Controlled release formulations of levodopa and uses thereof

WO 2009/085306 PCT/US2008/0140S0

CONTROLLED RELEASE FORMULATIONS OF LEVODOPA AND USES THEREOF

Throughout this application various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.

FIELD OF THE INVENTION

The present invention relates to controlled release pharmaceutical compositions of levodopa (LD), formulated with an acid and a decarboxylase inhibitor, to yield enhanced pharmaceutical pharmacokinetic attributes. These formulations are useful for the treatment of conditions such as neurological diseases associated with reduced or impaired dopamine levels.

BACKGROUND OF THE INVENTION

Combinations of levodopa (LD) and a decarboxylase inhibitor (typically carbidopa (CD)) to treat Parkinson’s disease (PD) are known in the pharmaceutical arts and are considered by many to be the ‘gold standard’ treatment for symptoms of PD. Currently, several formulations containing a combination of LD and CD are commercially available, e.g., S1NEMET®, STALEVO®, PARCOPA®, and ATAMET®. Nonetheless, a need remains for an oral LD formulation that provides steadier plasma concentrations of LD with minimal ‘peak-to-trough’ fluctuations during daily dosing and that yields a longer duration-of-effect than the currently available oral dosage forms of CDZLD.

Patients suffering from PD frequently have periods in which their mobility becomes difficult, often resulting in an inability to move. Abnormally low levels of dopamine, a neurotransmitter that affects mobility and control of the skeletal-muscular system, is commonly believed to be the cause of these motor symptoms in PD patients. However, administration of dopamine is not effective to treat Parkinson’s disease because dopamine 1 WO 2009/085306 PCT/US20O8/014080 does not cross the blood-brain barrier. To resolve this problem, PD patients are administered levodopa, the metabolic precursor of dopamine, but levodopa is not without its issues either.

While levodopa crosses the blood-brain barrier and is rapidly converted to dopamine, LD is problematic because of its rapid decarboxylation by tissues other than the brain. Thus, when LD is administered alone, large doses are required because only a small portion is transported to the brain unchanged. Furthermore, when levodopa is administered orally it is rapidly decarboxyl ated to dopamine in extracerebral tissues so that only a small portion of a given dose is transported unchanged to the central nervous system. Carbidopa inhibits the decarboxylation of peripheral levodopa and does not cross the blood-brain barrier. Since its decarboxylase inhibiting activity is limited to extracerebral tissues, administration of carbidopa with levodopa has been popular to make levodopa more available for transport to the brain.

In addition to these difficulties associated with absorption of LD, over time patients treated with LD also exhibit symptoms of “wearing off”. PD patients treated with LD may develop motor fluctuations characterized by end-of-dose failure, peak dose dyskinesia, and akinesia. The advanced form of motor fluctuations (also commonly referred to as the ‘on-off phenomenon) is characterized by unpredictable swings from mobility to immobility. Although the causes of these motor fluctuations are not completely understood, some patients may be attenuated by treatment regimens that produce steady plasma levels of LD, Thus, a void remains in the LD treatment of PD patients, as plasma concentration levels remain difficult to control.

Currently available controlled release formulations of CD/LD are meant to allow for a continuous release of drug over a prolonged period of time in an attempt to maintain tight LD plasma ranges. However, the use of these controlled release dosage forms are problematic in that many PD patients wake up in the morning having little or no mobility due to the wearing off of the dose taken the day/evening before. Once the previous dose has worn off, such patients are usually unwilling, or even unable, to wait for the extended 2 206756/2

, period of time required for a controlled release dosage form to deliver the necessary plasma levels of LD. While the use of an immediate release formulation of LD can reduce this 'wait time', the use of an immediate release formulation of LD require more frequent dosing and are associated with more fluctuating plasma LD 5 concentrations. DUODOPA®, an intraduodenal infusion therapy approved outside of the United States, demonstrates significantly reduced motor complications and reduced 'off time. The cumulative experiences from DUODOPA® and experimental infusion studies show that the maintenance of stable plasma LD concentrations and the avoidance of low trough levels appear to be effective in reducing 'off time, 10 increasing 'on' time without disabling dyskinesia, and reduce the severity of dyskinesia in comparison to the standard oral formulations. However, such infusion therapies are extremely inconvenient to the patient. EP 1262198 recites a formulation comprising levodopa and decarboxylase inhibitor. However, EP 1262198 does not describe separate IR granules, CR granules or 15 carboxylic acid granules for a single oral pharmaceutical preparation. US 2007/0148238 recites a pharmaceutical composition comprising levodopa, carbidopa and carboxylic acid. However, US 2007/148238 shows that under fasted conditions, their formulations, which include a bioadhesive layer, did not result in a steady and prolonged absorption of the active ingredients. 20 US 2005/0070608 recites a pharmaceutical composition comprising levodopa, carbidopa and carboxylic acid. However, US 2005/0070608 does not describe separate IR granules, CR granules or carboxylic acid granules for a single oral pharmaceutical preparation.

The results of infusion therapies, such as DUODOPA®, strongly suggest a rationale 25 for the development of a LD treatment that provide constant, or relatively steady, LD plasma concentrations to optimize relief of PD symptoms and to minimize 'off times and dyskinesias. Indeed, a need remains for a more convenient, i.e., oral, dosage form that will improve the administration of LD to PD patients by narrowing blood plasma ranges of LD, which in turn will result in reduced 'off times', prolonged 'on time', and 30 decreased time to 'on'. The present invention fills this void by providing a novel controlled release oral solid dosage form of LD that is formulated with a 206756/1 decarboxylase inhibitor and an acid, to yield the desired pharmacokinetic attributes, i.e., steadier plasma concentrations of LD over a prolonged period of time.

SUMMARY OF THE INVENTION 5 The current invention provides a controlled release oral solid formulation of levodopa comprising levodopa, a decarboxylase inhibitor, and a carboxylic acid that is not levodopa nor the decarboxylase inhibitor. Also provided by this invention is multiparticulate, controlled release oral solid formulations of levodopa comprising: i) a controlled release component comprising a mixture of levodopa, a decarboxylase 3a i WO 2009/085306 PCT/US2008/014080 inhibitor and a rate controlling excipient; ii) a carboxylic acid component; and iii) an immediate release component comprising a mixture of levodopa and a decarboxylase inhibitor.

The current invention additionally claims a controlled release oral solid formulation of levodopa providing a relative steady levodopa plasma or serum concentration profile over a prolonged period of time; this controlled release oral solid formulation of levodopa having a levodopa plasma or serum concentration profile comprising: a) a time of administration; b) a first concentration; and c) a second concentration, wherein, the first concentration is equal to the maximum concentration of said profile; the second concentration is the minimum concentration occurring at a time later than said first concentration and earlier than or equal to about six hours following the time of administration; and wherein the second concentration is greater than or equal to about fifty percent of the first concentration.

Further, the invention provides methods for making and using the pharmaceutical formulations of the invention. Methods for reducing motor fluctuations in a patient suffering from Parkinson’s disease, reducing off time in a patient suffering from Parkinson’s disease, increasing on time in a patient suffering from Parkinson’s disease, reducing time to ‘on’ in a patient suffering from Parkinson’s disease and otherwise enhancing dopamine levels in a subject suffering from a disease associated with reduced or impaired dopamine levels are provided by the presently claimed invention.

BRIEF DESCRIPTION OF DRAWINGS

Figure 1 is a graph showing that IPX066 formulations provide infusion-like plasma profile for longer than about six hours.

Figure 2 shows a graph illustrating the in vivo plasma concentration profiles for IPX066-B-05-07 Formulations A, B and C compared to a reference (Sinemet®), as described in Example 1, infra. 4 WO 2009/085306 PCT/US2008/014080

Figure 3 shows a graph illustrating the in vivo plasma concentration profiles for IPX066-B-06-02 Formulations A, S, D and E compared to a reference (Sinernet®), as described in Example 2, infra.

Figure 4 shows a graph illustrating the in vivo plasma concentration profiles for IPX066-B-07-01 Formulations A, B and C compared to a reference (Sinemet®), as described in Example 3, infra.

Figure 5 shows a graph illustrating the in vivo plasma concentration profiles for IPX066 formulations.

Figure 6 is a table showing the robust lower intrasubject variability of the IPX066 formulations compared to Sinemet® CR.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to controlled release pharmaceutical formulations comprising LD, a decarboxylase inhibitor and a carboxylic acid for treating neurological diseases or conditions associated with reduced or impaired dopamine levels. The pharmaceutical formulations of the invention provide steadier, or more constant, LD plasma concentrations in patients, resulting in decreased motor-fluctuations, reduced “off” time and increased “on” time in PD patients.

DEFINITIONS

All scientific and technical terms used in this application have meanings commonly used in the art unless otherwise specified. As used in this application, the following words or phrases have the meanings specified. 5 WO 2009/085306 PCT/US2008/014080

The term "acid” refers to a chemical compound that, when dissolved in water, gives a solution with a pH less than 7. The “acid” can be organic. It can have a pKa in the range of e.g., 2-5. Examples of acids suitable for the invention include, but are not limited to, tartaric acid, adipic acid, succinic acid, citric acid, benzoic acid, acetic acid, ascorbic acid, 5 edetic acid, fumaric acid, lactic acid, malic acid, oleic acid, sorbic acid, stearic acid, palmitic and boric acid or mixtures thereof.

The term ’’effective amount” means an amount of a compound/composition according to the present invention effective in producing the desired therapeutic effect. 10 A “tablet” or “pill” comprises a pharmaceutical formulation pressed into a form. The form can be in any shape, for example, round, oblong, triangular or other shapes. A “capsule” comprises a pharmaceutical formulation in which the pharmaceutical 15 formulation is encased in a hard or soft soluble container. The container can be in the form of gelatin or other material.

The term "modified release” (also known as MR) includes delayed release (also known as DR) and controlled release (also known as CR, sustained release (SR), prolonged release 20 (PR) or extended release (ER)).

The term “delayed release” (also known as DR) relates to a pharmaceutical formulation or component that releases the active ingredients after a period of delay. 25 The term “controlled release” (also known as CR) refers to a pharmaceutical formulation or component thereof that releases, or delivers, one or more pharmaceutical agents over a prolonged period of time, in this case over a period of more than one hour.

The term "immediate release" (also known as instant release or IR) refers to a 30 pharmaceutical formulation or component thereof which releases, or delivers, one or 6 WO 2009/085306 PCT/US2008/014080 more pharmaceutical agents substantially immediately upon administration and will result in substantially complete dissolution within about one hour (or less).

The terms “release excipient” or “rate controlling excipient” may be used interchangeably. Release excipients or rate controlling excipients include all excipients and/or polymers that control the release of a pharmaceutical agent(s), e.g,, LD, CD and in this case acid, after administration in a subject. Examples of release excipients or rate controlling excipients include, but are not limited to, hypromellose, hydroxypropyl cellulose, ethyl cellulose and prop-2-enoic acid. Delay release polymers, as a subset of release excipients or rate controlling excipients, are used to delay the release of a pharmaceutical agent(s) after administration to a subject. Examples of delay release polymers include, but are not limited to, enteric polymers and/or neutral methacrylic polymers such as Eudragit® L100-55, Eudragit® S100 or Eudragit® FS30D (Rohm).

The “USP paddle method” refers to the Paddle and Basket Method as described in United States Pharmacopoeia, Edition XXII (1990).

The term “peak-to-trough ratio” refers to a comparison of the values for a peak (e.g., a high point in a line graph) plasma level of active agent and a trough (e.g., a low point in a line graph) level over a set amount of time. For example, in a line graph with plasma LD values ranging from 400 ng/ml (peak) compared to 200 ng/ml (trough) over a four hour period, gives a peak-to-trough ratio of 2 for that time. More than one peak-to-trough ratio can be illustrated in a graph.

The term ’’about’* when used in connection with percentages means +/-1%.

The “Mean Plasma Concentration” of a substance (e.g., LD or CD) as used herein, refers to the mean concentration of the substance found in multiple plasma samples. The mean plasma concentration is obtained by adding the concentrations of the substance found in the plasma samples then dividing the sum by the number of plasma samples. 7 WO 2009/085306 PCT/US2008/014080

The “upper small intestine” refers to the portion closest to the stomach and includes the duodenum and jejunum.

The term “outer coat” refers to a covering or barrier applied to a pharmaceutical formulation or component thereof and may be an enteric coat.

Diseases associated with reduced or impaired dopamine levels includes neurological or movement disorders such as restless leg syndrome, Alzheimer’s disease, dystonia, schizophrenia, Parkinson's disease and secondary parkinsonism, Huntington’s disease, Attention-Deficit I Hyperactivity Disorder (ADHD), Shy-Drager syndrome and conditions resulting from brain injury, including carbon monoxide or manganese intoxication.

The term “treating” a disease associated with reduced or impaired dopamine levels, means to manage a disease with the pharmaceutical formulation of the invention. Treatment can decrease the symptoms of a disease, reduce the severity of a disease, alter the course of disease progression, ameliorate and/or cure a disease associated with the neurological or movement disorders associated with reduced or impaired dopamine levels.

COMPOSITIONS OF THE INVENTION A significant aspect of the invention relates to the unexpected discovery of the effect of carboxylic acid, in controlling the absorption of LD such that the resulting formulations yield ‘tighter,’ i.e. steadier, LD plasma concentrations.

The invention provides controlled release oral solid formulations of levodopa comprising Ievodopa, a decarboxylase inhibitor, and a carboxylic acid. In one embodiment of the invention, the carboxylic acid is neither levodopa nor a decarboxylase inhibitor. In accordance with the practice of the invention, the carboxylic acid may be physically separated from the levodopa and the decarboxylase inhibitor. The pharmaceutical formulations can include a single acid or a mixture of acids. 8 WO 2009/085306 PCT/US2008/014080

In one embodiment» the carboxylic acid may be a polycarboxylic acid. In another embodiment» the carboxylic acid may be a dicarboxylic acid. Suitable examples of carboxylic acids include, but are not limited to, tartaric acid, adipic acid, succinic acid, 5 citric acid, benzoic acid, acetic acid, ascorbic acid, edetic acid, fumaric acid, lactic acid, malic acid, oleic acid, sorbic acid, stearic acid, palmitic acid and boric acid or mixtures thereof. In a particular embodiment, the dicarboxylic acid is a tartaric acid. A suitable example of a decarboxylase inhibitor includes but is not limited to carbidopa. 10

In accordance with the practice of the invention, the formulation may be a tablet or a caplet. The tablet or caplet may be single-layered or multi-layered. The tablet or caplet may be a matrix tablet or caplet. 15 Also in accordance with the practice of the invention, the formulation may be a multiparticulate formulation. In an embodiment of the invention, the multiparticulates are encapsulated. In another embodiment, the multiparticulates are not encapsulated. The multiparticulates may be pressed into a tablet. Alternatively, the multiparticulates may be in a sprinkle form that can be sprinkled directly onto food or liquids for easy 20 ingestion.

In an embodiment of the invention, the formulation reduces intrasubject variability in levodopa absorption. The intrasubject variability may be calculated as the standard deviation of the levodopa concentration divided by the mean levodopa concentration 25 determined over the range of about 0.5 hours after administration to about six hours after administration for a single dose of the formulation to an individual subject which when averaged over at least 12 subjects; is less than or equal to about 0.40.

