A case of motor neuron involvement in Gaucher disease.

Gaucher disease (GD) is a genetic disorder characterized by an accumulation of glucosylceramide in cells in the monocyte-macrophage system. We describe a case of a 33-year-old man with a previous diagnosis of type 3 GD who displayed a progressive weakening of the limbs followed by upper motor neuron involvement. A diagnosis of definite Amyotrophic Lateral Sclerosis was made. This is the first reported case of concurrent Gaucher disease and the ALS phenotype in the same patient.

A comparative molecular field analysis model for 6-arylpyrrolo[2,1-d] [1,5]benzothiazepines binding selectively to the mitochondrial benzodiazepine receptor.

A series of 42 6-arylpyrrolo[2,1-d][1,5]benzothiazepines, which we have recently described as selective ligands of the mitochondrial benzodiazepine receptor (MBR) (Fiorini I.; et al. J. Med. Chem. 1994, 37, 1427-1438), have been investigated using the comparative molecular field analysis (CoMFA) approach. The resulting 3D-QSAR model rationalizes the steric and electronic factors which modulate affinity to the MBR with a cross-validation standard error of 0.648 pIC50 unit. A set of seven novel pyrrolobenzothiazepine congeners has successively been synthesized and tested. The CoMFA model forecasts the binding affinity values of these new compounds with a prediction standard error of 0.536.

Novel ligands specific for mitochondrial benzodiazepine receptors: 6-arylpyrrolo[2,1-d][1,5]benzothiazepine derivatives. Synthesis, structure-activity relationships, and molecular modeling studies.

A novel class of ligands specific for MBR receptors has been identified: 6-arylpyrrolo[2,1-d][1,5]benzothiazepine derivatives. The majority of newly synthesized esters 37-64 as well as some intermediate ketones showed micro- or nanomolar affinity for [3H]PK 11195 binding inhibition. A SAR study on 42 compounds and a molecular modeling approach led to a preliminary structural selectivity profile: the 6,7-double bond, the carbamoyloxy, alcanoyloxy, and mesyloxy side chains at the 7-position, and the prospective chloro substitution at the 4-position seemed to be the most important structural features improving affinity. Therefore, 7-[(dimethylcarbamoyl)oxy]- and 7-acetoxy-4-chloro-6-phenylpyrrolo[2,1-d][1,5]benzothiazepine (43 and 57) were synthesized. With 7-[(dimethylcarbamoyl)oxy]-6-(p-methoxyphenyl)pyrrolo[2,1- d][1,5]benzothiazepine (65), these were the most promising compounds with IC50s of respectively 9, 8, and 9 nM, under conditions where PK 11195 had an IC50 of 2 nM.

Pyrrolobenzothiazepinones and pyrrolobenzoxazepinones: novel and specific non-nucleoside HIV-1 reverse transcriptase inhibitors with antiviral activity.

Two novel classes of pyrrolobenzothiazepinones and pyrrolobenzoxazepinones were investigated as potential anti-AIDS drugs. These compounds were found to inhibit HIV-1 reverse transcriptase (RT) enzyme in vitro and to prevent HIV-1 cytopathogenicity in T4 lymphocytes, without appreciable activity on HIV-2 cytopathic effects, and against HBV as well as calfthymus DNA alpha-polymerase. Their potency is influenced by substituents at position 6 and on the fused aromatic ring. Specifically, small lipophilic substituents at C-6 were preferred, whereas substitutions on the benzo-fused ring were found to be detrimental to activity, with respect to the unsubstituted compounds. Modification of the pie-system at C-6 is well tolerated, although the replacement of the benzo-fused with a [2,3]naphtho-fused ring leads to a less active compound. Maximum potency and specificity is achieved with a phenyl and an ethyl group at position 6 of the pyrrolobenzoxazepinone system. In the enzymatic assay the oxazepinone derivative (+/-)-6-ethyl-6-phenylpyrrolo[2,1-d][1,5] benzoxazepin-7(6H)-one 16e (IC50 = 0.25 microM) was found to be more potent than nevirapine (IC50 = 0.5 microM), tested in the same experimental conditions using rC.dG as a template-primer. In cell culture assay benzoxazepine 16e was active against HIV-1, both wild type and AZT-sensitive, and HIV-1 (IIIB) strains, but not against HIV-2. In enzyme assay although 16e inhibited HIV-1 RT, it was inactive against the nevirapine-resistant recombinant RT Y181C at 50 microM. Molecular modeling studies suggest that these derivatives present a 3D pharmacophoric arrangement similar to that of other non-nucleoside inhibitors such as nevirapine.

