AU2018202552B2 - Methods of using sustained release aminopyridine compositions - Google Patents

Abstract

A pharmaceutical composition which comprises a therapeutically effective amount of a aminopyridine dispersed in a release matrix, including, for example, a composition that can be formulated into a stable, sustained-release oral dosage formulation, such as a tablet which provides, upon administration to a patient, a therapeutically effective plasma level of the aminopyridine for a period of at about 12 hours and the use of the composition to treat various neurological diseases, including multiple sclerosis. A method of selecting individuals based on responsiveness to a treatment, including, for example, identifying individuals who responded to treatment with a sustained release fampridine composition. See Figure 10. WO 2005/099701 PCT/US2005/012427 10/12 35- Treatment means over time ** 30 ---------------------------- -- - - -- - ----- - - - 25-- ------------------**------ 20 --2---5- - - - - 0V) S-- placebo (N=47) i 15 -------- -- Farypridine Non-responders (N=100) --- Farrpridine Responders (N=58) 10 - -- - 50 --- - - - - - - - - - - - - - - - - - - - - - - - - - E 0 -5 - -------------- - - -10 titration 1st stable 2nd stable 3rd stable *:Significantly better than placebo and fampridine non-responders (p <0.001 for each). *:Significantly better than fampridine non-responders. Fig. 10

Description

35- Treatment means over time ** 30 ---------------------------- -- - - -- - ----- - - -

25-- -- 2---5- 20 - ------------------**------ - - - 0V)

S-- placebo (N=47) i 15 -------- -- Farypridine Non-responders (N=100) --- Farrpridine Responders (N=58) 10 - -- - 50 --- - - - - - - - - - - - - - - - - - - - - - - - - -

E 0

-5 - -------------- - -

-10 titration 1st stable 2nd stable 3rd stable

*:Significantly better than placebo and fampridine non-responders (p <0.001 for each). *:Significantly better than fampridine non-responders.

Fig. 10

METHODS OF USING SUSTAINED RELEASE AMINOPYRIDINE COMPOSITIONS CROSS REFERENCES

[0001] This application claims priority to U.S. Provisional Application No.

60/560,894 filed April 9, 2004.

BACKGROUND

[0002] This invention relates a sustained release oral dosage form of an

aminopyridine pharmaceutical composition that can be used to treat individuals affected with

neurological disorders wherein said pharmaceutical composition maximizes the therapeutic

effect, while minimizing adverse side effects.

[0003] The sustained release oral dosage form of the present invention may be

utilized to treat neurological disorders such as multiple sclerosis, spinal cord injuries,

Alzheimer's disease and ALS.

[0004] Multiple sclerosis (MS) is a degenerative and inflammatory neurological

disease that affects the central nervous system, more specifically the myelin sheath. The

condition of MS involves demyelination of nerve fibers resulting in "short-circuiting" of

nerve impulses and thus a slowing or blocking of transmission along the nerve fibers, with

associated disabling symptoms. Treatment alternatives for promoting transmission along

affected nerves have thus far been limited.

[0005] Potassium channel blockers are a class of compounds that has been found to

improve the conduction of nerve impulses. As a result, they have become the focus of

attention in the symptomatic treatment of spinal cord injury, MS and Alzheimer's disease.

One sub-class of potassium channel blockers, aminopyridines have shown promise in the

treatment of neurological diseases. 4-aminopyridine (4-AP), a mono-aminopyridine known

as fampridine, has been found to reduce the potassium flow in nerve impulse transmission and, thereby, shows effectiveness in restoring conduction in blocked and demyelinated nerves.

[0006] Early studies of monoaminopyridines were conducted using an intravenous

composition, comprising 4-AP. This was followed by the development of an

immediate-release (IR) composition for oral administration of 4-AP, commonly known as

fampridine. The IR composition consisted of 4-AP powder in a gelatin-based capsule and

produced rapid peak plasma concentrations shortly after dosing with a time to maximum

concentration of about 1 hour and a plasma half life of about 3.5 hours. The rapid release and

short half life of fampridine makes it difficult to maintain effective plasma levels without

producing high peaks following each dose that may cause undesirable side effects such as

seizures and trembling.