The controlled release oral solid formulation of the invention may comprise carbidopa 30 and levodopa in a ratio of about 1:1 to about 1:10. In one embodiment, the ratio of carbidopa to levodopa is about 1:4. 9 WO 2009/085306 PCT/US2008/014080

The controlled release oral solid formulation of the invention may have a ratio of moles of dicarboxylic acid to levodopa of less than 4:1. In one embodiment, the ratio of moles of dicarboxylic acid to levodopa is greater than 1:4 and less than 3:2. In another 5 embodiment, the ratio of moles of dicarboxylic acid to levodopa is greater than 1:2 and less than 4:3. In yet another embodiment, the ratio of moles of dicarboxylic acid to levodopa is greater than 2:3 and less than 5:4. In a further embodiment, the ratio of moles of dicarboxylic acid to levodopa is greater than 1:1 and less than 4:3. 10 The controlled release oral solid formulation of the invention may comprise from about 25 mg to about 2000 mg levodopa. In one embodiment, the formulation comprises about 50 to 600m.gof levodopa.

The controlled release oral solid formulation of the invention may comprise from about 15 10 mg to about 300 mg carbidopa. In an embodiment of the invention, the controlled release oral solid formulation comprises about 10 to 80 mg of carbidopa.

The invention also provides multiparticulate, controlled release oral solid formulations of levodopa. A multiparticulate, controlled release oral solid formulation of the invention 20 comprises 1) a controlled release component comprising a mixture of levodopa, a decarboxylase inhibitor and a rate controlling excipient; 2) a carboxylic acid component; and 3) an immediate release component comprising a mixture of levodopa and a decarboxylase inhibitor. In an embodiment of the invention, the decarboxylase inhibitor is carbidopa. The multiparticulate, controlled release oral solid formulation of the 25 invention may reduce intrasubject variability in levodopa absorption.

In accordance with the practice of the invention, the controlled release component may be a distinct component (e.g., separate from the carboxylic acid and immediate release components). In an embodiment of the invention, the carboxylic acid component is a 30 distinct component (e.g., separate from the controlled release and immediate release components). In yet another embodiment, the immediate release component is a distinct 10 WO 2009/085306 PCT/US2008/014080 component (e.g., separate' from the controlled release and carboxylic acid components). In still another embodiment of the invention, the controlled release component, the immediate release component and the carboxylic acid component are each manufactured as distinct, separable beads.

In an embodiment of the invention, all of the components (namely, the controlled release, carboxylic acid, and immediate release components) of the multiparticulate, controlled release oral solid formulation are coformulated into a single component.

The multiparticulate, controlled release oral solid formulation of the invention may further comprise one or more controlled release carboxylic acid components comprising a carboxylic acid and a rate controlling excipient. Suitable examples of carboxylic acids include but are not limited to tartaric acid, adipic acid, succinic acid, citric acid, benzoic acid, acetic acid, ascorbic acid, edetic acid, fumaric acid, lactic acid, malic acid, oleic acid, sorbic acid, stearic acid, palmitic acid and boric acid or mixtures thereof. In a particular embodiment, the carboxylic acid is tartaric acid. Further, the controlled release carboxylic acid components may comprise a carboxylic acid core coated with one or more enteric polymers. In a particular embodiment, the multiparticulate, controlled release oral solid formulation has at least two controlled release carboxylic acid components that release the carboxylic acid at different rates.

In accordance with the practice of the invention, the rate controlling excipient of the multiparticulate, controlled release oral solid formulation may be an enteric polymer or a mixture of more than one type of enteric polymer. In one embodiment, the rate controlling excipient is a neutral methacrylic polymer.

In an embodiment of the multiparticulate, controlled release oral solid formulation of the invention, the controlled release component further comprises a carboxylic acid. The carboxylic acid may be a dicarboxylic acid. 11 WO 2009/085306 PCT/US2008/014080

In an embodiment of the multiparticulate, controlled release oral solid formulation of the invention, the controlled release component comprises a core of levodopa and decarboxylase inhibitor coated with one or more enteric polymers.

The multiparticulate, controlled release oral solid formulation provides an embodiment wherein carbidopa and levodopa is present in a ratio of about 1:1 to about 1:10. in one embodiment, the ratio of carbidopa to levodopa is 1:4.

The multiparticulate, controlled release oral solid formulation of the invention provides an embodiment wherein the ratio of moles of carboxylic acid to levodopa is less than 4:1. In one embodiment, the multiparticulate, controlled release oral solid formulation, has a ratio of moles of carboxylic acid to levodopa of greater than 1:4 and less than 3:2. In another embodiment, the multiparticulate, controlled release oral solid formulation has a ratio of moles of carboxylic acid to levodopa of greater than 1:2 and less than 4:3. In yet another embodiment, the multiparticulate, controlled release oral solid formulation has a ratio of moles of carboxylic acid to levodopa of greater than 2:3 and less than 4:3. In an additional embodiment, the multiparticulate, controlled release oral solid formulation has a ratio of moles of dicarboxylic acid to levodopa of greater than 1:1 and less than 4:3.

The multiparticulate, controlled release oral solid formulation of the invention provides an embodiment comprising from about 25 mg to about 1200 mg levodopa. In one embodiment, the multiparticulate, controlled release oral solid formulation comprises about 50 to about 600 mg of levodopa.

The multiparticulate, controlled release oral solid formulation of the invention provides an embodiment comprising from about 10 mg to about 300 mg carbidopa. In an embodiment, the multiparticulate, controlled release oral solid formulation comprises 20 mg to about 80 mg of carbidopa. 12 WO 2009/085306 PCT/US2008/014080

The pharmaceutical formulations of the invention can further comprise excipients including, but not limited to, surfactants (ionic and non-ionic), lipophilic vehicles, and hydrophilic vehicles.

In accordance with the practice of the invention, examples of a rate controlling excipient include, but are not limited to, hydroxypropyl cellulose, hypromellose, ethyl cellulose, and prop-2-enoic acid. One suitable example of a prop-2-enoic acid is Carbopol® (Noveon or Dow Chemical Co.). Examples of delay release polymers include a neutral methacrylic polymer such Eudragit® FS30D, Eudragit® S100, Eudragit® L100-55 and/or any mixture or combination thereof (Rohm). Eudragit® LI00-55 is an enteric polymer which can be used in coated dosage forms to target the drug release in the upper small intestine where the pH is above 5.5. Eudragit® S100 can be usedLto achieve targeted drug release in the lower small intestine to the colon, where the pH is above 7, The modified release components of the formulations of this invention can be formulated with any, and/or a mixture, of the above polymers, to achieve the desired LD plasma concentration profiles. The choice of the polymers that can be used in the invention includes, but is not limited to, Eudragit®, cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate LF, hydroxypropyl methylcellulose acetate succinate HF, and others.

The pharmaceutical formulations of the invention can further comprise other excipients commonly known and used by those of skill in the art, including a plasticizer agent (e.g., triethyl citrate), lubricant (such as talc and magnesium stearate), and disintegrant (such as croscarmellose sodium and crospovidone), or any combination thereof.

The invention provides a controlled release oral solid formulation of levodopa having or exhibiting a levodopa plasma or serum concentration profile, which profile comprises a time of administration, a first concentration and a second concentration. The controlled release oral solid formulation of levodopa may further comprise at least one carboxylic acid, for example, tartaric acid. 13 WO 2009/085306 PCT/US2008/014080

In accordance with the practice of the invention the first concentration may be equal to the maximum levodopa plasma or serum concentration of the profile, the second concentration may be the minimum concentration occurring at a time later than said first concentration and earlier than or equal to about six hours following the time of 5 administration. The second concentration may be greater than or equal to about fifty percent of said first concentration.

In one embodiment of the formulation, the levodopa plasma or serum concentration profile is a median levodopa plasma or serum concentration profile. In another 10 embodiment, the levodopa concentration profile is the mean levodopa plasma or serum concentration profile.

In a further embodiment, the levodopa plasma or serum concentration profile further comprises a third concentration. The levodopa profile in this third concentration may be 15 greater than or equal to fifty percent of the first concentration. . Further, the third concentration may occur a time earlier than said first concentration and within about ninety minutes of the time of administration. In a specific embodiment, levodopa in the third concentration may be greater than or equal to sixty percent of the first concentration and the second concentration may be greater than or equal to sixty percent of the first 20 concentration. in accordance with the practice of the invention, the second concentration may be the minimum concentration occurring between one hour after said time of administration and the second time. In one embodiment, the first concentration is between 825 and 1505 25 ng/mL for a 380 mg dose of levodopa.

The levodopa plasma or serum concentration profile may have a ratio of mean AUC which is measured in units of ng h/mL, to the mass of levodopa in the formulation, where said mass is measured in mg, is between 11:1 and 25:1. In one embodiment, the ratio is 30 between 14:1 and 19:1. Additionally, the levodopa plasma or serum concentration profile may have a ratio of mean AUC which is measured in units of ng h/mL, to said first 14 WO 2009/085306 PCT/US2008/014080 concentration, where said concentration is measured in units of ng/mL, of between 9:2 and 6:1.

The levodopa plasma or serum concentration profile may have a mean AUC of between 4330 and 8000 ng h/mL for a 380 mg dose of levodopa. In one embodiment, the mean AUC is between 5000 and 7000 ng h/mL for a 380 mg dose of levodopa.

The levodopa plasma or serum concentration profile may have a ratio . of the first concentration which is measured in units of ng/mL, to the mass of levodopa in the formulation, where said mass is measured in mg, of between 3:1 and 5:1. In one embodiment, the ratio is between 5:2 and 7:2. In another embodiment, the ratio is greater than or equal to about 3:1.

In one embodiment, the levodopa plasma or serum concentration profile comprises a time of administration, a first concentration at a first time that occurs within one hour of the time of administration; a second concentration at a second time, that occurs after said first time; and a third concentration at a third time, that occurs at least four hours after said second time. The second concentration may be equal to the maximum concentration of levodopa in the profile; the first concentration may be equal to about fifty percent of the second concentration; said third concentration may be equal to about fifty percent of the second concentration.

In another embodiment, the controlled release oral solid formulation has a levodopa plasma or serum concentration profile substantially the same as levodopa formulation IPX066 in Figure 1 for a 3 80 mg dose of levodopa, or having a levodopa plasma or serum concentration profile substantially proportional to said formulation in Figure 1 for a dose other than 380 mg.

In yet another embodiment, the controlled release oral solid formulation has a levodopa plasma or serum concentration profile such that the ratio of the maximum concentration 15 WO 2009/085306 PCT/US2008/014080 of the profile to the concentration at any time between one hour and seven hours after administration of the formulation is less than or equal to 4:1.

In a further embodiment, the controlled release oral solid formulation of levodopa has a median levodopa plasma or serum concentration profile comprising: a first concentration at a first time; a second concentration at a second time, that occurs within about one hour after said first time; a third concentration at a third time, that occurs at least four hours after said second time; and a maximum concentration. The second concentration may be equal to the maximum concentration of said profile; the first concentration is equal to fifty percent of said second concentration; the third concentration is equal to fifty percent of the second concentration.

Coating of the Pharmaceutical Formulation

Another aspect of the invention relates to the method of preparation of the enteric coating of the formulations containing an acid, e.g., tartaric acid. The acid causes a slow and variable drug release rate. The prolonged and slow drug release rate may be due to the interference, of the dissolution of the enteric coat, affected by the presence of the acid in the core. The interference can be significantly reduced by partially neutralizing the enteric polymers of the coating, for example, by adding a base (e.g., NH3 or NH4OH) to the coating formulation so as to increase the pH of the coating. The neutralizing technique can be equally effective for different enteric polymers including, but not limited to, Eudragit® L100, S100, and FS100.

METHODS OF THE INVENTION

The invention provides a method of enhancing dopamine levels in a subject suffering from a condition associated with reduced or impaired dopamine levels comprising administering to a subject an effective amount of a pharmaceutical formulation of the invention thereby enhancing dopamine levels in the subject suffering from a condition associated with reduced or impaired dopamine levels. 16 WO 2009/085306 PCT/US2008/014080

The invention also provides methods for enhancing or maintaining dopamine levels in a subject suffering from a condition associated with reduced or impaired dopamine levels comprising administering to the subject an effective amount of a pharmaceutical formulation of the invention thereby maintaining dopamine levels in the subject suffering from a condition associated with reduced or impaired dopamine levels.

Enhancing or maintaining dopamine levels in a subject can treat the subject suffering from a condition associated with reduced or impaired dopamine levels. Examples of a condition associated with reduced or impaired dopamine levels include, but is not limited to, Alzheimer’s disease, dystonia, schizophrenia, and Parkinson’s disease.

The invention further provides methods of reducing motor fluctuations in a patient suffering from Parkinson’s disease comprising administering to the patient an effective amount of any of the formulations of the invention thereby providing a plasma concentration of levodopa effective to reduce motor fluctuations in the patient. In one embodiment of the invention, the formulation is administered in six-hour intervals.

Additionally provided are methods of reducing off time in a patient suffering from Parkinson’s disease comprising administering to the patient an effective amount of any of the formulations of the invention thereby providing a plasma or serum concentration of levodopa effective to reduce off time in the patient.

Further, the invention provides methods of increasing “on” time in a patient suffering from Parkinson’s disease comprising administering to the patient an effective amount of any of the formulations of the invention thereby providing a plasma or serum concentration of levodopa effective to increase on time in the patient.

Also provided are methods of reducing time to ‘on’ (e.g., accelerating the efficacy of levodopa) in a patient suffering from Parkinson’s disease comprising administering to the patient an effective amount of any of the formulations of the invention thereby providing 17 WO 2009/085306 PCT/US2008/014080 a plasma or serum concentration of levodopa effective to reduce the time to on in the patient.

Determining “on” and “off” time can be based on measuring conventional parameters 5 such as the Uniform Parkinson’s Disease Rating Scale (UPDRS) motor exam, walking time, and/or tapping number. For each of these parameters, definitions of “on” may be based on change from predose measure, and the results analyzed in the standard manner. For example, for the tapping number measure an about 10% change from the average of the predose measurements, may define the time of “on”. For walking time, an about 15% 10 change may be used.

In accordance with the practice of the invention, the plasma or serum concentration of levodopa comprises: a controlled release oral solid formulation of levodopa having a levodopa plasma or serum concentration profile comprising: a time of administration; a 15 first concentration, and a second concentration. In one embodiment of the method, said first concentration is equal to the maximum concentration of said profile; said second concentration is the minimum concentration occurring at a time later than said first concentration and earlier than or equal to about six hours following the time of administration; and wherein the second concentration is greater than or equal to about 20 fifty percent of said first concentration. In another embodiment, the second concentration is the minimum concentration occurring between one hour after said time of administration and said second time.

In the method of the invention, the concentration profile may be the median plasma or 25 serum concentration profile. Further, the concentration profile may be the mean plasma or serum concentration profile.

In one embodiment, the concentration profile further comprises a third concentration, wherein said third concentration is greater than or equal to fifty percent of said first 30 concentration and said third concentration occurs at a time earlier than said first concentration and within about ninety minutes of said time of administration. In another 18 WO 2009/085306 PCT/US2008/014080 embodiment, the third concentration is greater than or equal to sixty percent of said first concentration and said second concentration is greater than or equal to sixty percent of said first concentration.

In accordance with the practice of the invention, the disease includes but is not limited to Alzheimer’s disease, dystonia, schizophrenia and Parkinson’s disease.