Pyrrolo[2,1-c][1,4]benzothiazines: synthesis, structure-activity relationships, molecular modeling studies, and cardiovascular activity.

The synthesis and pharmacological evaluation of a series of pyrrolo[1,4]benzothiazine derivatives are described. These compounds, related to diltiazem, have been shown to be representative of a novel series of calcium channel antagonists. The IC50S for inhibition of [3H]nitrendipine binding calculated by radioreceptor assay on rat cortex and rat heart homogenates showed that some of the described compounds possess an affinity equal to or higher than those of the reference calcium antagonists verapamil and cis-(+)-diltiazem. Furthermore, the alteration of the benzothiazepinone system of diltiazem to the pyrrolo[1,4]benzothiazine system of the title compounds resulted in a clear-cut selectivity for cardiac over vascular tissue, as shown in functional studies. In fact comparison of calcium antagonist activity on guinea pig aorta strips with the negative inotropic activity, determined by using an isolated guinea pig left atrium, revealed that the compounds examined displayed higher selectivity than the reference standard, within a wide variation of data. A number of structure-activity relationship trends have been identified, and possible explanation is advanced in order to account for the observed differences in selectivity. Prerequisite for in vitro calcium channel-blocking activity is the presence of two pharmacophores, namely, the substitution at C-4 and the substitution on the pyrrole ring. Two of the tested compounds, 8b and 28a, were identified as potent calcium antagonists selective for cardiac over vascular tissue.

A concerted study using binding measurements, X-ray structural data, and molecular modeling on the stereochemical features responsible for the affinity of 6-arylpyrrolo[2,1-d][1,5]benzothiazepines toward mitochondrial benzodiazepine receptors.

The 7-(acyloxy)-6-arylpyrrolo[2,1-d][1,5]benzothiazepine derivatives have been recently proposed as a new class of ligands specific for the mitochondrial benzodiazepine receptor (Fiorini et al. J. Med. Chem. 1994, 37, 1427-1438) (Greco et al. J. Med. Chem. 1994, 37, 4100-4108). In this paper we report the X-ray crystallographic structures of three potent (1-3) and two inactive (4 and 5) previously described benzothiazepines, as well as binding affinity constants for two newly assayed analogs in which the acyloxy side chain was replaced by a methoxy group (6) or removed (7). Structure-affinity relationships and molecular mechanics calculations performed using crystal structures as references have led to a revised 3D pharmacophore model accounting for all the data available up until now. Interestingly, the hypothetical receptor-bound conformations of 1-3 display a considerable degree of similarity with their crystal geometries. Additional calculations have confirmed that the poor affinities of benzothiazepines bearing an aroyloxy group (4 and 5) should be ascribed to the steric and/or electronic features of the side chain aryl moieties rather than to unfavorable conformational properties.

Thiazolothiazepine inhibitors of HIV-1 integrase.

A series of thiazolothiazepines were prepared and tested against purified human immunodeficiency virus type-1 integrase (HIV-1 IN) and viral replication. Structure-activity studies reveal that the compounds possessing the pentatomic moiety SC(O)CNC(O) with two carbonyl groups are in general more potent against purified IN than those containing only one carbonyl group. Substitution with electron-donating or -withdrawing groups did not enhance nor abolish potency against purified IN. By contrast, compounds with a naphthalene ring system showed enhanced potency, suggesting that a hydrophobic pocket in the IN active site might accommodate an aromatic system rather than a halogen. The position of sulfur in the thiazole ring appears important for potency against IN, as its replacement with an oxygen or carbon abolished activity. Further extension of the thiazole ring diminished potency. Compounds 1, 19, and 20 showed antiviral activity and inhibited IN within similar concentrations. These compounds inhibited IN when Mn(2+) or Mg(2+) was used as cofactor. None of these compounds showed detectable activities against HIV-1 reverse transcriptase, protease, virus attachment, or nucleocapsid protein zinc fingers. Therefore, thiazolothiazepines are potentially important lead compounds for development as inhibitors of IN and HIV replication.