[0007] Electrophysiological recordings from isolated spinal cord have shown

chronic failure of action potential conduction in surviving myelinated axons, following a

blunt contusion injury (Blight, A.R., "Axonal physiology of chronic spinal cord injury in the

cat: intracellular recording in vitro", Neuroscience. 10:1471-1486 (1983b)). Some of this

conduction block can be overcome, at the level of single nerve fibers, using the drug 4

aminopyridine (4-AP) (Blight, A.R., "Effect of 4-aminopyridine on axonal conduction-block

in chronic spinal cord injury", Brain Res. Bull. 22:47-52 (1989)). Intravenous injection of

this compound in animals with experimental or naturally occurring spinal cord injuries

produces significant improvements in electrophysiological (Blight, A.R. and Gruner, J.A.,

"Augmentation by 4-aminopyridine of vestibulospinal free fall responses in chronic spinal

injured cats," J. Neurol. Sci. 82:145-159, (1987)) and behavior function (Blight, A.R., "The

effects of 4-aminopyridine on neurological deficits in chronic cases of traumatic spinal cord

injury in dogs: a phase I clinical trial," J. Neurotrauma, 8:103-119 (1991)).

[0008] An initial study in spinal cord injury patients was organized by Dr. Keith

Hayes and indicated a potential for a therapeutic benefit, mostly at the electrophysiological

level, combined with a lack of serious side effects (Hayes et al, "Effects of intravenous 4

aminopyridine on neurological function in chronic spinal cord injured patients: preliminary

observations," Proc. IBRO World Conf. Neurosci., p. 345 1991).

[0009] A recent study of fampridine in patients with chronic incomplete SCI was

reported in Clinical Neuropharmacology 2003: Keith C. Hayes; Patrick J. Potter; Robert R.

Hansebout; Joanne M. Bugaresti; Jane T. C. Hsieh; Sera Nicosia; Mitchell A. Katz; Andrew

R. Blight; Ron Cohen 26(4):185-192.

SUMMARY OF THE INVENTION

[0010] One embodiment of the present invention relates to a pharmaceutical

composition which contains one or more potassium channel blockers and which can be used

in the effective treatment of various diseases, for example, spinal cord injury, multiple

sclerosis, Alzheimer's disease, and ALS. Embodiments of the present invention are directed

to compositions that include a matrix and a potassium channel blocker. The potassium

channel blockers may include aminopyridines, for example, 4-aminopyridine, 3,4

diaminopyridine and the like, most preferably 4-aminopyridine. The composition provides

for sustained-release of the aminopyridine from the matrix to maintain the efficacious and

safe plasma level of an aminopyridine. The aminopyridine dispersed in the matrix is capable

of providing, upon administration to a patient, a desired release profile. The composition

may be used to establish in patients in need of such treatment, a therapeutically effective

blood plasma level of the aminopyridine for a period of at least about 6 hours and preferably

up to at least about 12 hours in the patient in a twice-daily administration while avoiding

excessive peaks and troughs in the level of the aminopyridine. The composition may include a mono- or di-aminopyridine, preferably 4-AP or 3,4-DAP or a combination thereof, homogeneously dispersed in a rate-controlling polymer matrix, preferably including a hydrophilic polymer like hydroxypropylmethylcellulose (HPMC). The composition of the present invention may also include one or more additional active ingredients and/or one or more pharmaceutically acceptable excipients. These compositions can be used to treat various neurological diseases, for example, spinal cord injury, multiple sclerosis, Alzheimer's disease, and ALS.

[0011] Another embodiment of the present invention is a stable pharmaceutical

composition that comprises a therapeutically effective amount of an aminopyridine dispersed

in a matrix that provides a release profile of the aminopyridine to a patient that has a desired

Cmax to C, ratio. The composition may be used to establish and/or maintain in a patient, a

therapeutically effective level of the aminopyridine. Preferably the aminopyridine in the

composition is released over time so that a therapeutically effective level of the

aminopyridine in the patient can be achieved with twice daily dosing of the composition. In a

more preferred embodiment, undesirable spikes or peaks in the release of the aminopyridine

are avoided.

