MODULE 2.4 NONCLINICAL OVERVIEW · m2.4. Nonclinical Overview INTRODUCTION Ropinirole Prolonged...

CONFIDENTIAL

MODULE 2.4 NONCLINICAL OVERVIEW

CONFIDENTIAL m2.4. Nonclinical Overview

INTRODUCTION

Ropinirole Prolonged Release Tablets (2.0, 3.0, 4.0, and 8.0 mg)

Ropinirole is a potent selective dopamine D2/D3 receptor agonist that is effective as monotherapy in the symptomatic treatment of early stage Parkinson's disease and in combination with L-dopa in the later stages of the condition. Ropinirole is also an effective treatment for idiopathic/primary Restless Legs Syndrome (RLS).

Parkinsonism occurs as a result of the degeneration of dopaminergic neurones and a consequent deficiency in the neurotransmitter (dopamine) in part of the brain known as the basal ganglia. The disease is chronic and progressive with no known cure. The condition is characterised by poor movement, rigidity and tremor, leading to increasing disability including the inability to walk, mask-like expression, impairment of speech and sometimes dementia. For over 40 years L-dopa has been the mainstay of treatment for Parkinson's disease, but unfortunately this drug is associated with the development of erratic involuntary movements (dyskinesia) within a few years of initiation of treatment. As a dopamine receptor agonist ropinirole, exerts a direct postsynaptic effect, bypassing the degenerating presynaptic dopaminergic neurones, thus acting as a synthetic neurotransmitter.

Ropinirole has comparable efficacy to L-dopa in the early stages of Parkinson's disease and initiation of therapy with dopamine antagonists rather than L-dopa has been shown to delay the onset and reduce the prevalence of motor fluctuations. Even though therapy eventually needs to be supplemented with L-dopa, in most patients the dose of L-dopa required for an optimal therapeutic response is significantly lower than if L-dopa were administered alone. This strategy of combining a dopamine agonist and L-dopa seems to effectively inhibit the development of fluctuations in disability during long term treatment. In addition, unlike L-dopa, ropinirole has a low propensity to produce dyskinesias.

Ropinirole is available as an Immediate Release (IR) and a Prolonged Release (PR) Tablet formulation for the treatment of Parkinson’s disease. Ropinirole PR Tablets are available in strengths of 2, 3, 4, and 8 mg, containing ropinirole hydrochloride as the active ingredient.

Ropinirole has been in therapeutic use since 1996 and is available in over 60 countries worldwide for the treatment of Parkinson’s disease. It is estimated that the cumulative exposure to ropinirole from launch to January 2010 is approximately 1,046 million patient days (2,866,000 patient years).

This document describes the well established pharmacological and toxicological properties of ropinirole and presents a preclinical assessment supporting the use of ropinirole tablets for the proposed indication.


INDICATIONS AND DOSAGE

Indications

Ropinirole PR Tablets are indicated for the treatment of Parkinson's disease, either as a monotherapy or as an adjunctive therapy to L-dopa.

Dosage

Parkinson’s Disease

As a monotherapy

The recommended initial dose of Ropinirole PR Tablets is 2 mg once daily for one week. A guide for the titration regimen for the first four weeks of treatment is given in the table below:

Week 1 2 3 4

Total daily dose (mg) 2 4 6 8

After the initial titration, weekly increments of up to 4 mg once every one to two weeks may be given. If symptomatic control is not achieved, or maintained, the dose of ropinirole may be increased up to a maximum of 24 mg/day until an acceptable therapeutic response is established. The safety and efficacy of doses above 24 mg/day have not been established.

In conjunction with L-dopa

When Ropinirole PR Tablets are administered as an adjunct therapy to L-dopa, the concurrent doses of L-dopa may be reduced gradually by around 30%.

Switching from Ropinirole IR Tablets to Ropinirole PR Tablets

Patients may be switched overnight from ropinirole IR tablets to ropinirole PR tablets. The dose of ropinirole PR tablets should be based on the total daily dose of ropinirole IR tablets that the patient was taking and then adjusted depending on the therapeutic response.

Dosage in children

The safety and efficacy of ropinirole have not been substantiated in patients under 18 years of age, therefore ropinirole is not recommended for use in this age group.


Justification for Dosage

There are no major toxicological findings associated with ropinirole at doses of approximately 30 times the maximum proposed clinical dose of 0.48 mg/kg/day for Parkinson’s disease (24 mg/day assuming a 50 kg human body weight). Based on pharmacokinetic data, safety margins of at least 5-fold and 7-fold have been established in terms of Cmax for the IR and PR formulation, respectively, while an approximately equal or greater exposure was demonstrated in animal species in terms of AUC for both formulations.

Ropinirole is well tolerated in laboratory animals in the dose range of 15 to 50 mg/kg. The toxicity profile is principally determined by the pharmacological activity of the drug (behavioural changes, hypoprolactinaemia, decreases in blood pressure and heart rate, ptosis and salivation). Genotoxicity was not observed and carcinogenicity studies conducted in the mouse and rat at doses up to 50 mg/kg revealed no evidence of carcinogenicity in the mouse. In the rat, the only drug-related lesions were Leydig cell hyperplasia/adenoma in the testis resulting from the hypoprolactinaemic effect of ropinirole. These lesions are considered to be a species specific phenomenon and do not constitute a hazard with regard to the clinical use of ropinirole.


FORMULATION

The composition of Ropinirole PR Tablets is presented in the following tables:

Component

Quantity (mg/tablet) Function

Reference to

Standard Upper Barrier Layer1

Active Layer Lower Barrier Layer1 2 mg 3 mg 4 mg 8 mg

Core Active Ingredient Ropinirole HCl2 - 2.28 3.42 4.56 9.12 - Active GSK

Other Ingredients Hypromellose USP Type 2208 100,000 mPa s

62.83 61.50 61.50 61.50 61.50 76.29 Release controlling matrix polymer

PhEur/USP

Lactose Monohydrate

- 46.32 45.18 44.04 39.48 - Diluent PhEur/USNF

Glycerol Dibehenate 35.00 - - - - 42.50 Matrix permeability regulator

PhEur/USNF

Mannitol 33.04 - - - - 40.12 Diluent PhEur/USP

Carmellose Sodium - 15.00 15.00 15.00 15.00 - Viscosity regulating agent

PhEur/USP

Castor Oil, Hydrogenated

- 15.00 15.00 15.00 15.00 - Matrix strengthener

PhEur/USNF

Povidone K29-32 7.00 - - - - 8.50 Binder PhEur/USP

Maltodextrin - 7.50 7.50 7.50 7.50 - Binder PhEur/USNF

Magnesium Stearate 1.40 1.50 1.50 1.50 1.50 1.70 Lubricant PhEur/USNF

Silica, Colloidal Anhydrous

0.56 0.90 0.90 0.90 0.90 0.68 Glidant PhEur/USNF

Ferric Oxide (Yellow) E172, CI 77492

0.17 - - - - 0.21 Colourant USNF

Water, Purified3 - - - - - - Granulation solvent

PhEur/USP

Notes 1. The composition and weights of the barrier layers are identical for all dosage strengths. 2. Equivalent to nominal dose of ropinirole free base. 3. Used in the granulation process and removed during manufacturing. Does not appear in the final product. Key: PhEur = European Pharmacopoeia USP = United States Pharmacopoeia USNF = United States National Formulary


Formulation table continued:

Component

Quantity (mg/tablet) Function

Reference

to Standard

Active Layer

2 mg 3 mg 4 mg 8 mg

Film Coat1,2 Opadry® Pink OY-S-24900

(13.80 mg)

Opadry® Purple

03B20024 (13.80 mg)

Opadry® Tan OY-27207

(13.80 mg)

Opadry® Red

03B25227 (13.80 mg)

Colouring Supplier

Containing (%)

Hypromellose 2910 6,000 mPa s

66.00 62.50 62.50 62.50 PhEur

Titanium Dioxide (E171, CI 77891)

27.00 23.43 21.25 24.19 PhEur

Macrogol 400 6.60 6.25 6.25 6.25 PhEur

Ferric oxide (Red) (E172, CI 77491)

0.25 - - 6.14 USNF

Ferric oxide (Yellow) (E172, CI 77492)

0.15 - - 0.03 USNF

Carmine (E120, CI 75470) - 4.88 - -

Indigo Carmine Aluminium Lake , FD&C Blue No 2 (E132, CI 73015)

- 2.86 1.00 -

Sunset yellow FCF Aluminium Lake , FD&C Yellow No 6 (E110, CI 15985)

- 0.08 9.00 -

Ferric oxide (Black) (E172, CI 77499)

- - - 0.89

Notes: 1. Information as provided by supplier. 2. The weight of film coat applied may vary depending upon the efficiency of the film coating process. Figures

provided are for a 3% w/w weight gain, acceptable range 2 to 4% w/w. Key: PhEur = European Pharmacopoeia USP = United States Pharmacopoeia USNF = United States National Formulary Further information on the excipients is presented in 'Toxicology relating to the specific formulation.


ACTIVE INGREDIENT INFORMATION

Ropinirole

Pharmacology

Ropinirole is a potent and highly selective non-ergot dopamine D2/D3 receptor agonist which is used for the treatment of Parkinson’s disease and idiopathic/primary Restless Legs Syndrome (RLS). Parkinson’s disease occurs as a result of degeneration of dopaminergic neurones in the substantia nigra, resulting in a deficiency of dopamine in the neostratum. As a dopamine receptor agonist, ropinirole exerts a direct postsynaptic effect by bypassing the degenerating presynaptic dopaminergic neurones in the substantia nigra and acting directly as a synthetic neurotransmitter. In this way ropinirole relieves the poor movement, rigidity and tremor which are characteristic of Parkinsonism. The pathogenesis of RLS remains unknown, but a consistent positive response of the clinical symptoms to dopaminergic agents suggests some specific abnormality of the dopamine system.

The high affinity and functional potency of ropinirole for D2/D3 receptors has been demonstrated in a number of in vitro studies, and the drug has also been shown to restore motor function in animal models of Parkinson’s disease. To date, no preclinical studies have been undertaken specifically to investigate the use of ropinirole in RLS due to the lack of an ideal animal model for this disorder.

Ropinirole is active both centrally and peripherally, with demonstrable CNS and cardiovascular activity in several species. There is no evidence that tolerance develops to the therapeutic properties of ropinirole following long term treatment, whereas the peripheral effects of the compound show very rapid tolerance. Co-medication with antidepressants and anxiolytics in human seems unlikely to be problematic and there is no evidence that ropinirole has abuse liability or the potential to cause physical dependence. Unlike L-dopa, ropinirole also has only a low propensity to produce dyskinesias.

Primary Pharmacodynamics

Electrophysiological, electroencephalographic, biochemical and behavioural data from rat, mouse and marmoset support the evidence that ropinirole crosses the blood brain barrier.

Studies with human cloned receptors show that ropinirole binds to human D2 and D4 receptors with similar affinity but has a somewhat greater affinity for the human D3 receptor. The profile is similar for rat D2, D3 and D4 receptors. Ropinirole and its metabolite SK&F 89124 exhibit highly specific binding to the central D2 dopamine receptor family (D2, D3, D4) in human and rat tissue while another metabolite SK&F 104557 shows a lower affinity. Ropinirole and both metabolites have a low affinity for D1 dopamine receptors. Additional tests suggest, however, that neither metabolite is likely to contribute to the central D2 agonist activity of ropinirole in humans. The high binding affinity of ropinirole for the native rat D2 receptor is associated with high potency in a D2 functional assay. Ropinirole also binds with low affinity to opiate


receptors but not to 5-HT1, 5-HT2, benzodiazepine, GABA, α- or β-adrenoceptors or muscarinic receptors. Ropinirole has also shown moderate receptor activity at peripheral D2 prejunctional dopaminergic neurones.

