Structure of a nanobody-stabilized active state of the ß(2) adrenoceptor.

G protein coupled receptors (GPCRs) exhibit a spectrum of functional behaviours in response to natural and synthetic ligands. Recent crystal structures provide insights into inactive states of several GPCRs. Efforts to obtain an agonist-bound active-state GPCR structure have proven difficult due to the inherent instability of this state in the absence of a G protein. We generated a camelid antibody fragment (nanobody) to the human ß(2) adrenergic receptor (ß(2)AR) that exhibits G protein-like behaviour, and obtained an agonist-bound, active-state crystal structure of the receptor-nanobody complex. Comparison with the inactive ß(2)AR structure reveals subtle changes in the binding pocket; however, these small changes are associated with an 11 Å outward movement of the cytoplasmic end of transmembrane segment 6, and rearrangements of transmembrane segments 5 and 7 that are remarkably similar to those observed in opsin, an active form of rhodopsin. This structure provides insights into the process of agonist binding and activation.

Discovery of olodaterol, a novel inhaled beta2-adrenoceptor agonist with a 24 h bronchodilatory efficacy.

Compound 4p was identified from a series of 6-hydroxy-4H-benzo[1,4]oxazin-3-ones as potent agonist of the human beta2-adrenoceptor with a high beta1/beta2-selectivity. A complete reversal of acetylcholine-induced bronchoconstriction which lasted over the whole study period of 5h was demonstrated for 4p in a guinea pig in vivo model without any signs of cardiovascular effects up to 10-fold above the first dose reaching 100% bronchoprotection. The enantiomerically pure (R)-form of 4p exerted a bronchodilatory efficacy over 24 h in dogs and guinea pigs in the absence of systemic pharmacodynamic effects. Formoterol which was tested as comparator in the same in vivo models of acetylcholine-induced bronchoconstriction did not retain efficacy after 24 h. In summary, the preclinical profile of compound (R)-4p (olodaterol, also known as BI 1744 CL) suggests a potential for once-daily dosing in man accompanied with an improved safety profile.

Use of 5-hydroxy-4H-benzo[1,4]oxazin-3-ones as beta2-adrenoceptor agonists.

Novel beta(2)-agonists with a 5-hydroxy-4H-benzo[1,4]oxazin-3-one moiety as head group are described. Systematic chemical variations at the phenethylamine residue of these compounds lead to the discovery of compound 6m as potent, full agonist of the beta(2)-adrenoceptor with a high beta(1)/beta(2)-selectivity. Molecular modeling revealed an interaction between the carboxylic acid group of 6m and a lysine residue (K305) of the beta(2)-receptor as putative explanation for the high observed selectivity. Further, compound 6m displayed in a guinea pig in vivo model a complete reversal of acetylcholine induced bronchoconstriction which lasted over the complete study time of 5h.

Studies towards topical selective beta2-adrenoceptor agonists with a long duration of action.

Beta2-adrenoceptor agonists with basic and acidic groups attached via an alkyl linker to the phenyl ethanolamine core were prepared and investigated in vitro and in vivo. The compounds exhibited a high potency in a functional cellular assay and a bronchoprotective effect in a guinea pig model which lasted over the complete study period of 5h.

A convergent three-component total synthesis of the powerful immunosuppressant (-)-sanglifehrin a.

The potent immunosuppressive agent (-)-sanglifehrin A (5), initially discovered in a soil sample from Malawi, has been synthesized in a highly convergent and stereocontrolled manner. The enantioselective approach relies on initial construction of the iodovinyl carboxylic acid 14, which is coupled to tripeptide 59 in advance of a key macrolactonization step that generates 61a. An alternative protocol that involves the linkage of 14 to 46 for possible construction of the large ring failed due to an inability to bring about a corresponding macrolactamization maneuver. An efficient means for elaborating the C26-N42 spirolactam western sector of 5 is also detailed. This requisite fragment was assembled through the proper adaptation of consecutive aldol tactics for construction of the nine stereogenic centers, six of which are contiguous. The first aldol process consisted of the tin triflate-mediated reaction of the aldehyde derived from 72 with enantiopure ketone 73 to generate the syn C36-C37 relationship resident in 75. Once the conversion of 75 to 78 had been completed, the attachment to ketone 66 was effected with (+)-DIPCl, thereby setting the C33-C34 relationship as anti. Once functional group modifications had given rise to 62, spirolactamization was achieved to deliver predominantly 94, thereby setting the stage for the acquisition of vinyl stannane 13 and its subsequent palladium-catalyzed Stille coupling to 61b. Controlled acidic hydrolysis completed the synthesis of 5. Other important features of the present route are addressed where relevant.