[0012] Another embodiment of the present invention is a stable, sustained-release

oral dosage formulation of a composition which includes a therapeutically effective amount

of a 4-aminopyridine dispersed in a matrix that provides a release profile of 4-aminopyridine

in the blood plasma of the patient extending over a period of at least 6 hours, preferably at

least 8 hours, and more preferably, at least about 12 hours. In another embodiment, a stable,

sustained-release oral dosage formulation of a composition includes a therapeutically

effective amount of a 4-aminopyridine dispersed in a matrix that provides a therapeutically

effective blood plasma level of 4-aminopyridine in the patient extending over about 24 hours.

[0013] Preferably, the oral dosage formulation of the composition is a monolithic

tablet formed by compression of the pharmaceutical composition of the present invention. In

preferred embodiments, the oral dosage formulation includes a compressed tablet of a

therapeutically effective amount of 4-aminopyridine dispersed in matrix that includes a

hydrophilic polymer such as HPMC. The oral dosage form of the present invention may also

include one or more pharmaceutically acceptable excipients.

[0014] The dispersion of 4-aminopyridine throughout the matrix imparts chemical

and physical stability to the composition while providing a sustained-release profile. This

enhanced dosage stability is most notably observed in compositions and dosage forms of the

present invention having low concentrations of 4-aminopyridine, and stability is achieved

while maintaining the desired controlled-release profile. Specifically, the compressed tablet

formulation of the present invention exhibits superior resistance to moisture absorption by

ambient humidity and maintains a uniform distribution of the 4-aminopyridine throughout the

tablet while providing a release profile of 4-aminopyridine that permits establishment of a

therapeutically effective concentration of the potassium channel blocker with once daily or

twice daily dosing of the formulation. Preferably the therapeutically effective concentration

released by the formulation extends over at least about 6 hours, preferably at least about 8

hours, and more preferably at least about 12 hours. In addition, the homogeneity of the

dosage form renders it amenable to formation by simple and inexpensive manufacturing

processes as compared with the multi-layered structure of prior sustained-release dosage

formulations.

[0015] The compositions of the present invention may be used in the treatment of a

condition in a patient that includes establishing a therapeutically effective concentration of a

potassium channel blocker in the patient in need thereof. The compositions may be used for

building up a level and or maintaining a therapeutically effective concentration of an aminopyridine in the patient by twice daily dosing. The dosages of the present compositions can be made with a lower concentration of the aminopyridine to facilitate restful periods for the patient during the day or night, depending on desired results or dosage schedule. Where desirable, the compositions of the present invention may be formulated to avoid large peaks in initial release of the aminopyridine. The compositions of the present invention when administered to a patient in need thereof provide for the treatment of neurological diseases that are characterized by a degradation of nerve impulse transmission. Preferably, the compositions are a stable, sustained-release tablet of a therapeutically effective amount of a mono- or di-aminopyridine, dispersed in HPMC such that therapeutically effective blood plasma level of the mono- or di-aminopyridine is maintained in the patient for a period of at least 6 hours, preferably at least 8 hours, and more preferably at least about 10-12 hours in a once or twice daily administration.

[0016] One embodiment of the present invention relates to a method of increasing

walking speed comprising administering to a patient with multiple sclerosis an effective

amount of a sustained release aminopyridine composition twice daily, wherein said effective

amount is less than about 15 milligrams of aminopyridine. In a preferred embodiment, the

effective amount is about 10 to about 15 milligrams of aminopyridine.

[0017] In a further embodiment of the present invention a method of improving

lower extremity muscle tone comprising administering to a patient with multiple sclerosis an

effective amount of a sustained-release aminopyridine composition twice daily is provided.

In a preferred embodiment, said effective amount is less than about 15 milligrams of

aminopyridine.