The neurochemical selectivity of ropinirole has been confirmed in studies of its effects on neurotransmitter turnover in mouse brain. Oral doses of 0.03 to 10 mg/kg dose-dependently increased dopamine metabolites but did not affect noradrenaline or 5-HT. Bromocriptine (D2 agonist control) also increased dopamine metabolites but was approximately 3 to 10 times weaker.

Ropinirole was active in rodent models of hypo-dopaminergic function [0.01 to 87.7 mg/kg IP (mice), 0.05 to 6.4 mg/kg SC and 1.0 to 10 µg intrastriatum (rat)]. SK&F 89124 was found to be equipotent with ropinirole. The metabolite SK&F 104557 and two other metabolites (SK&F 96990 and SK&F 97930) proved not to be significantly active at doses up to 15 mg/kg SC.

Motor deficits resembling Parkinson's disease induced in marmosets by the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were reversed when ropinirole was given at 0.09 and 0.88 mg/kg SC. Ropinirole restored motor function in the trunk, limb, head and neck; additionally facial expression and movement co-ordination was improved. At a lower dose of 0.04 mg/kg, some marmosets demonstrated significant improvement in motor function while a few showed little or no response. In animals dosed at 0.09 and 0.88 mg/kg SC BID for seven days, there was no evidence of tolerance. Ropinirole was also active orally at doses of 0.44 and 0.88 mg/kg although a 'nausea response' and emesis was seen following both doses and hyperactivity at the higher dose. Ropinirole after an acute single dose of 0.88 mg/kg PO reversed MPTP-induced deficits but did not cause emesis or the associated 'nausea response'. Bromocriptine 1.0 mg/kg (D2 agonist control) had little effect on these animals. L-dopa produced qualitatively similar beneficial effects but at a dose 500 times higher than ropinirole.

Ropinirole at doses of 0.01 to 8.77 mg/kg IP in rats inhibited dyskinesias induced by dopamine antagonism. By comparison the neuroleptic tiapride, shown to inhibit dyskinesias in human, also inhibited the induced dyskinesias in this model. Ropinirole was shown to have a low potential to induce dyskinesia in drug-naive MPTP-lesioned marmosets, significantly less than L-dopa at equivalent anti-Parkinson doses [Pearce, 1996]. In the same model, combinations of L-dopa and ropinirole resulted in a lower incidence of dyskinesia than L-dopa alone, while still alleviating parkinsonian symptoms [Maratos, 1998]. Furthermore, a trend indicated that the ropinirole dominant therapy induced less dyskinesia than the L-dopa dominant therapy. In a 6-hydroxydopamine-lesioned rat model of Parkinson’s disease, ropinirole administered at doses of 5 mg/kg/day (IP) for 19 days produced a mild dyskinetic response characterised by sensitisation of contralateral turning (SCT) in the animals. In contrast L-dopa, at doses of 5 mg/kg/day (IP) for 19 days, induced both SCT and abnormal involuntary movements [Carta, 2008].

Ropinirole 0.2 to 2.0 mM did not inhibit the activity of DOPA-decarboxylase suggesting ropinirole will not affect the metabolism of endogenous or co-administered L-dopa.


Secondary Pharmacodynamics

Ropinirole (0.9 to 8.77 mg/kg) administered IP to mice inhibited spontaneous locomotion, while at the higher dose (87.7 mg/kg) a marked stimulation in locomotor activity was observed. Such a biphasic profile is characteristic of a centrally acting dopamine agonist. In rats the effect was similar but hypoactivity was only seen at the lowest dose of 0.26 mg/kg IP. SK&F 104557 the major metabolite in monkey and human reduced rat spontaneous locomotor activity at 26.32 mg/kg IP. Marmosets showed hyperactivity in response to a low dose of ropinirole (0.1 mg/kg). No evidence of hypoactivity was seen with SK&F 104557, although a small increase in locomotor activity was seen with the highest dose of 10 mg/kg. In contrast to apomorphine and amphetamine, ropinirole did not cause stereotypy in mice and rats. Unlike many dopamine agonists, the locomotor stimulant activities of ropinirole can be dissociated from the ability to elicit dyskinetic behaviour in mice and rats.

There is evidence to show that ropinirole (0.09 to 8.77 mg/kg IP or SC) unlike bromocriptine has anxiolytic-like actions similar to those of diazepam in mice, rat and marmoset. Additionally there was no tolerance to the anxiolytic effect in mice over a 14 day dosing period, and withdrawal of treatment did not produce rebound anxiogenesis (unlike diazepam) nor did it adversely interact with diazepam (a common adjunct therapy for anxiety in Parkinsons patients).

Unlike bromocriptine, ropinirole has antidepressant-like activity in rats and mice, and in mice ropinirole (0.88 and 8.77 mg/kg IP) is more potent than amitriptyline. The antidepressant effect of amitriptyline, a common adjunct therapy for depression in Parkinsons was not compromised by co-treatment with ropinirole.

In a series of in vitro tests ropinirole was shown to have cardiovascular (CV) effects typical of a drug of its class. SK&F 89124 caused CV effects similar to those seen following ropinirole administration. Dose related antihypertensive effects of ropinirole were seen in anaesthetised spontaneously hypertensive rats (SHR) at IV doses of up to 0.44 mg/kg (infused over 6 min). In the conscious SHR significant reductions in blood pressure (BP) were seen at doses of ropinirole above 20 mg/kg orally and 2.5 mg/kg IV; no effects were seen at doses of up to 30 mg/kg IP. SK&F 104557 showed antihypertensive activity in the anaesthetised SHR at bolus IV doses of 0.3 to 1.0 mg/kg (approximately 33 times less active than ropinerole). SK&F 89129 was active at doses 0.125 to 5.0 µg/kg IV (conscious rats) and 1.0 to 100 µg/kg IV (anaesthetised rats). Enhancement of the orthostatic hypotensive response was also seen in both rat [450 µg/kg (IV infusion)] and cat (0.44 mg/kg infused IV over 10 min). The effect produced by ropinirole in the cat was not affected by seven days TID pre-treatment with L-dopa and benserazide. Inhibition of electrically induced increases in heart rate (HR) has been shown in dog, cat and monkey (8.77 to 132 µg/kg IV or 43.9 to 263 µg/kg ID). In anaesthetised dogs ropinirole 10 µg/kg/min and SK&F 89124 1.0 µg/kg/min infused IV over 10 min decreased BP, HR, coronary flow (CF), myocardial oxygen consumption (MOC) and total peripheral resistance (TPR). In repeat studies, using bolus IV doses, ropinirole (8.77 µg/kg) caused reductions in BP and TPR while 87.7 µg/kg produced more sustained changes in BP, TPR and a slight non significant change in MOC. A dose-related tolerance to the CV effects developed rapidly (evident after one day of dosing at


35.1 mg/kg orally BID in the rat or after 7 and 14 days of pre-treatment with 4.39 mg/kg orally in the monkey). In both cases this tolerance could not be overcome by increasing the challenge dose of ropinirole 10-fold. Cross-tolerance to the CV effects of bromocriptine was demonstrated in both species, suggesting switching from bromocriptine treatment to ropinirole is unlikely to have any significant CV consequence.

The possibility that ropinirole may act as a neuroprotective agent, slowing the progression of Parkinson’s disease has been explored. In a rat model ropinirole protects against 1-methyl-4-phenylpyridinium (MPP+)-evoked hydroxyl radical production. Ropinirole had no concomitant effect on dopamine release [Rose, 1997]. The protective effect of ropinirole may be explained by direct antioxidant or hydroxyl radical scavenging action. Also it has been reported that D2 receptor binding is coupled with pathways that inhibit apoptosis. In the mouse, administration of ropinirole increases superoxide dismutase and it is likely that auto-oxidation was also suppressed by the activation of glutathione-regulating enzymes such as glutathione peroxidase, glutathione reductase and glutathione S-transferase [Tanaka, 2001]. Additionally protection of the nigro-striatal dopaminergic neurones (via dopamine D2 receptors) against 6-hydroxydopamine-induced destruction was seen [Ogawa, 1999].

Although ropinirole had a low binding affinity for opiate receptors there is no evidence that ropinirole possesses a physical dependence liability nor any clear evidence of a psychological dependence liability. Additionally ropinirole did not suppress barbiturate withdrawal in phenobarbitone dependent monkeys.

Safety Pharmacology

Overt Central and Peripheral Effects

Ropinirole 0.09 to 8.77 mg/kg IP was found not to improve cognitive function or reverse a scopolamine induced deficit in learning in rats. Ropinirole was administered orally to mice (at doses of 0.09, 8.77 or 87.7 mg/kg PO) in a series of tests designed to detect general changes in central nervous activity. These doses caused no change in central nervous system activity other than hypothermia (confirmed in rats) and a reduction in locomotor activity which are characteristic of a D2-dopamine receptor agonist. An increase in hexabarbitone-induced sleeping time was probably due to altered barbiturate metabolism rather than a direct CNS effect. A higher dose of 877 mg/kg was lethal in two of three mice. The metabolite SK&F 89124 at doses of 1.0 to 100 mg/kg orally also caused dose-related hypothermia, increases in hexabarbital-induced sleeping time and a reduction in locomotor activity but additionally (at the high dose) increased the latency to chemically induced convulsions and showed analgesic activity.

Other than the expected inhibition of reflex-mediated stimulation of the sympathetic nervous system, ropinirole 43.9 and 439 µg/kg IV infused over 10 min, had no effect on the ANS function in anaesthetised cats. Ropinirole at doses up to 10 µM had little effect on the spontaneous motility of rabbit isolated ileum. In rat isolated ileum at 10 µM increases in resting tension were seen but at a dose of 100 µM effects were non-specific.


Actions on Respiratory Systems

Ropinirole infused over a ten-minute period at 43.9 and 439 µg/kg, in spontaneously breathing, anaesthetised cats did not compromise the respiratory system.

Actions on Cardiovascular Systems

The haemodynamic effects of ropinirole, at a pharmacologically active dose, and at ten times that dose, (43.9 and 439 µg/kg infused over 10 min) were examined in anaesthetised cats. As expected there was a dose-related fall in BP, HR, aortic flow and total peripheral resistance. Stroke volume was increased at the low dose and to a lesser extent at the high dose. Effect on cardiac repolarisation

Ropinirole blocked hERG-mediated currents in hERG-transfected Chinese Hamster Ovary (CHO-K1) cells with an IC50 value of 1.2 µM [Hurst, 2003] and prolonged the duration of the cardiac action potential in canine [Hurst, 2003] and sheep Purkinje fibres. These findings suggest that ropinirole has the potential to prolong cardiac repolarisation and therefore the QT interval in vivo. In-house hERG binding and functional electrophysiology assays have confirmed that ropinirole blocks the hERG channel, but suggest that the potency (IC50 values of 10.5 µM and 44.7 µM, respectively) is somewhat lower than that reported by Hurst. Given the very different technologies used, these differences are neither large nor unexpected.

