| ln Vitro |
GP515 can efficiently and specifically inhibit isolated human cardiac adenosine kinase, with an IC50 value of 4 nM[2]. After 18 hours of normoxic culture, GP515 (2-20 μM; 18 hours) can increase the expression of VEGF mRNA in cultured rat cardiomyocytes by 1.67-fold and 1.82-fold, respectively[2]. After 18 hours of normoxic culture, GP515 (0.2-200 μM; 18 hours) can increase the expression of VEGF protein in cultured rat cardiomyocytes in a dose-dependent manner, with an increase of up to 54% at the highest concentration[2]. After 12 hours of normoxic culture, GP515 (1 μM; 12-24 hours) can significantly increase the expression of VEGF protein in cultured rat cardiomyocytes, and this effect can last up to 24 hours[2]. After 18 hours of co-incubation with adenosine deaminase, GP515 (2 μM; 18 h) completely blocked the induction of VEGF protein in cultured rat cardiomyocytes[2]. GP515 (20 μM; 24 h) stimulated the proliferation of human umbilical vein endothelial cells by 98% and increased the incorporation of [3H]thymidine by 82% after 24 hours of incubation, but had no effect on the proliferation of rat cardiomyocytes[2]. GP515 (20 μM; 18 h) increased the expression of VEGF protein in cultured rat cardiomyocytes by 37% under normoxic conditions and by an additional 27% under mild hypoxia (10% O2) conditions (after 18 hours of incubation), but had no effect under severe hypoxia (1% O2) conditions[2].
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|---|---|
| ln Vivo |
Following hemorrhagic shock, systemic administration of GP515 (0.25 mg/kg; intravenous injection; continuous infusion for 1 hour; starting 90 minutes after the onset of hemorrhagic hypotension) significantly improved hepatic microcirculatory parameters (sinusoidal blood flow, diameter and perfusion index) 2 days after shock, and these parameters returned to normal within 5 days in female Sprague-Dawley rats (200-250 g) [1].
|
| Cell Assay |
ELISA detection [2]
Cell Types: Cultured rat myoblasts (RMMs) Tested Concentrations: 0.2 μM; 2 μM; 20 μM; 200 μM Incubation Duration: 18 hours Experimental Results: At 0.2 μM, VEGF protein level increased to 1.99 ng/mg total cell protein (an increase of 8%). At 2 μM, VEGF protein level increased to 2.50 ng/mg total cell protein (an increase of 36%). At 20 μM, VEGF protein level increased to 2.56 ng/mg total cell protein (an increase of 39%). At 200 μM, VEGF protein level increased to 2.84 ng/mg total cell protein (an increase of 54%). The VEGF protein level in the control group was 1.84 ng/mg total cellular protein, and all increases were statistically significant. ELISA detection [2] Cell Types: Cultured rat cardiomyocytes (RMMs) Tested Concentrations: 1 μM Incubation Duration: 2 hours; 6 hours; 12 hours; 24 hours Experimental Results: After 2 hours or 6 hours of incubation, VEGF protein levels did not increase significantly. After 12 hours, VEGF protein levels increased significantly by 20%. VEGF protein levels continued to increase until 24 hours, at which point the levels were similar in quantity to those induced by equimolar adenosine. ELISA detection [2] Cell Types:Cultured rat myoblasts (RMMs) Tested Concentrations:2 μM (co-incubated with 10 U/mL adenosine deaminase) Incubation Duration:18 hours Experimental Results:Co-incubation with adenosine deaminase completely blocked the GP515-induced increase in VEGF protein, reducing its level to 0.75 ng/mg total cell protein, a 60% reduction compared to the control group's 1.84 ng/mg total cell protein. |
| Animal Protocol |
Animal/Disease Models:Sprague-Dawley mice (female, 200-250 g, induced by bloodletting to induce pressure-controlled hemorrhagic hypotension, maintaining mean arterial pressure at 40 mmHg for 90 minutes, followed by resuscitation with 60% blood loss and lactated Ringer's solution) [1]
Doses: 0.25 mg/kg Route of Administration: Intravenous injection; continuous infusion for 1 hour; started 90 minutes after the onset of hemorrhagic hypotension Experimental Results: Two days after shock, sinus blood flow increased to 40833 µm3/s, mean sinus diameter increased to 12.08 µm, and perfusion index increased to 91.5%, all of which were significantly higher than those in the placebo group. Five days after shock, sinus diameter, blood flow, and perfusion index returned to normal, with no significant difference compared to placebo-treated rats. |
| References |
|
| Molecular Formula |
C10H13BRN6O3
|
|---|---|
| Molecular Weight |
345.15
|
| CAS # |
144928-48-3
|
| Appearance |
Typically exists as solids at room temperature
|
| SMILES |
O[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(N)=C3C(Br)=N2)O[C@@H]1CN
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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 | 2.8973 mL | 14.4865 mL | 28.9729 mL | |
| 5 mM | 0.5795 mL | 2.8973 mL | 5.7946 mL | |
| 10 mM | 0.2897 mL | 1.4486 mL | 2.8973 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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