benzofurans and lazabemide

Topics: Benzofurans; Binding Sites; Clorgyline; Flavonoids; Humans; Inhibitory Concentration 50; Kinetics; Molecular Docking Simulation; Monoamine Oxidase; Monoamine Oxidase Inhibitors; Oxazolidinones; Picolinic Acids

This study has used receptor autoradiography to characterize imidazoline binding sites (I-BS) in monoamine oxidase (MAO) A knockout and wild-type mice. A comparison between MAO-A and MAO-B, binding of the endogenous beta-carboline [(3)H]harmane, and I-BS, has been made using sections from brain and kidney. The loss of binding to MAO-A in the knockout animals was confirmed using the selective radioligand [(3)H]Ro41-1049, with labelling reduced to background levels. The binding of [(3)H]Ro19-6327 to MAO-B was unaffected, indicating no change in this isoform in response to the loss of MAO-A. A reduction in binding to the I(2)-BS, as labelled by both [(3)H]idazoxan and [(3)H]2-BFI (2-(2-benzofuranyl)-2-imidazoline), was seen in the MAO-A knockout animals in both brain and kidney sections, whereas binding to the I(1)-BS in kidney sections remained unchanged. The loss of I(2) binding was found to be regionally dependent and was positively correlated with the relative expression of MAO-A in specific regions in the wild-type animals. Using the MAO-A knockout mice it was also possible to demonstrate a non-MAO-A population of binding sites labelled by the putative I-BS endogenous ligand, harmane.

Topics: Animals; Autoradiography; Benzofurans; Binding Sites; Binding, Competitive; Brain; Cell Membrane; Epithelial Cells; Harmine; Idazoxan; Imidazoles; Imidazoline Receptors; Imidazolines; Iodine Radioisotopes; Kidney; Mice; Mice, Inbred C3H; Mice, Knockout; Monoamine Oxidase; Neurons; Picolinic Acids; Radioligand Assay; Receptors, Drug; Thiazoles

This study was designed to determine the affinity and binding profile of beta-carbolines for imidazoline I2 receptors and catalytic sites of monoamine oxidase (MAO)-A/B in rat brain and liver. The aim was also directed to assess the in vivo effects of norharman (beta-carboline) and LSL 60101 (I2 ligand) on brain 3,4-dihydroxyphenylalanine (DOPA) synthesis in morphine-dependent rats. Competition experiments against [3H]2-BFI revealed that beta-carbolines recognize the high- and low-affinity components of the brain imidazoline I2 receptor with the rank order of potency (K(iH) in nM): noreleagnine (12)>norharman (20)>harmalol (82)>harmaline (177)>>harmine (630)>harman (700)>>FG-7142 (>100,000). In liver, this rank was different: harmine (51)>harmaline (103)=noreleagnine (103)>>harmalol (1290)>harman (2000)>>norharman (12,382)>>FG-7142 (>100,000). In brain and liver, competition curves for beta-carbolines against [3H]Ro41-1049 (MAO-A) and [3H]Ro19-6327 (MAO-B) were monophasic and resulted in different drug potencies for the two MAO isozymes (higher affinities for MAO-A) and in similar pharmacological profiles in both tissues. In morphine-dependent rats, naloxone (2 mg/kg, 2 h)-precipitated withdrawal increased the synthesis of DOPA in the cerebral cortex and hippocampus (50%). Pretreatment with norharman (20 mg/kg) or LSL 60101 (20 mg/kg) (30 min before naloxone) fully prevented the stimulatory effect of opiate withdrawal on DOPA synthesis. Norharman and LSL 60101 also attenuated the severity of the withdrawal syndrome. The results indicate that beta-carbolines bind with high affinity to imidazoline I2B receptors, and similarly to I2 ligands (LSL 60101) can block the behavioural and biochemical effects of opiate withdrawal.