A recent publication by Humphrey and coworkers showed that ropinirole could prolong the QTc interval in conscious dogs at a peak plasma drug concentration of 3.5 ng/mL, which is within the therapeutic range [Humphrey, 2006). However, there is no evidence of QTc interval prolongation from repeat dose toxicology studies with ropinirole. A review of the available clinical trial and post-marketing data, considered in the context of overall patient exposure to ropinirole (>960,000 patient years, estimate to March 2006), does not suggest an increase in treatment-related risk for adverse events possibly linked to QTc prolongation at therapeutic dosages. In addition, a thorough QT study in healthy volunteers with doses of ropinirole up to 4 mg/day did not show effects on cardiac repolarization, and a pharmacokinetic/pharmacodynamic analysis of data pooled from clinical trials at doses up to 24 mg/day has also shown no clinically relevant effects on QT prolongation.

Effects on the renal system

The effect of ropinirole (0.26, 2.63 and 26.3 µg/kg/min IV for 20 min) on renal haemodynamics and excretion of water and solutes by the kidneys was assessed in conscious normotensive rats. Ropinirole caused a dose dependent reduction in urine flow and potassium excretion and a non-significant fall in sodium excretion was observed at the highest dose.

Haematological and gastric effects

Ropinirole 1.0 to 100 mg/kg PO had no effect on haematological parameters in blood from anaesthetised rats. Charcoal transit time was unaffected by ropinirole at the same


doses in mice. In histological studies, 1.0 and 10 mg/kg PO caused no injury to the stomach or duodenum, although at 100 mg/kg some congestion was noted and one of six rats developed capillary haemorrhage in the glandular stomach.

Pharmacokinetics

Pharmacokinetics, distribution and metabolism studies have been carried out in mice, rats, cynomolgus monkeys and to a limited extent marmosets and dogs. Drug-related material was well absorbed and widely distributed throughout the body following oral and intravenous administration in all species. Ropinirole, and to a lesser extent SK&F 89124, were shown to cross the blood brain barrier in the rat and monkey and ropinirole related material was also shown to transfer into the milk of lactating rats and cross the placentae of pregnant rats. In the rat and monkey, ropinirole was subject to first pass metabolism leading to dose dependent kinetics. The major route of metabolism is via liver microsomal enzymes (predominantly CYP1A2). Plasma clearance of ropinirole was high, and very little of the drug was excreted unchanged. The major route of excretion of total drug-related material was via the urine in all species. Minimum accumulation occurred following once daily dosing to mouse and human.

Absorption

The extent of urinary excretion of drug-related material was virtually identical following oral or intravenous administration of ropinirole to mice, rats and monkeys indicating that, at the doses examined [8.8 mg/kg (mouse) up to 220 mg/kg (rat) and up to 13 mg/kg (monkey)] nearly all of the orally administered dose (>94%) was absorbed.

Plasma/Serum Concentrations

A summary of systemic exposure in the mouse, rat, monkey and human is given in the table overleaf.

Systemic exposure to ropinirole in male mice following single oral administration at 4.39, 13.2 and 43.9 mg/kg produced Cmax values for plasma of 49.0, 204 and 307 ng/mL. Cmax values occurred approximately 1 h post dose. The corresponding AUC0-inf values were 88.3, 212 and 497 ng h/mL. Similar data were obtained for the female animals. Pharmacokinetic parameters for ropinirole were similar after single and once daily repeat oral administration of the drug to the mouse for 60 days.

In rat and monkey, data obtained after intravenous administration indicated that the blood clearance of ropinirole was high in both species 3.12 and 1.4 (L/h)/kg, respectively. Since the major route of excretion of total drug-related material is via the urine, with ropinirole accounting for ≤ 1% of this material after oral administration in rat and monkey, the high clearance values would indicate that ropinirole undergoes substantial first-pass metabolism in both species. Apparent terminal phase plasma elimination half lives of ropinirole were about 0.5 and 1.3 h in rat and monkey, respectively, although there was evidence of a second elimination phase in the monkey with a mean half life of about 5 to 11 h.


In rat and monkey at acute oral doses of between 1.5 and 50 mg/kg and 1.5 and 15 mg/kg, respectively, the extensive first-pass metabolism was saturable leading to non-linear pharmacokinetics of ropinirole and its major metabolite in each species. Additionally in the monkey the mean ratio of ropinirole to metabolite decreased with increasing dose. Based on Cmax and AUC values, the systemic exposure to orally administered ropinirole increased more than proportionately to the increase in dose. Mean Cmax at 1.5, 5.0 and 15 mg/kg in monkeys was 1.94, 20.5 and 313 ng/mL with a corresponding AUC0-inf of 3.8, 43.8 and 856 ng.h/mL. Comparison of the AUC values with those obtained after IV infusion at 1.5 mg/kg indicated the oral bioavailability of ropinirole at 1.5 mg/kg was low (0.5%). Pharmacokinetic parameters for ropinirole and SK&F 104557 were similar after single and once daily repeat oral administration of the drug to the monkey for 12 months. In the rat, however, AUC values for ropinirole following once daily oral administration were up to 16-fold higher on Day 14 compared to those on Day 1.


Table: Systemic exposure to ropinirole, SK&F 104557 and SK&F 89124 following oral administration of ropinirole to mice, rats, cynomolgus monkeys and humans

Species Dose Systemic (plasma) exposure (mg/kg/day) Ropinirole SK&F 104557 SK&F 89124 Cmax

(ng/mL) AUC

(ng.h/mL)* Cmax

(ng/mL) AUC

(ng.h/mL)* Cmax

(ng/mL) AUC

(ng.h/mL)* Mouse 8.8 68.9 (1.9) 72.7 (0.2) NA NA NA NA 21.9 267 (7.3) 207 (0.4) NA NA NA NA 43.8 430 (11.7) 325 (0.6) NA NA NA NA Rat 1.5 ND ND ND ND 2.11 9.77 15 ND ND ND ND 10.3 53.5 50 479 (13.0) 1580 (2.8) 281 (9) 1320 (2) 55.1 (42) 198 (10) Monkey 1.5 5.58 (0.2) 15.9 (<0.1) 132 893 ND ND 5 24.8 (0.7) 69.3 (0.1) 464 2420 ND ND 15 184 (5.0) 511 (0.9) 2930 (89) 11500 (19) ND ND Human 0.48† 36.7 557 33.0 605 1.3 19.2 Data presented are for male and female animals and are after daily repeated oral administration (at the end of the 60-day mouse study, 14 day rat study, and 12 month monkey study. Data for humans extrapolated from dose normalised data obtained in male and female patients following tid regimen). * AUC0-6 in the mouse, AUC0-t in the rat and monkey, dose normalised AUC0-t x 24 in human. ND - Not detected. NA - SK&F 104557 and SK&F 89124 were not assayed in mouse plasma. † Calculated from a total daily dose (8mg tid) assuming a body weigh of 50 kg for man. Figures in parentheses represent ratios of exposure in animals to those in Parkinsonian patients at 0.48 mg/kg/day


Investigation of intravenous infusion to male anaesthetised beagle dogs of ropinirole and SK&F 89124A at 1.0 mg/kg showed apparent terminal phase elimination half life and plasma clearance of approximately 2h and 1.14 (L/h)/kg, respectively for ropinirole and approximately 2 h and 1.98 (L/h)/kg for the metabolite. Emesis prevented pharmacokinetic profiling of ropinirole following oral administration in the dog.

Investigation in the pregnant rabbit was limited, however, data showed that concentrations of SK&F 104557 were higher than ropinirole and that the concentrations of both compounds increased with increasing dose. SK&F 89124 was not generally detected.

Distribution

Ropinirole exhibited a large volume of distribution in rat and monkey (2.0 and 2.7 L/kg, respectively) indicating that the compound is distributed in excess of total body water. Ropinirole and, to a lesser extent, SK&F 89124 were shown to cross the blood brain barrier following intravenous administration to the rat and oral administration to the monkey. SK&F 104557, SK&F 97930 and SK&F 96990 were not detected in the CNS of the monkey following intravenous administration of ropinirole.

In the rat, following intravenous administration, drug-related material was rapidly taken up by all tissues including the CNS. The distribution after oral administration was similar to intravenous, although due to limitations of the radiometric techniques used, drug-related material was not detected in the CNS or the eye. Apart from the gut, highest concentrations were associated with some exocrine and endocrine glands after intravenous dosing, and in the liver and urinary bladder after oral dosing. In pigmented rats radioactivity following both intravenous and oral administration was higher in pigmented tissue (skin and eye). After either route of administration, radioactive material was generally cleared from the tissues (with exception of pigmented tissue) quite rapidly and by 168 h, only traces of drug-related material were detected in liver and the contents of the gut. Traces of drug related material were found in pigmented tissue up to 28 days after dosing.

Following oral administration of 14C ropinirole to pregnant rats, whole body autoradiography revealed drug-related material distributed throughout all maternal and fetal tissue at 2 h after dosing, indicating placental transfer had occurred. Ropinirole, SK&F 104557 and the glucuronide of SK&F 89124 were tentatively identified as the major drug-related components present in maternal plasma, amniotic fluid and fetal tissue. Ropinirole-related material was also shown to transfer into the milk of lactating rats in small amounts (approximately 0.01% of the dose per pup).

Plasma protein binding of ropinirole (9 to 5800 ng/mL) and SK&F 89124 (6 to 1300 ng/mL) in rat, dog and human and ropinirole (0.03 to 3000 ng/mL) and SK&F 89124 (70 to 2100 ng/mL) in the monkey was low (10 to 39%) and independent of concentration. Drug interactions due to displacement of drug from plasma proteins, and alterations in the pharmacokinetics of ropinirole as a result of changes in binding proteins in disease states would therefore not be expected. In vitro blood to plasma ratios for ropinirole and SK&F 89124 in rat, dog, monkey and human were essentially independent


of the concentration used and ranged from 0.94 in the rat to 1.25 in the primate. In an in vitro study, ropinerole (1 µM) was shown not to be a substrate for human P-glycoprotein.

Metabolism

Phase I metabolism comprised two pathways. In rat, the major metabolic pathway was via hydroxylation at position 7 to form SK&F 89124. In mouse, cynomolgus monkey and marmoset the major pathway was via N-depropylation to form SK&F 104557, which was further metabolised to a limited extent to the pharmacologically inactive metabolites SK&F 96690 (7-hydroxy SK&F 104557) and SK&F 97930 (carboxylic acid derivative of SK&F 104557). In rat and monkey, as the dose increased, the alternative phase I pathway became more significant (i.e. N-depropylation for the rat and hydroxylation for the monkey). The metabolites formed by either pathway were in some cases metabolised further by glucuronidation (Phase II metabolism) in all species.

In vitro studies in human microsomes have demonstrated that the main enzyme responsible for the metabolism of ropinirole is the cytochrome P450 (CYP) enzyme CYP1A2, with CYP3A making a minor contribution.

Ropinirole was shown not to inhibit cytochrome P450 activities in human liver, with the exception of CYP2D6 for which 50% inhibition occurred at concentrations approximately 130 fold greater than the Cmax at the maximum clinical dose (8.0 mg TID). In the rat at doses more than 100-fold higher than the maximum clinical dose, the compound caused some induction of drug metabolising enzyme activity, but the level of induction was considerably less than that of classic inducers of cytochrome P450, such as phenobarbitone and beta-napthoflavone. Overall it is unlikely that ropinirole will inhibit or induce these enzymes in man.