New antipsychotic agents with serotonin and dopamine antagonist properties based on a pyrrolo[2,1-b][1,3]benzothiazepine structure.

The development of a synthetic approach to the novel pyrrolo[2, 1-b][1,3]benzothiazepine and its derivatives and their biological evaluation as potential antipsychotic drugs are described. In binding studies these compounds proved to be potent 5-HT2, D2, and D3 receptor ligands. The more potent benzothiazepine (+/-)-3b was resolved into its enantiomers by using HPLC techniques. In vitro testing confirmed that (-)-3b is a more potent D2 receptor ligand, maintaining high affinity for 5-HT2 receptors. In contrast, the (+)-3b enantiomer presents a 35 times higher affinity for 5-HT2 than for dopamine D2 receptors with a similar dopamine D1 receptor affinity to that of (-)-3b. Overall, (+)-3b shows an "atypical" neuroleptic binding profile, while (-)-3b has a more "classical" profile. Furthermore pharmacological and biochemical testing displayed that the novel benzothiazepine (+/-)-3b is able to increase the extracellular levels of dopamine in the rat striatum and causes a dose-related suppression of apomorphine-induced locomotor activity. At low doses (+/-)-3b does not induce catalepsy, showing atypical antipsychotic properties similar to those of olanzapine. These heterocyclic compouds represent new leads for the development of novel antipsychotic drugs with atypical properties.

Synthesis, biological activity, and SARs of pyrrolobenzoxazepine derivatives, a new class of specific "peripheral-type" benzodiazepine receptor ligands.

The "peripheral-type" benzodiazepine receptor (PBR) has been reported to play a role in many biological processes. We have synthesized and tested a novel series of PBR ligands based on a pyrrolobenzoxazepine skeleton, in order to provide new receptor ligands. Several of these new compounds proved to be high affinity and selective ligands for PBR, and benzoxazepines 17f and 17j were found to be the most potent ligands for this receptor to have been identified to date. The SAR and the molecular modeling studies detailed herein delineated a number of structural features required for improving affinity. Some of the ligands were employed as "molecular yardsticks" to probe the spatial dimensions of the lipophilic pockets L1 and L3 in the PBR cleft and to determine the effect of occupation of L1 and L3 with respect to affinity, while other C-7 modified analogues provided information specifically on the hydrogen bonding with a putative receptor site H1. The new pyrrolobenzoxazepines were tested in rat cortex, a tissue expressing high density of mitochondrial PBR, and exhibited IC50 and Ki values in the low nanomolar or subnanomolar range, as measured by the displacement of [3H]PK 11195 binding. A subset of the highest affinity ligands was also found to have high affinities for [3H]PK 11195 and [3H]Ro 5-4864 binding in rat adrenal mitochondria. All the ligands in this subset are stimulators of steroidogenesis having similar potency and extent of stimulation as PK 11195 and Ro 5-4864 of steroidogenesis in the mouse Y-1 adrenocortical cell line.

Cardiovascular characterization of pyrrolo[2,1-d][1,5]benzothiazepine derivatives binding selectively to the peripheral-type benzodiazepine receptor (PBR): from dual PBR affinity and calcium antagonist activity to novel and selective calcium entry blockers.