Topics: Animals; Benzofurans; Binding, Competitive; Brain; Carbolines; Cerebral Cortex; Dihydroxyphenylalanine; Dose-Response Relationship, Drug; Harmine; Hippocampus; Imidazoles; Imidazoline Receptors; Liver; Male; Monoamine Oxidase; Morphine; Morphine Dependence; Naloxone; Norepinephrine; Picolinic Acids; Radioligand Assay; Rats; Rats, Sprague-Dawley; Receptors, Drug; Substance Withdrawal Syndrome; Thiazoles; Tritium; Tyrosine 3-Monooxygenase

The imidazoline I2 binding sites in the central nervous system have previously been described in several different species including rat, mouse, rabbit and frog. The present study has investigated the imidazoline I2 binding site, and its relationship to the monoamine oxidase isoforms, in pig whole brain and compared the results obtained with data from other species. Results from saturation binding studies revealed that the imidazoline I2-selective ligand, [3H]2BFI (2-(2-benzofuranyl)-2-imidazoline) labelled a single saturable population of sites with a KD=6.6 nM and Bmax=771.7 fmol/mg protein. The pharmacological characterisation of the sites was similar to that previously reported with a rank order of potency for the imidazoline I2 ligands of 2BFI>BU224>Idazoxan>BU226. Displacement by the imidazoline I1 ligands was low affinity and the monoamine oxidase inhibitors displaced with micromolar affinity. The majority of compounds displaced the binding in a monophasic manner, however, displacement by the putative endogenous ligand, harmane was biphasic. The relative populations of the two monoamine oxidase isoforms revealed a 10 fold greater expression of monoamine oxidase B relative to monoamine oxidase A. These data confirm the presence of imidazoline I2 binding sites in pig brain and show that their pharmacology is characteristic of that seen in other species. The proportion of monoamine oxidase A and B expressed in the pig brain is similar to that seen in the human brain therefore, given the association between imidazoline I2 binding sites and monoamine oxidase, the pig may provide a more useful model for human imidazoline I2 binding sites than other species such as the rat.

Topics: Animals; Benzofurans; Binding Sites; Binding, Competitive; Brain; Carbolines; Clonidine; Dose-Response Relationship, Drug; Female; Idazoxan; Imidazoles; Imidazoline Receptors; Kinetics; Male; Monoamine Oxidase Inhibitors; Picolinic Acids; Radioligand Assay; Receptors, Drug; Swine; Thiazoles; Tritium

The distribution and relative densities of imidazoline-receptor binding sites (I-RBS) in bovine adrenal gland were determined using [3H]clonidine, [3H]2-(2-benzofuranyl)-2-imidazoline ([3H]2-BFI), and [3H]rilmenidine. In light of strong evidence that I-RBS and monoamine amine oxidase enzymes are linked, the selective radioligands [3H]RO41-1049 and [3H]RO19-6327 were used to label the distribution of MAO-A and -B enzymes, respectively. [3H]Clonidine (12 nM) labeled sites in two discrete regions of the bovine adrenal gland, the zona glomerulosa (39 +/- 7 fmol/mg tissue equivalent) and inner medulla (34 +/- 1 fmol/mg tissue). Binding was nonadrenergic (i.e., not inhibited by 100 nM methoxyidazoxan) and inhibited by 60-70% by 100 nM 2-BFI, the selective I2-RBS, suggesting binding predominantly to an I2-RBS. [3H]2-BFI (5 nM), the selective I2-RBS ligand, also labeled a high density of binding sites in the zona glomerulosa (57 +/- 9 fmol/mg) and chromaffin cells in the inner medulla (53 +/- 4 fmol/mg). These sites, however, were insensitive to clonidine (100 nM). By contrast, [3H]rilmenidine (40 nM) labeled I-RBS in all regions of the adrenal gland, that is, the zonae glomerulosa (59 +/- 10 fmol/mg), fasciculata (78 +/- 10 fmol/mg) and reticularis (63 +/- 7 fmol/mg), and outer and inner medullary chromaffin cells (42 +/- 1 and 55 +/- 2 fmol/mg, respectively). Binding to sites in the zona glomerulosa was partially inhibited (16%) by 100 nM 2-BFI. These results are consistent with previous studies indicating that [3H]rilmenidine labels an I2-RBS and additional I-RBS in rat brain and kidney. The distribution of [3H]RO19-6327 (5 nM) binding resembled that of [3H]2-BFI and [3H]clonidine binding with high densities of MAO-B enzyme located in the zona glomerulosa and chromaffin cells of the inner medulla (55 +/- 7 and 76 +/- 6 fmol/mg tissue, respectively), suggesting the colocalization of MAO-B enzyme with I2-RBS. [3H]RO41-1049 (20 nM) binding to MAO-A was highest in the zona reticularis (196 +/- 7 fmol/mg tissue) compared to the zonae glomerulosa and fasciculata (90 +/- 12 and 116 +/- 14 fmol/mg tissue) and inner medulla (149 +/- 38 fmol/mg tissue). Although the existence of I-RBS in bovine adrenal chromaffin cells is well established, this is the first description of I-RBS in the adrenal cortex. Further investigations are now required to determine whether imidazolines can affect adrenal function via actions at these sites.