Excretion

The major route of excretion of drug-related material was renal after either oral or intravenous administration of ropinirole to mice (86 to 89% of dose), rats (57 to 72% of dose), dogs (40 to 50% of dose) and monkeys (72% of dose). The majority of the remaining dose was excreted in the faeces, and total recoveries of material were in excess of 92% of the administered dose. In bile duct cannulated rats, 76%, 24% and <1% of the dose was recovered in bile, urine and faeces, respectively over 48 hours. The majority of the recovered radioactivity was excreted within 48 hours. Following oral administration, unchanged ropinirole accounted for only a small proportion of the dose (≤1%) in the urine of the mouse and the monkey. Ropinirole was not detected in the urine of rats at low oral doses.

Toxicology

The toxicity profile of ropinirole has been investigated in a range of single- and repeat-dose oral, intraperitoneal and/or intravenous studies of up to 12 months duration in the rat, mouse and monkey. Effects seen in these studies consisted primarily of exaggerated


pharmacological changes related to central and peripheral dopamine receptor stimulation. These included hyperactivity, stereotypic behaviour, convulsions and, in rodents at oral dosages ≥50 mg/kg, deaths.

Genotoxicity was not observed in a battery of in vitro and in vivo tests. Two-year carcinogenicity studies did not reveal any carcinogenic effects in the mouse or rat; findings were limited to an increased incidence of benign endometrial polyps in female mice, and species-specific Leydig cell hyperplasia and adenoma of the testes in rats resulting from the hypoprolactinaemic effect of ropinirole. In reproductive toxicology studies in the rat, ropinirole had no effect on male or female fertility; embryotoxicity was seen at ≥60 mg/kg/day and digit malformations at 150 mg/kg. In the rabbit there were no embryotoxic effects at up to 20 mg/kg/day, and no effects on peri- and post-natal parameters were seen in the rat at up to 10 mg/kg/day.

These studies reveal no major toxicological liabilities that should preclude the clinical use of ropinirole in a controlled manner.

Single dose toxicity

Single oral or intravenous doses of ropinirole given to mice and rats produced clinical signs associated with central D2 agonist effects, i.e. hyperactivity, abnormal locomotion, stereotypy, tremors and convulsions. The approximate median lethal dose was 862 mg/kg (rat) and 657 mg/kg (mouse) orally and approximately 66 mg/kg (rat) and 46 mg/kg (mouse) intravenously. These studies did not reveal any histological evidence of toxicity at 14 days post dose. Effects of the metabolites SK&F 104557, SK&F 89124 and SK&F 97930 at doses up to 10 mg/kg IV were also investigated in the mouse or rat. There were no clinical signs or target organ toxicity with SK&F 104557 or SK&F 97930 in the mouse. Transient clinical signs consistent with centrally and peripherally mediated D2 agonist activity were observed with SK&F 89124 in the rat.

Following acute oral dosing in the monkey, the minimum lethal dose of ropinirole was 702 mg/kg. Clinical signs were primarily of CNS stimulation, and there were no drug-related macroscopic findings in any of the treated animals.

Repeat dose toxicity

Oral administration of ropinirole at doses of up to 600 mg/kg (oral sighting study) and 219.3 mg/kg (two and three month studies) in mice revealed pharmacologically-mediated D2-receptor agonist associated clinical observations at doses above 25 mg/kg. These observations included ptosis, subdued behaviour, tremors, convulsions and death. Convulsions often preceded death, which occurred at doses of ≥43.5 mg/kg. Longer duration of treatment was associated with deaths at lower doses. There were no drug induced macroscopic or microscopic lesions associated with treatment.

Repeat dose oral toxicity studies in the rat were performed using doses of up to 625 mg/kg in dose sighting studies (14 and 14 to 57 day), doses of up to 548 mg/kg in a 14 day study and doses of up to 250 mg/kg, 125 mg/kg and 100 mg/kg in 1, 6 and 12 month studies, respectively. Clinical signs included kneading, toe-walking,


hyperactivity, ptosis, salivation and stereotypic preening/biting behaviour. Convulsions were dose-exposure and age-dependent. In the 12 month study, drug related mortality was seen at 100 mg/kg and was preceded by convulsions in approximately 30% of cases. Overall, doses of 50 mg/kg and below were well tolerated in the 12 month study (although there were 4/40 deaths at 50 mg/kg during the 6 month study). In rat studies ranging from 14 days to 12 months increases in liver weight of up to 26%, slightly higher alkaline phosphatase and alanine aminotransferase values (1.5- to 2 – fold, transient in the six month and absent in the 12 month study) and liver cell hypertrophy (associated with an increase in smooth endoplastic reticulum in hepatocytes) were observed. These changes were considered to represent an adaptive change attributable to enzyme induction. The above changes were seen at doses of ≥50 mg/kg in the 12 month study and above 250 mg/kg in studies of shorter duration. Another liver effect noted in the 12 month study was an increased incidence of foci of hepatocellular alteration in drug-treated females. The significance of this finding in relation to ropinirole treatment is questionable, as there was no evidence of a drug-related increase in the incidence of hepatocellular tumours in the 12 month study or in the carcinogenicity study.

Gastric erosion/ulceration was infrequently seen in various rat studies at doses of ≥50 mg/kg, irritation being the most likely explanation for this finding. Reactive hyperplasia and ulceration of the urinary bladder epithelium were observed infrequently in rats treated with doses of 250 mg/kg ropinirole and above. This finding is considered a high dose effect in the rat and not relevant to humans.

Ropinirole induced dose-related hypoprolactinaemia in the rat. The inhibitory effect of ropinirole on prolactin secretion resulted in a species specific increase in ovarian weight associated with an increased number of corpora lutea (≥10 mg/kg). There was also a higher frequency of uteri in pro-oestrus, acidophilic glandular acini in female mammary glands, adrenocortical hypertrophy in females and prominent gonadotrophs in male pituitaries (all these changes regressed or were absent after a 6 week recovery period). In the 12 month study an increase in Leydig cell hyperplasia (LCH) of the testes was again thought to be due to the prolactin reducing effect of ropinirole in males [Sharpe, 1979]. Unlike rat, prolactin receptors are not present on human Leydig cells, and so LCH is not a risk to man [Wahlström, 1983]. Females of the 50 and 100 mg/kg group showed an increased oestrogen/progesterone ratio, confirmed by histological changes comprising increased ovarian follicle/corpus luteum ratio, endometrial hyperplasia and vaginal cornification. Increased adrenal weights in both sexes (≥50 mg/kg) may have resulted from adrenocorticotrophic hormone release as a consequence of the increase in oestrogen/progesterone ratio.

Repeat dose oral studies (10 day, 1, 6 and 12 month) in the cynomolgus monkey resulted in clinical signs ranging from hyperresponsiveness and stereotypy to ataxia, formication, convulsions and self-mutilation. Convulsions were not seen at or below 60 mg/kg, but other signs were seen in both sexes at all doses ranging from 5.0 to 240 mg/kg. In the 12 month study with dosing up to 15 mg/kg/day observations were limited to centrally-mediated dopaminergic action of ropinirole, i.e. stereotypy, post-dose salivation, formication and emesis. Limiting CNS-related toxicity, consistent with the dopaminergic properties of ropinirole were observed at 30 mg/kg/day [6 month oral toxicity study (dose increased from 15 mg/kg during week 8)]. High-dose males in the 12 month study


(15 mg/kg/day) gained less weight than the controls (24% versus 38%) which may have been due to increased activity as often seen with dopamine agonists. Mean serum prolactin concentration was decreased in 5.0 and 15 mg/kg/day females and significantly decreased in 15 mg/kg/day males. There was no evidence of a drug-related ophthalmologic or electrocardiographic effect. An increase in organ weights was recorded for adrenals in high dose 12 month study animals and testes and epididymes in high-dose males. These weight increases were not associated with any morphological alteration. There were no drug-related macroscopic or microscopic changes.

Intravenous repeat dose 14 day studies at doses of 0.5, 2.0 and 10 mg/kg/day in rats and 0.5, 1.0 and 2.0 mg/kg in monkeys resulted in CNS-related clinical signs without any histological evidence of target organ toxicity.

The following table shows the animal to human exposure ratios obtained at well tolerated dose levels in the longer-term toxicity studies, 15, 25 and 50 mg/kg/day in the monkey, mouse and rat, respectively. Comparisons are made at the maximum clinical dose for both Parkinson’s disease (8 mg TID or 0.48 mg/kg/day) and RLS (4 mg/day or 0.08 mg/kg/day).

Species Well-tolerated Dose

(mg/kg/day)

AUC (ng.h/mL)

Cmax (ng/mL)

Animal to Human Exposure Ratio* PD / RLS

AUC Cmax

Mouse 25 207 267 0.4 / 3.5 7.3 / 46

Rat 50 1580 479 2.8 / 27 13 / 82

Monkey 15 511 184 0.9 / 8.8 5.0 / 32

* Calculated using predicted human AUC and Cmax values for the IR tablet formulation at the maximum clinical dose for Parkinson’s disease of 8 mg TID (557 ng.h/mL and 36.7 ng/mL, respectively), or actual human data following repeat dosing with the IR formulation at the maximum clinical dose for RLS of 4 mg/day (58.4 ng.h/mL and 5.83 ng/mL, respectively).

Genotoxicity

In vitro studies showed ropinirole was not genotoxic in an Ames test with or without metabolic activation at concentrations up to 5000 µg/plate. Ropinirole did not cause point (gene) mutation in two strains of Escherichia coli with or without metabolic activation up to 5000 µg/plate. In a chromosome aberration test in human lymphocytes ropinirole did not induce aberrations in the absence or presence of metabolic activation when tested up to a concentration of 5000 µg/mL. In a gene mutation test in mouse lymphoma cells, ropinirole failed to induce reproducible increases in mutation either in the absence or presence of metabolic activation at doses up to 5000 µg/mL.


In a mouse micronucleus test at oral doses of 100, 200 and 400 mg/kg, ropinirole was not able to induce micronuclei in the bone marrow of mice, despite the highest dose being clearly toxic (since a number of animals died).

Carcinogenicity

In rat and mouse carcinogenicity studies ropinirole was administered at doses of 1.5, 15, and 50 and 5.0, 15, and 50 mg/kg/day, respectively. No carcinogenic effect was observed in the mouse. Increased incidences of Leydig cell hyperplasia and adenoma were noted in the testes of drug-treated rats. Although Leydig cell hyperplasia was observed at all doses, the no effect dose for Leydig cell adenomas was 1.5 mg/kg. The proliferative effect on Leydig cells as seen in rats treated with ropinirole is believed to be a rat-specific epigenetic phenomenon resulting from the prolactin-lowering effect of ropinirole. It is well established that hypoprolactinaemia in rats is associated with a reduction in the number of luteinizing hormone (LH) receptors on the Leydig cell. With regard to the potential risk for induction of proliferative Leydig cell lesions in humans by ropinirole, it has been shown that the Leydig cell of man does not express the prolactin receptor. Therefore, it is extremely unlikely that the mechanism described above for the rat can operate in humans.

