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The parasympathetic system and its associated muscarinic receptors have been the subject of a renaissance of interest for the following two main reasons: (1) the association of endothelial muscarinic receptors and the nitric oxide (NO) pathway; (2) the discovery of several muscarinic receptor subtypes and drugs interacting with them. In the present survey modern insights into the subdivision of muscarinic receptors have been dealt with as the basis for a description of the muscarinic receptor agonists and antagonists thus far known. There are at least four pharmacologically defined M receptors (M1, M2, M3, M4) in primary tissues, and five muscarinic receptors have been cloned (m1, m2, m3, m4, m5). Selective agonists for M-receptor subtypes hardly exist, and all classical agonists (acetylcholine, carbachol, etc.) are clearly nonselective. A few selective antagonists for M1 (pirenzepine) and M2 receptors (AF-DX 116) have been introduced, although selective M3 receptors are hardly available. Finally, the potential therapeutic use of M-receptor agonists (myocardial ischemia, hypertension) and muscarinic antagonists (certain forms of bradycardia, coronary spasm) has been critically discussed. Although only in a preliminary stage, this development appears to be promising and at least of great fundamental interest.

The present study examined the effects of a series of tricyclic muscarinic receptor antagonists on muscarinic receptors present in the guinea-pig ileum, both in vitro and in vivo. The selectivity profiles of these antagonists and that of atropine were determined by their affinity for cortical muscarinic M1, cardiac M2 and submandibular M3 receptors and for m4 receptors expressed in CHO cells. The compounds pirenzepine, UH-AH 37, AQ-RA 391 and AQ-RA 618 possessed high affinity (pKi 7.94-8.22) for muscarinic M1 receptors. Pirenzepine exhibited the most pronounced muscarinic M1 selectivity. AF-DX 384 and AQ-RA 741 possessed an approximately 10-fold higher affinity for the cardiac muscarinic M2 receptor than AF-DX 116. However, both compounds also exhibited high affinity for muscarinic m4 receptors. High affinity for muscarinic M3 and m4 receptors was observed for UH-AH 37, AQ-RA 391 and AQ-RA 681. The antagonists were then tested for their interaction with the muscarinic receptors which are responsible for the methacholine-induced contraction of longitudinal muscle in vitro, circular muscle in vivo and muscarinic receptors which mediate the distension-evoked ascending reflex contraction of circular muscle in vitro. Compounds showing high affinity for muscarinic M3 receptors (e.g. AQ-RA 618) were the most potent antagonists in the functional experiments. Comparison of the binding displacement data with the functional results indicates that the effects of methacholine on the longitudinal and circular muscle of the guinea-pig ileum were predominantly mediated by muscarinic M3-type receptors. In contrast, the correlation between muscarinic M2 receptor affinity and antagonism of muscarinic receptors in the ileum was very weak.

The discovery of the M1-selective receptor antagonist pirenzepine was the impetus for a research project directed towards the development of selective muscarinic antagonists. In the pursuit of this objective, compounds with different selectivity profiles have been found. AF-DX 116 was the first cardioselective antagonist synthesized. Subsequently novel M2 receptor antagonists have been discovered with higher potency and selectivity. Moreover, a pirenzepine-type compound UH-AH 37 has been identified that, in contrast to pirenzepine, shows a higher affinity for ileal than for atrial muscarinic receptors. Among tricyclic muscarinic receptor antagonists three different selectivity profiles have been identified, namely: M1 greater than M3 greater than M2, Msm for pirenzepine; M2 greater than M1 greater than M3, Msm for AF-DX 116, AF-DX 384, AQ-RA 741; and Msm congruent to M1 greater than M2, M3 for UH-AH 37 and its (+) enantiomer.

The novel antimuscarinic compound UH-AH 37 (6-chloro-5,10-dihydro-5-[(1-methyl-4-piperidyl)acetyl]-11H- dibenzo-[b, e][1,4]diazepine-11-one hydrochloride) showed a 14-fold higher affinity for ileal than for atrial muscarinic receptors. In receptor binding studies UH-AH 37 showed no marked selectivity for either atrial, glandular or ileal muscarinic binding sites. Moreover, it did not reveal binding heterogeneity in membranes from ileal smooth muscle. These result indicate that UH-AH 37 possesses a unique and novel selectivity profile.