Topics: Adrenal Cortex; Adrenal Medulla; Adrenergic beta-Agonists; Animals; Autoradiography; Benzofurans; Binding Sites; Cattle; Clonidine; Imidazoles; Imidazoline Receptors; Isoenzymes; Monoamine Oxidase; Monoamine Oxidase Inhibitors; Oxazoles; Picolinic Acids; Rats; Receptors, Drug; Rilmenidine; Thiazoles; Tritium

The distribution and relative densities of imidazoline-receptor binding sites (I-RBS) and monoamine oxidase (MAO)-A and -B enzyme(s) in rat and rabbit kidney were compared autoradiographically using fixed nanomolar concentrations of [3H]rilmenidine and [3H]2-(benzofuranyl)-2-imidazoline ([3H]2-BFI) to label I-RBS, and [3H]RO41-1049 and [3H]RO19-6327 to label MAO-A and -B isoenzymes, respectively. In rat kidney, high densities of I-RBS labelled by [3H]rilmenidine were observed in the cortex and outer stripe (120-280 fmol/mg tissue), in contrast to low I-RBS densities labelled by [3H]2-BFI (<4 fmol/mg). A relatively high density of [3H]RO41-1049 binding to MAO-A enzyme was present in all regions of the rat kidney (160-210 fmol/mg) compared with a low density of [3H]RO19-6327 binding to MAO-B (< 25 fmol/mg). Comparison of MAO-A and -B distributions with that of [3H]rilmenidine-labelled I-RBS strongly suggests a lack of association in rat kidney. Similarly, the extremely low densities of [3H]2-BFI-labelled I2-RBS in rat kidney contrasts with the density of MAO-A, but is consistent with the low density of MAO-B. Rabbit kidney cortex and outer stripe contained high relative densities of [3H]rilmenidine-labelled I-RBS (200-215 fmol/mg) and [3H]2-BFI-labelled I2-RBS (45-60 fmol/mg) with lower densities in the inner stripe and inner medulla (< or = 100 and 30 fmol/mg respectively). A high density of MAO-A binding was observed in the inner stripe (515 fmol/mg) with lower levels in the cortex and outer stripe (100-240 fmol/mg), while high densities of MAO-B binding were observed in the cortex and outer stripe (290-450 fmol/mg) with lower levels in the inner stripe (65 fmol/mg). The correlation between the localization of [3H]rilmenidine-labelled I-RBS and [3H]RO19-6327-labelled MAO-B in rabbit kidney (r = 0.87, P = 0.057) suggest that [3H]rilmenidine may label a binding site co-existent with MAO-B, but not MAO-A (n.s.), in this tissue, but rilmenidine did not inhibit [3H]RO41-1049 or [3H]RO19-6327 binding. The distribution of [3H]2-BFI-labelled I2-RBS overlapped the combined distributions of both MAO-A and -B isoenzymes, suggesting that [3H]2-BFI may label sites on both enzymes in the rabbit, but [3H]2-BFI binding only correlated with [3H]RO19-6327 (r = 0.84, P = 0.07), not [3H]RO41-1049 binding (n.s.). Moreover, 2-BFI only inhibited [3H]RO19-6327, not [3H]RO41-1049 binding. These data are consistent with reports that I2-RBS are located on MAO-B and allosterically i

Topics: Adrenergic alpha-Agonists; Animals; Autoradiography; Benzofurans; Imidazoles; Imidazoline Receptors; In Vitro Techniques; Isoenzymes; Kidney; Male; Monoamine Oxidase; Monoamine Oxidase Inhibitors; Oxazoles; Picolinic Acids; Rabbits; Rats; Rats, Inbred WKY; Receptors, Drug; Rilmenidine; Species Specificity; Thiazoles