Simvastatin reduces insulin-like growth factor-1 signaling in differentiating C2C12 mouse myoblast cells in an HMG-CoA reductase inhibition-independent manner.

Inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase occasionally cause myopathy characterized by weakness, pain, and elevated serum creatine phosphokinase (CK). In this study, we investigated the effects of simvastatin, an HMG-CoA reductase inhibitor, on the viability and insulin-like growth factor-1 (IGF-1) signaling in differentiating C2C12 mouse myoblast cells. Simvastatin decreased cell viability and CK activity, a marker of myogenesis, in differentiating cells in a dose-dependent manner. Although the simvastatin-induced decrease in viability in proliferating and differentiated cells was completely abolished by mevalonate or geranylgeranyl-pyrophosphate, the inhibitory effects of simvastatin in differentiating cells were not abolished by mevalonate or isoprenoid derivatives of mevalonate. Moreover, the sensitivity of differentiating cells to simvastatin regarding cell viability was about 7 times higher than that of proliferating cells. After induction of differentiation in the presence of 1 microM simvastatin for 2 days, IGF-1-induced activation of ERK1/2 and Akt was significantly decreased. Although mRNA expression of the IGF-1 receptor beta-chain (IGF-1R beta) did not change, protein level of the 200 kDa IGF-1Rbeta precursor was significantly increased by simvastatin in a dose-dependent manner. Mevalonate did not abolish the effect of simvastatin on IGF-1Rbeta expression. These results suggest that simvastatin decreases IGF-1 signaling via a regulation of the post-translational modification of IGF-1Rbeta in an HMG-CoA reductase inhibition-independent manner.

Comparison of the binding affinity of some newly synthesized phenylethanolamine and phenoxypropanolamine compounds at recombinant human beta- and alpha1-adrenoceptor subtypes.

We evaluated six new compounds, SWR-0065HA ([4-[2-[3-[[(3,4-dihydro-4-oxo-[1,2,4]-triazino(4,5-a)indol)-lyl]oxy]-2-hydroxypropylamino]ethoxy]phenyl]acetic acid methyl ester hydrochloride), SWR-0098NA ((R*R*-UE)-(E)-[4-[3-[(2-phenyl-2-hydroxyethyl)amino]-1-butenyl]phenoxy]acetic acid sodium salt), SWR-0315NA ((E, Z)-[4[[1-[2-[(3-phenoxy-2-hydroxy propyl)]amino]ethyl]-1-propenyl]phenoxy]acetic acid sodium), SWR-0338SA ((E)-[4-[5-[(2-phenyl-2-hydroxyethyl)amino]-2-pentene-3-yl]phenoxy] acetic acid ethanedioic acid), SWR-0342SA ((S)-(Z)-[4-[[1-[2-[(2-hydroxy-3-phenoxypropyl)]amino] ethyl]-1-propenyl]phenoxy]acetic acid ethanedioic acid) and SWR-0345HA ((E)-2-methyl-3-[4-[2-(2-phenyl-2-hydroxyethylamino)ethoxy]phenyl]-2-propenoic acid ethyl ester hydrochloride) for their potencies as selective ligands at human beta-adrenoceptors expressed in COS-7 cells and compared the binding affinities for human alpha(1)-adrenoceptors expressed in Chinese hamster ovary (CHO) cells using a radioligand-binding assay. Phenoxypropanolamine derivatives SWR-0315NA and SWR-0342SA showed higher binding affinities for beta-adrenoceptor subtypes; SWR-0065HA, however, showed a higher affinity for only beta-adrenoceptors, accounting for 3-fold and 6-fold selectivity against beta(1)- and beta(3)-adrenoceptors. Compounds SWR-0315NA and SWR-0342SA did not show any binding selectivity for any of the subtypes. However, functionally these two compounds are selective for beta(3)-adrenoceptors. Among the phenylethanolamine derivatives, SWR-0338SA and SWR-0345HA showed 9-fold and 16-fold higher binding selectivity for beta(3)-adrenoceptors against beta(1)-adrenoceptors, respectively, whereas they both showed a 7-fold higher binding selectivity for beta(3)-adrenoceptors against beta(2)-adrenoceptors. SWR-0098NA did not show any significant binding affinity for any of the beta-adrenoceptor subtypes. These compounds, except for SWR-0098NA, were not found to possess any significant binding affinity for alpha(1)-adrenoceptor subtypes over that for beta-adrenoceptor subtypes. However, SWR-0098NA has about a 3-fold to 22-fold higher binding selectivity for alpha(1)-adrenoceptor subtypes against beta-adrenoceptor subtypes, making it difficult for use in a beta-adrenoceptor receptor study. Compounds SWR-0315NA and SWR-0342SA have similar binding potency for alpha(1)-adrenoceptors as adrenaline (epinephrine), proving the finding of this manuscript that this phenoxypropanolamine group of beta-adrenoceptor ligands could also be used as alpha(1)-adrenoceptor ligands. Functional assays have to be performed to confirm their agonistic activity.

Molecular characterization of pharmacological properties and selectivity of SWR-0315NA for beta3-adrenoceptors.

The pharmacological properties of SWR-0315NA, (E,Z)-[4[[1-[2-[(3-phenoxy-2-hydroxy propyl)]amino]ethyl]-1-propenyl]phenoxy]acetic acid sodium, were compared with those of (-)-isoproterenol. In the radioligand binding studies of [(125)I]iodocyanopindolol with COS-7 cell membranes that transiently expressed human beta-adrenoceptor (beta-AR) subtypes, SWR-0315NA exhibited 1-fold and 2-fold greater binding affinities for beta(3)-AR than those for beta(1)- and beta(2)-ARs, respectively. The maximal stimulatory effects of SWR-0315NA on cAMP accumulation in CHO cells expressing all the beta-AR subtypes were 79%, 3% and 93% for beta(1)-, beta(2)- and beta(3)-ARs of those produced by (-)-isoproterenol, respectively. SWR-0315NA has 26.3-fold and more than 630-fold greater selectivity for beta(3)-AR than those for beta(1)- and beta(2)-ARs in potency, respectively. These results indicate that although SWR-0315NA has lower binding selectivity towards beta-AR subtypes, it is a selective agonist with high intrinsic activity for beta(3)-AR as compared with (-)-isoproterenol.