Dose Of Disulfiram Research Articles

Abstract- Poster

Disulfiram has been most widely used in patients suffering from alcohol dependence. When taken along with alcohol it can cause various side effects like flushing, drowsiness, rashes, hyperventilation, palpitations etc. Mania due to Disulfiram is an uncommon side effect and there are only a few reports of it. We hereby report a case of development of Mania in an individual with alcohol dependence following 2 months of treatment with therapeutic dose of Disulfiram. Before the onset of mania, the patient was abstinent from alcohol for about 2 months, which made substance-induced mania unlikely. The possible mechanism for this being the dopamine hypothesis which suggests that Disulfiram inhibits dopamine-Beta-hydroxylase which is responsible for the conversion of dopamine to Nor-adrenaline and increases the dopamine level which is responsible for psychotic and mania symptoms. This possibility of Disulfiram induced mania should be assessed for whenever clinicians encounter patients with dual diagnosis, as this might change the management as such.

Combination Therapy with Disulfiram, Copper, and Doxorubicin for Osteosarcoma: In Vitro Support for a Novel Drug Repurposing Strategy

Although many cancer cells have significantly higher copper concentrations compared with normal cells and tissues, the role of copper in cancer biology and metastatic disease remains poorly understood. Here, we study the importance of copper in osteosarcoma, which frequently metastasizes to the lungs and is often chemoresistant. K12 and K7M2 are murine OS cells with differing metastatic phenotypes: K7M2 is highly metastatic, whereas K12 is much less so. Intracellular copper levels were determined using atomic absorption. Copper transporters were quantified by qPCR. Cytotoxicity of doxorubicin, disulfiram, and copper(II) chloride was assessed with a cell viability fluorescence stain. Additionally, K7M2 viable cell counts were determined by trypan blue exclusion staining after 72 hours of treatment. Copper levels were found to be significantly higher in K12 OS cells than in K7M2 cells. qPCR showed that K12 cells upregulate the copper influx pump CTR1 and downregulate the copper efflux pump ATP7A compared to K7M2 OS cells. Combination treatment of copper chloride (50 nM) with disulfiram (80 nM) was only cytotoxic to K12 cells. Triple treatment with doxorubicin, disulfiram, and copper displayed potent and durable cytotoxicity of highly metastatic K7M2 cells. We demonstrate here that murine OS cell lines differing in metastatic potential also vary in endogenous copper levels and regulation. Additionally, these differences in copper regulation may contribute to selective cytotoxicity of K12 cells by extremely low doses of copper-potentiated disulfiram. The combination of doxorubicin, disulfiram, and copper should be explored as a therapeutic strategy against OS metastases.

A TITE-CRM phase I/II study of disulfiram and copper with concurrent radiation therapy and temozolomide for newly diagnosed glioblastoma.

2033 Background: Disulfiram (DSF) has shown promising activity against glioblastoma in preclinical studies and is more effective when combined with copper (Cu). Our previous phase I study established the maximum tolerated dose (MTD) of DSF when combined with adjuvant temozolomide (TMZ). This phase I/II study aims to establish the MTD when disulfiram and copper are combined with concurrent radiation therapy (RT) and TMZ for newly diagnosed glioblastoma and to explore preliminary efficacy. Methods: Eligible patients were treated with standard RT and TMZ plus escalating doses of DSF (250 mg - 375 mg PO QD) and Cu (2 mg PO TID), followed by adjuvant TMZ plus DSF (500 mg/day) and Cu. The time-to-event continual reassessment method (TITE-CRM) was used to continuously estimate the probability of dose-limiting toxicity (DLT) and to assign patients to doses with an estimated DLT probability of approximately 20% with a margin of 5%. Tumor mutations were evaluated with next-generation sequencing for all patients. Results: Eighteen glioblastoma patients were treated with the study therapy: 8 with DSF of 250 mg/day and 10 with 375 mg/day. Three DLTs were observed: 1 with 250 mg/day (grade 2 urinary incontinence and ataxia), and 2 with 375 mg/day (both grade 3 elevated liver enzymes). DSF had an estimated DLT probability of 10% (95% CI: 3-29%) at 250 mg/day, and 21% (95% CI: 7-42%) at 375 mg/day. After a median follow-up of 12.3 months, 1-year progression-free survival (PFS) was 57%, and 1-year overall survival (OS) was 69%. There was no significant difference in PFS/OS when stratified by DSF doses, surgical extent, or MGMT methylation status. However, glioblastomas with IDH1 (n = 6), BRAF (n = 2), or NF1 (n = 1) mutations had significantly better PFS and OS than those without the mutations: 1-year PFS: 100% vs 22%, respectively, p = 0.001; 1-year OS: 100% vs 42%, respectively, p = 0.006. Conclusions: The MTD of DSF with RT/TMZ/Cu for glioblastoma is 375 mg/day, and the recommended phase II dose is 250 mg/day. Although confirmation with larger sample size is needed, the combination demonstrates promising preliminary efficacy for the subset of glioblastoma with IDH1, BRAF, and NF1 mutations. Clinical trial information: NCT02715609.

