Preclinical evaluation of the abuse potential of Pitolisant, a histamine H3 receptor inverse agonist/antagonist compared with Modafinil.

BACKGROUND AND PURPOSE: Pitolisant, a histamine H3 receptor inverse agonist/antagonist is currently under Phase III clinical trials for treatment of excessive daytime sleepiness namely in narcoleptic patients. Its drug abuse potential was investigated using in vivo models in rodents and monkeys and compared with those of Modafinil, a psychostimulant currently used in the same indications. EXPERIMENTAL APPROACH: Effects of Pitolisant on dopamine release in the nucleus accumbens, on spontaneous and cocaine-induced locomotion, locomotor sensitization were monitored. It was also tested in three standard drug abuse tests i.e. conditioned place preference in rats, self-administration in monkeys and cocaine discrimination in mice as well as in a physical dependence model. KEY RESULTS: Pitolisant did not elicit any significant changes in dopaminergic indices in rat nucleus accumbens whereas Modafinil increased dopamine release. In rodents, Pitolisant was without any effect on locomotion and reduced the cocaine-induced hyperlocomotion. In addition, no locomotor sensitization and no conditioned hyperlocomotion were evidenced with this compound in rats whereas significant effects were elicited by Modafinil. Finally, Pitolisant was devoid of any significant effects in the three standard drug abuse tests (including self-administration in monkeys) and in the physical dependence model. CONCLUSIONS AND IMPLICATIONS: No potential drug abuse liability for Pitolisant was evidenced in various in vivo rodent and primate models, whereas the same does not seem so clear in the case of Modafinil.

[Pulsed dye laser treatment for Jessner's lymphocytic infiltration of the skin]. / Infiltrat lymphocytaire de Jessner et Kanof traité par laser à colorant pulsé

BACKGROUND: Jessner's lymphocytic infiltration of the skin (LIS) is a chronic, benign T-cell infiltrative disorder, usually manifesting as erythematous papules or plaques on the face, neck and back. PATIENTS AND METHODS: five patients presented LIS with numerous skin lesions on the face and back characteristic of this disease. Histological examination showed a lymphocytic infiltrate in the dermis without any modification of the epidermis. Direct immunofluorescent study was negative in all cases. Response to dermocorticoids proved inconsistent or negative in all patients. An excellent outcome was achieved in all five patients with pulsed dye laser. In one case, further skin lesions appeared at 1 year and responded to the same treatment. Following a single treatment session with 6-8 J/cm(2), three of five patients showed normal skin. Regression occurred in the other two cases after two to three sessions. Pulsed dye laser appeared to be the best treatment for Jessner-Kanof disease for three patients at 4-8 years of follow-up. DISCUSSION: only one case of Jessner-Kanof disease treated by pulsed dye laser has been reported. Pulsed dye laser has been used in cutaneous lupus and annular granuloma. Selective photothermolysis allows photocoagulation of dilated vessels. CONCLUSION: pulsed dye laser at 595nm could offer a valuable therapeutic alternative, and even a first-line treatment with no side effects.

Influence of tropolone on Poria placenta wood degradation.

Fenton reactions are believed to play important roles in wood degradation by brown rot fungi. In this context, the effect of tropolone (2-hydroxycyclohepta-2,4,6-trienone), a metal chelator, on wood degradation by Poria placenta was investigated. Tropolone (50 micro M) strongly inhibits fungal growth on malt agar, but this inhibition could be relieved by adding iron salts. With an experimental system containing two separate parts, one supplemented with tropolone (100 micro M) and the other not, it was shown that the fungus is able to reallocate essential minerals from the area where they are available and also to grow in these conditions on malt-agar in the presence of tropolone. Nevertheless, even in the presence of an external source of metals, P. placenta is not able to attack pine blocks impregnated with tropolone (5 mM). This wood degradation inhibition is related to the presence of the tropolone hydroxyl group, as shown by the use of analogs (cyclohepta-2,4,6-trienone and 2-methoxycyclohepta-2,4,6-trienone). Furthermore, tropolone possesses both weak antioxidative and weak radical-scavenging properties and a strong affinity for ferric ion and is able to inhibit ferric iron reduction by catecholates, lowering the redox potential of the iron couple. These data are consistent with the hypothesis that tropolone inhibits wood degradation by P. placenta by chelating iron present in wood, thus avoiding initiation of the Fenton reaction. This study demonstrates that iron chelators such as tropolone could be also involved in novel and more environmentally benign preservative systems.