The invention further provides methods of providing a therapeutically effective and stable median and/or mean blood plasma level of levodopa in a subject comprising administering to the subject a therapeutically effective amount of any of the formulations of the invention. In one embodiment, the blood plasma level does not fluctuate more than 40% between 0.5 hours after administration and six hours after administration,

ADVANTAGES OF THE INVENTION

Optimally, after administration to a patient suffering from a condition associated with reduced or impaired dopamine levels, a pharmaceutical formulation of the invention releases LD into the plasma of the patient at a steady or near constant level, i.e., with a low peak-to-trough ratio of LD plasma concentration, without any significant decrease or fluctuation for an extended amount of time, for example, mimicking infusion administration; thereby reducing motor fluctuations or the “on-off” effect associated with fluctuations in plasma LD levels caused by currently available oral dosage forms of CD/LD.

The pharmaceutical formulations of the invention provide a superior plasma LD profile to a patient than currently available oral pharmaceutical formulations. The formulations of the invention are able to provide a significantly smaller peak-to-trough LD plasma concentration ratio (i.e., narrowing the blood plasma ranges of LD after the initial peak) with e.g., Q6h, dosing. Additionally, some embodiments of the pharmaceutical formulations of the invention provide for improvement by increasing the plasma level of LD. The pharmaceutical formulations of the invention also provide narrow ranges of 19 WO 2009/085306 PCT/US2008/014080 plasma LD levels thereby minimizing the “on-off’ effect in some patients. The sustained, steady LD plasma profile of this invention is expected to provide a superior and consistent disease control.

The following examples are presented to illustrate the present invention and to assist one of ordinary skill in making and using the same. The examples are not intended in any way to otherwise limit the scope of the invention.

EXAMPLE 1: A TABLET OF CARBIDOPA AND LEVODOPA WITH

TARTARIC ACID

The bioavailability/pharmacokinetic results of a tablet foimulation of the present invention, using 50-200 mg of CD-LD with 215 mg of tartaric acid, were compared to the controlled release version of Sinemet®.

Preparation of a tablet of CD-LD with tartaric acid (IPX066-BQ5-Q7) CD, LD, and hydroxypropyl were charged into a blender and mixed uniformly. The powder mix was then charged in a high shear granulator and granulated with purified water. Drying of the granules was done overnight in an oven at 60 ± 10°C. Dried granules were passed through a 25 mesh screen, then charged and blended with magnesium stearate in a blender.

To prepare the tartaric acid final blend, granular tartaric acid was passed through a 20 mesh screen. The tartaric acid, microcrystalline cellulose, and hypromellose were charged and blended in a high shear mixer and granulated with ethyl alcohol. The resultant granules were dried in fluid bed processor at 55 ± 10°C . The dry granules were passed through a 25 mesh screen then charged and blended with magnesium stearate in a blender.

The final CD/LD and tartaric acid blends were compressed into a tablet. 20 WO 2009/085306 PCT/US2008/014080

Resulting Effect of Tartaric Acid on the Pharmacokinetics of Carbidopa-Levodopa 50-200 mg Formulations in Human Subjects

Table 1

Product Strength (mg) Dissolution in SGF ΪΡΧ066 CD/LD Tablets containing tartaric acid 50-200 Release over 4 hr 1PX066 CD/LD Tablets without tartaric acid 50-200 Release over 6 hr ΪΡΧ066 CD/LD Tablets with tartaric acid 50-200 Release over 6 hr Sinemet® CR Tablets3 50-200 Release over 3 hr 5 a Merck & Co., Inc., expiration date Aug. 2007 in vitro dissolution profiles of study drugs are listed below. Formulation tables for IPXO66-B05-O7 A, B and C are shown at the end of this example. 10 Table 2

Test Formulation Tartaric Acid (mg) A bi-layer 215 B ER 0 C bi-layer 215 Drug release (%); SGF, 50 rpm, USP Apparatus II w/basket sinker Test Compound 30 60 120 180 240 360 A CDP 164: 1.8 3O±3.5 554:6,2 73±7.8 86±7.5 974:3,0 LDP I6±1.8 304:3,6 544:6.3 724:7.9 844:7.5 954:3.0 B CDP 134:0.6 22±1.3 384:2.5 52±3,5 644:4.3 844:4.7 LDP 13=1=0.7 224:1.5 3 94:2.9 53=1=4.1 664:5.0 874:5.2 C CDP 124:0.9 21=1=2.1 39±4.6 554:6.3 684:6.7 864:5.3 LDP 12=1=1.1 21=1=2.3 39±5,0 554:6.7 684:7.2 87±5.9 21 WO 2009/085306 PCT/US2008/014080

Results and Discussion: Figure 2 shows a graph illustrating the in vivo plasma concentration profiles of three formulations of CD, LD with tartaric acid (referred to herein as IPX066-B05-07 formulations A, B and C) compared to Sinemet® after oral administration.

Pharmacokinetics of1PX066 Test B and Sinemet® CR

Following administration of one IPX066 Test B tablet, multiple peaks appeared in the LD plasma profiles, with a maximum plasma concentration (Cmax) occurring at approximately 2.5 hours post dose (Figure 2). In contrast, CD was slowly absorbed, with median Cmax occurring at approximately 4 hours post dose. Due to extended dissolution rate, IPX066 Test B with the dissolution rate of 6 hours as opposed to the Reference Sinemet® CR with the dissolution rate of 3 hours decreased by 46% in Cmax and 44% in AUC of LD, and 38% of Cmax and 41% in AUC of CD.

Pharmacokinetics of IPX066 Test C and ΪΡΧ066 Test B

Following administration of one IPX066 Test C tablet, multiple peaks also appeared in the LD plasma profiles, with Cmax occurring at approximately 3 hours post dose (Figure 2). In contrast, CD was slowly absorbed, with median Cmax occurring at approximately

4.5 hours post dose. Even with the extended dissolution rate, IPX066 Test C with the addition of 215 mg of tartaric acid, when compared to ΪΡΧ066 Test B without the addition of tartaric acid, increased by 50% in Cmax, 41% in AUC, 119% in C6h, and 65% in C8h of LD, and 32% of Cmax and 35% in AUC of CD

Pharmacokinetics of IPXQ66 Test C and IPX066 Test A

Following administration of one IPX066 Test A tablet, multiple peaks also appeared in the LD plasma profiles, with Cmax occurring at approximately 2 hours post dose (Figure 2). In contrast, CD was slowly absorbed, with median Cmax occurring at approximately 22 WO 2009/085306 PCT/US2008/014080 4.5 hours post dose. In comparison with IPX066 Test A, IPX066 Test C contained the same amount of tartaric acid and yet a slower dissolution rate by 2 hours. As a result, the LD Cmax, LD AUC, LD C6h, LD C8h, CD Cmax, and CD AUC were reduced by approximately 20%, 14%, 26%, 4%, 22%, and 18%, respectively. 5

Pharmacokinetics ofIPX066 Test A and Sine met® CR

Following administration of one IPX066 Test A tablet, multiple peaks also appeared in the LD plasma profiles, with Cnax occurring at approximately 2 hours post dose (Figure 10 2). in contrast, CD was slowly absorbed, with median Cmax occurring at approximately

4.5 hours post dose. BE assessment demonstrated that IPX066 Test A with the dissolution rate of 4 hours and tartaricacidincluded in formulation is bioequivalent to the Reference Sinemet® CR with the dissolution rate of 3 hours with regard to the Cmax and AUC values for both LD and CD. In addition, the C6h and C8h of LD of IPX066 Test A 15 were lower than those of Reference Sinemet® CR by approximately 25% and 4%, respectively.

The data herein demonstrated that decreasing dissolution rate decreases the exposure of LD and CD, and addition of tartaric acid increases the Cmax and AUC of LD and CD. 20

ί PXO66-BO507 Formulation A ingredients Per Tablet %(w/w) Mg Carbidopa 16.8 54.0 Levodopa 25.47 200.0 Hydroxypropyl cellulose (Klucel-LF) 12.63 99.2 Tartaric Acid 27.38 215.0 Macrocrystalline cellulose (Avicel PHI01) 21.63 169.8 Hypromellose (Methocel K100LV) 5.48 43.0 Magnesium Stearate 0.53 4.2 Total 100 785.2

Note. 53.09mg CD, USP is equivalent to 50.0 mg CD anhydride 23 WO 2009/085306 PCT/US20O8/014080

ΪΡΧ066-Β05-07 Formulation B

Ingredients Per Tablet %(w/w) Mg Carb tdopa 16,8 54.08 Levodopa 62.2 200.00 Hydroxypropyl cellulose 20.0 64.3 Magnesium Stearate 1.0 3.2 Purified water - - Total 100.0 321.5

5 IPX066-B05-07 Formulation C

Ingredients Per Tablet %(w/w) mg Carbidopa 6.30 54.0 Levodopa 23.34 200.00 Hydroxypropyl cellulose (KIucel-LF) 19,94 170.9 Tartaric Acid 25,09 215.0 Microcrystalline cellulose (Avicel PH101) 17.82 152,7 Hypromellose (Methocel K100LV) 7.03 60.2 Magnesium Stearate 0.48 4.1 Total 100 856.9 Note. 53.09mg CD, USP is equivalent to 51 3.0 mg CD anhyc ride EXAMPLE 2 10

Preparation of IPX066-B06-02 Formulations A and B

The following steps were performed to prepare enteric coated pellets containing CD and LD. 24 WO 2009/085306 PCT/US2008/014080 CD, LD. and microcrystalline cellulose (Avicel PH-101) were charged and mixed uniformly. The powder mix was charged into a high shear granulator and granulated with purified water. The granulated wet mass was extruded in an extruder with 1.0 mm hole size screen. The extrudate was charged and spheronized in a spheronizer equipped with 3 mm cross-hatch disc. The CD/LD pellets were dried at 60 ± 10°C in fluid bed processor. The CD/LD pellets were passed through different size screens. The pellets collected were retained on 18 and 25 mesh screens.

The following steps were performed to prepare the enteric coating solution for the CD-LD pellets.

For Formulation IPX066-B06-02 A, Eudragit® SI00 and Eudragit® LI00 (at the weight ratio of 2:1) and triethyl citrate were dissolved in isopropyl alcohol and acetone solution. The mixture was mixed until dissolved. Talc was dispersed into the polymer solution and mixed continuously throughout the coating process.

For Formulation 1PX066-B06-02 B, Eudragit® SI00 and triethyl citrate were dissolved in isopropyl alcohol and acetone solution. The mixture was mixed until dissolved. Talc was dispersed into' the polymer solution and mixed continuously throughout the coating process.

For both formulations, the CD/LD pellets were spray-coated using the coating dispersion prepared in Glatt GPCG-1 coater. The coated pellets were dried. The dried, coated CD/LD pellets were screened through a 16 mesh screen. The screened CD/LD pellets were charged and blended with talc in a blender.

The following steps were performed to prepare enteric coated pellets containing tartaric acid (TA). 25 WO 2009/085306

PCT/US2OO8/O1408O TA was passed through a 20 mesh screen. The screened TA and microcrystalline cellulose (Avicel PHI01) were charged in a high shear mixer and granulated with purified water. The granules were extruded in an extruder with 1.0 mm hole size screen. The extrudate was charged and spheronized into a spheronizer equipped with a 3 mm cross hatch disc. The seeds were dried overnight in an oven at 60 ± 10°C. The dried pellets were passed through 16, 18 and 25 mesh screens. The pellets retained were collected on 18 and 25 mesh screens. A seal coat solution was prepared by charging hypromellose (Pharmacoat 606) and ethylcellulose into an alcoholic solution. The mixture was mixed until dissolved. The dried TA pellets were charged in a coater and spray coated with the seal coat solution prepared. The seal coated CD/LD pellets were dried, and passed through a 14 mesh screen.

The following steps were performed to prepare the enteric coating solution for the TA pellets.

For Formulation IPX066-B06-02 A, Eudragit® SI00 and Eudragit® LI00 (at the weight ratio of 2:1) and triethyl citrate were dissolved in isopropyl alcohol and acetone solution. The mixture was mixed until dissolved. Talc was dispersed into the polymer solution and mixed continuously throughout the coating process.

For Formulation IPX066-B06-02 B, Eudragit® SI00 and triethyl citrate were dissolved in isopropyl alcohol and acetone solution. The mixture was mixed until dissolved. Talc was dispersed into the polymer solution and mixed continuously throughout the coating process.

For both formulations, the sealed coated TA pellets were charged in a Glatt GPCG-1 coater and spray coated with the enteric coating solution prepared. The coated TA pellets were dried. The dried, coated TA pellets were passed through a 12 mesh screen. The coated TA pellets were charged and blended with talc in a blender. 26 WO 2009/085306 PCT/US2008/014080

The following steps were performed to encapsulate the enteric coated drug pellets and enteric coated TA pellets.

The coated CD/LD pellets prepared as described above and coated TA pellets prepared as described above were encapsulated into hard gelatin capsules. The filled capsules contained 50 mg carbidopa anhydride, 200 mg LD and 215 mg TA.

Preparation of IPX066-B06-02 Formulations D and E

The following steps were performed to prepare the carbidopa (CD) and levodopa (LD) final blend. CD, LD, microcrystalline cellulose, and croscarmellose sodium were charged into a blender and mixed uniformly into a powder. Com starch was dispersed in purified water and stirred for 15 minutes, transferred to boiling water, and stirred continuously until it became starch paste. The spray rate of the peristaltic pump was verified using the starch paste. The powder mix prepared above was charged in a high shear granulator and granulated with starch paste at a flow rate of 50 ~ 1000 g/min. The granules were dried in an oven at 60 ± 10°C until LOD was lower than 3.0%. The dried granules were passed through a 25 mesh screen. The screened CD/LD granules, crospovidone and magnesium stearate were charged and blended in a blender.

To prepare the TA final blend, granular TA was passed through a 20 mesh screen. The TA and microerystalline cellulose were charged and blended in a high shear mixer and granulated with purified water. The resulting granules were dried in an oven at 60 ± 10°C until L.O.D, measured by a moisture analyzer was lower than 2,0%. The dry granules were passed through a Fitzmill equipped with a 24 mesh screen. The dried TA granules and magnesium stearate were charged and blended in a blender.

The final blends were compressed into core tablets as follows. 27 WO 2009/085306 PCT/US2008/014080

The CD/LD final blend and TA final blend were weighed, mixed and compressed into a tablet. The tablet contained 50 mg carbidopa anhydride, 200 mg LD and 215 mg TA. A seal coat was applied to the core tablet by spray coating the core tablet with 5 hypromellose dissolved in the mixture of isopropyl alcohol and purified water solution in

Pan Coater. The tablets were dried in the coating pan at 60 ± 10°C until the L.O.D. was lower than 3.0%.