Disulfiram-induced psychosis at a therapeutic dose and in clear sensorium: Two case demonstrations

The literature on disulfiram induced psychosis showed, it was either associated with delirium or preceded by a prescription of a higher dose. We report two cases of psychosis in clear sensorium developing in individuals with alcohol dependence without any familial loading following a short duration of a therapeutic dose of disulfiram. Before the onset of psychosis, the patients were abstinent from alcohol for about a month, which made substance-induced psychosis unlikely. Psychosis resolved following discontinuation of disulfiram. Hence, the diagnosis of disulfiram-induced psychosis was considered. A plausible mechanism of such association has been discussed. Disulfiram-induced psychosis though unusual could be disruptive and severe. Close surveillance following disulfiram is important for early identification and management of such condition.

Protein S-glutathionylation lowers superoxide/hydrogen peroxide release from skeletal muscle mitochondria through modification of complex I and inhibition of pyruvate uptake.

Protein S-glutathionylation is a reversible redox modification that regulates mitochondrial metabolism and reactive oxygen species (ROS) production in liver and cardiac tissue. However, whether or not it controls ROS release from skeletal muscle mitochondria has not been explored. In the present study, we examined if chemically-induced protein S-glutathionylation could alter superoxide (O2●-)/hydrogen peroxide (H2O2) release from isolated muscle mitochondria. Disulfiram, a powerful chemical S-glutathionylation catalyst, was used to S-glutathionylate mitochondrial proteins and ascertain if it can alter ROS production. It was found that O2●-/H2O2 release rates from permeabilized muscle mitochondria decreased with increasing doses of disulfiram (100–500 μM). This effect was highest in mitochondria oxidizing succinate or palmitoyl-carnitine, where a ~80–90% decrease in the rate of ROS release was observed. Similar effects were detected in intact mitochondria respiring under state 4 conditions. Incubation of disulfiram-treated mitochondria with DTT (2 mM) restored ROS release confirming that these effects were associated with protein S-glutathionylation. Disulfiram treatment also inhibited phosphorylating and proton leak-dependent respiration. Radiolabelled substrate uptake experiments demonstrated that disulfiram inhibited pyruvate import but had no effect on carnitine uptake. Immunoblot analysis of complex I revealed that it contained several protein S-glutathionylation targets including NDUSF1, a subunit required for NADH oxidation. Taken together, these results demonstrate that O2●-/H2O2 release from muscle mitochondria can be altered by protein S-glutathionylation. We attribute these changes to the protein S-glutathionylation complex I and inhibition of mitochondrial pyruvate carrier.