Oligoribonucleotide-based gene-specific transcription inhibitors that target the open complex.

We have demonstrated that oligoribonucleotides that lack a 3'-OH group and cannot be extended by RNA polymerase can hybridize to the single-stranded DNA formed inside the transcription initiation bubble (or open complex) and inhibit transcription. Using the lacUV5/Escherichia coli RNA polymerase or trpEDCBA/E. coli RNA polymerase transcription system as a model, we have found that effective inhibitors are five nucleotides in length and must be complementary to the DNA template strand in the region from -5 to +2 about the transcription start site (designated +1). We have used the DNA cleavage activity of 1,10-phenanthroline-copper to confirm that the mechanism of inhibition is via oligoribonucleotide hybridization to the open complex and have used this cleavage chemistry to demonstrate that these oligonucleotide inhibitors hybridize in an antiparallel orientation to their DNA target. Systematic modification of the parent phosphodiester oligoribonucleotide pentamer revealed that the phosphorothioate backbone-containing analogs have increased open complex binding affinity and are more effective transcription inhibitors than their phosphodiester counterparts.

Tomorrow's anticancer agents: inhibitors of Ras farnesylation.

The purpose of this chapter is to concentrate on what can be considered as definite milestones on the way from examples of inhibitors of farnesyl-protein transferase (FPTase) to candidate drugs actually being considered for or already being evaluated in clinical trials. Emphasis will be placed on results obtained using experimental tumour models in vivo, with a detailed discussion of these results and of the questions which remain to be studied or are still unanswered. The data discussed here are almost exclusively based on published reports, with only brief reference, in the chapter "use of the FPTase inhibitors in the clinic", to some of the newer compounds reported on during recent meetings, details of which have not yet appeared in the peer-reviewed literature. For those requiring a more extensive review of the catalogue of FPTase inhibitors now discovered, some excellent reviews have been committed to this purpose [1-3].

Fate of Terpene Compounds in Activated Sludge Wastewater Treatment Systems.

Terpene-based cleaners are being widely used in industrial cleaning formulations because of their ability to replace suspected ozone-depleting chemicals such as 1,1,1-trichloroethane and 1,1,2-trichloro-1,2,2-trifluoroethane (CFC-113). Substitution of chlorinated solvents with ter-pene-based cleaners, however, is expected to result in increased discharges to wastewater from industrial operations. A pilot-scale study was conducted at the U.S. Environmental Protection Agency's (EPA) Test & Evaluation Facility in Cincinnati, OH, to quantify the fate of specific terpene compounds in the activated sludge wastewater treatment process. Biodegradation rates of terpenes were estimated from the difference between the influent terpene mass flow rates and the amounts volatilized to air, partitioned to waste sludge, and passed through the treatment process unchanged. Any chemical transformation of the terpene compounds studied was attributed to biodegradation. Analytical methods were developed to determine ter-pene concentrations in aqueous and gaseous media. The fate of two common terpene compounds (d-limonene and terpinolene) were evaluated in three identical pilot-scale systems: (1) a system with a high target spike range (2-10 mg/L), (2) a system with a low target spike range (0.5-2 mg/L), and (3) a control system (no spike). The study showed that the primary removal mechanism for the terpene compounds in the activated sludge process is biodegradation. Typically, greater than 90% of the mass of terpenes entering the aeration basin of the activated sludge process biodegrades to other compounds; volatilization from the reaction basin accounts for less than 10%, while loss to waste activated sludge and the secondary clarifier effluent accounts for less than 1%.

Optimizing the targeted chemical nuclease activity of 1,10-phenanthroline-copper by ligand modification.