The enteric coating solution was prepared as follows, 10

For IPX066-B06-02 Formulation D, Eudragit® S100 and Eudragit® L100 (at the weight ratio of 0.25:1) and triethyl citrate were dissolved in isopropyl alcohol and acetone solution. The mixture was mixed until dissolved. Talc was dispersed into the polymer solution and mixed continuously throughout the coating process. 15

For IPX066-B06-02 Formulation E, Eudragit® LI00 and triethyl citrate were dissolved in isopropyl alcohol and acetone solution. The mixture was mixed until dissolved. Talc was dispersed into the polymer solution and mixed continuously throughout the coating process. 20

The sealed coated tablet was charged into a Pan Coater and spray coated with the enteric coating solution prepared. The coated tablet was dried in the coating pan at 40 £ 10°C for at least 30 minutes. 25 Resulting Effect of Tartaric Acid and pH of Enteric Coating on the Pharmacokinetics of

Carbidopa-Levodopa 50-200 mg Formulations in Human Subjects

This study shows the effect of acid addition and of various pH of enteric-coating to carbidopa (CD)/levodopa (LD) 50-200 mg formulation on the PK of CD and LD. 30 28 WO 2009/085306 PCT/US2008/014080

Table 3

Product Strength (mg) Enteric Coated pH IPX066 CD/LD Capsule with tartaric acid 50-200 6.5 IPX066 CD/LD Capsule with tartaric acid 50-200 7.0 Sinemet® CR Tablet3 50-200 N/A IPX066 CD/LD Tablet with tartaric acid 50-200 6.5 ΪΡΧ066 CD/LD Tablet with tartaric acid 50-200 6.0 a Merck & Co,, Inc., expiration date Aug. 2007

Formulation information of study drugs are listed below. Formulation tables for IPX066-5 B06-02 A, B, D and E are shown at the end of this example.

Table 4

Test Formulation Particle size (pm) Tartaric Acid (mg) A Capsule D(v,0.9) =36.22 D(v,0.9) =139.48 215 B Capsule D(v,0.9) =36.22 D(v,0.9) =139.48 215 D Tablet D(v,0.9) =36.22 D(v,0,9) =139.48 215 E Tablet D(v,0.9) =36.22 D(v,0,9) =139.48 215

Results and Discussion: Figure 3 show a graph illustrating the in vivo plasma 10 concentration profiles of four formulations of CD, LD and TA (referred to herein as IPX066-B06-02 formulations A, B, D and E) compared to Sinemet® after oral administration. 29 WO 2009/085306 PCT/US2008/014080

Pharmacokinetics of IPX066 Test A and Sinemet® CR

Following administration of one IPX066 Test A capsule, one peak appeared in the LD median plasma profiles, with a maximum plasma concentration (Cmax) occurring at approximately 5.0 hours post dose (Figure 3). In contrast, CD was slowly absorbed, with median Cmax occurring at approximately 6.0 hours post dose. Due to delayed dissolution rate (enteric coated pH~6.5), no LD concentration was observed before 1 hr for IPX066 Test A and the LD concentration of ΪΡΧ066 Test A between 5 and 8 hr was higher than that of the Reference Sinemet® CR based on the median profile suggesting that addition of TA (215 mg) increase the absorption of LD in the later part of intestine. The AUC of LD for 1PX066 Test A was only 58% of that of reference Sinemet® CR.

Pharmacokinetics of IPXQ66 Test B and Sinemet® CR

Following administration of one IPX066 Test B capsule, one peak also appeared in the LD median plasma profiles, with delayed and lower Cmax occurring at approximately 4.0 hours post dose (Figure 3). In contrast, CD was slowly absorbed, with median Cmax occurring at approximately 4.75 hours post dose. No LD concentration was observed before 2 hr for IPX066 Test B. Even with the addition of TA (215 mg), IPX066 Test B did not have higher LD concentration between 5 and 8 hr when compared to Sinemet® CR. This could be due to the dissolution rate of TA is slower than that of LD. The AUC of LD for IPX066 Test B was only 23% of that of reference Sinemet® CR.

Pharmacokinetics of IPX066 Test D and Sinemet® CR

Following administration of one IPX066 Test D tablet, one peak also appeared in the LD median plasma profiles, with delayed and lower Cmax occurring at approximately 4,25 hours post dose (Figure 3). In contrast, CD was slowly absorbed, with median Cmax occurring at approximately 6.0 hours post dose. No LD concentration was observed before 2 hr for IPX066 Test D. Even with the addition of TA (215mg), LD AUC of IPX066 Test D was only 24% of that of Reference Sinemet® CR and LD concentration 30 WO 2009/085306 PCT/US2008/014080 between 5 and 8 hr was lower than that of Sinemet® CR. This is due to the highly variable in vitro dissolution profile and the faster release rate of TA when compared with that of LD.

5 Pharmacokinetics of IPX066 Test E and Sinemet® CR

Following administration of one IPX066 Test E tablet, one peak also appeared in the LD median plasma profiles, with delayed and lower Cmax occurring at approximately 4.0 hours post dose (Figure 3). In contrast, CD was slowly absorbed, with median Cmax 10 occurring at approximately 5.0 hours post dose. No LD concentration was observed before 2 hr for IPX066 Test E, Even with the addition of tartaric acid (215mg), LD AUC of IPX066 Test E was only 20% of that of Reference Sinemet® CR and LD concentration between 5 and 8 hr was lower than that of Sinemet® CR. This is also due to the highly variable in vitro dissolution profile and the faster release rate of TA when compared with 15 that of LD.

The data herein demonstrated that delayed dissolution rate decreases the exposure of LD and CD, and addition of TA which has similar dissolution rate to LD and CD has higher LD concentration between 5 and 8 hr when compared to Sinemet® CR.

IPX066-B06-02 Formulation A

Ingredients % (w/w) Amount (mg) Carbidopa 14.06 53.98 Levodopa 52.10 200,0 Microcrystalline Cellulose, (Avicel PH-101) 28.36 108.85 Eudragit® L100 1.14 4.39 Eudragit® SI00 2.34 8.98 Triethyl Citrate 1.00 3.82 Talc 1.00 3.83 Total 100.0 383.85 31 WO 2009/085306 PCTZUS2008/014080

Tartaric Acid 57.6 215.0 Microcrystalline Cellulose, NF (Avicel PH- 101) 14.4 - 53.8 Ethylcellulose 6.6 24.8 Hypromellose, Type 2910 1.4 5.1 Eudragit® L100 4.6 17.2 Eudragit® SI00 9.4 35.1 Triethyl Citrate 4.0 14.9 Talc 2.0 7.5 Total 100.0 373.4

IPX066-B06-02 Formulation B

Ingredients % (w/w) Amount (mg) Carbidopa 12.92 53.98 Levodopa 47.88 200.00 Microcrystalline cellulose (Avicel PHI01) 26.06 108.85 Eudragit® SI00 8.85 36.95 Triethyi Citrate 2.53 10.55 Talc 1.76 7.37 Total 100.0 417.70

Tartaric Acid 57.6 215.0 Microcrystalline cellulose (Avicel PHI01) 14.4 53.8 Ethycellulose 6.6 24.8 Hypromellose, 1.4 5,1 Eudragit® SI 00 14.0 52.3 Triethyl Citrate 4.0 14.9 Talc 2.0 7.5 Total 100.0 373.4 32 WO 2009/085306 PCT/US2008/014080

IPX066-B06-02 Formulation D ingredients % (w/w) mg Carbidopa 8,57 53,98 Levodopa 31,77 200.00 Microcrystailine cellulose (Avicel FH101) 15,17 95.5 Com Starch, 1.43 8.98 Croscarmellose Sodium 1.43 8,98 Crospovidone, 0.93 5,88 Magnesium Stearate 1.13 7.11 Tartaric Acid 34,15 215.0 Hypromeltose, type 2910 2.93 18.42 Eudragit^ L100 1.40 8.82 Eudragit® S 100 0.35 2.20 Tri ethyl Citrate 0.50 3.15 Talc 0.25 1.57 Total 100.0 629.59 33 WO 2009/085306 PCT/US2008/014080

ΪΡΧ066-Β06-02 Formulation E

Ingredients % (w/w) Amount (mg) Carbidopa 8.57 53.98 Levodopa 31.77 200.0 Avicel PH-101 15.17 95.5 Com Starch 1.43 8.98 Croscarmellose Sodium 1.43 8.98 Crospovidone 0.93 5.88 Magnesium Stearate 1.13 7.11 Tartaric Acid 34.15 215.00 Hypromellose, Type 2910 2.93 18.42 Eudragit^LlOO 1.75 11.02 Triethyl Citrate 0.5 3.15 Talc 0.25 1.57 Total 100.0 629.59 EXAMPLE 3

5 Preparation of IPX066-B07-01 Formulations A, B and C

The following steps were performed to prepare enteric coated pellets containing Carbidopa-Levodopa (CD-LD). 10 CD, LD, and microcrystalline cellulose (Avicel PH-101) were blended together. The mixture was charged into a high shear granulator and granulated with purified water. The granulated wet mass was extruded in an extruder with 1.0 mm hole size screen. The extrudate was charged and spheronized in a spheronizer equipped with 3 mm cross-hatch disc. The spheres obtained from the spheronizer were dried at 60+10°C in a Glatt GCPG- 15 1 coater. Drying the pellets in the Glatt GPCG-1 eliminated discoloration of the pellets and also reduced the amount of degradation product DHP. The drug-loaded pellets were 34 WO 2009/085306 PCT/US2008/014080 screened with 16, 18 and 25 mesh screens and the collected pellets retained on 18 and 25 mesh screens. The core pellets were then coated with hypromellose (Pharmacoat 606) aqueous solution in a Glatt GCPG-1 coater. The coated pellets were screened with an 18 mesh screen after they were dried in GPCG-1. 5

The enteric coating solution was prepared as follows. For formulation IPX066-B07-01 A, Eudragit® SI00 and Eudragit® LI00 were dispersed separately at the weight ratio of 5:1 into purified water and mixed until a uniform dispersion. IN NH4OH solution was then added drop-wise to both solutions until the pH reached 5.5, The two solutions were 10 combined together and mixed thoroughly. A talc suspension was prepared by dispersing Talc into triethyl citrate solution in purified water and then stirred for 1 hour. The above mixture of LI00 and SI00 was then combined with the Talc dispersion and mixed thoroughly. The dispersion was screened through a 140 mesh screen before starting the coating process. For IPX066-B07-01 formulation B, the same procedure was followed 15 for formulation A except using only Eudragit® S100 polymer. For IPX066-B07-01 formulation C, the same procedure was followed for formulation A, except using only Eudragit® FS30D polymer.

The hypromellose coated seeds were spray coated with the enteric coating dispersion 20 preparation. The coated CD/LD pellets were dried in an oven. The dried CD/LD pellets were passed through a 14 mesh screen. The final blend was prepared by blending the screened CD/LD pellets with talc.

The following steps were performed to prepare enteric coated pellets containing TA. 25 TA was passed through a 20 mesh screen. The screened TA and microcrystalline cellulose (Avicel PH101) was charged in a high shear mixer and granulated with purified water. The wet mass was extruded in an extruder with 1.0 mm hole size screen. The extrudate was charged and spheronized in a spheronizer equipped with a 3 mm cross 30 hatch disc. The pellets obtained were dried overnight in an oven at 60 ± I0°C. The dried pellets were passed through 16, 18 and 25 mesh screens and the collected pellets retained 35 WO 2009/085306 PCT/US2008/014080 on 18 and 25 mesh screens. The seal coat solution was prepared by dissolving hypromellose (Pharmacoat 606) and ethylcellulose in alcoholic solution. The seal coat solution was applied to the TA pellets in a Glatt GPCG-1 coater. The pellets were dried in a GPCG-1 and passed through a 14 mesh screen.

The enteric coating solution was prepared as follows. For IPX066-B07-0! formulation A, Eudragit® S100 and Eudragit® L100 were dispersed separately at the weight ratio of 5:1 into purified water and mixed until a uniform dispersion. IN NH4OH solution was then added drop-wise to both solutions until pH reached 5.5. The two solutions were combined together and mixed thoroughly. A talc suspension was prepared by dispersing Talc into triethyl citrate solution in purified water and then stirred for 1 hour. The above mixture of L100 and S100 was then combined with the Talc dispersion and mixed thoroughly. The dispersion was screened through a 140 mesh screen before starting the coating process. For IPX066-B07-01 formulation B, the same procedure was followed for formulation A, except using only Eudragit® S100 polymer. For IPX066-B07-01 formulation C, the same procedure was followed for formulation A except using only Eudragit® FS30D polymer. The screened tartaric acid pellets were charged in a Glatt GPCG-1 coater and spray coated with the enteric coating dispersion. The coated seeds were dried in oven. The dried, coated pellets were screened through a 14 mesh screen. The final blend was prepared by blending the screened pellets and talc.

The coated CD/LD pellets and coated TA pellets were encapsulated into hard gelatin capsules. The filled capsules contained 50 mg carbidopa anhydride, 200 mg LD and 215 mg TA. 36 WO 2009/085306 PCT/US2008/014080

Resulting Effect of Tartaric Acid and pH of Enteric Coating on the Pharmacokinetics of Carbidopa-Levodopa 50-200 mg Formulations in Human Subjects

Objectives: 5 This study tested the effect of acid addition and of various pH of enteric-coating (Eudragit® S100/L100=5, Eudragit® S100 and Eudragit® FS 30D) to carbidopa (CD)/levodopa (LD) 50-200 mg formulation on the PK of CD and LD.

Table 5

Product Strength (mg) Enteric Coated Polymer ΪΡΧ066 CD/LD Capsule with tartaric acid 50-200 Eudragit®1 S1Q0/L 100=5 1PX066 CD/LD Capsule with tartaric acid 50-200 Eudragit® SI00 ΪΡΧ066 CD/LD Capsule with tartaric acid 50-200 Eudragit® FS 30D Sinemet® CR Tablet3 50-200 N/A 10 a Merck & Co., inc., expiration date 02/2009

Formulation information and in vitro dissolution profiles of study drugs are listed below. Formulation tables for IPX066-B07-01 A, B and C are shown in the end of this example. 15 Table 6

Test Formulation Particle size (pm) Tartaric Acid (mg) A Capsule D(v,0.9) =36.22 D(v,0.9) =139.48 215 B Capsule D(v,0.9) =36.22 D(v,0.9) = 139.48 215 C Capsule D(v,0.9) =36.22 D(v,0.9) =139.48 215 37 WO 2009/085306 PCT/US2008/014080

Results and Discussion: Figure 4 shows a graph illustrating the in vivo plasma concentration profiles of three formulations of CD, LD and TA (referred to as IPX066-B07-01 formulations A, B and C) compared to Sinemet® after oral administration.

5 Pharmacokinetics of ΪΡΧ066 Test A and Sinemet® CR

Following administration of one IPX066 Test A capsule, one peak appeared in the LD median plasma profiles, with a maximum plasma concentration (Cmax) occurring at approximately 2.75 hours post dose (Figure 4), and the LD concentration remains at the 10 same level from 2.0 to 3.5 hrs. In contrast, CD was slowly absorbed, with median Cmax occurring at approximately 4.0 hours post dose. Due to delayed dissolution rate (enteric coated with Eudragit® S100/L100=5), the absorption of LD (the ascending phase of plasma profile) from IPX066 Test A was slow compared with Sinemet® CR and the LD concentration of IPX066 Test A between 5 and 10 hr was higher than that of the 15 Reference Sinemet® CR based on the median profile suggesting that addition of TA (215 mg) increase the absorption of LD in the lower part of intestine. The AUC of LD for IPX066 Test A was only 87.72% of that of reference Sinemet® CR.

Pharmacokinetics of IPX066 Test B and Sinemet® CR 20

Following administration of one IPX066 Test B capsule, one peak also appeared in the LD median plasma profiles, with delayed and lower Cmax occurring at approximately 5.0 hours post dose (Figure 4). In contrast, CD was slowly absorbed, with median Cmax occurring at approximately 4.5 hours post dose. No LD concentration was observed 25 before 0.75 hr for IPX066 Test B. Because of tartaric acid (215 mg) in the formulation, IPX066 Test B also has higher LD concentration between 5 and 8 hr when compared to Sinemet® CR. However, the AUC of LD for IPX066 Test B was only 56.5% of that of reference Sinemet® CR. 38 WO 2009/085306 PCT/US2008/014080

Pharmacokinetics of IPX066 Test C and Sinemet® CR

Following administration of one 1PX066 Test C tablet, only 4 subjects have detectable LD concentration (Figure 4). This is due to the slow in vitro release profile for the 5 CD/LD core seed.

