| Protocol | In Vitro | In vitro activity: Bucladesine sodium (also known as Dibutyryl-cAMP) is a cell-permeable PKA activator and a cAMP analog that mimics the action of endogenous cAMP. It is a cyclic nucleotide derivative (structurally similar to cAMP) and is also a phosphodiesterase inhibitor. Dibutyryl-cAMP preferentially activates cAMP-dependent protein kinase. The products releaes butyrate due to intracellular and extracellular esterase action. Butyrate was shown to have distinct biological effects. The compound is used in a wide variety of research applications because it mimics cAMP and can induce normal physiological responses when added to cells in experimental conditions. Kinase Assay: Bucladesine sodium (also known as Dibutyryl-cAMP) is a cell-permeable PKA activator and a cAMP analog that mimics the action of endogenous cAMP. It is a cyclic nucleotide derivative (structurally similar to cAMP) and is also a phosphodiesterase inhibitor. Dibutyryl-cAMP preferentially activates cAMP-dependent protein kinase. |
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| In Vivo | Experimentally naïve male Albino Swiss mice (Laboratory Animals Breeding, Słaboszów, Poland) weighing 20–30 g were used in all experiments. The animals were housed in polycarbonate cages at a controlled temperature (23–25°C) and humidity (50–60%) with 12 h light/dark cycle (lights on at 6 h). Tap water and food pellets were available ad libitum. All experiments were performed after at least 7 days of acclimatization. The experimental protocols were approved by the Ethical Committee of the Medical University in Lublin (license number 40/2010). To allow for skin penetration of the cream formulations and based on preliminary experiments with ketoprofen cream in this model (data not presented), a pre-treatment time of 3 h was selected. To evaluate the effect of 0.5 or 1.5% bucladesine cream, an amount of 10 ± 1 mg cream each was spread onto the outer surface of both, left and right ears using small spatula. A third group of mice was administered with 2.5% ketoprofen gel. Corresponding vehicle treatment groups were administered either the respective cream base without bucladesine or, as reference for ketoprofen, a cosmetic night cream. To evaluate the effect of repeated administration, four separate groups of mice were dosed twice, i.e., 7 and 3 h before administration of arachidonic acid with the 0.5 and 1.5% cream or corresponding vehicle. Arachidonic acid or actetone were administered 3 h after the second cream administration onto left and right ears, respectively. The ear thickness was measured with a precise spring-loaded caliper (Mitutoyo, type 7309, Kawasaki, Japan) before and 60 min after application of arachidonic acid or vehicle. |
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| Animal model | Naïve male Albino Swiss mice |
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These protocols are for reference only. InvivoChem does not
independently validate these methods.
Preparing Stock Solutions
| Solvent volume to be added |
Mass (the weight of a compound) |
| Mother liquor concentration |
1mg | 5mg | 10mg | 20mg |
| 1mM | 2.1304 mL | 10.6521 mL | 21.3042 mL | 42.6085 mL |
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| 5mM | 0.4261 mL | 2.1304 mL | 4.2608 mL | 8.5217 mL |
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| 10mM | 0.2130 mL | 1.0652 mL | 2.1304 mL | 4.2608 mL |
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| 20mM | 0.1065 mL | 0.5326 mL | 1.0652 mL | 2.1304 mL |
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Quality Control Documentation
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