[0018] Another embodiment of the present invention relates to a method of

improving lower extremity muscle strength comprising administering to a patient with multiple sclerosis an effective amount of a sustained-release aminopyridine composition twice daily, wherein said effective amount is less than about 15 milligrams of aminopyridine.

[0019] One embodiment of the present invention relates to a method of selecting

individuals based on responsiveness to a treatment. The method comprises identifying a

plurality of individuals; administering a test to each individual prior to a treatment period;

administering a treatment to one or more of the individuals during the treatment period;

administering the test a plurality of times to each individual during the treatment period; and

selecting one or more individuals, wherein the selected individuals exhibit an improved

performance during a majority of the tests administered during the treatment period as

compared to the test administered prior to the treatment period. In certain embodiments, the

method may further comprise administering the test to each individual after the treatment

period, wherein the selected individuals further exhibit an improved performance during a

majority of the tests administered during the treatment period as compared to the test

administered after the treatment period.

[0020] A further embodiment relates to a method of selecting individuals based on

responsiveness to a treatment, the method comprising identifying a plurality of individuals;

administering a test to each individual prior to a treatment period; administering a treatment

to one or more of the individuals during the treatment period; administering the test a

plurality of times to each individual during the treatment period; administering the test to

each individual after the treatment period; and selecting one or more individuals, wherein the

selected individuals exhibit an improved performance during a majority of the tests

administered during the treatment period as compared to the better performance of the test

administered prior to the treatment period and the test administered after the treatment period.

BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 is a histogram to show the number of treatment visits at which

subjects showed faster walking speed on the timed 25 foot walk than at all of the five non

treatment visits.

[0022] Figure 2 is a graph of the average walking speeds (ft/sec) by study day

(observed cases, ITT population).

[0023] Figure 3 is a histogram of the percent change in average walking speed

during the 12-week stable dose period (observed cases, ITT population).

[0024] Figure 4 is a histogram of the percentage of protocol specified responders

(subjects with average changes in walking speed during the 12-week stable dose period of at

least 20%) by treatment group [(observed cases, ITT population]).

[0025] Figure 5 is a graph of LEMMT by study day (observed cases, ITT

population).

[0026] Figure 6 is a histogram of change in LEMMT during the 12-week stable dose

period (observed cases, ITT population).

[0027] Figure 7 is a histogram of the percentage of post hoc responders by treatment

group (ITT population) according to a responder analysis of the present invention.

[0028] Figure 8 is a histogram of the percentage of responders for placebo subjects

vs. fampridine subjects pooled (ITT population) according to a responder analysis of the

present invention.

[0029] Figure 9 are histograms of the validation of the post hoc responder variable

using subjective scales (observed cases, ITT population).

[0030] Figure 10 is a graph of percent change in walking speed at each double-blind

visit by responder analysis grouping (observed cases, ITT population).

[0031] Figure 11 is a graph of the change in LEMMT at each double-blind visit by

responder analysis grouping (observed cases, ITT population).

[0032] Figure 12 is a graph of change in overall Ashworth Score at each double

blind visit by responder analysis grouping (observed cases, ITT population).

DETAILED DESCRIPTION OF THE INVENTION

[0033] Before the present compositions and methods are described, it is to be

understood that this invention is not limited to the particular molecules, compositions,

methodologies or protocols described, as these may vary. It is also to be understood that the

terminology used in the description is for the purpose of describing the particular versions or

embodiments only, and is not intended to limit the scope of the present invention which will

be limited only by the appended claims.

[0034] The terms used herein have meanings recognized and known to those of skill

in the art, however, for convenience and completeness, particular terms and their meanings

are set forth below.

[0035] It must also be noted that as used herein and in the appended claims, the

singular forms "a", "an", and "the" include plural reference unless the context clearly dictates

otherwise. Thus, for example, reference to a "spheroid" is a reference to one or more

spheroid and equivalents thereof known to those skilled in the art, and so forth. Unless

defined otherwise, all technical and scientific terms used herein have the same meanings as

commonly understood by one of ordinary skill in the art. Although any methods and

materials similar or equivalent to those described herein can be used in the practice or testing

of embodiments of the present invention, the preferred methods, devices, and materials are

now described. All publications mentioned herein are incorporated by reference. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0036] "Local administration" means direct administration by a non-systemic route

at or in the vicinity of the site of affliction, disorder, or perceived pain.

