| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Targets |
P2Y12 Receptor
2-Methylthio-AMP specifically targets the P2Y12 receptor, a Gi-coupled GPCR expressed on platelets. It functions as a selective and direct antagonist, blocking the binding of the endogenous agonist ADP. By antagonizing this receptor, it inhibits the downstream signaling events that lead to platelet activation, shape change, and aggregation. It has no effect on P2Y1, the other ADP receptor on platelets. |
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| ln Vitro |
2-Methylthio-AMP (2-MeSAMP) prevents platelets from activating αIIbβ3 when triggered by an agonist[1]. Thrombin, PAR1-AP, PAR4-AP, and ADP-mediated platelet Ca2+ mobilization are not significantly inhibited by 2-methylthio-AMP (50 μM; for 5 minutes). 2-methylthio-AMP does not increase cAMP or cause vasodilator-stimulated phosphoprotein phosphorylation in wild-type platelets, nor does it block Ca2+ mobilization in P2Y12-deficient mouse platelets[1].
In vitro, 2-Methylthio-AMP acts as a direct antagonist of the P2Y12 receptor. At a concentration of 50 uM, it inhibits agonist-mediated alphaIIbbeta3 activation in platelets, which is the final step in platelet aggregation. It effectively blocks ADP-induced platelet activation without affecting the activity of other platelet receptors (e.g., P2Y1, thrombin receptors). Its mechanism is competitive antagonism. |
| ln Vivo |
In vivo, 2-Methylthio-AMP is used to study the effects of acute P2Y12 receptor blockade. When administered intravenously to animal models, it effectively inhibits ex vivo ADP-induced platelet aggregation. While not used clinically, it serves as a tool to validate the antithrombotic effects of P2Y12 antagonism. It can be used to study the role of P2Y12 in tumor metastasis and inflammatory responses, as platelet activation is known to facilitate these processes.
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| Enzyme Assay |
The specific protocol for assessing P2Y12 receptor binding uses a radioligand binding assay with [33P]-2-methylthio-ADP. Human platelet membranes are prepared from platelet concentrates via washing and lysis. The membranes are incubated with 0.1 nM of the radioligand and increasing concentrations of 2-Methylthio-AMP (0.1 nM to 100 uM) in binding buffer. Non-specific binding is determined with excess unlabeled ADP (10 uM). After a 60-minute incubation at 4degC, the reaction is terminated by vacuum filtration. The filters are washed, and bound radioactivity is measured. The Ki is calculated from the competition curve.
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| Cell Assay |
The gold standard in vitro assay for 2-MeSAMP is platelet aggregation. Human whole blood is collected via venipuncture into 3.8% sodium citrate tubes. Platelet-rich plasma (PRP) is prepared by centrifugation at 200g for 15 minutes. The platelet count is adjusted to 2-3e8/mL. 2-Methylthio-AMP (50-100 uM) is pre-incubated with PRP at 37degC for 5 minutes. Aggregation is induced by the addition of ADP (5 uM) or 2-methylthio-ADP (100 nM), and the increase in light transmission is recorded in an aggregometer. The extent of aggregation is measured as the percentage increase in light transmission over 5 minutes.
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| Animal Protocol |
An in vivo protocol for 2-Methylthio-AMP would involve a mouse model of pulmonary thrombosis. Male BALB/c mice (8-10 weeks old) are pre-treated with 2-Methylthio-AMP via intravenous injection (tail vein) at doses of 1-10 mg/kg. Five minutes later, a mixture of collagen (0.5 mg/kg) and epinephrine (60 ug/kg) is administered intravenously to induce acute pulmonary thromboembolism. The survival time of the mice is recorded. Alternatively, lungs are harvested 15 minutes after the thrombotic challenge, and the number of thrombi in the pulmonary arteries is counted under a dissecting microscope. Inhibition of P2Y12 should improve survival and reduce thrombus count.
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| ADME/Pharmacokinetics |
2-Methylthio-AMP, being a nucleotide analog, has a very short half-life in vivo due to rapid degradation by ectonucleotidases on vascular endothelium and in plasma. It is expected to have poor oral bioavailability. To achieve systemic effects, it is typically administered intravenously. For in vitro studies, it is stable in aqueous buffers and plasma for the duration of typical assays. Its phosphate group prevents cellular uptake, ensuring it acts only on surface P2Y12 receptors.
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| Toxicity/Toxicokinetics |
Specific toxicity data for 2-Methylthio-AMP is not available. As an inhibitor of the P2Y12 receptor on platelets, the primary toxicological effect is an increased risk of bleeding. High systemic doses could lead to prolongation of bleeding time and potential hemorrhage. It is generally non-cytotoxic to cells at concentrations up to 100 uM. Safety handling for laboratory use involves standard precautions to avoid inhalation or skin contact. It is for research use only.
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| References |
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| Additional Infomation |
2-Methylthio-AMP (2-MeSAMP) is a research-grade chemical and is not approved for clinical use. Its molecular formula is C11H16N5O7PS, with a molecular weight of 393.31. It is soluble in DMSO. It is often used as a selective P2Y12 antagonist, distinguishing the role of P2Y12 from P2Y1. Compared to clinical drugs like clopidogrel, which are prodrugs requiring metabolism, 2-MeSAMP acts directly and rapidly, making it a superior tool for acute ex vivo and in vitro experiments.
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| Molecular Formula |
C11H16N5O7PS
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|---|---|
| Molecular Weight |
393.31
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| Exact Mass |
393.051
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| CAS # |
22140-20-1
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| Related CAS # |
2-Methylthio-AMP diTEA;1227193-98-7
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| PubChem CID |
189762
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| Appearance |
White to off-white solid powder
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| LogP |
-2.7
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
12
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
25
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| Complexity |
529
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| Defined Atom Stereocenter Count |
4
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| SMILES |
CSC1=NC(=C2C(=N1)N(C=N2)[C@H]3[C@@H]([C@@H]([C@H](O3)COP(=O)(O)O)O)O)N
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| InChi Key |
XVTFTCNRRAQHEQ-KQYNXXCUSA-N
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| InChi Code |
InChI=1S/C11H16N5O7PS/c1-25-11-14-8(12)5-9(15-11)16(3-13-5)10-7(18)6(17)4(23-10)2-22-24(19,20)21/h3-4,6-7,10,17-18H,2H2,1H3,(H2,12,14,15)(H2,19,20,21)/t4-,6-,7-,10-/m1/s1
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| Chemical Name |
[(2R,3S,4R,5R)-5-(6-amino-2-methylsulfanylpurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methyl dihydrogen phosphate
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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: 100 mg/mL (254.25 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.36 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.36 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.36 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5425 mL | 12.7126 mL | 25.4252 mL | |
| 5 mM | 0.5085 mL | 2.5425 mL | 5.0850 mL | |
| 10 mM | 0.2543 mL | 1.2713 mL | 2.5425 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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