GP515
GP515 is a potent and selective adenosine kinase inhibitor with a human IC50 of 4 nM. GP515 exerts tissue protective effects, produces long-lasting hepatic microcirculation effects after hemorrhagic shock, and induces dose- and time-related VEGF mRNA and protein expression in normoxic rat myocardial myoblasts, with additive VEGF increases during mild hypoxia and no effect during severe hypoxia. GP515 suppresses IFNγ synthesis and CD69 expression in DSS-induced colitis. GP515 also shows a dose-dependent suppression of TNF-α production with an IC50 of 80 μM and can be reversed in the presence of the cAMP antagonist (Rp)-cAMPS. Combinations of GP515 with either adenosine or rolipram led to an additive inhibition of TNF-α synthesis. GP515 can be used for the research of hemorrhagic shock.
For research use only. We do not sell to patients.
- CAS No.: 144928-48-3
- Formula: C10H13BrN6O3
- Molecular Weight:345.15
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
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Biological Activity
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Chemical Information
- Protocols
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
In Vitro
GP515 potently and specifically inhibits isolated human cardiac adenosine kinase with an IC50 of 4 nM[2].
GP515 (2-20 μM; 18 h) increases VEGF mRNA expression in cultured rat myocardial myoblasts by 1.67-fold and 1.82-fold, respectively, after 18 h of normoxic incubation[2].
GP515 (0.2-200 μM; 18 h) induces a dose-related increase in VEGF protein expression in cultured rat myocardial myoblasts after 18 h of normoxic incubation, reaching up to a 54% increase at the highest concentration[2].
GP515 (1 μM; 12-24 h) significantly increases VEGF protein expression in cultured rat myocardial myoblasts after 12 h of normoxic incubation, with effects persisting through 24 h[2].
GP515 (2 μM; 18 h) co-incubated with adenosine deaminase completely blocks the VEGF protein-inducing effect in cultured rat myocardial myoblasts after 18 h of normoxic incubation[2].
GP515 (20 μM; 24 h) stimulates human umbilical vein endothelial cell proliferation by 98% and increases [3H]thymidine incorporation by 82% after 24 h of incubation, with no effect on rat myocardial myoblast proliferation[2].
GP515 (20 μM; 18 h) increases VEGF protein expression in cultured rat myocardial myoblasts by 37% under normoxia and by an additional 27% under mild hypoxia (10% O2) after 18 h of incubation, but has no effect under severe hypoxia (1% O2)[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:cultured rat myocardial myoblasts (RMMs)
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Concentration:0.2 μM; 2 μM; 20 μM; 200 μM
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Incubation Time:18 h
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Result:Increased VEGF protein levels to 1.99 ng/mg total cell protein (8% increase) at 0.2 μM.
Increased VEGF protein levels to 2.50 ng/mg total cell protein (36% increase) at 2 μM.
Increased VEGF protein levels to 2.56 ng/mg total cell protein (39% increase) at 20 μM.
Increased VEGF protein levels to 2.84 ng/mg total cell protein (54% increase) at 200 μM; control VEGF protein levels were 1.84 ng/mg total cell protein, with all increases statistically significant.
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Cell Line:cultured rat myocardial myoblasts (RMMs)
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Concentration:1 μM
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Incubation Time:2 h; 6 h; 12 h; 24 h
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Result:Did not significantly increase VEGF protein levels after 2 or 6 h of incubation.
Caused a significant 20% increase after 12 h.
Continued to increase levels such that by 24 h, levels were quantitatively similar to those induced by equimolar adenosine.
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Cell Line:cultured rat myocardial myoblasts (RMMs)
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Concentration:2 μM (co-incubated with 10 U/mL adenosine deaminase)
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Incubation Time:18 h
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Result:Co-incubation with adenosine deaminase completely blocked the GP515-induced increase in VEGF protein, reducing levels to 0.75 ng/mg total cell protein, a 60% decrease compared to control levels of 1.84 ng/mg total cell protein.
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Cell Line:human umbilical vein endothelial cells (HUVECs), rat myocardial myoblasts (RMMs)
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Concentration:20 μM
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Incubation Time:24 h
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Result:Increased HUVEC cell number by 98% (from control 8.5 × 105 cells/well).
Increased [3H]thymidine incorporation by 82% (from control 11.91 × 103 cpm/well), with both changes statistically significant.
Had no effect on RMM proliferation.
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Cell Line:cultured rat myocardial myoblasts (RMMs)
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Concentration:20 μM
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Incubation Time:18 h (under normoxia, mild hypoxia, severe hypoxia)
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Result:Increased VEGF protein levels by 37% (to 2.79 ng/mg total cell protein) compared to normoxic control levels of 2.04 ng/mg total cell protein under normoxia.