Restrained management of copper level enhances the antineoplastic activity of imatinib in vitro and in vivo

The present study was designed to investigate if elevated copper level can be targeted to enhance the efficacy of a significant anticancer drug, imatinib (ITB). The antineoplastic activity of this drug was assessed in the HepG2, HEK-293, MCF-7 and MDA-MD-231 cells targeting elevated copper level as their common drug target. The cell lines were treated with the different doses of copper chloride (Cu II) and disulfiram (DSF) alone as well as in their combinations with the drug for 24 h in standard culture medium and conditions. The treated cells were subjected to various assays including MTT, PARP, p-53, caspase-7, caspase-3, LDH and single cell electrophoresis. The study shows that DSF and Cu (II) synergizes the anticancer activity of ITB to a significant extent in a dose-specific way as evidenced by the combinations treated groups. Furthermore, the same treatment strategy was employed in cancer-induced rats in which the combinations of ITB-DSF and ITB-Cu II showed enhanced antineoplastic activity as compared to ITB alone. However, DSF was more effective than Cu (II) as an adjuvant to the drug. Hence, restrained manipulation of copper level in tumor cells can orchestrate the redox and molecular dispositions inside the cells favoring the induction of apoptosis.

ACTR-21. UPDATED RESULTS OF A PHASE I DOSE-ESCALATION, DOSE-EXPANSION STUDY OF DISULFIRAM AND ADJUVANT TEMOZOLOMIDE FOR NEWLY DIAGNOSED GLIOBLASTOMA

BACKGROUND Disulfiram has promising preclinical activity against glioblastoma (GBM) as well as synergy with temozolomide. In a phase I study for newly diagnosed GBM after chemoradiotherapy, we have previously shown the maximum tolerated dose (MTD) of disulfiram with adjuvant temozolomide is 500mg per day (QD). Here we report the updated results of the phase I study with a dose-expansion cohort with the addition of concurrent copper gluconate supplement.

Toxicological Evaluation of Disulfiram, Copper Gluconate and Disulfiram/Copper Gluconate Combination on Renal Function in Rodents

This research work investigated and compared the chronic renal toxicological profile of disulfiram, copper gluconate and disulfiram/copper gluconate combination, in a 90-day time- and dose-dependent study in rodents. 88 rats weighing an average of 280 g divided into eleven groups consisting of 8 rats each were used for this experiment. The control groups received normal saline as placebo and 99.5% dimethyl sulfoxide (DMSO) (solvent control). Three oral doses (low, medium and high) of disulfiram (18.65 mg/kg, 37.3 mg/kg and 74.6 mg/kg), copper gluconate (3.75 mg/kg, 7.5 mg/kg and 15 mg/kg) and both drugs in combination were administered daily with those of the combination given 12 hours apart. Blood samples were collected via cardiac puncture in heparinised bottles and centrifuged, and the serum was decanted on 30, 45, 60 and 90 days for analysis. Renal function parameters—electrolytes (Na+, K+), urea and creatinine were evaluated. Results showed significant (p < 0.05) dose- and time-dependent increase in electrolyte level (Na+, K+), blood urea and creatinine respectively. The results are all pointers to the development of renal failure. It therefore appears that the DSF/CG combination is nephrotoxic and this effect is dose-dependent and synergistic.

False negativity to carbohydrate-deficient transferrin and drugs: a clinical case

Introduction:In this work we report on the possible effect of the medical therapy on CDT concentration in a chronic alcohol abuser, with known medical history (July 2007 – April 2012) and alcohol abuse confirmed by relatives.Case history:At the end of 2007, patient displayed the following laboratory results: AST 137 U/L, ALT 120 U/L, GGT 434 U/L, MCV 101 fL and CDT 3.3%. On December 2007, after double coronary artery bypass surgery, he began a pharmacological treatment with amlodipine, perindopril, atorvastatin, isosorbide mononitrate, carvedilol, ticlopidine and pantoprazole. In the next months, until may 2011, the patient resumed alcohol abuse, as confirmed by relatives; however, CDT values were repeatedly found negative (0.8% and 1.1%) despite elevated transaminases and GGT, concurrent elevated ethyl glucuronide concentration (> 50 mg/L) and blood alcohol concentration (> 1 g/L). Alcohol consumption still continued despite increasing disulfiram doses ordered by an Alcohol Rehab Center. On May 2011, the patient was transferred to a private medical center where he currently lives.Conclusions:This study suggests the possibility that a medical therapy including different drugs may hamper the identification of chronic alcohol abusers by CDT.