The data herein demonstrated that delayed dissolution rate decreases the exposure of LD and CD, and addition of TA to LD and CD formulation has higher LD concentration between 5 and 8 hr when compared to Sinemet® CR.

ΪΡΧ066-Β07-01 Formulation A

Ingredients % (w/w) Amount (mg) Carbidopa 11.24 53.98 Levodopa 41.65 200,0 Microciystalline cellulose (Avicel PH101) 22.67 108.85 HPMC 3.98 19,10 Eudragit® LI 00 1.70 8.17 Eudragit® SI 00 8.51 40.85 Triethyl Citrate 7.13 34.26 Talc 3.04 14.58 Ammonia Solution 0.08 0.38 Total 100.0 480.17 39 WO 2009/085306 PCT/US2008/014080

Tartaric Acid 42.27 215.0 Microcrystalline cellulose (Avicel PH101) 10.58 53.8 Ethylcellulose 4.88 24.8 HPMC 1.00 5.1 Eudragit® LI 00 3.48 17.7 Eudragit® SI00 17.36 88.3 Triethyl Citrate 14.57 74.1 Talc 5.70 29.0 Ammonia Solution 0.16 0.8 Total 1 100.0 508.6

IPX066-B07-01 Formulation B

Ingredients % (w/w) Amount (mg) Carbidopa 12.65 53.98 Levodopa 46.87 200.0 Microcrystalline cellulose (Avicel PHI01) 25.51 108.85 Hypromellose, Type 2910 4.48 19.10 Eudragit® S100 6.42 27.38 Triethyl Citrate 3.21 13.71 Talc 0.80 3.40 Amonia Solution 0.06 0.27 Total 100.0 426,69 40 WO 2009/085306 PCT/US2008/014080

Tartaric Acid 42.38 215.0 Microcry stall inc cellulose (Avicel PH101) 10.61 53.8 Ethylcellulose 4.89 24.8 Hypromellose, Type 2910 1.01 5.1 Eudragit® SI00 26.01 131.95 Triethyl Citrate 13,01 65.98 Talc 1.81 9.16 Ammonia Solution 0.29 1.46 Total 100.0 507.25

ΪΡΧΟ66-Β07 Ο1 Formulation C

Ingredients % (w/w) Amount (mg) Carbidopa 8.57 53.98 Levodopa 31.74 200.0 Microcrystalline cellulose (Avicel PH-101) 17.28 108.85 Hypromellose, Type 2910 3,03 19.10 Eudragit® FS 30D 30.31 190.97 Triethyl Citrate 0.91 5.71 Talc 8.07 50.86 Ammonia Solution 0.10 0.63 Total 100.0 630.10 41 WO 2009/085306 FCT/US2008/014080

Tartaric Acid 38.60 215.0 Microcrystalline cellulose (Avicel PH-101) 9.66 53.8 Ethylcellulose 4,45 24.8 Hypromellose, Type 2910 0,92 5.1 Eudragit® FS 30D 35.75 199.1 Triethyl Citrate 1.08 6.0 Talc 9.44 52.6 Ammonia Solution 0.11 0.6 Total 100.0 557.0 EXAMPLE 4 5

The data herein shows the bioavailability/pharmacokinetic results of an enteric coated tablet CD/LD formulations, using 50-200 mg of CD-LD with 0-430 mg of tartaric acid, compared to the controlled release version of Sinemet®. 10 Four formulations of IPX-066-AH1 were evaluated for PK parameters. Information of the study drugs are listed below in Table 7.

Formulation A is a capsule containing IR beads exhibiting a rapid dissolution profile. 15 Formulation B is a capsule containing ER CD/LD beads and ER TA beads. The ER CD/LD beads were formulated by coating the IR beads with Eudragit® polymers (S100:L100 ratio of 2:1). The ER TA beads were coated with a sealed coat and a Eudragit® coat (S100:L100 ratio of 2:1) exhibiting a dissolution profile similar to the CD/LD beads. 20

Formulation C is a capsule containing ER CD/LD beads and ER TA beads. The ER CD/LD beads were formulated by coating the IR beads with Eudragit® polymers 42 WO 2009/085306 PCT/US2008/014080 (S1OO:L1O0 ratio of 5:1). The ER TA beads were coated with a sealed coat and a Eudragit® coat (S10O:L1OO ratio of 5:1) exhibiting a dissolution profile similar to the CD/LD beads. 5 Formulation D is similar to formulation B except that the quantity of the TA beads is twice the amount in formulation B.

Formulation E is the reference product, Sinemet® CR 200mg Tablet. 10 Table 7 IFX066- AHl(A-D)

Product CD-LD Strength (mg) Enteric Coated pH Formulation A ΪΡΧ066 CD/LD IR Capsule 50-200 — Formulation B IPX066 CD/LD Capsule with 215 mg tartaric acid 50-200 Formulation C IPX066 CD/LD Capsule with 215 mg tartaric acid 50-200 6.5 (SL5)b Formulation D IPX066 CD/LD Capsule with 430 mg tartaric acid 50-200 6.5 (SL2)b Formulation E (Reference) Sinemet® CR Tablet3 50-200 — a Merck & Co., Inc.; b: SUFEudragi? SIOO/L100 is’2:i; SL5: Eudragit®’ST 00/t 100 is 5:1.

The qualitative and quantitative compositions for these formulations are summarized 15 below in Table 8. 43 WO 2009/085306 PCT/US2008/014080

Table 8 Qualitative and Quantitative Composition of Formulation

Formulation

A B C

Ingredients mg/capsule

Component I Component Π Component IV Componen II Carbidopa, USP 27 27 27 27 Levodopa, USP 100 100 100 100 Microcrystalline Cellulose, NF (Avicel PH-101) 18.4 18-4 18.4 18.4 Lactose Monohydrate, NF 18-4 18-4 18.4 18.4 Sodium Starch Glycolate, NF 9.2 9.2 9.2 9.2 Sodium Lauryl Sulfate, NF 9.2 9-2 9.2 9.2 Povidone, USP 1.84 1.85 1.85 1.85 Talc, USP 1.95 1-95 4.2 1.95 Methacrylic acid copolymer, Type A, NF (Eudragit® L100) 2.25 2.15 2.25 Methacrylic acid Copolymer, Type B, NF (Eudragit4 SI00) 4.55 10.75 4.55 Triethyl Citrate, NF — 1.95 9.00 1.95 IN NIL,OH solution — — 0.1 — Purified Water, USP — — .N/A* — Acetone, NF N/A* N/A* — N/A* Isopropyl Alcohol, USP N/A* N/A* — N/A* Component in Component y Componen III Tartaric acid, NF — 107.5 107.5 215 Microcrystailine Cellulose, NF (Avicel PH-101) 26.95 26.9 53.9 Ethylcellulose, NF (Ethocel Standard- 10FP Premium) 11.2 11.3 22.4 Hypromeliose, USP Type2910 (Pharmacoat 606, 6 cps) 20.35 20.25 40.7 Methacrylic acid copolymer, Type A, NF (Eudragit® LI00) — 9.75 6.1 19.5 Methacrylic acid Copolymer, Type B, NF (Eudragit® SI00) 19.3 30.35 38.6 Triethyl Citrate, NF — 8.3 25.55 16.6 Talc, USP — 5.2 10.3 10.4 INNH4OH — — 0.3 — Purified Water, USP — — N/A* — Acetone, NF N/A* N/A* — N/A* Isopropyl Alcohol, USP N/A* N/A* — N/A* 44 WO 2009/085306 PCT/US2008/014080

Hard Gelatin Capsule 1 unit 1 unit 1 unit Total Capsule Fill Weight 184.97 403.3 448.4 * evaporated in drying process

Manufacture of IPX066- AHI Formulations

Four formulations of IPX066- AHI

Manufacture of Component J - CD-LD fast release beads

Carbidopa, levodopa, and microcrystalline cellulose, Lactose Monohydrate, Sodium Starch Glycolate, Sodium Lauryl Sulfate, and Povidone were blended and charged in a high shear granulator and granulated with purified water. The granulated wet mass was extruded in an extruder with. 1.0 mm hole size screen. The extrudate was charged and spheronized in a spheronizer equipped with 3 mm cross-hatch disc. The spheres obtained from the spheronizer were dried in and the drug-loaded pellets were screened with 16, 18 and 25 mesh screens. The pellets retained on 18 and 25 mesh screens were collected. The final blend was prepared by blending the screened CD/LD pellets with talc.

Manufacture of Component // - CD-LD fast release beads coated with Eudragit® SlOOtLIOO (2:1) CD/LD core seeds (also referred to as pellets) were prepared as Component I seeds except that there was no final blend with Talc. The enteric coating solution was prepared by dissolving Eudragit® S100 and Eudragit® L100 at the weight ratio of 2:1 and triethyl citrate in isopropyl alcohol and acetone solution. Talc was then dispersed into the polymer solution and mixed continuously throughout the coating process. The CD/LD pellets were spray coated using the coating dispersion in a coater. The coated pellets were dried and then screened through a 16 mesh screen. The final blend was prepared by blending the screened CD/LD pellets with talc. 45 WO 2009/085306 PCT/US2008/014080

Manufacture of Component III - Tartaric acid fast release beads coated with Eudragit® S1OO:L1QO (2:1)

Tartaric acid was passed through a 20 mesh screen. Screened tartaric acid and 5 microcrystalline cellulose was charged in a high shear mixer and granulated with purified water. The wet mass was extruded in an extruder with 1.0 mm hole size screen. The extrudate was charged and spheronized in a spheronizer equipped with a 3 mm cross hatch disc and the resulting pellets dried overnight in an oven at 60 ± 10°C. The dried pellets were passed through 16, 18 and 25 mesh screens and the pellets retained on 18 and 10 25 mesh screens were collected. A seal coat solution was prepared by dissolving hypromellose (Pharmacoat 606) and ethylcellulose in alcoholic solution. The seal coat solution was applied to the tartaric acid pellets in a coater. The pellets were dried and the dried pellets passed through a 14 15 mesh screen.

An additional seal coat was produced by dissolving hypromellose (Pharmacoat 606) in alcoholic solution. The seal coat solution was applied to the tartaric acid pellet coater, and then the pellets were dried in GPCG-1. The dried pellets were passed through a 14 mesh 20 screen.

An enteric coating solution was prepared by dissolving Eudragit® SI00 and Eudragit® LI00 at the weight ratio of 2:1 and triethyl citrate in isopropyl alcohol and acetone solution. Mix until dissolved. Talc was dispersed into the polymer solution and mixed 25 continuously throughout the coating process. The screened tartaric acid pellets were charged in a coater and spray coated with the enteric coating dispersion. The coated seeds were dried, and the dried, coated pellets screened through a 14 mesh screen. The final blend was prepared by blending the screened pellets and talc. 46 WO 2009/085306 PCT/US2008/014080

Manufacture of Component IV - CD-LD fast release beads coated with Eudragit® S100. L100 (5:1)

Method of manufacture for this component is the same as those of the component Π beads, except that the preparation of the enteric coating dispersion was changed. Specifically, the enteric coating dispersion is prepared by charging the required amount of a first portion of purified water in a stainless steel container, and to which while stirring, Methacrylic Acid Copolymer, Type A, NF were charged and dispersed, thereby making a dispersion. The required amount of IN NH4OH solution was added drop wise to the dispersion. A required amount of a second portion of purified water in a separate stainless steel container was charged. While stirring, Methacrylic Acid Copolymer, Type B, NF were charged and dispersed. Then a required amount of IN NH4OH solution was added drop wise to the dispersion. A required amount of a third portion of purified water was charged in a separate stainless steel container. Triethyl citrate was charged and dissolved and talc was added to the solution.

The first dispersion above was added and mixed to the second dispersion and to which was then added the third dispersion above. The mixed solutions were screened through 140 mesh screen.

Manufacture of Component V - TA fast release beads coated with Eudragit® SI00:LI00 (5:1)

Method of manufacture for component V beads is the same as those of the component HI beads, except that the preparation of the enteric coating dispersion was changed. 47 WO 2009/085306 PCT/US2008/014080

Specifically the enteric coating dispersion was prepared by charging the required amount of a first portion of purified water in a stainless steel container and, to which, while stirring, Methacrylic Acid Copolymer, Type A, and NF were charged and dispersed to yield a dispersion; then the required amount of IN NH4OH solution was added drop wise 5 to the above dispersion.

In a separate stainless steel container, the required amount of the second portion of purified water was charged and while stirring, Methacrylic Acid Copolymer, Type B, and NF were charge and dispersed to yield a dispersion; then the required amount of IN 10 NH4OH solution was added to the above dispersion.

In a separate stainless steel container, the required amount of the third portionof purified water was charged and to which triethyl citrate was added and charged; then talc was added to the above solution. 15

The first dispersion above was added and mixed to the second dispersion and to which was then added the third dispersion above. The mixed solutions were screened through 140 mesh screen. 20 Manufacture of the IPX066 Capsules ' The required amounts of the component beads were filled into hard gelatin capsules according to the specified fill weights in the Table 9 below. The in-process fill weight is controlled at target ± 10% of the target weights according to Table 9. 48 WO 2009/085306 PCT/US2008/014080

Table 9 Target Fill Weights of IPX066 Capsule Test Formulations

Components Formulation IPXO66-AH1(A) mg/capsule IPX066- AH1(B) mg/capsule 1PX066- AH1(C) mg/capsule IPX066- AH1(D) mg/capsule Component 1 Beads 184.97 — — — Component H Beads — 194.75 — 194.75 Component ΙΠ Beads — 208.55 417.1 Component IV Beads — — 210.25 — Component V Beads — — 238.55 — Total Capsule Fill weight 403.3 448.8 611.85

Pharmacokinetics result for biostudy IPX066- AHI 5 The Pharmacokinetics parameters of CD and LD for four tested formulations compared to Sinemet® CR after oral administration are summarized in Table 10.

Table 10 Mean ratio of Ln-transformed Cmax, AUC of Levodopa and Carbidopa

Parameter Test/Ref Ratio (%) LD Ln(Cmax) A/E 192.54 Ln(Cmax) B/E 127.05 Ln(Cmax) C/E 123.22 Ln(Cmax) D/E 111.99 Ln(AUC) A/E 126.53 Ln(AUC) B/E 115.03 Ln(AUC) C/E 127.15 Ln(AUC) D/E 108.74 CD Ln(Cmax) A/E 144.73 Ln(Cmax) B/E 88.97 Ln(Crnax) C/E '124.35 Ln(Croax) D/E 81.06 Ln(AVC) A/E 161.73 Ln(AUC) B/E 91.24 Ln(AUC) C/E 139.79 Ln(AUC) D/E 88,03 49 WO 2009/085306 PCTZUS2008/014080

Results and Discussion:

The data herein demonstrated that formulation A, with the inclusion of water soluble filler such as lactose and a surfactant such as sodium lauryl sulfate» exhibits a rapid LD 5 absorption with a rmax of 30 min. The PK profiles of Formulation B and D are desirable. Both formulations exhibit a significantly higher LD plasma concentration at 6 hours postdose relative to the Sinemet® CR Tablet. The absorption of LD from formulation B and D are also similar as indicated by a relative AUC of 115% and 109%, respectively. Since the PK profiles of formulation B and D are similar, formulation D with double the 10 quantity of TA beads does not offer any additional benefit over formulation B. Formulation C, with coating consisting of Eudragit® S100:L100 ratio of 5:1, exhibits a PK profile suggesting a faster in-νίνο release relative to formulation C and D. This is further supported by an AUC of 127% relative to Sinemet® CR Tablet indicating LD was rapidly released and absorbed in the upper G1 tract. 15 EXAMPLE 5

The data herein shows the bioavailability/pharmacokinetic results of an enteric coated tablet CD/LD formulations, using 50-300 mg of CD-LD with 0-270 mg of tartaric acid 20 compared to the controlled release version of Sinemet®. information of study drugs are listed below in Table 11.

