Bay 65-1942, an ATP-competitive inhibitor that selectively targets IKKbeta kinase activity, was administered intraperitoneally either prior to ischemia, at reperfusion, or 2 h after reperfusion. Compared with untreated animals, mice treated with IKKbeta inhibition had significant reduction in left ventricular infarct size. Cardiac function was also preserved following pretreatment with IKKbeta inhibition. [1]

IKKβ inhibition decreases size of infarction.
Delivery of Bay 65-1942 prior to ischemia significantly decreased left ventricular infarct size compared with animals receiving vehicle . Compared with sham animals, animals receiving vehicle had a significant increase in the infarct-to-AAR ratio . This ratio was significantly reduced by treatment with Bay 65-1942 at each time point (prior to ischemia 42.7 ± 4.1%, at reperfusion 42.7 ± 7.5%, 2 h of reperfusion 29.4 ± 5.2%; each group P < 0.05 vs. vehicle). The differences in this ratio between those pretreated with the IKKβ inhibitor and those that received the inhibitor in a delayed fashion were not significant. No significant differences in AAR existed between sham, vehicle, and treatment groups .[1]

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IKKβ inhibition attenuates myocardial injury.
Myocardial injury was assessed with the measurement of serum CK-MB levels 1 h following reperfusion. The CK-MB fraction was significantly elevated in the vehicle group (n = 3) compared with the sham group (n = 4) (30,530 ± 371.2 vs. 9,675 ± 608.4 units, P < 0.05). Animals pretreated with Bay 65-1942 (n = 3) had significantly attenuated CK-MB levels compared with those animals without treatment prior to IR (14,170 ± 3,219 units, P < 0.05 vs. vehicle).[1]

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IKKβ inhibition preserves cardiac function.
We assessed cardiac function by comparing pressure-volume recordings in mice at baseline and following IR with or without Bay 65-1942. Bay 65-1942 administration alone did not improve myocardial function above baseline hemodynamic parameters. Ejection fraction (EF) and dP/dt (the first derivative of left ventricular pressure) were significantly lower in the mice that underwent 30 min of LAD occlusion followed by 3 days of reperfusion when compared with the baseline group and the group administered Bay 65-1942 without surgery. The treatment group had a significantly improved EF and dP/dt from the IR with vehicle group, whereas no difference existed when compared with the baseline groups.[1]

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IKKβ inhibition decreases NF-κB-associated protein expression.
Western blots on left ventricular homogenates from sham, vehicle, and pretreatment groups (n = 3 for each group) killed 30 min and 1 h after reperfusion were performed to observe the effects of IKKβ inhibition on the NF-κB pathway . Expression of phospho-IκBα, the direct downstream product of IKKβ activation, was significantly elevated in vehicle animals compared with sham animals 30 min after reperfusion (P < 0.05). This difference between sham and vehicle groups was statistically lost 1 h following reperfusion, suggesting that initial IKKβ activation is at its height within 1 h of reperfusion injury. Animals treated with Bay 65-1942 had lower levels of phospho-IκBα expression compared with the vehicle group at both 30 min and 1 h following reperfusion (P < 0.05, Bay 65-1942 vs. vehicle).[1]

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IKKβ inhibition decreases TNF-α and IL-6 expression.
TNF-α and IL-6 are NF-κB-dependent cytokines activated in response to myocardial injury. One hour after reperfusion, levels of TNF-α and IL-6 were significantly elevated in the vehicle groups compared with the sham animals (TNF-α: 248.6 ± 25 vs. 35.1 ± 35.1 pg/ml, P < 0.05; IL-6: 5,974 ± 1,976 vs. 433.9 ± 83.1 pg/ml, P < 0.05). When administered Bay 65-1942 prior to ischemia, the amount of serum TNF-α dropped significantly compared with the vehicle group (22.4 ± 7.3 pg/ml, P < 0.05 vs. vehicle). IL-6 was also significantly lower in animals that received the IKKβ inhibitor compared with vehicle animals (417.8 ± 118.2 pg/ml, P < 0.05 vs. vehicle). There were no differences in cytokine concentrations between sham and treatment animals.[1]

Am J Physiol Heart Circ Physiol. 2007 Oct;293(4):H2248-53.;PLoS One. 2013 Jun 24;8(6):e66755.

C22H25N3O4

395.4516

395.185

758683-21-5

600734-02-9; 600734-06-3 (HCl salt)

136091530

White to gray solid

3.55

3

6

5

29

586

1

OC1=CC=CC(OCC2CC2)=C1C3=NC(NC(OC4)=O)=C4C([C@H]5CCCNC5)=C3

IGJVFGZEWDGDOO-AWEZNQCLSA-N

InChI=1S/C22H25N3O4/c26-18-4-1-5-19(28-11-13-6-7-13)20(18)17-9-15(14-3-2-8-23-10-14)16-12-29-22(27)25-21(16)24-17/h1,4-5,9,13-14,23,26H,2-3,6-8,10-12H2,(H,24,25,27)/t14-/m0/s1

7-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-5-[(3R)-piperidin-3-yl]-1,4-dihydropyrido[2,3-d][1,3]oxazin-2-one

BAY 65-1942 R-isomer; BAY-65-1942; BAY65-1942; BAY 651942; BAY-651942; BAY651942

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)

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

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.)