Non Peptide Ligands at the Opioid Receptor Like-1 (ORL-1)Mini Reviews in Medici nal Chemistry, 2001 , Vol. 1, No.

4 371 Table 9.Structure and Binding Affinity of Compounds 6. a CompdXRKi (nM) a 6a CH 2 H79.9 10.1 6b OH50.7 7.3 6c OCH 3 7.0 1.4 6d OC 2 H 5 8.2 0.3 6e OOCH 3 11.8 1.2 6f OCF 3 1.8 0.2 6g ONO 2 2.3 0.4 6h OBr2.6 0.3 6i OCl2.2

0.3 6j ONH 2 82.4 6.5 6k OOH46.7 4.9 a) Displacement of [ 3 H]nociceptin (0.5 nM) binding from human ORL-1 receptors expressed in HeLa cells . Data are given as mean SE of at least three experiments. (a) Methanol, reflux; (b) Dowtherm A, 280 C; (c) Me 2 SO 4 , toluene, reflux; (d) AcONH 4 , 135 C; (e) HCl; (f) benzoyl chlorides. Scheme 3.Chart 3. NNHNH 2 NNHNH 2 XR 6a-k6 OONH 2 NR OMeaNMeORNHbNNOHRc,d,eNNH 2 NH 2 Rf NNRR 6 O OOOOOOHHHNH 2 HOONNNOH Spiroxatrine NNNOOO 5 372 Mini Reviews in Medicinal Chemistry, 2001 , Vol. 1, No. 4Barlocco et al. (a) SOCl 2 , 0 C; (b) NEt 3 , 0 C; (c) LiAlH

4 , Et 2 O, 0 C; (d) NaOH, water; (e) CrO 3 , H 2 SO 4 . Scheme 4. general synthetic approach). In particular, compounds 6a and 6b were selected for further investigation. First, moving of the phenethyl group of 6a was studied, but the ortho positionproved to be specific. Then, introduction of varioussubstituents on the terminal phenyl ring of 6b was examined.The best result was obtained with compound 6c , whichbecame the new lead. Several different groups (ethyl,methoxy, trifluorome thyl, nitro, bromo, or chloro) wereintroduced instead of the methyl group in para position of 6c . All the compounds ( 6d-i ) showed significant bindingaffinity, independently from the electronic properti es of thesubstituent, although the effect of the electron-withdrawinggroups was stronger than the electron-donating groups.However, introduction of either an am ino ( 6j ) or a hydroxygroup ( 6k ) at the para position decreased the affinity, thoughby different degrees. These results indic ated that lipophilic,but not hydrophilic, substituents at this position played a nimportant role in the enhancement of binding. In addition,the position of the s ubstituent proved to be important, and itsmoving to the meta or ortho position resulted detrimental.The SAR on the quinoline ring showed that both the primary 4-amino- and the 2-methyl- groups are essential forthe activity. Inserti on of an isoquinoline instead of aquinoline moiety led to a weaker but still act ive derivative,thus indicating that the nitrogen atom is not directly involvedin binding. Finally, no substitution at the amide-nitrogen norits inversion were t olerated.Compound 6d , though not the most active in the bindingassay, was chosen for further in vivo studies, because itshowed the best bioavailability. It did not inhibit forskolin -stimulated cyclic AMP accumulation in human ORL-1receptor-expressing HeLa cells , but it prevented nociceptin-induced inhibition of cyclic AMP accumulation, thu

sindicating that it possesses full antagonistic activity. Oraladministration of 6d decreases allodinya induced byintrathecal injection of nociceptin. The analgesic activity of 6d was verified on the hot-plate test (mice) and the biphasicpaw-licking response i nduced by formalin injection in rats.In both cases, the analgesic effect was not reversed bynaloxone, thus ruling out the involvement of the opioidreceptors. In fact, the selectivity of 6d for ORL-1 over , , and receptors is about 12.5, 129, an 1055-fol .Researchers from SmithKline Beecham in collaborationwith the University of Milan [39] have recently patente aseries of compoun s of general formula 7 , which are sai tobe either agonists or antagonists at the ORL-1 receptor. Thei rsynthetic approach an some of the most representativecompoun s are epicte in Scheme 4 an Chart 4,respectively. As constantly seen for ORL-1 agents, also fo rthis series the affinity was greatly affecte by stereochemicalfactors, when th e substrate presente chiral atoms an /ormultiple bon s. Therefore, enantiomers an geometricalisomers were isolate when possible an teste incomparison with their racemates an iastereoisomericmixtures. As alrea y seen for other ORL-1 l igan s, normallyone enantiomer prove to be more potent at the ORL-1receptor tha n the other. On the contrary, the opioi receptorswere less influence by steric eman s, what resulte in abetter ORL-1/opioi receptor selectivity. Their affi nity at theORL-1 receptor range from 1 to 1000 nM. [ 3 H]-Nociceptinwas use as ra ioligan .Several other patents [40-46] have recently appeare , ealing with the ORL-1 non-pepti ic agonists an /or COOHRnaCOClRnNHR 1 OHRnNR 1 RnNR 1 RnNR 1 bec, OOOOO Non Pepti e Ligan s at the Opioi Receptor Like-1 (ORL-1)Mini Reviews in Me ici nal Chemistry, 2001 , Vol. 1, No. 4 373 antagonists. A selection of some representative structures isreporte in Chart 5 .Finally, it shoul be note that a few opioi ligan s (e.g.etorphine, Mr 2266, TRK-820) were reporte to have someactivity on the ORL-1 receptor. However, it s houl be a e that the concentrations require are extremely high(micromolar ran ge) with respect to those which are effectiveon the opioi receptors (nanomolar range). The onlyexception seems to be buprenorphine, which was in icate as a ful l agonist of the ORL-1 receptor in several stu ies[47,48]. It showe pEC 50 values ranging from 7.40 [49] to8.07 [50]. CONCLUSION In conclusion, the new non-pepti ic ligan s at the ORL-1receptor will be of grea t utility to better un erstan thephysiological roles of the nociceptin/ORL-1 sy

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