IPX066-AH2 Formulation A (IPX066-AH2(A)) is a capsule containing 5 different component beads. Component I is an immediate-release CD/LD bead type. Component H 25 is an ER CD/LD bead type with a fast ER release profile. Component ill is an ER CD/LD bead type with a slow ER release profile. Component IV is a TA bead type with a release profile similar to those of Component Π. Component V is a TA bead type with a release profile similar to those of Component III. 50 WO 2009/085306 PCT/US2008/014080 IPX066-AH2 Formulation B (IPX066-AH2(B)) is a capsule containing ER CD/LD beads formulated with Cremophor RH40 and Poloxamer 188. TA is not included in the formulation. 5 1PX066-AH2 Formulation C (IPX066-AH2(C)) is a capsule containing ER CD/LD beads formulated with Cremophor RH40 and Poloxamer 188, and TA. IPX066-AH2 Formulation D (IPX066-AH2(D)) is a capsule containing ER CD/LD beads formulated with TA. The capsule does not contain Cremophor RH40 nor Poloxamer 188. 10

The reference product is Sinemet® CR Tablet 200 mg.

Table Π, ΪΡΧ066-ΑΗ2

Product CDLD Strength (mg) Enteric Coated pH A IPX066 CD/LD Combo Capsule with 270 mg tartaric acid (IR + fast release core +slow release core) 75-300 6.5 (SL2) and slow core for fast release B EPX066 CD/LD Capsule with surfactants 50-200 6.5 (SL2) C IPX066 CD/LD/tartaric acid Capsule with 215 mg tartaric acid and surfactants 50-200 6.5 (SL2) D IPX066 CD/LD/tartaric acid Capsule with 215 mg tartaric acid 50-200 6.5 (SL2) E Sinemet® CR Tablet3 50-200 - a Merck & Co., Inc. 15

The qualitative and quantitative compositions for these formulations are summarized below in Table 12 and Table 13. 51 WO 2009/085306 PCT/US2008/014080

Table 12. Formulation A (IPX066-AII2(A))

Formulation A Ingredients mg/capsule Component I Carbidopa USP 6.75 Levodopa USP 25 Microcrystal line Cellulose NF 4.6 Lactose Monohydrate NF 4.5 Sodium Starch Glycolate NF 2,3 Sodium Lauryl Sulfate NF 2.3 Povidone USP 0.46 Talc USP 0.23 Purified Water, USP N/A* Component Π Carbidopa USP 6.75 LevodopaUSP 25 Microcrystalline Cellulose NF 4.6 Lactose Monohydrate NF 4.6 Sodium Starch Glycolate NF 2.3 Sodium Lauryl Sulfate NF 2.3 Povidone USP 0.46 Methacrylic acid copolymer, Type A NF (Eudragit® LI 00) 0.55 Methacrylic acid Copolymer, Type B NF (Eudragit® SI00) L15 Triethyl Citrate NF 0.48 Talc USP 0.49 Acetone, NF N/A* Isopropyl Alcohol, USP N/A* Purified Water, USP N/A* Component III Carbidopa USP 26.99 Levodopa USP 100 Microcrystalline Cellulose NF 54,42 Methacrylic acid copolymer, Type A NF (Eudragit® L100) 2.18 Methacrylic acid Copolymer, Type B NF (Eudragit® SI00) 4.5 Triethyl Citrate NF 1.91 Talc USP 1.91 Acetone, NF N/A* Isopropyl Alcohol, USP N/A* Purified Water, USP N/A* Component IV 52 WO 2009/085306 PCT/US2008/014080

Tartaric acid NF 27 Microcrystalline Cellulose NF 6.79 Ethylcellulose NF 2.81 Hypromellose, Type2910 USP 5.01 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 2.44 Methacrylic acid Copolymer, Type B NF (Eudragit® SI 00) 4.84 Triethyl Citrate NF 2.08 Talc USP 1.31 Acetone, NF N/A* isopropyl Alcohol, USP N/A* Purified Water, USP N/A* Component V Tartaric acid NF 107.5 Microcrystalline Cellulose NF 26.85 Ethylcellulose NF 12.38 Hypromellose Type 2910 USP 2.63 Methacrylic acid copolymer, Type A NF (Eudragit® L100) 8.62 Methacrylic acid Copolymer, Type B NF (Eudragit® SI00) 17.5 Triethyl Citrate NF 7.45 Talc USP 4.69 Acetone, NF N/A* Isopropyl Alcohol, USP N/A* Purified Water, USP N/A* Hard Gelatin Capsule 1 unit Total Capsule Fill weight 526.63 *evaporated in drying process 53 WO 2009/085306 PCT/US2008/014080

Table 13. Formulation B, C and D (ΪΡΧ066-ΑΗ2(Β), IPX066-AH2(C), IPX066-AH2(D))

Ingredients Formulation B C D mg/capsule mg/capsule mg/capsule Carbidopa USP 26.99 26.99 26.99 Levodopa USP 100 100 100 Microcrystal line Cellulose, NF 118.75 65.00 30.00 Lactose Monohydrate NF 53.75 — — Mannitol, USP — — 50.00 Povidone, USP 17.35 — — Talc, USP 23.99 11.11 9.97 Methacrylic acid copolymer, Type A, NF (Eudragit® LI 00) 48.98 20.13 18.09 Methacrylic acid Copolymer, Type B, NF (Eudragit® S100) 98.19 40.35 36.17 Triethyl Citrate, NF 42.18 17.32 15.55 Poloxamer 188. NF 33.5 33.5 — Polyoxyl 40 Hydrogenated Castor Oil, NF (Cremophor RH 40) 17.5 17.5 Tartaric acid, NF — 107.5 107.5 Hypromellose, USP Type 2910 52.58 47,18 Acetone, NF N/A* N/A* N/A* Isopropyl Alcohol, USP N/A* N/A* N/A* Purified Water, USP N/A* N/A* N/A* Hard Gelatin Capsule 1 unit 1 unit 1 unit Total Capsule Fill Weight 581.18 491.98 441.45 *evaporated in drying process 54 WO 2009/085306 PCT/US2008/O14080

Manufacture of Formulation Λ (IPX066-AII2(A))

Manufacture of Component Bead I- CD/LD Fast release beads 5 Method of manufacture for this component beads is the same as to that for component I beads in ΓΡΧ066-ΑΗ1. as discussed in Example 4 above.

Manufacture of Component ΙΪ - CD-LD fast release beads coated with Eudragit® SIOQ.LIOO (2:1) 10

Method of manufacture for this component beads is the same as that for component II beads.in IPX066-AH1, as discussed in Example 4 above.

Manufacture of Component 111 - CD-LD slow release beads coated with Eudragit® 15 S100:L100 (2:1)

Method of manufacture for this component beads is the same as that for enteric coated CD-LD beads in formulation A tested in 1PX066-B06-O2, as discussed in Example 2 above. 20

Manufacture of Component IV - TA fast release beads coated with Eudragit® SI00:LI00 (2:1)

Method of manufacture for this component beads is the same as that of component III 25 beads in biostudy IPX066-AH1, as discussed in Example 4 above. 55 WO 2009/085306 PCT/US2008/014080

Manufacture of Component V -- TA slow release beads coated with Eudragit® S100.TJ00 (2:1)

Method of manufacture for this component beads is the same as that for enteric coated 5 tartaric acid beads in formulation A tested in IPX066-BO6-02 as discussed in Example 2 above.

Manufacture of the 1PX066 Capsules for Formulation A 10 The required amounts of the component beads were filled into hard gelatin capsules according to the specified target fill weights ± 10%.

Table 14. Target Fill Weights of IPX066 Capsule Test Formulations.

Components IPX066»AH2(A) mg/capsule Component I Beads 46.2 Component II Beads 48.7 Component ΙΠ Beads 191.9 Component IV Beads 52.1 Component V Beads 187.7 Total Capsule Fill weight 526.6

15 Manufacture of Test Formulation B fIPX066-AH2(B)J

To make formulation B. a suitable amount of ethanol and purified water were mixed and to which Cremophor RH40 was dissolved and charged in a first solution; then povidone was charged and mixed in the solution. A separate granulating fluid was prepared by 20 dissolving the required amount of povidone in the required amount of purified water.

Suitable amounts of carbidopa, levodopa, microcrystalline cellulose, lactose monohydrate and poloxamer were charged into a suitable granulator and mixed until a uniform powder is formed. 56 WO 2009/085306 FCT/US2008/014080

The mixed powder was granulated by adding the first solution above followed by continued granulation by adding the separate granulating fluid above thereby resulting in a wet mass. The wet mass was extruded through a suitable extruder equipped with a screen size of 1.0 mm. The extrudate was spheronized in a spheronizer at an appropriate speed and the wet extruded beads were dried in a fluidized bed dryer. The dried beads were then passed through a US #16 mesh screen, US #18 mesh screen, US #25 mesh screen and pan. Only the beads that pass through 18 mesh but retained on 25 mesh screen are collected.

The enteric coating dispersion was prepared by dispensing and mixing a suitable amount of acetone and isopropyl alcohol and while mixing the solution, triethyl citrate was charged and dissolved in the solution. The mixing was continued until the material was fully dissolved. While mixing, Methacrylic Acid Copolymer, Type A, NF was charged to the solution. The solution was mixed until the material was fully dissolved then again while mixing, Methacrylic Acid Copolymer, Type B, NF was charged to the solution. Mixing continued until the material was fully dissolved. While mixing, talc was charged and dispersed in the solution. Mixing continued throughout the coating process.

The collected beads above were charged into a suitable fluidized-bed coater equipped with a Wurster insert and were spray coated using the enteric coating dispersion above. The coated beads were dried, and the dried beads were passed through a US #14 mesh screen. The resulting screened material was charged and a suitable amount of talc was added to it in a suitable blender and mixed until uniform.

Manufacture of Test Formulation C (IPX066-AH2(C)) A suitable amount of ethanol and purified water was dispensed and mixed in a stainless steel container to which was charged and dissolved a suitable amount of Cremophor RH40 resulting in a granulating solution. 57 WO 2009/085306 PCT/US2008/014080 A suitable amount of carbidopa, levodopa, tartaric acid, microcrystalline cellulose, and poloxamer was charged into a suitable granulator and mixed until uniform. The mixed powder was granulated with the granulating solution above thus creating a wet mass. The wet mass was extruded through a suitable extruder equipped with a screen size of 1.0 mm 5 and the extrudate was spheronized in a suitable spheronizer at an appropriate speed.

The wet extruded beads were dried in the fluidized bed dryer. The LOD was measured using a Moisture Analyzer and the drying process was stopped when the LOD value was not more than a target value 3%. The dried beads were then passed through a US #16 10 mesh screen, US #18 mesh screen, US #25 mesh screen and pan. Only the beads retained on 18 mesh and on 25 mesh screen were collected and blended.

The seal coat solution was prepared as follows. An ethanol solution was prepared by mixing the required amount of Alcohol, USP and purified water in a stainless steel 15 container. A suitable amount hypromellose was charged and dissolved in the ethanol solution.

The collected beads retained on 18 mesh and on 25 mesh screen were charged into a fluidized bed coater equipped with a Wurster insert and spray coated using the seal coat 20 solution prepared above. The coated beads were dried in the fluidized bed and the dried beads were passed through a US #14 mesh screen. The beads that pass through the screen were collected.

The enteric coating dispersion was prepared as follows. A suitable amount of 25 Methacrylic Acid Copolymer, Type A, NF, Methacrylic Acid Copolymer, Type B, NF, triethyl citrate and talc were dispensed. A suitable amount of acetone and isopropyl alcohol were mixed well and during the mixing, triethyl citrate was charged and mixed into the solution. Methacrylic Acid Copolymer, Type A, NF was further charged into the solution. Mixing continued until the material was fully dissolved. Methacrylic Acid 30 Copolymer, Type B, NF was additionally charged into the solution. Mixing continued 58 WO 2009/085306 PCT/US2008/014080 until the material was fully dissolved. Talc was then charged and dispersed in the solution. Mixing occurred throughout the process.

The collected beads were coated with the enteric coating dispersion as follows. The collected beads were charged in a suitable fluidized-bed coater equipped with a Wurster insert and spray coated with the enteric coating dispersion above. The coated beads were then dried and the dried beads passed through a US #14 mesh screen. The screened material was mixed and charged with a suitable amount of talc in a blender.

Manufacture of Test Formulation D (1PXO66-AH2(D)) A granulating solution was prepared by dissolving polyoxyl in a solution of ethanol and purified water. A suitable amount of carbidopa, levodopa, screened tartaric acid, microcrystalline cellulose, lactose monohydrate and poloxamer were charged and mixed in a granulator. The mixed material was granulated with the granulating solution thereby creating a wet mass. The wet mass was extruded through a suitable extruder equipped with a screen size of 1.0 mm. The resulting extrudates were spheronized in a suitable spheronizer. The wet extruded beads were dried in a fluidized bed dryer and the LOD was measuring using a Moisture Analyzer. The drying process was stopped when the LOD value is not more than a target value 3%.

The dried beads were passed through a US #16 mesh screen, US #18 mesh screen, US #25 mesh screen and pan. Only the beads retained on 18 mesh and on 25 mesh screen were collected and retained.

The dried beads were coated with a seal coat. The seal coat may be prepared as follows. An ethanol solution was prepared by mixing the required amount of Alcohol, USP and purified water in a stainless steel container to which a suitable amount hypromellose was charged. The dried beads were charged into a fluidized bed coater equipped with a 59 WO 2009/085306 PCT/US2008/014080

Wurster insert and spray coated using the seal coat solution. The coated beads were then dried in a fluidized bed and the dried coated beads passed through a US #14 mesh screen. The beads that passed through the screen were collected. 5 The collected beads were coated with an enteric coating dispersion. The enteric coating dispersion was prepared as follows. A suitable amount of Methacrylic Acid Copolymer, Type A, NF, Methacrylic Acid Copolymer, Type B, NF, triethyl citrate and talc was dispersed. A suitable amount of acetone and isopropyl alcohol was mixed well. During the mixing, triethyl citrate was charged and dissolved in the acetone and isopropyl alcohol 10 solution. Then while mixing, Methacrylic Acid Copolymer, Type A, and NF were charged into the solution. Mixing continued until the material was fully dissolved. During the mixing, Methacrylic Acid Copolymer, Type B, and NF were charged into.the. solution. Mixing continued until the material was fully dissolved. Then talc was charged and dissolved in the solution. Mixing continued throughout the coating process. 15

The seal coated beads were charged into a suitable fluidized-bed coater equipped with a Wurster insert and spray coated using the enteric coating dispersion. The resulting coated beads were dried and the dried beads were passed through a US #14 mesh screen. The screened material were charged and mixed with a suitable amount of talc in a blender 20 until uniform.