Effects of Disulfiram on QTc Interval in Non–Opioid-Dependent and Methadone-Treated Cocaine-Dependent Patients

Methadone and cocaine are each known to prolong the QTc interval, a risk factor for developing potentially fatal cardiac arrhythmias. Disulfiram, often administered in the context of methadone maintenance to facilitate alcohol abstinence, has been shown to have some efficacy for cocaine dependence. Disulfiram has differential effects on cocaine and methadone metabolism, but its impact on methadone- or cocaine-induced changes in QTc interval is unclear. Thus, the effects of disulfiram on QTc interval in a subset of cocaine-dependent patients participating in a 14-week, randomized, double-blind, placebo-controlled clinical trial of disulfiram were prospectively determined. Opioid-dependent participants were inducted onto methadone (weeks 1-2; MT) and both MT and non-opioid-dependent (UT) participants were randomized to receive disulfiram (weeks 3-14) at one of the following doses: 0, 250, 375, or 500 mg/d. Electrocardio-grams were obtained before study entry and during weeks 2 and 4. Complete QTc-interval data in 23 MT and 18 UT participants were analyzed. QTc interval tended to be higher in MT participants relative to UT participants, regardless of disulfiram dose and time point, but disulfiram did not differentially alter QTc interval. QTc interval was, however, significantly greater in participants with recent cocaine use than in those with no recent use. These results suggest that cocaine use and possibly MT status, but not disulfiram, are risk factors for QTc prolongation.

A study of cardiovascular complications of disulfiram-ethanol reaction

Background: Alcohol abuse is a chronic behavioral problem that can further develop as a progressive disease. Disulfiram is the most extensively used deterrent drug for clinical management of alcoholism. A supervised disulfiram-ethanol reaction (DER) test is carried out in patients who are willing for it, and who are skeptical of the effect of disulfiram in causing a DER. Objective: The aim of this work was to study the possible cardiovascular changes and complications during disulfiram-ethanol reaction in persons fulfilling the inclusion and exclusion criteria. Materials and Methods: After written informed consent was obtained, 60 subjects underwent alcohol detoxification and received disulfiram 250 mg BD orally for 5 days. On 5th day, alcohol challenge was carried out with constant monitoring of all the vital parameters. Cardiovascular changes were monitored and recorded during the DER. Results: Disulfiram was generally well tolerated and DER started to occur within 10-15 minutes and lasted for 90-240 minutes. Cardiovascular changes observed were blood pressure changes [hypotension (n=16, 26.67%) and hypertension (n=3, 5%)] and ECG changes [sinus tachycardia (n=22, 36.67%), transient ischemic changes (n=13, 21.67%), occasional atrial ectopics (n=1, 1.67%) and junctional rhythm (n=1, 1.67%)]. Only 23 subjects (38.33%) did not show any significant ECG changes. All subjects recovered with no residual ECG changes except those who developed hypotension (n=16) who required intervention with IV fluids. Conclusion: Cardiovascular complications can occur even in properly selected subjects having no baseline cardiovascular impairment, which could be serious and potentially life threatening. Based on the results of the present study, disulfiram-ethanol challenge test appears to be a relatively safe procedure if carried out under standardized conditions and using a safe maintenance dose of disulfiram.

The Impact of Disulfiram Treatment on the Reinforcing Effects of Cocaine: A Randomized Clinical Trial