Incidences of retinal atrophy were also increased at a dose of 50 mg/kg/day. This was considered to be light induced and attributable to an indirect effect of dopaminergic stimulated hyperactivity and mydriasis rather than a result of direct toxicity to the retina. A follow up study investigating the effect of different light intensities on the retina of both pigmented and albino rats showed no retinal atrophy in pigmented rats with or without ropinirole treatment (100 mg/kg/day) for three months. Retinal atrophy in the albino rats was intensity related but there was no statistically significant effect of ropinirole on the severity of light-induced retinal atrophy.

Reproductive and Developmental Toxicity

Fertility and early embryonic development

As the rat depends on prolactin prior to and after implantation, rat studies were specially designed to avoid interruption of pregnancy and lactation as a consequence of the hypoprolactinaemia induced by ropinirole. Doses up to 100 mg/kg (maternally toxic) were used in the female fertility study but sub or low pharmacological doses were used during the prolactin-dependent phase [pregnancy days 0 to 8 (5 mg/kg) and lactation (5, 10 and 20 mg/kg)]. Doses were determined from various oral reproductive preliminary studies. Using such a dosing regimen, ropinirole at doses up to 100 mg/kg did not adversely affect fertility, mating performance or pregnancy rate. At doses of ≥50 mg/kg, drug-related decreases in neonatal body weight gain and a delay in some aspects of physical and behavioural development was observed. No effects were seen on male fertility at doses up to 125 mg/kg, although the expected dose-related CNS toxicity was seen and treatment-related mortalities were seen in male rats treated with 125 mg/kg.


Embryofetal development

In the main rat embryofetal development study, a dose of 20 mg/kg was administered to all groups on days 6 and 7 of pregnancy and doses of 20, 60, 90 120 and 150 mg/kg on days 8 to 15 of pregnancy. Doses were again determined from preliminary studies and were chosen to limit the species specific hypoprolactinaemia effect. Embryotoxicity, considered to be due to maternal toxicity, was indicated by lower fetal weight at 60 mg/kg/day and above and increased post-implantation loss at 90 mg/kg/day and above. Digit malformations (cause unclear) were observed at 150 mg/kg/day.

Due to maternal deaths occurring during a preliminary study in the 50, 75, 100 and 150 mg/kg/day dose groups (33, 50, 50 and 100% respectively) and increased post-implantation death from 25 mg/kg/day, doses for the main developmental toxicology study in the rabbit were limited to 1.0, 5.0 and 20 mg/kg, administered from Days 6 to 18 of gestation. Maternal but no developmental toxicity was observed in rabbits at oral doses up to 20 mg/kg.

Systemic exposure data were not generated during the embryofetal development studies with ropinirole, and it is therefore not possible to correlate effects on the fetus directly with systemic exposure. However, for the rat, estimates of the minimum safety margins for systemic exposure can be obtained by extrapolation of data generated from a 14 day repeat dose pharmacokinetic study in non-pregnant females, assuming no effect of pregnancy on systemic exposure. These data are shown in the following table.

Dose (mg/kg/day)

AUC (ng.h/mL)

Animal to Human Exposure Ratio* PD / RLS

60 1800 1.2 / 16 90 2700 1.9 / 25

120 3600 2.5 / 33 150 4500 3.1 / 41

* AUC values in rats have been extrapolated from data obtained at 50 mg/kg/day in non-pregnant females in a 14 day repeat dose pharmacokinetic study. Human AUC data are the maximum predicted or achieved following the highest clinical dose for Parkinson’s disease and RLS of 8 mg TID and 4 mg/day respectively, using the IR formulation (1446 and 110 ng.h/mL, respectively).

In a further study, ropinirole (10 mg/kg), L-dopa (250 mg/kg) or the combination of these were given once daily by oral gavage to pregnant rabbits on Days 6 to 20 of gestation. The combination of the two compounds was maternally toxic in the rabbit as evidenced by abortion and death and produced a higher incidence of behavioural alterations, particularly motor disturbances, altered startle response and unkempt appearance when compared to treatment with ropinirole or L-dopa alone. L-dopa also produced growth retardation and fetal malformations, particularly of the digits. Ropinirole did not produce developmental toxicity and did not augment the effects of L-dopa on embryo-fetal development in the rabbit.


Prenatal and Postnatal development

In the pre and postnatal rat study dosing was limited to 0.1, 1.0 and 10 mg/kg based on data from a preliminary dose range study in pregnant and lactating rats. Maternal effects consistent with the drug's dopaminergic activity were seen at 10 mg/kg/day. Offspring growth retardation during the pre-weaning period was attributed to maternal hypoprolactinaemia/reduced lactation at 10 mg/kg/day. Female offspring's startle response to auditory and tactile stimulation at age 29 days was reduced by the maternal dose of 1.0 and 10 mg/kg/day. Only females from the 10 mg/kg/day group had the reduced startle reflex at sexual maturity. Ropinirole had no adverse effects on parturition, offspring number or viability or offspring's physical development, learning, memory, motor activity or reproductive performance at doses up to 10 mg/kg/day.

Local tolerance

Local tolerance studies revealed that an IV dose of ropinirole in the dog, at the maximum dose of 5 µg/mL proposed for use in human did not cause vascular or perivascular irritation at the injection site. In addition, ropinirole was a non-sensitiser to guinea pig skin and classified according to the EC labelling regulations as non-irritant to rabbit skin and eye.

Other toxicity Studies

A 10 mg/mL solution of ropinirole did not cause haemolysis when mixed with human blood in vitro. Furthermore, ropinirole did not cause platelet aggregation in vitro and inhibited stimulated platelet aggregation at a dose of 0.9 mg/mL.


TOXICOLOGY RELATING TO THE SPECIFIC FORMULATION


To provide a nonclinical rationale for use of the ropinirole PR formulation in Parkinson’s disease, an MPTP marmoset study was conducted to assess whether continuous release of ropinirole via an osmotic mini-pump provided an advantage over oral pulsatile administration of ropinirole on measures of anti-parkinsonian activity and dyskinesia. Drug-naïve marmosets were treated with MPTP, and once motor deficits had been verified and stabilised they were randomly allocated to two treatment groups. One received ropinirole (0.4 mg/kg BID) administered by oral gavage for 14 days, the other ropinirole (0.8 mg/kg/day) administered via subcutaneous osmotic mini-pump for 14 days. Following this initial treatment period, the marmosets were switched to the other treatment in order to assess the effect of mode of administration on individual response. For the first 14 day phase, MPTP marmosets assigned to the continuous administration of ropinirole demonstrated significant sustained improvements in locomotor activity and reductions in disability compared to oral pulsatile administration. No significant dyskinesias were noted in the continuous ropinirole-treated group, while mild or moderate dyskinesias were seen in some animals treated with oral pulsatile ropinirole. Following the switching of administration for a further 14 days, continuous ropinirole treatment administered to those previously treated orally, resulted in a sustained improvement in disability scores, while dyskinesias that were present in a few animals following oral treatment disappeared after 1 week of constant infusion. For those animals switched from continuous to oral pulsatile ropinirole, locomotor activity was sustained but improvements in disability scores were not maintained. No significant dyskinesias were demonstrated when switching from continuous to oral pulsatile ropinirole.

This study showed that continuous release of ropinirole could significantly improve locomotor activity, and reduce disability and the incidence of dyskinesias in drug naïve MPTP marmosets. The data provided indirect evidence that the continuous dopaminergic stimulation offered by the ropinirole PR formulation would improve efficacy in Parkinson’s disease patients relative to pulsatile ropinirole IR administration.

The manufacturing process for ropinirole provides material of high purity (>98%). There are five main organic impurities, all of which were present in batches of compound used in the toxicological evaluation of ropinirole. The levels of these impurities are routinely monitored, as are those of other organic compounds recognised as potential impurities. Three of these main impurities are also degradation products detected on storage of the tablet form. For the PR formulation, the main degradation product is SB-270341, a formaldehyde adduct, which is not present in the IR formulation. The end-of-shelf-life specification limit for this impurity is in line with the ICH qualification threshold of 0.5%. Thus the impurity and degradation profiles of ropinirole and the limits defined for impurities give no cause for concern regarding the safety of the final product.

All of the excipients used in the formulation for Ropinirole PR Tablets (with the exception of the film coating) are generally recognised constituents of pharmaceutical preparations and are of pharmacopoeial grade. The formulations for the four colour film coats are prepared to company specifications which are of a suitable grade for


pharmaceutical preparations. Additionally all individual constituents used in production of the film coatings are of pharmacopoeial grade. All of the excipients are considered safe for use in the preparation of products for oral administration and there are no toxicological implications associated with their use in this product.

CONCLUSIONS

The dopamine agonist, ropinirole, is an effective treatment for Parkinson's disease both as monotherapy in the early stages of the disease or as adjunctive treatment with L-dopa in later stage development. Ropinirole has also been shown to be effective in the treatment of Restless Legs Syndrome (RLS). Importantly, ropinirole appears to have a remarkably low propensity to produce dyskinesias, and in preclinical studies, dyskinesia is even reduced or reversed when switching from L-dopa monotherapy to ropinirole monotherapy. Extensive nonclinical safety studies with ropinirole show the adverse event profile to be similar to that of other dopaminergic drugs, producing no additional unexpected problems. The nonclinical safety data therefore support the safe clinical use of Ropinirole PR Tablets for the treatment of Parkinson’s disease. This has been substantiated by the clinical safety profile of ropinirole, since its launch in 1996, which shows that ropinirole has an acceptable safety profile when used in the treatment of both Parkinson's disease and RLS. In addition, ropinirole does not appear to produce the ergot-associated side effects that occur rarely with some other currently used dopamine agonists.

In the last 5 years, additional nonclinical studies have been performed to summarise the distribution effects of ropinirole in a 6-hydroxydopamine rat model of Parkinson’s disease, and to determine whether ropinirole is a substrate for the human P-glycoprotein transporter. These additional investigations have generated no further information that would alter the original conclusions regarding the safety of Ropinirole Immediate Release Tablets in the clinic or altered the risk-benefit evaluation for this formulation when prescribed under the recommended therapeutic dosage regimen.

Recommendations for Nonclinical Statements to be Addressed in the Labelling.

Pharmacodynamics

Mechanism of Action

Ropinirole is a potent non-ergoline D2/D3 dopamine agonist.

Parkinson's Disease is characterised by a marked dopamine deficiency in the nigral striatal system. Ropinirole alleviates this deficiency by stimulating striatal dopamine receptors.


Pharmacodynamic Effects

Ropinirole acts in the hypothalamus and pituitary to inhibit the secretion of prolactin.

Pregnancy and Lactation

Fertility

See ‘Nonclinical Information’.

Pregnancy

It is recommended that ropinirole is not used during pregnancy unless the potential benefit to the patient outweighs the potential risk to the fetus (see Non-clinical Information).

Lactation

Ropinirole should not be used in nursing mothers as it may inhibit lactation.

Nonclinical Information

Carcinogenesis, mutagenesis

Two year studies have been conducted in the mouse and rat at dosages up to 50 mg/kg. The mouse study did not reveal any carcinogenic effect. In the rat, the only drug related lesions were Leydig cell hyperplasia/adenoma in the testes resulting from the hypoprolactinaemic effect of ropinirole. These lesions are considered to be a species specific phenomenon and do not constitute a hazard with regard to the clinical use of ropinirole.

Genotoxicity was not observed in a battery of in vitro and in vivo tests.

Reproductive toxicology

In fertility studies in rats, effects were seen on implantation due to the prolactin-lowering effect of ropinirole. In humans, chorionic gonadotropin, not prolactin, is essential for implantation in females. No effects were seen on male fertility.