Pharmacokinetics result for biostudy IPXQ66-AH2

The Pharmacokinetics parameters of CD and LD for four tested formulations compared to 25 Sinemet® CR after oral administration are summarized in Table 15. 60 WO 2009/085306 PCT/US2008/014080

Table 15. Mean ratio of Ln-transformed AUC of Levodopa and Carbidopa

Parameter Test/Ref Ratio (%) LD Ln(Cmax) A/E 80.73 Ln(Cmax) B/E 16.19 Ln(Cmax) C/E 18.15 Ln(Cmax) D/E 16.55 Ln(AUC) A/E 108.65 Ln(AUC) B/E 10.54 Ln(AUC) C/E 15,08 Ln(AUC) D/E 15.79 CD Ln(Cmax) A/E 80.12 Ln(Cmax) B/E 7.03 Ln(Cmax) C/E 13.85 Ln(Cmax) D/E 14.41 Ln(AUC) A/E 94,93 Ln(AUC) B/E 2.69 Ln(AUC) C/E 12.54 Ln(AUC) D/E 11.06

Results and Discussion 5 The data herein demonstrated that formulation A achieved a desirable in vivo plasma concentration profile. By combining two components of tartaric acid beads (with enteric coating on either fast or slow release core) with three components of CD/LD drug seeds/beads (instant release portion, and enteric coated portion with coating on either fast or slow release core), formulation A showed a significantly flatter plasma concentration 10 profile with a lower Cmax (80,7%) and comparable AUC (109%) relative to the reference drug Sinemet® CR. A PK simulation study was conducted to calculate the peak to trough plasma concentration (PT) ratio assuming a three times daily (every 6 hours) dosing regimen. 15 The PT ratio is defined on steady state 18 hours post-dose plasma concentration. The 61 WO 2009/085306 PCT/US2008/014080 simulation results are summarized in Table 16, showing that formulation A has a significantly lower, 1.7, PT ratio relative to Sinemet® CR, 3.4.

The addition of surfactants in formulation significantly decreased LD bioavailability. 5 Moreover, it is more advantageous to have separate drug and tartaric acid seeds in formulation, and the incorporation of tartaric acid into same drug seeds significantly decreased LD bioavailability.

Table 16. Simulation of Steady State Plasma Concentration, AUC, and Peak-to-10 trough Concentration Ratio.

Formulation CD-LD Dose (mg) Css,max Css, 18hr Peak to Trough Plasma Cone ratio A (IPX066- AH2(A)) 75-300 1030 591 1.7 E (reference) 50-200 1212 349 3.4 EXAMPLE 6

Formulation with separate CD/LD containing surfactant seeds and tartaric acid seeds for 15 IPX-066 was evaluated for PK parameters. ΙΡΧ066-ΛΗ3

Information of study drugs are listed below in Table 17. Formulation A (IPX066-20 AH3(A))is a capsule containing ER CD/LD beads formulated with Cremophor RH40 and

Poloxamer 188 and ER TA beads with a similar dissolution profile. 62 WO 2009/085306 PCT/US2008/014080

Table 17. IPX066-AH3

Product CD-LD Strength (mg) Enteric Coated pH A (IPX066- AH3(A)) IPX066 CD/LD containing surfactant Capsule with 215 mg tartaric acid 50-200 6.5 (SL2) B "SinemeFcR Table? 50-200 -- * Merck & Co., Inc. 5 The qualitative and quantitative compositions for these formulations are summarized below in Table 18.

Table 18. Qualitative and Quantitative Composition of Test Formulation

Ingredients Formulation A (IPX066-AH3(A)) mg/capsuie Bead I Carbidopa, USP.................................................... 17.99 Levodopa, USP 66.67 Microcrystalline Cellulose, NF (Avicel PH-101) 79.17 Lactose Monohydrate, NF 35.83 Poloxamer 188, NF (Lutrol F-68, NF) 22.33 Polyoxyl 40 Hydrogenated Castor Oil, NF (Cremophor RH 40) 11,67 Povidone, USP 11.57 Methacrylic acid copolymer, Type A, NF (Eudragit® Li00) 32.65 Methacrylic acid Copolymer, Type B, NF (Eudragit® SI 00) 65.46 Triethyl Citrate, NF 28.12 Talc, USP 15.99 Acetone, NF N/A* Isopropyl Alcohol, USP N/A* Purified Water, USP N/A* Bead II Tartaric acid, NF 71.67 Microcrystalline Cellulose, NF (Avicel PH-101) 17.93 63 WO 2009/085306 PCT/US2008/014080

Ethylcellulose, NF (Ethocel Standard-10FP Premium) 7.53 Hypromellose, USP Type2910 (Pharmacoat 606,6 cps) 13.50 Methacrylic acid copolymer, Type A, NF (Eudragit® LI 00) 14.77 Methacrylic acid Copolymer, Type B, NF (Eudragit® SI 00) 29.50 Tri ethyl Citrate, NF 12.63 Talc, USP 7.20 Acetone, NF N/A* Isopropyl Alcohol, USP N/A* Purified Water, USP N/A* Hard Gelatin Capsule 1 unit Total weight 562.18 * evaporated in drying process

Manufacture of Test Formulation 5 Manufacture of Component Bead 1 - CD/LD beads

Method of manufacture for this component beads is the same as to that for formulation B of IPX066-AH2, as discussed above in Example 5.

Manufacture of Component Bead Π - TA beads

Method of manufacture for this component beads is the same as that of component III 10 beads in IPX066-AH1, except the content for enteric coating is higher in formulation, as discussed in Example 4 above.

Pharmacokinetics result for IPX066-AH3;

The Pharmacokinetics parameters of CD and LD for tested formulations compared to 15 Sinemet® CR after oral administration are summarized in Table 19. 64 WO 2009/085306 PCT/US2008/014080

Table 19. Mean ratio of Ln-transforraed Cmax» AUC of Levodopa and Carbidopa

Parameter Test/Ref Ratio (%) LD Ln(Cmax) Ln(AUC) A/B A/B 14,89 9.88 CD Ln(Cmax) Ln(AUC) A/B A/B 4.87 7.11

Results and Discussions 5 The addition of surfactants in formulation significantly decreased LD bioavailability even when the formulation contains separate drug and tartaric acid seeds.

ΪΓΥΑΜΪΠ Ϊ? *7 MrzkAlYLJr JL/JtL· Z 10 Formulations for IPX066-AH4

Four formulations of IPX-066 were evaluated for PK parameters. Information of study drugs are listed below in Table 20. 15 Table 20. IPX066-AH4

Formulation CD-LD Dose (mg) Molar Ratio of ΠΓA .T ΤΛ A A. «jLfXA Amount of Capsule/dose A (IPX066- AH4(A)) 75-300 1.4:1 2 B (IPX066-AH4(B)) 90-360 1.4:1 3 C (IPX066-AH4(C)) 90-360 0.5 :1 2 D (IPX066- AH4(D)> 90-360 0.75 :1 2 Sinemet® CR Tablet0 50-200 — — a Merck & Co., Inc. 65 WO 2009/085306 PCT/US2008/014080

These formulations contain CD/LD beads and LD beads with different release characteristics. Similar to formulation A of IPX066-AH2, these beads are coated with Eudragit® L100 and S100 polymer in the ratio of 1:2. The quantities of CD/LD and TA in each of the bead type are shown in Table 20. 5

Component I is a CD/LD bead type which exhibits an immediate-release dissolution profile.

Component ΪΪ is a CD/LD bead type which exhibits a faster extended-release dissolution 10 profile.

Component ΙΠ is a CD/LD bead type which exhibits a slower extended-release dissolution profile. 15 Component IV is a TA bead type which exhibits a dissolution profile which mimics those of component Π.

Component V is a TA bead type which exhibits a dissolution profile which mimics those of component III. 20

Component VI is a TA bead type which exhibits an intermediate dissolution profile between those of component II and III.

The qualitative and quantitative compositions for these formulations are summarized 25 below in Table 21. 66 WO 2009/085306 PCT/US2008/014080

Table 21. Qualitative and Quantitative Composition of Formulation

Formulation A Formulation B Formulation C Formulation » (IPX066- AH4(A)) (IPX066- AH4(B)) (IPX066- AH4(Q) (IPX066- AH4(D)) Ingredients mg/capsule mg/capsule mg/capsule mg/capsule Component I Carbidopa USP 6.75 6.48 9.72 9.72 LevodopaUSP 25 24 36 36 Macrocrystalline Cellulose NF 4.6 4.42 6.62 6.62 Lactose Monohydrate NF 4.6 4.42 6.62 6,62 Sodium Starch Glycolate NF 2.3 2.21 3.31 3.31 Sodium Lauryl Sulfate NF 2.3 2.21 3.31 3.31 . Povidone USP 0.46 0.44 0.66 0.66 Talc USP 0.23 0.22 0.33 0.33 Purified Water, USP N/A* N/A* N/A* N/A*

Component Π

Carbidopa USP 6.75 5.4 8.10 8.10 LevodopaUSP 25 20 30 30 Microcrystalline Cellulose NF 4.6 3.68 5.52 5.52 Lactose Monohydrate NF 4.6 3.68 5.52 5.52 Sodium Starch Glycolate NF 2.3 1.84 2.76 2.76 Sodium Lauryl Sulfate NF 2.3 1.84 2.76 2.76 Povidone USP 0.46 0.37 0.55 0.55 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 0.56 0.45 0.67 0.67 Methacrylic acid Copolymer, Type B NF (Eudragit® SI 00) 1.14 0.91 1.36 1.36 Triethyl Citrate NF 0,49 0.39 0.58 0.58 Talc USP 0.49 0.39 0.58 0.58 Acetone, NF N/A* N/A* N/A* N/A* Isopropyl Alcohol, USP N/A* N/A* N/A* N/A* Purified Water, USP N/A* N/A* N/A* N/A*

Component HI

Carbidopa USP 26.99 20.51 30.77 30.77 LevodopaUSP 100.00 76.00 114.00 114.00 Microcrystalline Cellulose NF 54.43 41.36 62.04 62.04 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 2,19 1.67 2.50 2.50 Methacrylic acid Copolymer, Type B NF (Eudragit® S100) 4.49 3.41 5.12 5,12 Triethyl Citrate NF 1.91 1,45 2.18 2.18 Talc USP 1.92 1.46 2.18 2.18 Acetone, NF N/A* N/A* N/A* N/A* Isopropyl Alcohol, USP N/A* N/A* N/A* N/A* 67 WO 2009/085306 PCT/US2008/014080

Purified Water, USP N/A* N/A* N/A* N/A* Component IV Tartaric acid NF 27.00 21.50 11.40 - Microcrystahtne Cellulose NF 6.74 5.37 2.85 - Ethylcellulose NF 2.81 2.24 1.19 Hypromellose, Type2910 USP 0.94 0.75 0.40 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 2,16 1.72 0.91 Methacrylic acid Copolymer, Type B NF (Eudragit® S100) 4.41 3,51 1.86 - Triethyl Citrate NF 1,87 1.49 0.79 - Talc USP 1.18 0.94 0,50 - Acetone, NF N/A* N/A* N/A* - Isopropyl Alcohol, USP N/A* N/A* N/A* - Purified Water, USP N/A* N/A* N/A* - Component V Tartaric acid NF 107.50 81.70 43.4 - Microcry stall ine Cellulose NF 26.90 20.44 10.86 - Ethylcellulose NF 12.40 9.42 5.00 - Hypromellose Type 2910 USP 2.55 1.94 1.03 - Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 8.60 6.54 3.47 - Methacrylic acid Copolymer, Type B NF (Eudragit® SI00) 17.55 13.34 7.09 - Triethyl Citrate NF 7.45 5.66 3.01 - Talc USP 4.70 3.57 1.90 - Acetone, NF N/A* N/A* N/A* - Isopropyl Alcohol, USP N/A* N/A* N/A* - Purified Water, USP N/A* N/A* N/A* Component VI Tartaric acid NF - - - 82.20 Microcrystalline Cellulose NF - - 20.57 Ethylcellulose NF - - - 9.14 Hypromellose Type 2910 USP - - 2.29 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 6.58 Methacrylic acid Copolymer, Type B NF (Eudragit® S100) - - 13.42 Triethyl Citrate NF - 5.70 Talc USP - - - 3.59 Acetone, NF N/A* N/A* N/A* N/A* Isopropyl Alcohol, USP N/A* N/A* N/A* N/A* Purified Water, USP N/A* N/A* N/A* N/A* Hard Gelatin Capsule 1 unit 1 unit 1 unit 1 unit Total Capsule Fill weight 521.62 409.34 439.41 487.25 * evaporated in drying process 68 WO 2009/085306 PCT/US2008/014080

Manufacture of Test Formulations

Manufacture of Component I- CD-LD fast release beads 5

Method of manufacture for this component beads is the same as to that for component I beads for formulation A in IPX066-AH2, as discussed in Example 5 above.

Manufacture of Component II - CD-LD fast release beads coated with Eudragit® 10 SIOO.LIOO (2:1)

Method of manufacture for this component beadsuis the same as to that for component Π beads for formulation A in IPX066-AH2, as discussed in Example 5 above. 15 Manufacture of Component III — CD-LD slow release beads coated with Eudragit® S100:L100 (2:1)

Method of manufacture for this component beads is the same as to that for component ΙΠ beads for formulation A in IPX066-AH2, as discussed in Example 5 above. 20

Manufacture of Component IV — TA fast release beads coated with Eudragif SI 00:LI00 (2:1)

Method of manufacture for this component beads is the same as to that for component IV 25 beads for formulation A in IPX066-AH2. as discussed in Example 5 above, except no additional seal coat was introduced between seal coating layer and enteric coating layer. 69 WO 2009/085306 PCT/US2008/014080

Manufacture of Component V TA slow release beads coated with Eudragit® SI 00: LI 00 (2:1)

Method of manufacture for this component beads is the same as to that for component V 5 beads for formulation A in IPX066-AH2, as discussed in Example 5 above.

Manufacture of Component VI - TA medium release beads coated with Eudragit® S100:L100 (2:1) 10 Method of manufacture for this component beads is the same as to that for component V beads except the content ratio of ethylcellulose/hypromellose in seal coating is 4/1 instead of 5/1.

Manufacture of the IPX066 Capsules 15

The required amounts of the component beads were filled into hard gelatin capsules according to the specified fill weights in the Table 22 below. The in-process fill weight is controlled at target ± 10% of the target weights according to Table 22. 20 Table 22. Target Fill Weights of ΪΡΧ066 Capsule Test Formulations

Components Formulation Code 1PXO66-AH4(A) mg/capsule IPX066- AH4(B) mg/capsule ΪΡΧ066- AH4(C) mg/capsule ΪΡΧ066- AH4(D) mg/capsule Component 1 Beads 46,24 44.4 66.57 66.57 Component Π Beads 48.69 38.95 58.40 58.40 Component III Beads 191.93 145,86 21879 218.79 Component IV Beads 47.11 37.52 19.89 Component V Beads 187.65 142.61 75.76 Component VI Beads - - 143.49 Total Capsule Fill weight 521.62 409.34 439.41 487.25 70 WO 2009/085306 PCT/US2008/014080

Pharmacokinetics result for ΪΡΧ066-ΑΗ4

The Pharmacokinetics parameters of CD and LD for four tested formulations compared to Sinemet® CR after oral administration are summarized in Table 23.