BackgroundClinical trials indicate that disulfiram (250 mg/d) reduces cocaine use, though one study found that treatment with lower doses of disulfiram (62.5 and 125 mg/d) increased cocaine use. We conducted the present study to better understand how disulfiram alters the reinforcing effects of cocaine in cocaine users.MethodsSeventeen non-treatment seeking, cocaine-dependent volunteers participated in this double-blind, placebo-controlled, laboratory-based study. A cross-over design was utilized in which participants received placebo in one phase and disulfiram (250 mg/d) in the other. Following three days of study medication participants completed two choice sessions. In one they made 10 choices between receiving an intravenous infusion of saline or money that increased in value (US$ 0.05–16) and in the other cocaine (20 mg) or money.ResultsParticipants chose cocaine more than saline under both disulfiram and placebo conditions (p<0.05). Unexpectedly, disulfiram increased both the number of cocaine and saline infusion choices (p<0.05). We next examined the relationship between disulfiram dose and cocaine choices. Disulfiram dose (mg/kg bodyweight) was negatively correlated with number of choices for cocaine (p<0.05). Disulfiram also enhanced cocaine-induced increases in cardiovascular measures (p's<0.05–0.01).ConclusionsDisulfiram's impact on the reinforcing effects of cocaine depends on dose relative to body weight. Our results suggest that the use of weight-based medication doses would produce more reliable effects, consistent with weight-based dosing used in pediatrics and in preclinical research.Trial RegistrationClinicaltrials.gov NCT00729300

Disulfiram metabolite S-methyl-N,N-diethylthiocarbamate quantitation in human plasma with reverse phase ultra performance liquid chromatography and mass spectrometry

Disulfiram has been used extensively for alcohol abuse and may have a role in treatment for cocaine addiction. Recent data suggest that disulfiram may also reactivate latent HIV in reservoirs. Disulfiram has complex pharmacokinetics with rapid metabolism to active metabolites, including S-methyl-N,N-diethylthiocarbamate (DET-Me) which is formed from cytochrome P450 (CYP450). Assessing disulfiram in HIV-infected individuals with a CYP450 inducing drug (e.g., efavirenz) or a CYP450 inhibiting drug (e.g., HIV-1 protease inhibitors) requires an assay that can measure a metabolite that is formed directly via CYP450 oxidation. Therefore, an assay to measure concentrations of DET-Me in human plasma was validated. DET-Me and the internal standard, S-ethyldipropylthiocarbamate (EPTC) were separated by isocratic ultra performance liquid chromatography using a Waters Acquity HSS T3 column (2.1mm×100mm, 1.8μm) and detection via electrospray coupled to a triple quadrupole mass spectrometer. Multiple reaction monitoring in positive mode was used with DET-Me at 148/100 and the internal standard at 190/128 with a linear range of 0.500–50.0ng/mL with a 5min run time. Human plasma (500μL) was extracted using a solid phase procedure. The interassay variation ranged from 1.86 to 7.74% while the intra assay variation ranged from 3.38 to 5.94% over three days. Representative results are provided from samples collected from subjects receiving daily doses of disulfiram 62.5mg or 250mg.

Biochemical and histopathological investigation of the protective effect of disulfiram in ischemia-induced ovary damage

It was biochemically and histopathologically investigated whether disulfiram has protective effects on ischemia-induced ovary damage. For this purpose, levels of tGSH, superoxide dismutase (SOD), malondialdehyde (MDA), and 8-OH Gua/Gua were investigated in ischemic rat ovary tissue. Results show that used doses of disulfiram (10, 25, and 50 mg/kg) prevent MDA, a product of ischemia-induced lipid peroxidation, formation in female rat ovary tissue and prevent decrease of enzymatic and non-enzymatic (SOD, GSH) antioxidant parameters. Additionally, all doses of disulfiram significantly prevent DNA damage when compared to control group. Fewer histopathological findings were observed in tissues with higher antioxidant levels and lower oxidant and DNA damage levels.

A double-blind, placebo-controlled assessment of the safety of potential interactions between intravenous cocaine, ethanol, and oral disulfiram