Administration of ropinirole to pregnant rats at maternally toxic doses resulted in decreased foetal body weight at 60 mg/kg, increased foetal death at 90 mg/kg and digit malformations at 150 mg/kg. There was no teratogenic effect in the rat at 120 mg/kg and no indication of an effect on development in the rabbit. There have been no studies of ropinirole in human pregnancy.


Animal toxicology and/or pharmacology

Ropinirole caused no serious or irreversible toxicity in laboratory animals at 15 mg/kg (monkey), 20 mg/kg (mouse) or 50 mg/kg (rat). The toxicology profile is principally determined by the pharmacological activity of the drug (behavioural changes, hypoprolactinaemia, and decrease in blood pressure and heart rate, ptosis and salivation).


LIST OF LITERATURE CITATIONS

Carta AR, Frau L, Pontis S, Pinna A, Morelli M. Direct and indirect striatal efferent pathways are differentially influenced by low and high dyskinetic drugs: Behavioural and biochemical evidence. Parkinsonism and Rel Dis. 2008;14: S165-S168.

Humphrey SJ, Turman CN, Curry JT, Wheeler GJ. Cardiovascular and electrocardiographic effects of the dopamine receptor agonists ropinirole, apomorphine, and PNU-142774E in conscious beagle dogs. J Cardiovasc Pharmacol 2006;47 :337-47.

Hurst RS, Higdon NR, Lawson JA, Clark MA, Rutherford-Root KL et al. Dopamine receptor agonists differ in their actions on cardiac ion channels. Eur J. Pharmacol 2003; 482: 31-37.

Maratos E, Jackson MJ, Pearce RKB, Jenner P and Marsden CD. Dyskinesia induction in MPTP-treated common marmosets following repeated treatment with combinations of L-DOPA and ropinirole. Mov Disord 1998; 13: 159.

Ogawa N, Miyazaki I, Tanaka K, Iida M and Asanuma M. Dopamine D2 receptor mediated antioxidant and neuroprotective effects of ropinirole. Parkinsons Dis Rel Disord 1999; 5: S81.

Pearce RKB, Banerji T, Jenner P and Marsden CD. Effects of repeated treatment with L-DOPA, bromocriptine and ropinirole in drug-naive MPTP-treated common marmosets. Br J Pharmacol 1996; 188: 37P.

Rose S, Warren D, Walgama O and Jenner P. Ropinirole and bromocriptine prevent MPP+-evoked hydroxyl radical formation in the rat striatum in vivo. Br J Pharmacol 1997; 122: 280P.

Sharpe RM and McNeilly AS. The effect of induced hyperprolactinaemia on leydig cell function and LH-induced loss of LH-receptors in the rat testis. Mol Cell Endocrinol 1979; 16: 19-27.

Tanaka K, Miyazaki I, Fujita N, Haque ME, Asanuma M and Ogawa N. Molecular mechanism in activation of glutathione system by ropinirole, a selective dopamine D2 agonist. Neurochem Res 2001; 26: 31-6.

Wahlström T, Huhtaniemi I, Hovatta O and Seppälä M. Localization of luteinizing hormone, follicle-stimulating hormone, prolactin and their receptors in human and rat testis using immunohistochemistry and radioreceptor assay. J Clin Endocrinol Metab 1983; 57: 825-30.


LIST OF NONCLINICAL STUDIES

Ropinirole Hydrochloride = SKF-101468

Report No. Study No. Title

M4.2.1 PHARMACOLOGY

M4.2.1.1 Primary Pharmacodynamics

PH-1004/SKF-101468/2

- Characterisation of the binding of ropinirole, its major metabolites and other D agonists to rat and human D2 and D3 receptors.

PH-1005/SKF-101468/2

- Inhibition by ropinirole of radioligand binding to a synthetic version of the human D4.4 receptor expressed in 293 cells.

PW 023 BA - Interaction of SK&F 101468-A and other DA-agonists with receptor binding sites.

PW 019 BA - Receptor binding studies with SK&F 101468 in rat and bovine brain membranes: an assessment of data provided by Nova Pharmaceutical Corporation, Baltimore, USA.

PH/0001/SKF-101468

- Effects of SK&F 101468 and its major metabolites SK&F 89124 and SK&F 104557 on CNS, dopamine receptors in vitro, assessed using D1 and D2-receptor binding assays and D2-receptor regulation of acetylcholine release.

FP/0001/SKF-101468

- Effect of SK&F 101468 (ropinirole) and SK&F 89124 on rat caudate adenylate cyclase as an index of central D1-agonist activity

PH-0005/SKF-101468/2

- A behavioural analysis of the dopamine agonists, amphetamine, SKF 81297, SKF 83565, SKF 38393, ropinirole (SKF 101468) and PHNO, and the dopamine antagonists, SCH 23390 and BRL 34778 in rats trained to discriminate (+)-amphetamine from active or SKF 81297 from active.

PP 011 BA - Interaction of SK&F 101468-A, SK&F 89124-A and SK&F 104557-A with central adrenergic receptors.



PW 033 BA - Interaction of SK&F 101468-A and other dopamine agonists at opioid binding sites.

PW 005 BA - A centrally acting dopamine agonist having anti-Parkinson, antidepressant and anxiolytic activity.

PW 026 BA - Potential of SK&F 101468-A to interact with amitriptyline and diazepam.

PW 018 BA - The anxiolytic and antidepressant activities of SK&F 101468-A: Comparison with bromocriptine.

PWO30BA - Maintenance of the anxiolytic action of SK&F 101468-A on chronic treatment, and lack of problems on withdrawal from chronic treatment.

SKF-101468/RSD-100L4R/1

A study of the functional potency and selectivity of ropinirole determined by microphysiometry, at human dopamine D2(long), D3, and D4.4 receptors.

PW 032 BA - Dopamine agonist action of SK&F 101468-A in mouse striatum demonstrated after unilateral 6-OHDA lesions of the substantia nigra: lack of tolerance on chronic treatment.

PW 015 BA - Action of SK&F 101468-A in mice with unilateral 6-OFIDA lesions of the substantia nigra.

PH- 1003/SKF-101468/2

- A study of the effect of ropinirole, its major metabolites, and other D-agonists on free radical-induced lipid peroxidation.

PH-1001/SKF-101468/1

- The effects of ropinirole (acute and chronic administration - 0, 0.03, 0.1, 0.3, 1, 3, 10 mg/kg p.o. and i.p.) and bromocriptine (acute administration - 0, 0.1.0.3,1,3, l0 and 30mg/kgi.p) on neurotransmitter turnover in vivo in mice.

PW 034 BA - The effects of SK&F 101468-A on locomotor activity, stereotyped behaviour and anxiety in the rat in the presence and absence of naloxone.

PH-1002/SKF-101468/2

- The effect of ropinirole (SK&F 101468) and its metabolites SK&F 89124, SK&F 104557, SK&F 97930 and SK&F 96990 in the in vivo circling rat



model of Parkinson's disease.

TF-1002/SKF-1014681/1

- Effects of acute, oral administration of SK&F 101468A on core temperature in the conscious rat.

PW 027 BA - The ability of SK&F 101468-A to inhibit dyskinesias induced by 2-di-n-propylamino-5,6-dihydroxytetralin in the rat.

PW 016 BA - Action of SK&F 101468-A in a model of Parkinson’s disease induced by 1 -methyl-~phenyl-1,2,3,6-tetrahydropyridine in the common marmoset.

PW 022 BA - The effects of SK&F 101468-A on behavioural deficits induced by I -methyl-4-phenyl- 1,2,3,6-tetrahydropyridine (MI~) in the common marmoset (Caliithrix jacchus). A comparison with L-Dopa and bromocriptine.

PH/0002/SKF-101468/2

- The effects of SKF 104557A on locomotor behaviour in the rat and marmoset.

PW 028 BA - The effects of chronic administration of SK&F 101468-A in the MHP-treated common marmoset (Callithrix jacchus).

PW 029 BA - The effect of SK&F 101468-A in the human threat test of anxiety in the common marmoset (Callithrix jacchus).

PW 017 BA - Action of SK&F 101468-A on the motor impairment caused by MPTP in the common marmoset

(subcutaneous dose response relationship).

PH-1006/SKF-101468/1

- Effect of ropinirole on nigrostriatal dopamine neurons in the rat: an extracellular single cell study in vivo.

PP-1001/SKF-10146811

- Japanese General Pharmacology Studies.

M4.2.1.2 Secondary Pharmacodynamics

PP 008 BA - Activity of SK&F 8912~A in several in vitro



receptor assays.

PP 006 BA - Effect of SK&F 89124-A on neurotransmitter release in canine vascular tissue.

PP 009 BA - Effect of SK&F 101468-A on atrial contractility in vitro.

PW 008 BA - A comparison of the effects of SK&F 101468-A with propranolol on cardiac function in vitro.

PW 013 BA Pharmacological studies, in vitro and in vivo with SK&F 89124-A, an active metabolite of SK&F 101468-A.

SKF-101468/RSD-1004W7/1

The effects of SK&F 101468-A (ropinirole) in the Geller-Seifter model of anxiety

HH2003/00010/00 An evaluation of ropinirole (SKF-101468-A) in the SOD-1 transgenic mouse model of ALS.

HH2003/00009/00 An investigation of the ability of ropinirole to reverse L-DOPA induced dyskinesia in MPTP-treated common marmosets.

HH2004/00105/00 Investigation of the antiparkinsonian activity and dyskinetic potential of continuous administration of ropinirole.

HH2004/00106/00 Statistical analysis of antiparkinsonian activity and dyskinetic potential of continuous administration of ropinirole

HH2004/00107/00 The effect of ropinirole in a model of capsaicin-induced spinal sensitization in the rat.

PW 020 BA - An investigation into the development of tolerance to the cardiovascular effects of SK&F 101468-A in spontaneously hypertensive rats.

TF-1001/SKF-104557/2

- Investigation of the effects of acute, intravenous administration of SK&F 104557 on cardiovascular parameters in the anaesthetised, spontaneously hypertensive rat.

PW 006 BA - The effect of SK&F 101468-A on blood pressure and heart rate in the conscious spontaneously



hypertensive rat.

PW 007 BA - The dose-related effects on blood pressure and heart rate of the intravenous inflision of SK&F 101468-A to anaesthetised spontaneously hypertensive rats.

PP 007 BA - The effect of SK&F 101468-A on tilt-induced hypotension and tachycardia in anesthetized normotensive and spontaneously hypertensive rats.

PW 014 BA - The inhibition of isolated guinea-pig kidney dihydroxyphenylalanine decarboxylase by SK&F 101468A, a novel D2~opamine receptor agonist.

PP 005 BA - Prejunctional dopaminergic activity of SK&F 101468-A in the isolated perfused rabbit ear artery.

PP 0l0 BA - Comparison of the neuroinhibitory activity of SK&F 101468-A and two of its metabolites (SK&F 8912~A and SK&F 104557-A) in the isolated perfused rabbit ear artery.

PW 021 BA - An investigation of the effects of SK&F 101468-A on cardiovascular responses to partial occlusion of the abdominal vena cava in the anaesthetised cat following chronic administration of L-Dopa and benserazide.

PW 010 BA - The effect of SK&F 101468-A on the tachycardia following stimulation of the cardiac aecelerans nerve in the anaesthetised cat.