The synthesis and cardiovascular characterization of a series of novel pyrrolo[2,1-d][1,5]-benzothiazepine derivatives (54-68) are described. Selective peripheral-type benzodiazepine receptor (PBR) ligands, such as PK 11195 and Ro 5-4864, have recently been found to possess low but significant inhibitory activity of L-type calcium channels, and this property is implicated in the cardiovascular effects observed with these compounds. In functional studies both PK 11195 (1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinolinecarboxa mide) and Ro 5-4864 (4'-chlorodiazepam) did not display selectivity between cardiac and vascular tissue. Therefore, several 7-(acyloxy)-6-arylpyrrolo[2,1-d][1,5]benzothiazepines, potent and selective peripheral-type benzodiazepine receptor ligands recently developed by us (3, 7-20), were subjected to calcium channel receptor binding assay. Some of these compounds showed an unexpected potency in displacing the binding of [3H]nitrendipine from L-type calcium channels, much higher than that reported for PK 11195 and Ro 5-4864 and equal to or higher than that of reference calcium antagonists such as verapamil and (+)-cis-diltiazem. Specifically, in rat cortex homogenate, our prototypic PBR ligand 7-acetoxy-6-(p-methoxyphenyl)pyrrolo[2,1-d][1,5]benzothiazepine (3) showed an IC50 equal to 0.13 nM for inhibition of [3H]nitrendipine binding. Furthermore, in functional studies this compound displayed a clear-cut selectivity for cardiac over vascular tissue. Comparison of calcium antagonist activity on guinea pig aorta strips with the negative inotropic activity, determined by using isolated guinea pig left atria, revealed that 3 displayed higher selectivity than the reference (+)-cis-diltiazem. Thus, the pyrrolobenzothiazepine 3 might represent a new tool for characterizing the relationship between the PBR and cardiac function. Furthermore, we have also investigated the structural dependence of binding to PBR and L-type calcium channels, and this study allowed us to identify a new class of potent calcium channel blockers selective for cardiac over vascular tissue, with no affinity for PBR. A number of structure-activity relationship trends have been identified, and a possible explanation is advanced in order to account for the observed differences in selectivity. Three structural features, namely, (i) the saturation of the C(6)-C(7) double bond, with a consequent higher molecular flexibility, (ii) the presence of a substituent in the benzofused ring, and (iii) a basic side chain at C-10 of the pyrrolobenzothiazepine ring system, were found to be responsible for potent L-type calcium channel antagonism and clear-cut selectivity for cardiac over vascular tissue. Among the synthesized compounds the pyrrolobenzothiazepine 62 was found to be the most promising selective calcium channel blocker. Additionally, the molecular structure determination of the key intermediate 48 by X-ray diffraction, molecular modeling, and NMR analysis is reported.

Comparison between plasma concentrations of testosterone, nitric oxide and endothelin 1-2 in penile and brachial venous blood: preliminary results in men with psychogenic impotence.

In the present study venous plasma concentrations of testosterone (T), nitric oxide (NO) and endothelin 1-2 (ET1-2) in the flaccid penis and brachial blood were measured in men with psychogenic impotence. T and NO were significantly lower in the penile venous blood, while ET1-2 showed no statistical difference. These data support the hypothesis of testosterone dependence of penile nitric oxide synthesis (NOS).

Immunologic disturbances and levels of parathyroid hormone in uremic patients in replacement dialysis therapy.

We studied the changes in cellular immunity in patients in replacement dialysis therapy (RDT) and examined the relationship between T-lymphocyte function and plasma levels of parathyroid hormone (PTH). In a preliminary study we found that increased plasma levels of PTH were associated with a decrease of T-lymphocytes and CD4, an increase in CD8 and a reduction in the ratio of CD4 to CD8. In the present study we examined the relationship between plasma levels of PTH, interleukin 2 receptors (IL-2R) and soluble human CD8 (S-CD8). We studied 54 patients divided into two groups: 26 patients with normal levels of PTH and 28 patients with increased levels of PTH. We found a significant reduction in total T-lymphocytes in both groups as compared to controls, with an inverse correlation between total T-lymphocytes and plasma PTH in the second group (R = -0.52). There was an increase in IL-2R in the group II as compared to the controls and also in the total population of uremic patients with a linear correlation between levels of IL-2R and PTH (R = 0.6). The levels of S-CD8 showed a significant increase in both groups with a linear correlation between levels of SC-D8 and PTH (R = 0.63). No specific differences were seen between patients treated with and without 1,25-dihydroxyvitamin D3. The elevated levels of PTH affect the lymphocyte function and are associated with change in cellular immunity with reduction in total number of T cells, and increases in levels of CD8, S-CD8 and IL-2R.

Benzothiazine and benzothiazepine derivatives: synthesis and preliminary biological evaluation.

Several tricyclic benzothiazines and benzothiazepines have been synthesized by different intramolecular cyclization reactions. Their functionalization led to biologically active compounds. Some stereochemical aspects as well as biological responses have been outlined.

Research on compounds with psychotropic activity. IX--Synthesis of 6-p-methoxyphenylpyrrolo[2,1-d][1,5]benzothiazepines and evaluation of their affinities for BDZ and GABA receptor subtypes.