BackgroundA majority of cocaine addicts have a comorbid alcohol use disorder. Previous studies demonstrated efficacy of disulfiram in the treatment of cocaine dependence among patients with comorbid alcohol use disorder or opioid dependence. However, the cardiac risks of a disulfiram–ethanol reaction (DER) in individuals who drink, when coupled with the cardiac effects of cocaine, could result in significant toxicity or lethality due to the 3-way drug interaction. AimsThis study examined the safety of combining cocaine (30mg i.v.) and ethanol (0.4g/kg i.v.) in disulfiram-treated (0, 250, and 500mg/d, p.o.) cocaine-dependent research volunteers. ResultsThe results showed that disulfiram did not enhance the cardiovascular effects of cocaine and may have reduced the subjective high from cocaine. In contrast, ethanol produced adverse ECG changes including QTc prolongation and a DER consisting of hypotension, tachycardia, nausea, and flushing in disulfiram-treated subjects. The severity of the DER was related to disulfiram dose and the trial with 500mg/d was stopped prematurely due to safety concerns. The DER-related hypotension and tachycardia seen with ethanol infusion alone in disulfiram-treated subjects, was not exacerbated when combined with cocaine. In fact, cocaine tended to counteract the ethanol-related hypotension though it did exacerbate the tachycardia in two of seven subjects. ConclusionsThough conclusions are limited by the moderate doses of cocaine, ethanol, and disulfiram tested, the data do suggest that the risks of the moderate use of cocaine and ethanol in individuals treated with moderate doses of disulfiram (≤250mg/d) may not be as problematic as some may assume.

Determination of GABA, glutamate, dopamine and carbamathione in brain microdialysis samples by micellar electrokinetic chromatography and laser-induced fluorescence (MEKC-LIF)

A micellar electrokinetic chromatography and laser-induced fluorescence (MEKC-LIF) method based on derivatization with naphthalene-2,3-dicarboxyaldehyde (NDA) was optimized to separate and detect glutamate (Glu), gamma-aminobutyric acid (GABA), dopamine (DA) and carbamathione in brain microdialysis samples. The background electrolyte (BGE) consisted of 22.5 mmol L−1lithium tetraborate, 25 mmol L−1lithium dodecylsulfate and 10% methanol. The separation was carried out on a 65 cm × 50 μm id fused-silica capillary (50 cm effective) at +15 kV voltage with a run time of 25 min. The detection limits for GABA, Glu, DA and carbamathione were 6, 10, 5 and 10 nmol L−1, respectively. This method was used to monitor GABA, Glu, DA and carbamathione in brain microdialysis samples from the nucleus accumbens after the administration of an intra-peritoneal dose of disulfiram (200 mg kg−1) and revealed a disulfiram-induced change in GABA, Glu and DA levels. This method demonstrates a simple, rapid and accurate measurement of three amino acid neurotransmitters and carbamathione for in vivo monitoring in the brain using microdialysis sampling.

AVERSIVE THERAPY OF ADDICTIVE DISORDERS: PROBLEMS AND PERSPECTIVES

There was studied the contemporary state of aversive therapy in narcology. Effective disulfiram doses in alcoholic therapy were analyzed. There were given investigation results of effectiveness and safety of cyanamide (COLME) in therapy of alcoholism, including adolescents and women. There were shown perspectives of usage of aversive means during alcoholism in combination with naltrexone and comorbide pathology.

Urinary Ethylglucuronide Assessment in Patients Treated With Disulfiram

Disulfiram has been shown to be efficacious and safe in the treatment of alcohol relapse prevention. However, drinking alcohol while taking disulfiram can be harmful because of the resulting alcohol-disulfiram reaction/acetaldehyde reaction. Alcohol consumption of patients receiving a low disulfiram dose or low alcohol consumption in normal disulfiram dose can result in subclinical alcohol-disulfiram reactions. This undermines the learning of alcohol-associated punishment, might increase the risk to raise alcohol consumption, and can result in chronic acetaldehyde exposure, which has carcinogenic, neurotoxic, and cardiotoxic properties. Therefore, the use of an alcohol marker monitoring retrospective alcohol use is tested in this study. A total of 51 patients being treated with supervised disulfiram were unheralded measured for ethylglucuronide (EtG) if they attended at least for 2 weeks in the outpatient treatment program. Ethylglucuronide was measured with liquid chromatography-tandem mass spectrometry analysis (LC-MS/MS). Detection limit was 0.1 mg/L. Urinary EtG was found positive in 5.9% (3/51) of the patients. Regularly conducted breathalyzer tests had been continuously negative in these patients. Moreover, vegetative withdrawal symptoms had not been found in these patients. Two of the positive EtG tests could be classified as covered relapses, whereas the third remained unclear but showed a negative EtG in a repetition of the test few days later. Unheralded urinary EtG monitoring improved verification of abstinence in patients treated with disulfiram, was helpful in detecting covered consumption of alcoholic beverages or hidden alcohol exposition (eg, fruit juice or personal care products), and thereby improved safety by preventing chronic acetaldehyde reaction.

