents the blood from clotting during episodes of chest pain or a heart attack, or while the patient is undergoing a procedure to treat a blocked coronary artery. It is a non-peptide antagonist of the platelet glycoprotein (GP) IIb/IIIa receptor, and inhibits platelet aggregation. When administered intravenously, tirofiban inhibits ex vivo platelet aggregation in a dose- and concentration-dependent manner. When given according to the recommended regimen, >90% inhibition is attained by the end of the 30-minute infusion. Tirofiban has been recently shown in patients with unstable angina to reduce ischemic events at 48 hours following infusion when compared to standard heparin therapy.

C22H36N2O5S

440.6

440.234

144494-65-5

Tirofiban hydrochloride monohydrate;150915-40-5;Tirofiban hydrochloride;142373-60-2;Tirofiban-d9;1332075-40-7

60947

White to off-white solid powder

1.2±0.1 g/cm3

611.7±65.0 °C at 760 mmHg

223-225ºC

323.7±34.3 °C

0.0±1.8 mmHg at 25°C

1.532

4.14

3

7

14

30

579

1

CCCCS(=O)(=O)N[C@@H](CC1=CC=C(C=C1)OCCCCC2CCNCC2)C(=O)O

COKMIXFXJJXBQG-NRFANRHFSA-N

InChI=1S/C22H36N2O5S/c1-2-3-16-30(27,28)24-21(22(25)26)17-19-7-9-20(10-8-19)29-15-5-4-6-18-11-13-23-14-12-18/h7-10,18,21,23-24H,2-6,11-17H2,1H3,(H,25,26)/t21-/m0/s1

(2S)-2-(butylsulfonylamino)-3-[4-(4-piperidin-4-ylbutoxy)phenyl]propanoic acid

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)

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.

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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).

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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)

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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).

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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

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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.

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