6-p-Methoxyphenylpyrrolo[2,1-d][1,5]benzothiazepin-7(6H)-one (IV), cis-7-acetoxy-6,7-dihydro-6-p-methoxyphenylpyrrolo[2,1-d] [1,5]benzothiazepine (V) and some significant 7-acyloxy-6-p-methoxyphenylpyrrolo[2,1-d][1,5]benzothiazepines (VI a-g) were synthesized and tested in vitro for inhibition of the specific binding of 3H-Flunitrazepam, 3H-PK 11195, 3H-Muscimol and 3H-(-)Baclofen to central and peripheral benzodiazepine, GABA-A and GABA-B receptors, respectively. The compounds (IV), VI a) and (VI c) were active on the peripheral benzodiazepine receptor; in particular (VI a) and (VI c) were very active. The compound (VI g) showed an affinity, even though scanty, for the central benzodiazepine receptor.

Pyrazolo [4,3-c] quinolines synthesis and specific inhibition of benzodiazepine receptor binding (Note I).

A series of 2-arylpyrazolo[4,3-c] quinolin-3-one derivatives, bearing different substituents in the two aromatic rings, were prepared and tested for their ability to displace [3H] flunitrazepam from rat brain membranes. Some compounds have shown an affinity for receptors comparable and sometimes higher than that of CGS series.

[Relation between a rapid polyvalent test and single antistreptococcal tests]. / Relazione tra un test rapido polivalente e i singoli tests antistreptococcici.

ASO and Streptozyme tests have been comparatively tried on 11,200 sera. The former detected 11.96% positive and the latter 13.5%. The difference was due to a greater sensitivity of the Streptozyme test since the most of the Streptozyme positive and ASO negative sera resulted positive in one or more of the single antistreptococcal tests (SK, ADN, SJ, DNA). Only few of the sera which give rise to a positive Streptozyme reaction were otherwise negative and the positive result could be considered aspecific.

[Compounds with psychotropic activity. VIII. Synthesis and sedative activity of various 9-substituted derivatives of 5-phenylpyrrolo[2,1-d][1,5] benzothiazepine and cis-4,5-dihydro-4-hydroxy-5-phenylpyrrolo[2,1-d][1,5]benzothiazepine]. / Ricerche su composti ad attività psicotropa. Nota VIII - Sintesi ed attività sedativa di alcuni derivati 9-sostituiti della 5-fenilpirrolo [2,1-d] [1,5]benzotiazepina e della cis-4,5-diidro-4-idrossi-5-fenilpirrolo [2,1-d] [1,5]benzotiazepina.

Syntheses of 9-chloro-, 9-trifluoromethyl- and 9-methoxy-5- phenylpyrrolo [2,1-d] [1,5] benzothiazepine [II a-c] and of cis-9-chloro- and cis-9-trifluoromethyl-4,5-dihydro-4-hydroxy-5- phenylpyrrolo [2,1-d] [1,5] benzothiazepine with the respective acetyl derivatives (III a-d), according to previously restated routes, are described. The sedative activity was tested against the anti-amphetamine activity in the rat. The 1-[5-trifluoromethyl-2-(alpha- hydroxycarbonylbenzyl ) thiophenyl + ++]-pyrrole ( NF34 ) and the pyrrolo [2,1-d] [1,5] benzothiazepine -5-carboxamide ( NF44 ) showed sedative activity similar to that of diazepam.

[Research on a compound with psychotropic activity. V - a new derivative of pyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide: synthesis and sedative activity]. / Ricerche su composti ad attività psicotropa. Nota V - nuovi derivati del pirrolo[2,1-d][1,5]benzotiazepin-6,6-diossido: sintesi ed attività sedativa.

The following compounds have been synthesized according to a previously tested route, of the 9-chloro-, 9-trifluoromethyl- and 9-methoxy-5-phenylpyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide (II a-c), 9-chloro- and 9-trifluoromethyl-5-p-nitrophenylpyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide (II d, e), 5-(4-pyridyl)pyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide (III) and 5-(3-pyridyl)pyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide (IVa) with the 9-chloro- and 9-trifluoromethyl- derivatives (IV b, c) are reported. The sedative action in the rat was tested against the motor activity induced by amphetamine. The 9-chloro-5(3-pyridyl)pyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide was particularly active and showed sedative action superior to that of diazepam. Marked sedative activity was observed with 9-methoxy-5-phenylpyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide (NF19) and 9-chloro-5-p-nitrophenylpyrrolo[2,1-d][1,5]benzothiazepin-6,6-dioxide (NF20).