Our interest in improving the efficiency of targeted scission reagents has prompted us to study the influence of ring substituents on the nuclease activity of 1,10-phenanthroline-copper conjugated to oligonucleotides and DNA-binding proteins. Since methyl substitution at all but the 2 and 9 positions enhances the copper-dependent chemical nuclease activity of 1,10-phenanthroline, we have compared the activity of conjugates prepared from 5-(aminomethyl)-1,10-phenanthroline (MOP) to those of conjugates prepared from 5-amino-1,10-phenanthroline (amino-OP). Tethering MOP derivatives to the Escherichia coli Fis protein enhances DNA scission several-fold at the weaker cleavage sites initially observed with conjugates prepared from amino-OP. However, scission efficiency is not increased at the stronger cleavage sites, or when scission is targeted to single-stranded DNA by a complementary oligonucleotide. These results are consistent with a change in the rate-determining step for cleavage associated with the differential accessibility of the DNA-bound coordination complex to solvent and reductant. Although the free bis cuprous complex of 2,9-dimethyl-1,10-phenanthroline (neocuproine) is redox-inactive, an oligonucleotide tethered to neocuproine through C5 of the phenanthroline ring efficiently cleaves a complementary DNA sequence. These results establish that the nucleolytic species in targeted scission is the 1:1 cuprous complex and suggest that the oxidative reaction proceeds through a copper-oxo intermediate rather than a metal-coordinated peroxy species. However, substituents at the 2 and 9 positions of the ligand will often hinder close approach of the phenanthroline-copper moiety to the oxidatively sensitive ribose as shown by the preference of the oligonucleotide-targeted chimera for cleavage of single-stranded regions and the failure of neocuproine-DNA-binding protein chimeras and a C2-tethered chimera to cleave DNA.

Hybridization of a complementary ribooligonucleotide to the transcription start site of the lacUV-5-Escherichia coli RNA polymerase open complex. Potential for gene-specific inactivation reagents.

An ribooligonucleotide, UGGAA, complementary to the template strand of the lacUV-5 promoter can hybridize to the transcription "bubble" of the open complex formed by Escherichia coli RNA polymerase. Its site-specific binding, measured by gel retardation, enzyme inhibition, and chemical nuclease footprinting, is dependent on catalysis by RNA polymerase and the sequence of the hybridizing ribooligonucleotide. When UGGAA is linked to the chemical nuclease 1,10-phenanthroline copper, site-specific scission of the template strand of the transcriptionally active gene is observed. The formation of single-stranded DNA at transcription start sites by RNA polymerases provides a target for antigene strategies.

Chronic copper toxicity in a dairy herd.

The addition of excessive copper to a commercially prepared dairy ration caused chronic copper toxicity in a dairy herd. A formulation error by a feed company resulted in copper levels of 800 to 1,000 mg/kg in the "as fed concentrate," amounting to about 400-500 mg copper/kg of the whole ration. Five animals died with typical signs of acute copper toxicity, including intravascular hemolysis and methemoglobinemia. A further 39 cows died on the farm from a combination of debilitation and secondary infectious causes, and 215 were sent to slaughter because of debilitation and poor milk production. The mortality of calves born to dams that had been fed the toxic concentrate was approximately 50%.We postulate that dairy cows, particularly pregnant cows, may be more susceptible to copper toxicity than other cattle, and suggest reexamination of the presently allowable maximum levels of copper supplementation of diets for dairy cattle.

Dissolution of urinary stones by calcium-chelating agents: A study using a model system.

Pellets made by dispersing microcrystals of calcium oxalate monohydrate throughout an organic matrix have served as models for kidney stones in studies of factors governing their dissolution by calcium-chelating agents. These factors include pH, ionic strength, concentration of chelating agent, and addition of other acids and bases. The method shows good reproducibility. Results have been applied to improving a clinical procedure for kidney stone dissolution.

Computer-based approach to chelation therapy: a theoretical study of some chelating agents for the selective removal of toxic metal ions from plasma.

COMICS is a computer programme for calculating equilibrium concentrations of metal complexes and reactive species in multi-metal-multi-ligand systems. Its usefulness for analysing metal ion equilibria in blood plasma has been improved by including albumin as a ligand. Using this model system the distribution and removal of copper(II) and zinc ions in histidinaemia, lead poisoning and Wilson's disease have been examined. The efficacy of TRIEN in removing excess copper(II) is shown. The use of specific tripeptides such as Gly-Gly-His methyl ester for the selective removal of copper(II) is suggested. A possible chemoprophylaxis of influenza based on complexation of zinc is discussed. Calculations confirm that thiosemicarbazones such as methisazone and 2-acetylpyridine thiosemicarbazone are effective competitors for heavy metal ions under physiological conditions.