N6022

Alcohol dehydrogenase (ADH) Enzyme Assays

For ADH IB, ADH II, and ADH IV assays, enzyme activity was determined by measuring the increase in A340 due to reduction of NAD+ to NADH using conditions similar to those described by Sanghani et al. N6022 was dissolved and diluted in a PBS solution containing 1 N NaOH (1%). The IC50 values for N6022 versus ADH IB and ADH II were determined with ethanol as the substrate. The final assay conditions for ADH IB were 50 mM sodium phosphate (pH 7.4), 20 μg/mL ADH IB, 2 mM NAD+, and 2 mM ethanol, and the final assay conditions for ADH II were 90 mM sodium pyrophosphate (pH 8.9), 4.4 μg/mL ADH II, 23.6 mM NAD+, and 14.4 mM ethanol. The IC50 value of N6022 against ADH IV was determined using hexanol as a substrate. The hexanol was first dissolved in DMSO so that it could be dissolved in assay buffer. The final assay conditions were 50 mM sodium phosphate (pH 7.4), 1.25 μg/mL ADH IV, 2 mM NAD+, 400 μM hexanol, and 1% DMSO.

Animal models

Female BALB/c mice model

Dosage form

0.1 mg/kg to 10 mg/kg, i.v. for 1-48 h

Applications

N6022 dose-dependently decreased enhanced pause (Penh) and Bronchoalveolar lavage fluid (BALF) eosinophils, increased bronchoalveolar lavage fluid (BALF) nitrite and plasma cyclic guanosine monophosphate (cGMP) in ovalbumin (OVA)-sensitized mice. Moreover, N6022 attenuated the OVA-induced increase in nuclear factor kappa B (NFκB) activation and decreased methacholine (MCh)-induced contraction in isolated rat tracheal rings.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

1. Green, L. S., Chun, L. E., Patton, A. K., Sun, X., Rosenthal, G. J. and Richards, J. P. (2012) Mechanism of inhibition for N6022, a first-in-class drug targeting S-nitrosoglutathione reductase. Biochemistry. 51, 2157-21682

2. Blonder, J. P., Mutka, S. C., Sun, X., Qiu, J., Green, L. H., Mehra, N. K., Boyanapalli, R., Suniga, M., Look, K., Delany, C., Richards, J. P., Looker, D., Scoggin, C. and Rosenthal, G. J. (2014) Pharmacologic inhibition of S-nitrosoglutathione reductase protects against experimental asthma in BALB/c mice through attenuation of both bronchoconstriction and inflammation. BMC Pulm Med. 14, 3