Table 4.
Functional cAMP AlphaScreen® assays were performed to determine in vitro potency of compounds to induce cAMP signaling in five cell categories (Figure 6): (A) stable mMC3R cells, (B) stable mMC4R cells, (C) stable mMC3R cells transiently transfected with mMC4R, (D) stable mMC4R cells transiently transfected with mMC3R, and (E) an equal mixture of stable mMC3R cells and mMC4R cells. All assays and cell categories were run in parallel. The reported standard error of the mean (SEM) was determined from at least three independent experiments.
| Agonist cAMP Based Alpha Screen Results EC50 (nM) | |||||
|---|---|---|---|---|---|
| Compound | mMC3R | mMC4R | mMC3R Trans mMC4R | mMC4R Trans mMC3R | mMC3R Mixed with mMC4R |
| Mean±SEM | Mean±SEM | Mean±SEM | Mean±SEM | Mean±SEM | |
| NDP-MSH | 0.13±0.02 | 0.46±0.07 | 0.19±0.03 | 0.12±0.01 | 0.13±0.02 |
| α-MSH | 0.21±0.02 | 3.1±0.7 | 0.27±0.05 | 0.20±0.03 | 0.19±0.03 |
| MTII | 0.10±0.02 | 0.11±0.03 | 0.09±0.02 | 0.06±0.01 | 0.08±0.02 |
| CJL1-14 | 23±3 | 8.5±1 | 7.7±1 | 4.0±0.5 | 9.9±2 |
| CJL1-87 | 2.6±0.4 | 1.2±0.3 | 1.4±0.3 | 0.63±0.1 | 2.0±0.4 |