PW 021 BA - An investigation of the effects of domperidone, given subsequently to SK&F 101468-A, on cardiovascular responses to partial occlusion of the abdominal vena cava, in the anaesthetised cat.

PW 025 BA An investigation of the effects of SK&F 101468-A on cardiovascular responses to partial occlusion of the abdominal vena cava in the anaesthetised cat following chronic administration of L-dopa and benserazide.

PP 002 BA The effect of SK&F 101468-A on the pressor responses evoked by sympathetic nerve



stimulation in the perfused hindlimb of the dog.

PP 004 BA - Inhibitory effect of SK&F 89l2~A, SK&F 101468-A and SK&F 85378-H on cardiac neurotransmission in the anaesthetised dog.

PW 012 BA - Effects of SK&F 101468-A, a novel D2-dopamine receptor agonist, on myocardial oxygen consumption in anaesthetised dogs.

PP 003 BA - Hemodynamic profile of the selective D~-agonists SK&F 89124-A, SK&F 101468-A and SK&F 85738-H in the anesthetised dog.

PW 024 BA - An investigation into the development of tolerance to the cardiovascular effects of SK&F 101468-A in cynomolgus monkeys.

PW 001 BA - The effect of SK&F 101468-A on tachycardia following stimulation of the cardiac accelerans nerve in the anaesthetised cynomolgus monkey.

M4.2.1.3 Safety Pharmacology

PH-1007/SKF-101468/1

- The effects of ropinirole on cognitive function in the rat water maze experiment.

TF-1001/SKF-101468/1

- The effect of a single oral administration of SK&F 101468-A on temperature in the conscious mouse.

PW 009 BA - The effect of SK&F 101468-A, administered by mouth, on the central nervous system in mice.

PW 004 BA - The effect of SK&F 101468-A, administered intravenously, on autonomic nervous system function in anaesthetised cats.

PW 011 BA - The effect of acute intravenous administration of SK&F 101468-A (50 or 500 uglkg) on respiratory function in anaesthetised cats.

RH2004/00121/00 - The effects of the dopamine receptor agonist ropinirole (SKF-101468) on the hERG cardiac potassium channel.

PW 003 BA - Haemodynamic studies with SK&F 101468-A, administered intravenously to anaesthetised cats.



PW 002 BA - The effects of SK&F 101468-A on action potential characteristics of sheep Purkinje fibres in-vitro.

PP 001 BA - Effects of intravenous administration of SK&F 101468-A on renal function in conscious rats.

M4.2.2 PHARMACOKINETICS

M4.2.2.1 Analytical Methods and Validation Reports

BP 001 BA - Analysis of SK&F 101468 in plasma by high pressure liquid chromatography.

BP 002 BA - Analysis of SK&F 89124 and SK&F 85738 in plasma by high pressure liquid chromatography with electrochemical detection.

BP-1003/SKF-101468/1

- Direct and simultaneous LCIEC(UV determination of ropinirole and its metabolites (SK&F 89124 and SK&F 104557) in rat plasma utilizing semi-permeable surface guard columns and automated column switching devices for sample enrichment.

CP-1OOI/SKF-101468/1

- Summary report, preparation of radiolabeled SK&F 101468-A.

BP-0013/SKF-101468/l

- Quantification of concentrations of SK&F 89124 (a metabolite of SK&F 101468) in human and monkey plasma by HPLC with electrochemical detection.

M4.2.2.2 Absorption

BW02OBA - Absorption, metabolism and excretion in the male mouse following intravenous and oral administration at a target dose of 10 mg/kg.

BP/0011/SKF-101468/2

- A study to investigate systemic exposure of ropinirole after single, oral (5, 15 and 50 mg/kg) administration of a solution to male and female mice.

BP 003 BA - Absorption, mass balance and biliary secretion of 14C-SK&F 101468-A in male rats after a 20 mg/kg oral dose.

BF-1013/SKF-101468/1

14C-SK&F 101468A (ropinirole): Blood concentrations and excretion of radioactivity



following single and repeat oral administration at 0.5 mg pure free base/kg to male rats

BP 008 BA Oral absorption and intravenous pharmacokinetics of SK&F 101468 and SK&F 89124 in conscious male rats and pentobarbital-anaesthetised dogs.

BF-l004/SKF-101468/1

- Absorption of drug-related material following single administration of 1~-SK&F 101468-A into the lumen of Sections of the gastrointestinal tract of the male rat at a nominal dose level of 0.5 mg pure free base/kg.

BF-1OO5ISKF-10146811

- Blood concentrations and excretion of radioactivity following single oral administration to male and female rats.

TF-1007/101468/1 - 14 Day oral repeat dose pharmacokinetic study in rats.

BW 021 BA - Effect of oral administration (50 and 125 mg/kg, 14 days, p.o. and 250 mg/kg, 3 days, p.o.) on hepatic mixed function oxidase enzymes in the rat.

BP0010/SKF-101468/2

- Pharmacokinetics of SK&F 101468 following single oral (1.5, 5 and 15 mg/kg) and intravenous (1.5 mg/kg) administration of a solution to male cynomolgus monkeys.

BF-1001/SKF-10146812

- The pharmacokinetics of SK&F 101468 in male cynomolgus monkeys after single intravenous administration of SK&F 101468-A.

BW 005 BA - Concentrations of radioactivity in whole-blood and the excretion of radioactivity in male cynomolgus monkeys following (oral 15 mg/kg bodyweight) or intravenous (1 mg/kg bodyweight) administration of 14C-SK&F 101468-A.

BWO1OBA - Blood radioactivity and plasma SK&F 101468 and SK&F 89124 concentrations after oral and intravenous administration of 1~-SK&P 101468-A to the cynomolgus monkey (target doses 15 and 1 mg/kg, respectively.

BP 005 BA - Concentrations of SK&F 101468 and SK&F 89124 in a study of the acute toxicity of SK&F



101468-A in cynomolgus monkeys.

BF-1016/SKF-101468/1

14C-SK&F 101468A (ropinirole). Plasma concentrations and excretion of radioactivity following single and repeat oral administration at 0.5 mg pfb/kg to the cynomolgus monkey

016/SKF-101468/2 Amendment

14C-SK&F 101468A (ropinirole). Plasma concentrations and excretion of radioactivity following single and repeat oral administration at 0.5 mg pfb/kg to the cynomolgus monkey

BF-1029/SKF-101468/1

Pharmacokinetics of SK&F 101468 (ropinirole) and SK&F 104557 following a single and repeated daily oral administration of 14C-SK&F 101468A to the cynomolgus monkey for 21 days at 0.5 mg pfb/kg/day

BF-1015/SKF-101468/1

Bioequivalence study in cynomolgus monkeys (Pilot study). Dose administration and sample collection

BF-1030/SKF-101468/1

Preliminary pharmacokinetics of ropinirole (SK&F 101468) in the cynomolgus monkey following a single oral administration of 3 mg ropinirole using the 0.25 mg and 1.0 mg strength tablet

BF-1025/SKF-101468/1

SK&F 101468A: Bioequivalence study in cynomolgus monkeys. Dose administration and sample collection

BF-1031/SKF-101468/1

Bioequivalence of ropinirole (SK&F 101468) in the cynomolgus monkey following a single oral administration of 3 mg ropinirole using 0.25 mg and 1.0 mg tablet strengths

BP 007 BA - Concentration of SK&F 101468 in a toxicity study of SK&F 101468-A in cynomolgus monkeys after repeated intravenous administration for 14 days.

BP 006 BA - Concentration of SK&F 101468 in cynomolgus monkeys during a 30 day oral toxicity study of SK&F 101468-A/.

BP/0012/SKF-101468

- Plasma concentrations of SK&F 101468 and SK&F 104557 from Day 1 of a 52-week gavage



chronic toxicity study with SK&F 101468-A in cynomolgus monkeys.

M4.2.2.3 Distribution

BP 004 BA - Plasma protein binding and blood-to-plasma partition ratio of SK&F 101468 and SK&F 89124 in rat, dog and human blood.

BP-l00l/SKF-101468/1

- In vitro plasma protein binding and blood cell partitioning of 3H-SK&F 101468 in monkey and human blood.

CD2009/00540/0008DMM066

An in vitro investigation of the transport via heterologously expressed human P-glycoprotein of SKF-101468 in MDCKII-MDR1 cells.

BW 004 BA - 14C-SK&F 101468-A and 3H-SK&F 89124: binding to monkey and human serum proteins.

BW 003 BA - Preliminary investigation of the distribution of radioactivity into the brain after intravenous administration to the conscious male rat.

BWO14BA - Investigation of the distribution of 14C-SK&F 101468-A and 3H-SK&F 89124 in the brain of the conscious male rat.

BWOO2BAI2 - The distribution of radioactivity in male Wistar rats after repeated oral or single intravenous administration by whole body autoradiography at a target dose of 2 mg/kg.

BF-1018/SKF-101468/1

14C-SK&F 101468A (ropinirole): Tissue distribution of radioactivity following single and repeat oral administration to the rat at a nominal dose level of 0.5 mg pfb/kg

BW 006 BA - Tissue distribution of radioactivity after single oral and intravenous bolus administration to male albino rats. Target dose 2 mg/kg.

BW 007 BA - Assessment of melanin binding of radioactivity after single oral and intravenous bolus administration to male, Hooded Lister rats. Target



dose 2 mg/kg.

BWO22BA - Quantitative and qualitative investigation of the maternal/foetal distribution and composition of radioactivity following administration to pregnant rats (target dose 150 mg/kg free base).

BF-1OO3ISKF-101468/1

- Secretion of drug-related material into milk following single oral administration to the rat at a nominal dose level of 0.5 mg pfb/kg body weight.

BF-1011/SKF-101468/2

- Urinary radiometabolite patterns following single oral administration of [1~)-SK&F 101468-A to male and female rats at nominal dose level of 2 mg free base/kg.

BWO11BA - Distribution and composition of radioactivity in selected tissues following intravenous infusion of 14C-SK&F 101468-A in cynomolgus monkeys (target dose 1 mg/kg).

BF-1014/SKF-101468/1

14C-SK&F 101468A: Ex vivo binding of drug related material to rat and cynomolgus monkey plasma proteins

M4.2.2.4 Metabolism

BWO13BA - In vitro metabolism by freshly isolated rat, dog and monkey hepatocytes.

BF- 1002/SKF-101468/2

- An investigation of the in vitro metabolism of SK&F 101468 and the potential for drug interactions involving SK&F 101468 and the human cytochrome P450 enzymes lAl, iM, 2A6, 2C8-9, 2C19, 2D6, 2E1, 3A and 4A.

BW 008 BA - The interaction of SK&F 101468, SK&F 89124 and SK&F 104557 with rat liver cytoclirome P450 in vitro

BF-1017/SKF-101468/1

SK&F 101468A: Urinary excretion of radioactivity and urinary radiometabolite patterns following single and repeated oral administration to male rats at a target dose level of 0.5 mg pure free base/kg



BF-1026/SKF-101468/1

SK&F 101468A (ropinirole): The biliary excretion of radioactivity and radiometabolite patterns following single oral administration of 14C-SK&F 101468A to male rats at a target dose level of 0.5 mg pure free base/kg

BF-1023/SKF-101468/1

14C-SK&F 101468A: Pre-hepatic metabolism following a single oral administration at 0.5 mg free base/kg to male rats

BP009BA - Metabolism of SK&F 101468 and SK&F 89124 in conscious rats and pentobarbital anaesthetised dogs.