Disulfiram Attenuates Drug-Primed Reinstatement of Cocaine Seeking via Inhibition of Dopamine β-Hydroxylase

The antialcoholism medication disulfiram (Antabuse) inhibits aldehyde dehydrogenase (ALDH), which results in the accumulation of acetaldehyde upon ethanol ingestion and produces the aversive 'Antabuse reaction' that deters alcohol consumption. Disulfiram has also been shown to deter cocaine use, even in the absence of an interaction with alcohol, indicating the existence of an ALDH-independent therapeutic mechanism. We hypothesized that disulfiram's inhibition of dopamine β-hydroxylase (DBH), the catecholamine biosynthetic enzyme that converts dopamine (DA) to norepinephrine (NE) in noradrenergic neurons, underlies the drug's ability to treat cocaine dependence. We tested the effects of disulfiram on cocaine and food self-administration behavior and drug-primed reinstatement of cocaine seeking in rats. We then compared the effects of disulfiram with those of the selective DBH inhibitor, nepicastat. Disulfiram, at a dose (100 mg/kg, i.p.) that reduced brain NE by ∼40%, did not alter the response for food or cocaine on a fixed ratio 1 schedule, whereas it completely blocked cocaine-primed (10 mg/kg, i.p.) reinstatement of drug seeking following extinction. A lower dose of disulfiram (10 mg/kg) that did not reduce NE had no effect on cocaine-primed reinstatement. Nepicastat recapitulated the behavioral effects of disulfiram (100 mg/kg) at a dose (50 mg/kg, i.p.) that produced a similar reduction in brain NE. Food-primed reinstatement of food seeking was not impaired by DBH inhibition. Our results suggest that disulfiram's efficacy in the treatment of cocaine addiction is associated with the inhibition of DBH and interference with the ability of environmental stimuli to trigger relapse.

The Effects of Alcoholism Pharmacotherapy on Immune Responses in Alcohol-Dependent Patients

Chronic alcohol use has profound modulatory effects on the immune system. Both the innate and the acquired immunity are compromised. The use of pharmacotherapy is increasingly applied to enhance the percentage of success in maintaining alcoholic patients in remission. Disulfiram, naltrexone and gamma hydroxybutiric acid are the drugs used for this purpose in Italian Addiction Services. In this study we analyze the effect of pharmacotherapy of alcohol dependence on immune responses in alcoholics. Six groups were studied. Group A included 10 patients who were still using alcohol. Group B consisted of 10 patients abstinent from alcohol in treatment only with group therapy. Groups C, D and E were composed of 10 patients each, treated for at least 6 months with oral doses of gamma hydroxybutiric acid, naltrexone or disulfiram respectively. Ten age- and sex-matched healthy volunteers who never misused alcohol were included as a control group. Lymphoproliferation and peripheral mononuclear cell production of the Th1 cytokines IL-2 and IFN-gamma, the Th2 cytokine IL-4, and of the pro-inflammatory cytokines IL-1 and TNF-alpha were evaluated in all the patients and controls. The level of activity of the hypothalamus pituitary adrenal axis was assessed. Both ACTH and cortisol levels in plasma were elevated in alcoholic patients with no treatment. In this group a significant alteration of cytokine production was observed. TNF and IFN-gamma were lower than controls, while the Th2 cytokine IL-4 was increased. These altered levels state for a Th1/Th2 unbalance characterized by decreased Th1 response in the presence of Th2 predominance. In patients undergoing pharmacological treatment, none of the immune parameters were different from those observed in healthy controls, independently of the type of drug administered. These data indicate that pharmacotherapy more than group therapy treatment is able to ameliorate the immune system functioning in alcoholic patients.