Table 23. Mean ratio of Ln-transformed Cmax, AUC of Levodopa and Carbidopa

Parameter Test/Ref Ratio (%) LD Ln(Cmax) A/E 92.29 Ln(Cmax) B/E 119.02 Ln(Cmax) C/E 132.54 Ln(Cmax) D/E 133.78 Ln(AUC) A/E 105.14 Ln(AUC) B/E 136.15 Ln(AUC) C/E 143,73 Ln(AUC) D/E 154.34 CD Ln(Ctnax) A/E 92.76 Ln(Cmax) B/E 116.37 Ln(Cmax) C/E 126.66 Ln(Cmax) D/E 135.66 Ln(AUC) A/E 111.12 Ln(AUC) B/E 149.98 Ln(AUC) C/E 145.70 Ln(AUC) D/E 164.13

Results and Discussion 10 Test formulation A (IPX066-AH4(A)) showed a consistent in vivo plasma concentration profile as those of formulation A (IPX066-AH2(A)) in IPX066-AH2. Formulation IPX066-AH2(A) and IPX066-AH4(A) differs only in the filler used, i.e. lactose in IPX066-AH2(A) and mannitol in IPX066-AH4(A). The relative AUC (relative to Sinemet® CR Tablet) is 109% and 105% for formulation IPX066-AH2(A) and IPX066- 15 AH4(A), respectively. The relative Cmax (relative to Sinemet® CR Tablet) is 80.7% and 71 WO 2009/085306

PCT/US2OO8/01408O 92.3% for formulation 1PXO66-AH2(A) and IPX066-AH4(A), respectively. Therefore, the in-vivo performance and bioavailability of the formulation is reproducible and consistent. 5 Based on a simulation study, the steady state peak to trough plasma concentration (PT) ratio was calculated assuming a three times daily, every 6 hours, dosing. The simulated PT ratios, defined as ratio of Cmax and Css,18 hours, are summarized in Table 24. The results showed that PT ratio of formulation A (IPX066-AH4(A)), B (IPX066-AH4(B)), C (IPX066-AH4(C)), and D (fPX066~AH4(D)) are 1.8, 2.3, 2.8, and 2.0, respectively. The 10 PT ratio of Sinemet® CR Tablet is 3,9. Therefore, all IPX066 test formulations have a lower PT ratio relative to the reference, Sinemet® CR Tablet.

The data herein also demonstrated the optimum molar ratio of TA:LD to be 0.75, when comparing PT ratio of formulation B, C, and D. 15

The PK results also indicated that replacing the TA beads of fast and slow release profiles with TA beads with an intermediate release profile in formulation did not change the overall in vivo LD plasma profile significantly, as evidenced by the PT ratio. 20 Table 24. Simulation of Steady State Plasma Concentration, AUC, and Peak-to-trough Concentration Ratio.

Test Formulation CD-LD Dose (mg) TA;LD Molar Ratio Css,max Css,18hr AUCss,24 Peak to Trough Plasma Cone ratio Al 75-300 1.4 920 511 13215 1.8 B 90-360 1.4 1238 549 17048 2.3 C 90-360 0.5 1337 477 18204 2.8 D 90-360 0.75 1384 679 19848 2.0 E (reference) 50-200 0 984 254 12348 3.9 72 WO 2009/085306 PCT/US2008/014080 EXAMPLE 8 IPX066-AH5(A) (47.5mg CD/190 mg LD) was evaluated for PK parameters in a three way cross over PK study using Sinemet® CR (200 mg) as reference. The qualitative and 5 quantitative compositions for this formulation are summarized below.

Table 25, Quantitative Composition of Formulation IPX066~AH5(A)

Dosage strength % 190 mg Ingredients mg/capsule Carbidopa 10.21 51.29* Levodopa 37.84 190.00 Tartaric acid 17.63 88.52 Microcrystalline Cellulose 19.23 96.55 Mannitol 1.01 5.07 Ethylcellulose 2.03 10.2 Hypromellose, Type2910 0,43 2.16 Sodium Starch Glycolate 0.50 2.53 Sodium Lauryl Sulfate 0.50 2.53 Povidone 0.34 1.73 Talc 1.36 6.84 Methacrylic acid copolymer, Type A (Eudragit® L100) 2.09 10.51 Methacrylic acid Copolymer, Type B (Eudragit® SI00) 4.25 21.33 Triethyl Citrate 1.82 9.16 Croscarmellose Sodium 0.69 3.46 Magnesium Stearate 0.05 0.25 Purified water N/A** N/A** Isopropyl alcohol N/A** N/A** Acetone N/A** N/A** Ethyl alcohol N/A** N/A** Total 100.00 502.13 Hard Gelatin Capsules 1 unit (size 00) * Carbidopa is supplied as a monohydrate; the quantity is equivalent to 47.50 mg 10 carbidopa anhydrous accordingly, ** Evaporated during drying process. 73 WO 2009/085306 PCT/US2008/014080

Manufacture of Formulation

Four different component beads were manufactured for formulation IPX066-AH5(A). 5 The qualitative and quantitative compositions for each component seed are summarized in Table 26.

Table 26. Qualitative and Quantitative Composition for 4 Component Beads in ΪΡΧ066 Capsule Test Formulation

Ingredients Formulation Formulation Code IPX066-AH5(A) mg/capsule Component I Carbidopa USP 9.45* Levodopa USP 35.00 Croscarmeliose Sodium 3.46 Povidone 1.23 Magnesium Stearate 0.25 Purified Water, USP N/A** Component II Carbidopa USP 7.43* Levodopa USP 27.50 Microcrystalline Cellulose NF 5.07 Mannitol NF 5.07 Sodium Starch GlycolateNF 2.53 Sodium Lauryl Sulfate NF 2,53 Povidone USP 0.50 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 0.62 Methacrylic acid Copolymer, Type B NF (Eudragit® SI 00) 1.24 Triethyl Citrate NF 0.53 Talc USP 0.54 Acetone, NF N/A** Isopropyl Alcohol, USP N/A** Purified Water, USP N/A** Component ΠΙ Carbidopa USP 34.41* Levodopa USP 127.50 MicrocrystaUtne Cellulose NF 69.39 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 2.79 Methacrylic acid Copolymer, Type B 5.72 74 WO 2009/085306 PCT/US2008/014080 NF (Eudragit® SI 00) Triethyl Citrate NF 2.45 Talc USP 2.44 Acetone, NF N/A** Isopropyl Alcohol, USP N/A** Purified Water, USP N/A** Component IV Tartaric acid NF 88.52 Microcrystalline Cellulose NF 22.09 Ethylcellulose NF 10.20 HypromeHose Type 2910 USP 2.16 Methacrylic acid copolymer, Type A NF (Eudragit® LI 00) 7.10 Methacrylic acid Copolymer, Type B NF (Eudragit® SI00) 14.37 Triethyl Citrate NF 6.18 Talc USP 3.86 Acetone, NF N/A** Isopropyl Alcohol, USP N/A** Purified Water, USP N/A** Hard Gelatin Capsule 1 unit Total Capsule Fill weight 502.13 * Carbidopa is supplied as a monohydrate; the quantity is equivalent to 8.75, 6.88, 31.87 mg carbidopa anhydrous in component I, II, HI, respectively. **evaporated in drying process 5 Manufacture of Component Bead I- CD/LD Ultra Fast release granules

The required amounts of carbidopa, levodopa, croscarmellose sodium, and povidone were charged into a high shear granulator and mix until uniform. The mixed powder was granulated by adding the purified water. The granules were discharged and dried in the oven at 60 ± 10°C. The LOD was measured using a Moisture Analyzer and the drying 10 process was stopped when the LOD value is not more than a target value. The granules were crushed by using mortar and pestle. The crushed granules were passed through a US #18 mesh screen, and collect the granules that pass through 18 mesh screen. The collected granules were blended with magnesium stearate. 75 WO 2009/085306 PCT/US2008/014080

Manufacture of Component II - CD-LD fast release beads coated with Eudragit® SIOO.LIOO (2:1)

Method of manufacture and process flow diagram of this component beads is the same as 5 those of component 2 beads of formulation A in IPX066-AH4 except that mannitol is replaced by lactose.

Manufacture of Component III - CD-LD slow release beads coated with Eudragil® S1OQ:L1QO (2:1) 10 Method of manufacture and process flow diagram for this component beads is the same as those of the component 3 beads of formulation A in IPX066-AH4.

Manufacture of Component IV - TA slow release beads coated with Eudragil® SI 00: LI 00 (2:1) 15

Method of manufacture and process flow diagram for this component beads is the same as those of the component 5 beads of formulation A in IPX066-AH4.

Manufacture of the IPX066 Capsules for IPX066-AH5(A) 20

The required amounts of the component beads were filled into hard gelatin capsules according to the specified target fill weights ± 10%. 25 Table 27. Target Fill Weights of EPX066 Capsule Test Formulation.

Components IPX066~AH5(A) mg/capstile Component I Beads 49.39 Component II Beads 53.56 Component III Beads 244.70 Component IV Beads 154,48 Total Capsule Fill weight 502.13 76 WO 2009/085306 PCT/US2008/014080 EXAMPLE 9

The example describes the formulations IPX066-AH6(A), and IPX066-AH6(B) and methods for making them.

Table 28

Formulation % ΙΡΧΘ66- AH6(A) IPX066- AH6(B) Dosage strength 245 mg 195 mg Ingredients mg/capsule mg/capsule Carbidopa 10,21 66.14* 52.64* Levodopa 37.84 245.00 195.00 Tartaric acid 17.63 132.53 105.48 Microcrystalline Cellulose 19.23 124.63 99.20 Mannitol 1.01 6.43 5.12 Ethylcellulose 2.03 15.27 12,15 Hypromellose, Type2910 0.43 3.23 2.57 Sodium Starch Glycolate 0.50 3.21 2.55 Sodium Lauryl Sulfate 0.50 3.21 2.55 Povidone 0.34 2.52 2.01 Talc 1.36 9.45 7.52 Methacrylic acid copolymer, Type A (Eudragit® LI 00) 2.09 14.84 11.81 Methacrylic acid Copolymer, Type B (Eudragit® S100) 4.25 30.11 23.97 Triethyl Citrate L82 12.93 10.29 Croscarmellose Sodium 0.69 5.31 ’ 4.23 Magnesium Stearate 0.05 0.38 0.30 Purified water N/A** N/A** N/A** Isopropyl alcohol N/A** N/A** N/A** Acetone N/A** N/A** N/A** Ethyl alcohol N/A** N/A** N/A** Total 100.00 675.19 537.39 Hard Gelatin Capsules 1 unit (size 00) 1 unit (size 0EL) * Carbidopa is supplied as a monohydrate; the quantity is equivalent to 61.25 mg, 48.75 mg carbidopa anhydrous accordingly. 77 WO 2009/085306 PCT/US2008/014080 ** Evaporated during drying process.

Manufacture of Test Formulation 5 Four different component beads were manufactured for test formulation IPX066-AH6(A) and AH6(B). The qualitative and quantitative compositions for each component seed are summarized in Table 29.

Table 29. Qualitative and Quantitative Composition for 4 Component Beads 10 in ΪΡΧ066 Capsule Formulation

Ingredients IPX066-AH6(A) IPX066- AH6(B) mg/capsule mg/capsule Component I Carbidopa USP 14.50* 11.54* Levodopa USP 53.71 42.75 Croscartnellose Sodium 5.31 4.23 Povidone 1.89 1.50 Magnesium Stearate 0.38 0.30 Purified Water, USP N/A** N/A** Component Π Carbidopa USP 9.42* 7.50* Levodopa USP 34.87 27.75 Microcrystalline Cellulose NF 6.43 542 Mannitol NF 6.43 542 Sodium Starch Giycolate NF 3.21 2.55 Sodium Lauryl Sulfate NF 3.21 2.55 Povidone USP 0.63 0.51 Methacrylic acid copolymer, Type A NF (Eudragit® LI 00) 0.79 0,63 Methacrylic acid Copolymer, Type B NF (Eudragit® S100) 1.57 1.25 Triethyl Citrate NF 0.67 0.53 Talc USP 0.68 0.54 Acetone, NF N/A** N/A** isopropyl Alcohol, USP N/A** N/A** Purified Water, USP N/A** N/A** Component III Carbidopa USP 42.22* 33.60* Levodopa USP 156.42 124.50 Microcrystalline Cellulose NF 85.13 67.76 Methacrylic acid copolymer, Type A NF (Eudragit® LI00) 3.42 2.72 78 WO 2009/085306 PCT/US2008/014080

Methacrylic acid Copolymer, Type B NF (Eudragit® SI00) 7.02 5,59 Triethyl Citrate NF 3.01 2.39 Talc USP 2.99 2.38 Acetone, NF N/A** N/A** isopropyl Alcohol, USP N/A** N/A** Purified Water, USP N/A** N/A** Component IV Tartaric acid NF 132.53 105.48 Microcrystal line Cellulose NF 33.07 26.32 Ethylcellulose NF 15.27 12.15 Hypromellose Type 2910 USP 3.23 2.57 Methacrylic acid copolymer, Type A NF (Eudragit® LI 00) 10.63 8.46 Methacrylic acid Copolymer, Type B NF (Eudragit® S100) 21.52 17.13 Triethyl Citrate NF 9.25 7.37 Talc USP 5.78 4.60 Acetone, NF N/A** N/A** Isopropyl Alcohol, USP N/A** N/A** Purified Water, USP N/A** N/A** Hard Gelatin Capsule 1 unit 1 unit Total Capsule Fill weight 675.19 537.39 * Carbidopa is supplied as a monohydrate. **evaporated in drying process .

Manufacture of Component Bead I ~ CD/LD Ultra Fast release granules 5 Method of manufacture and process flow diagram of this component beads is the same as those of component 1 beads of formulation A in IPX066-AH5 except that granules were dried by GPCG-1 instead of in an oven, and the granules were milled by Fitzmill instead of crushed by motor and pestle.

Manufacture of Component Π ~ CD-LD fast release beads coated with Eudragit® 10 S100:L100(2:l)

Method of manufacture and process flow diagram of this component beads is the same as those of component 2 beads of formulation A in IPX066-AH5. 79 WO 2009/085306 PCT/US2008/014080

Manufacture of Component III CD-LD slow release beads coated with Eudragit® S100:L100 (2:1)

Method of manufacture and process flow diagram for this component beads is the same 5 as those of the component 3 beads of formulation A in IPX066-AH5.

Manufacture of Component IV - TA slow release beads coated with Eudragit® S100:LI 00 (2:1) Ϊ 0 Method of manufacture and process flow diagram for this component beads is the same as those of the component 4 beads of formulation A in ΪΡΧ066-ΑΗ5.

Manufacture of the ΪΡΧ066 Capsules for 1PXO66-AH6(A) and (B) 15 The required amounts of the component beads were filled into hard gelatin capsules according to the specified target fill weights ± 10%.

Table 30. Target Fill Weights of IPX066 Capsule Test Formulation.

Components IPX066-AH6(A) mg/capsule IPX066-AH6(B) mg/capsule Component I Beads 75.79 60.32 Component Π Beads 67.91 54.05 Component ΠΪ Beads 300.21 238.94 Component IV Beads 231.28 184.08 Total Capsule Fill weight 675.19 537.39 80