Increased VEGF protein levels by an additional 27% (to 4.17 ng/mg total cell protein) compared to mild hypoxia control levels of 3.29 ng/mg total cell protein under mild hypoxia.
Had no effect on VEGF protein levels (5.84 ng/mg total cell protein vs.
severe hypoxia control levels of 6.06 ng/mg total cell protein) under severe hypoxia.
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:Sprague-Dawley (female, 200-250 g, pressure-controlled hemorrhagic hypotension induced by blood withdrawal to maintain mean arterial pressure at 40 mmHg for 90 minutes, followed by resuscitation with 60% shed blood and lactate Ringer’s solution)[1]
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Dosage:0.25 mg/kg
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Administration:i.v.; continuous infusion over 1 hour; starting 90 minutes after onset of hemorrhagic hypotension
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Result:Increased sinusoidal blood flow to 40833 µm3/s, mean sinusoidal diameter to 12.08 µm, and perfusion index to 91.5% at 2 days post-shock, with all values significantly higher than placebo.
Normalized sinusoidal diameters, blood flow, and perfusion index by 5 days post-shock, with no significant difference from placebo-treated rats.
Chemical Information
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CAS No. 144928-48-3
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Molecular Weight 345.15
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Formula C10H13BrN6O3
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SMILES
O[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(N)=C3C(Br)=N2)O[C@@H]1CN
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
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Western Blot
Western blotting (WB) is a commonly used experimental method in molecular biology, biochemistry, and immunogenetics for identifying and quantifying target proteins. It combines gel electrophoresis with immunoassay, enabling researchers to analyze protein expression, post-translational modifications, and molecular weight.
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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DSS-Induced Colitis
Dextran sulfate sodium (DSS)-induced colitis is generated by administering DSS in mouse drinking water, producing epithelial injury, barrier disruption, weight loss, diarrhea, fecal blood, colon shortening, histologic mucosal damage, and inflammatory mediator changes; the model is mainly used to study acute or chronic intestinal inflammation resembling selected features of ulcerative colitis. DSS injury is interpreted through clinical and tissue readouts rather than a single molecular endpoint: daily body weight, stool consistency, and bleeding are combined into a disease activity index, while colon length, histology, cytokines, myeloperoxidase activity, intestinal permeability, and tight-junction markers provide complementary measures of inflammation and barrier damage.
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TNBS-Induced Colitis
TNBS-induced colitis is produced by intrarectal delivery of 2,4,6-trinitrobenzene sulfonic acid in ethanol, where ethanol disrupts the mucosal barrier and TNBS haptenates colonic proteins, generating immune-mediated colonic inflammation with weight loss, diarrhea, ulceration, transmural injury, inflammatory-cell infiltration, and cytokine responses. The model is used as an experimental intestinal inflammation model with Crohn’s disease–like features, especially when Th1-type responses, IL-12–dependent inflammation, chronic relapsing inflammation, or fibrosis-related endpoints are studied.
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Protocol For Protein Expression And Purification
Recombinant protein expression in Escherichia coli followed by purification of a His-tagged soluble protein by immobilized metal affinity chromatography (IMAC), with optional MBP fusion and TEV tag removal when the construct includes these elements. The biological readout is production of the encoded target protein, detected as an inducible band at the expected molecular mass by SDS-PAGE and quantified by total protein assay or chromatographic absorbance; the purification readout is enrichment of the target protein in elution fractions after selective binding of polyhistidine residues to immobilized Ni2+/metal-chelate resin and elution by imidazole-containing buffer. Expression is driven by an inducible bacterial expression system, commonly T7/lac-based, in which IPTG or lactose/auto-induction activates transcription and translation of the cloned gene; lower induction temperature, lower inducer concentration, induction timing, and solubility-enhancing fusion tags can influence the frac
Purity & Documentation
References
[2]. Gu JW, et al. Inhibition of adenosine kinase induces expression of VEGF mRNA and protein in myocardial myoblasts. Am J Physiol Heart Circ Physiol. 2000;279(5):H2116-H2123. [Content Brief]
[3]. Siegmund B, Rieder F, Albrich S, Wolf K, Bidlingmaier C, Firestein GS, Boyle D, Lehr HA, Loher F, Hartmann G, Endres S, Eigler A. Adenosine kinase inhibitor GP515 improves experimental colitis in mice. J Pharmacol Exp Ther. 2001 Jan;296(1):99-105. [Content Brief]
[4]. Eigler A, Matschke V, Hartmann G, Erhardt S, Boyle D, Firestein GS, Endres S. Suppression of TNF-alpha production in human mononuclear cells by an adenosine kinase inhibitor. J Leukoc Biol. 2000 Jul;68(1):97-103. [Content Brief]
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
GP515 Related Classifications
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