BW 024 BA - Metabolism in the rat and cynomolgus monkey.

BW 015 BA - Metabolism in the marmoset following oral administration (0.75 mg/kg).

BF-1012/SKF-101468/1

- Plasma radiometabolite patterns following single oral administration of (1~]-SK&F 101468-A to male rats at a nominal dose level of 0.5 mg free base/kg.

BW009BA - Metabolism of 1~-SK&F 101468-A following single and multiple oral administration to cynomolgus monkeys at a solution at a target dose of 1.5 mg/kg.

BF-1021/SKF-101468/2

14C-SK&F 101468A: Plasma radiometabolite patterns following single and 21 days repeat oral administration to male cynomolgus monkeys at a nominal dose level of 0.5 mg pure free base/kg

SKF-101468/RSD-1004LL/1

14C-SK&F 101468A: The isolation and identification of metabolites "D" and "K" of SK&F 101468 following oral dosing with 14C-SK&F 101468A to male cynomolgus monkeys at a nominal dose level of 15 mg pure free base/mg.

M4.2.2.5 Excretion

BW 001 BA - Excretion balance study in the male rat following intravenous bolus administration (2 mg/kg) and oral gavage administration (2 and 250 mg/kg).



BF-1027/SKF-101468/1

14C-SK&F 101468A: Plasma concentrations and the excretion of drug-related material in the urine of male cynomolgus monkeys after a single oral administration at 15 mg free base/kg

M4.2.2.6 Pharmacokinetic Drug Interactions

BF-1024/SKF-101468/1

An in vitro investigation of the inhibitory potential of ropinirole (SK&F 101468) on the human liver cystolic enzymes, dihydropyrimidine dehydrogenase and xanthine oxidase

M4.2.3 TOXICOLOGY

M4.2.3.1 Single Dose Toxicity

TW 001 BA - Acute oral and intravenous toxicity study in rats and mice.

TW 005 BA - Second acute oral and intravenous toxicity study in rats and mice.

TP-1001/SKF-104557/1

- Single dose intravenous toxicity study in mice.

TP-1007/SKF-101468/1

- Single dose intravenous toxicity study in mice.

TP-1001/SKF-089124/1

- Single dose intravenous toxicity study in rats.

TP 001 BA - Acute toxicity study of ropinirole-A in cynomolgus monkeys.

M4.2.3.2 Repeat Dose Toxicity

TW 019 BA - A Seven-day oral sighting study in the mouse.

TP-1008/SKF-101468/1

- Impurity evaluation in a 14 day oral toxicity study in mice.

TP 005 BA - 60-day dose-ranging study of ropinirole-A in mice.

TW 027 BA - 90 day oral toxicity study in the mouse.

TW 003 BA - 14-day oral sighting study in the rat.

TW 006 BA - 14 day intravenous toxicity study in the rat.



TP-1001/SKF-101468/2

- 14- to 57-day oral sighting study of SK&F 101468-A in Sprague Dawley rats.

TW 004 BA - 30 day oral toxicity study in rats.

TW 007 BA - A 30 day oral low dose toxicity study in female rats.

SKF-101468/RDS-100ZTN/1

- Evaluation of the retinal effects using various light intensities with or without ropinirole (SK&F-101468-A) in a 3 month oral study in albino and pigmented rats.

TW 012 BA - 6 month oral toxicity study in rats.

TP-1003/SKF-101468/2

- 1-year oral toxicity study of SK&F 101468 in rats.

TW 002 BA - Toxicity to cynomolgus monkeys by repeated intravenous administration for 14 days.

TP 002 BA - 30 day oral toxicity study of ropinirole-A in cynomolgus monkeys.

TW 015 BA - 26(34) week oral toxicity study in cynomolgus monkeys.

TP/0010/SKF-101468

- 52-Week gavage chronic toxicitY study of SK&F 101468-A in cynomolgus monkeys (Hazieton -terminated).

TP-0015/SKF-101468/1

- 1-year oral toxicity study of SK&F 101468-A in cynomolgus monkeys.

M4.2.3.3 Genotoxicity

TW 008 BA - Ames metabolic activation test to assess the potential mutagenic effects of SK&F 101468 and dopamine hydrochloride.

TG-1003/SKF-101468/1

- Mutation testing with Escherichia coli WP2 Pkm101 and WP2 uvrA pKM101 agar plate assay.

TW 010 BA - Study to determine the ability of SK&F 101468-A and dopamine hydrochloride to induce mutations to 6-thioguanine resistance in mouse lymphoma L5178Y cells using a fluctuation assay.



TW 009 BA - Study to evaluate the chromosome damaging potential of SK&F 101468-A and dopamine hydrochloride by their effects on cultured human lymphocytes using an in vitro cytogenetics assay.

TW 011 BA - Study to evaluate the potential of SK&F 101468-A to include micronuclei in the polychromatic erythrocytes of CD-i mice.

M4.2.3.4 Carcinogenicity

TP-1004/SKF-101468/2

- An oral carcinogenicity study of SK&F 101468-A in CD-1 mice.

TP-1005/SKF-10146811

- Two year carcinogenicity study of SK&F 101468-A in the Sprague-Dawley rat.

M4.2.3.5 Reproductive and Developmental Toxicity (including Juvenile Studies)

TP 006 BA - Dose-Range male fertility study of SK&F 101468-A in rats.

TP 007 BA - Male fertility study of SK&F 101468-A in rats (Segment 'I/A of the FDA Guidelines for Reproductive Studies).

TW 014 BA - Oral reproductive sighting study in Rats.

TW 022 BA - An investigation of abortifacient action in the rat.

TW 021 BA - A preliminary study of fertility and general reproductive performance in the female rat.

TW 025 BA - Fertility study in the female rat.

M4.2.3.5.1 Embryofetal development

TW 020 BA - Preliminary oral teratology study in the rat.

TW 023 BA - Oral teratology study in the rat.

TW 024 BA - Investigative teratology study in the rat.

TW 026 BA - Second investigative teratology study in the rat.

TP 003 BA - Dose-range study of SK&F 101468-A in pregnant rabbits (For a Segment II Developmental Toxicology Study).



TP 004 BA - Development Toxicology Study of SK&F 101468-A in rabbits (Segment II of the t'FDA Guidelines for Reproduction Studies").

TP-1010/SKF-101468/1

- Oral study for toxicological and embryofetal developmental effects in rabbits.

M4.2.3.5.2 Prenatal and postnatal development, including maternal function

TP 009 BA - Dose-range study of SK&F 101468-A in pregnant and lactating rats (for Segment Ill of the FDA guidelines for reproductive studies).

TP-0016/SKF-101468/1

- Perinatal/postnatal study of SK&F 101468-A in female rats.

M4.2.3.6 Local Tolerance

TP/0012/SKF-10l468

- Acute dermal irritation test in the rabbit.

TP/0013/SKF-101468

- Acute eye irritation test in the rabbit.

TP/0014/SKF-101468

- Guinea pig skin sensitisation test.

TW 016 BA - Acute intravenous and perivenous irritancy in the dog.

TW 013 BA - In vitro human red cell haemolysis study.

M4.2.3.7 Other Toxicity Studies

M4.2.3.7.1 Immunotoxicity

BW 017 BA - Improved radioimmunoassay for SK&F 101468 in human and monkey plasma.

BW 018 BA - A radioimmunoassay for SK&F 104557 (a metabolite of SK&P 101468) in human and monkey.

BW 018 BA/Addendum 1

- Validation of the radioimmunoassay method for SK&F 104557 (a metabolite of SK&F 101468) in human plasma and urine and monkey plasma.

BP-0014/SKF-101468/2

- Revalidation of the radioimmunoassay method for SK&F 101468 in human plasma and urine and monkey plasma.



BW 0016 BA - Radioimmunoassay for SK&F 101468 in human plasma.

M4.2.3.7.2 Mechanistic Studies

SKF-101468/RSD-100ZTN/1

SK&F-101468-A: Evaluation of retinal effects using various light intensities with or without ropinerole (SK&F-101468-A) in a 3 month oral study in albino and pigmented rats

M4.2.3.7.3 Dependence

TP-1002/SKF-101468/2

- Assessment of the physical dependence liability in naive cynomolgus monkeys following two 28~ay periods of oral administration.

TF-1004/SKF-101468/1

- Assessment of the effects on the morphine withdrawal syndrome in cynomolgus monkey.

TF-1005/skf-101468/1

- Assessment of the effects on the phenobarbitone withdrawal syndrome in cynomolgus monkeys.

TF-1006/SKF-101468/1

Assessment of the psychological dependence in cynomolgus monkeys

M4.2.3.7.4 Other

TW 018 BA - In vitro human platelet aggregation study.

TW 017 BA - The effect of caging density (n=2 or 5 on rat lethality).

TP-1006/SKF-101468/1

- Influence of SK&F 101468-A on Leydig Cell LH receptor profile in rats.

CONFIDENTIAL

MODULE 1.4.2 INFORMATION ABOUT THE EXPERT - NONCLINICAL

CONFIDENTIAL

1.4.2. INFORMATION ABOUT THE EXPERT - NONCLINICAL


According to his respective qualifications the undersigned expert declares hereby to have performed the duties set out in the Article 12 and in accordance with Annex I, Part I 1.4 of Directive 2001/83/EC, as amended. NONCLINICAL:

Name of the Expert:

Dr D.R. Newall Signature:

Position Director, Worldwide Non-Clinical Safety Projects, Established Products

Address: GlaxoSmithKline Research & Development Park Road Ware Hertfordshire, SG12 0DP

Date:

According to the Annex I of Directive 2001/83/EC as amended, brief information on the educational background, training and occupational experience is attached.

CONFIDENTIAL

CURRICULUM VITAE

Name: Derek R Newall

Education: BSc, Zoology, Newcastle upon Tyne 1968

PhD, Faculty of Medicine, Newcastle upon Tyne 1987

Appointments: Demonstrator in Zoology, Newcastle upon Tyne 1968 – 1972

Research Associate in Surgery, Cleft Palate Research Unit, Newcastle upon Tyne

1972 – 1983

Reproductive Toxicologist, Life Science Research, Suffolk

1984 – 1986

Reproductive Toxicologist, Glaxo Group Research

1986 – 1989

Head, In Vitro Reproductive Toxicology Unit, Glaxo Group Research

1989 – 1995

Group Toxicologist, Genetic and Reproductive Toxicology, Medicines Safety Evaluation Division, Glaxo Wellcome

1995 – 1997

Group Toxicologist, International Preclinical Safety Projects, Medicines Safety Evaluation Division, Glaxo Wellcome

1997 – 1999

Head, Full Development Projects (UK), International Preclinical Safety Projects, Medicines Safety Evaluation Division, Glaxo Wellcome

1999 – 2001

Director, Worldwide Non-Clinical Safety Projects, Full Development US, Safety Assessment Division, GlaxoSmithKline

2001 – 2004

Director, Worldwide Non-Clinical Safety Projects, Established Products, Safety Assessment Division, GlaxoSmithKline

2004 to date

Membership of Professional Bodies and Societies:

British Toxicology Society European Teratology Society In Vitro Toxicology Society Drug Information Association

MODULE 2.4 NONCLINICAL OVERVIEW · m2.4. Nonclinical Overview INTRODUCTION Ropinirole Prolonged...

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