Abuse liability assessment in preclinical drug development: predictivity of a translational approach for abuse liability testing using methylphenidate in four standardized preclinical study models.

OBJECTIVES: Preclinical abuse liability assessment of novel clinical CNS-active candidates involves several tests, addressing different aspects characteristic for abuse potential, which are considered predictive for substance abuse of these candidates, thus ensuring an appropriate translational approach. To demonstrate how such a strategy could work, a known drug of abuse, methylphenidate was evaluated in a full rodent test battery, comprising four test models, and in accordance with the requirements of the FDA, ICH and EMA guidelines. METHODS: Methylphenidate was tested orally at 2.5, 5 or 10mg/kg for its physical dependence potential in a repeated dose non-precipitated withdrawal test, for its drug profiling in a drug discrimination learning procedure (single escalating doses), and for its reinforcing properties in a conditioned place preference test (alternate dosing days) and an intravenous self-administration procedure (0.05 to 1mg/kg/IV infusion during 5 daily 1-h test sessions). The stimulant d-amphetamine served as positive control and was administered subcutaneously at 0.8mg/kg in the first three test models. In the intravenous self-administration procedure rats were habituated to intravenously self-administer d-amphetamine at 0.06mg/kg/IV infusion prior to methylphenidate substitution. RESULTS: Cessation of subchronic dosing up to 10mg/kg methylphenidate led to sustained or even exacerbated effects on locomotion and behavior, body temperature, body weight, food consumption, and alteration of the diurnal rhythm during withdrawal. Clear generalization to d-amphetamine was obtained in the drug discrimination test at 5 and 10mg/kg. Distinct reinforcing properties were present in the conditioned place preference test at 10mg/kg and in the intravenous self-administration study from 0.05mg/kg/IV infusion onwards. The maximum plasma exposure after oral administration of methylphenidate over the dose ranges tested in the present rat studies covered at least 1.9-fold to 18.9-fold the recommended human therapeutic exposure of 10ng/ml, a plasma level that is considered representative of the human efficacious methylphenidate dose. The ratio Cmax Hu/rat calculated from the intravenous self-administration data ranged from 14.9 to 576.5. Consequently the regulatory requirements, stating that preclinical drug abuse liability studies should include high doses that produce plasma levels that are multiples of the therapeutic dose were fulfilled (FDA, EMA, ICH). DISCUSSION: The presented preclinical models, implemented within a drug development environment, were considered highly predictive to assess the abuse potential of methylphenidate, and in accordance with the regulatory requirements of drug licensing authorities in terms of appropriate methods, dose selection and subsequent plasma exposure.

Effects of long-term resistance training and simultaneous electro-stimulation on muscle strength and functional mobility in multiple sclerosis.

BACKGROUND: Resistance training studies in multiple sclerosis (MS) often use short intervention periods. Furthermore, training efficiency could be optimized by unilateral training and/or electrical stimulation. OBJECTIVE: To examine the effect(s) of unilateral long-term (20 weeks) standardized resistance training with and without simultaneous electro-stimulation on leg muscle strength and overall functional mobility. METHODS: A randomized controlled trial involving 36 persons with MS. At baseline (PRE) and after 10 (MID) and 20 (POST) weeks of standardized (ACSM) light to moderately intense unilateral leg resistance training (RES(O), n = 11) only or resistance training with simultaneous electro-stimulation (RES(E), n = 11, 100 Hz, biphasic symmetrical wave, 400 µs), maximal isometric strength of the knee extensors and flexors (45°, 90° knee angle) and dynamic (60-180°/s) knee-extensor strength was measured and compared with a control group (CON, n = 14). Functional mobility was evaluated using the Timed Get Up and Go, Timed 25 Foot Walk, Two-Minute Walk Test, Functional Reach and Rivermead Mobility Index. RESULTS: Maximal isometric knee extensor (90°, MID: +10 ± 3%, POST: +10 ± 4%) in RES(O) and knee flexor (45°, POST: +7 ± 4%; 90°, POST: +9 ± 5%) in RES(E) strength increased (p < 0.05) compared with CON but RES(O) and RES(E) did not differ. Also, impaired legs responded positively to resistance training (unilateral leg strength analysis) and functional reaching increased significantly in RES(O) (+18%) compared with CON. Dynamic muscle strength and the remaining functional mobility tests did not change. CONCLUSION: Long-term light to moderately intense resistance training improves muscle strength in persons with MS but simultaneous electro-stimulation does not further improve training outcome.

Tovaxin, radiation-attenuated, patient-specific T-cells for the therapeutic vaccination of multiple sclerosis.

Opexa Pharmaceuticals Inc is developing Tovaxin, a trivalent formulation of attenuated myelin-peptide-reactive T-cells, for the potential treatment of multiple sclerosis. Tovaxin is being evaluated in phase II clinical trials. Opexa was previously investigating Tovaxin for the potential treatment of rheumatoid arthritis; however, no development has been reported for this indication since December 2002.

Interferon-gamma-induced calcium influx in T lymphocytes of multiple sclerosis and rheumatoid arthritis patients: a complementary mechanism for T cell activation?

Autoreactive T lymphocytes are considered to play a crucial role in orchestrating a chronic inflammation in the central nervous system (CNS) of multiple sclerosis (MS) patients and in the joints of rheumatoid arthritis (RA) patients. However, it has been suggested that the majority of T cells in the immune infiltrate are nonspecifically recruited into the CNS and into the inflamed joint. In addition, several lines of evidence suggest an important role for interferon-gamma (IFN-gamma) in the pathogenesis of MS and RA. We have studied whether peripheral blood T cells from patients with autoimmune diseases are more susceptible to activation in the presence of IFN-gamma. The results indicate that IFN-gamma mediates a sustained elevated [Ca(2+)](i) in T cells of (active) MS and RA patients as compared to healthy controls and patients with common viral infections. No [Ca(2+)](i) increase was observed in Ca(2+)-free medium, excluding an effect of IFN-gamma on Ca(2+)-release from intracellular stores. Although the IFN-gamma-activated Ca(2+)-influx is insufficient to induce T cell proliferation in vitro, our data indicate a significantly augmented proliferation in response to suboptimal doses of PHA in the presence of IFN-gamma. This study suggests that the IFN-gamma-induced Ca(2+)-influx can act as a complementary mechanism in the activation of blood T lymphocytes from MS and RA patients.