[0037] The terms "patient" and "subject" mean all animals including humans.

Examples of patients or subjects include humans, cows, dogs, cats, goats, sheep, and pigs.

[0038] The term "pharmaceutically acceptable salts, esters, amides, and prodrugs"

as used herein refers to those carboxylate salts, amino acid addition salts, esters, amides, and

prodrugs of the compounds of the present invention which are, within the scope of sound

medical judgment, suitable for use in contact with the tissues of patients without undue

toxicity, irritation, allergic response, and the like, commensurate with a reasonable

benefit/risk ratio, and effective for their intended use, as well as the zwitterionic forms, where

possible, of the compounds of the invention.

[0039] The term "prodrug" refers to compounds that are rapidly transformed in vivo

to yield the parent compounds of the above formula, for example, by hydrolysis in blood. A

thorough discussion is provided in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery

Systems," Vol. 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug

Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press,

1987, both of which are incorporated herein by reference.

[0040] The term "salts" refers to the relatively non-toxic, inorganic and organic acid

addition salts of compounds of the present invention. These salts can be prepared in situ

during the final isolation and purification of the compounds or by separately reacting the

purified compound in its free base form with a suitable organic or inorganic acid and

isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride,

sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate, lactobionate and laurylsulphonate salts, and the like.

These may include cations based on the alkali and alkaline earth metals, such as sodium,

lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium,

tetramethylammonium, tetramethylammonium, methlyamine, dimethlyamine,

trimethlyamine, triethlyamine, ethylamine, and the like. (See, for example, S.M. Barge et al.,

"Pharmaceutical Salts," J. Pharm. Sci., 1977, 66:1-19 which is incorporated herein by

reference.).

[0041] A "therapeutically effective amount" is an amount sufficient to decrease or

prevent the symptoms associated with a medical condition or infirmity, to normalize body

functions in disease or disorders that result in impairment of specific bodily functions, or to

provide improvement in one or more of the clinically measured parameters of the disease.

Preferably, improvement in symptoms associated with the disease including walking speed,

lower extremity muscle tone, lower extremity muscle strength, or spasticity. As related to the

present application, a therapeutically effective amount is an amount sufficient to reduce the

pain or spasticity associated with the neurological disorder being treated, or an amount

sufficient to result in improvement of sexual, bladder or bowel function in subjects having a

neurological disorder which impairs nerve conduction, which hinders normal sexual, bladder

or bowel functions.

[0042] "Treatment" refers to the administration of medicine or the performance of

medical procedures with respect to a patient, for either prophylaxis (prevention), to cure the

infirmity or malady in the instance where the patient is afflicted refers, or amelioration the

clinical condition of the patient, including a decreased duration of illness or severity of

illness, or subjective improvement in the quality of life of the patient or a prolonged survival

of the patient.

[0043] In addition, the compounds of the present invention can exist in unsolvated

as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol,

and the like. In general, the solvated forms are considered equivalent to the unsolvated forms

for the purposes of the present invention.

[0044] One aspect of the invention is a sustained-release pharmaceutical

composition comprising an aminopyridine dispersed in a sustained release matrix such as a

rate-controlling polymer. The composition of the present invention is capable of providing,

upon administration to a patient, a release profile of the aminopyridine extending over at least

6 hours, preferably least about 12 hours, and more preferably at least 24 hours or more.

Preferably the aminopyridine concentration in the composition is a therapeutically effective

amount, and preferably the aminopyridine is dispersed uniformly throughout the release

matrix. A therapeutically effective amount is an amount of a potassium channel blocker,

preferably an aminopyridine compound, that when administered to a patient or subject,

ameliorates a symptom of a neurological disease.

[0045] When the compositions of the present invention are administered to a patient,

the concentration of the aminopyridine in the patient's plasma over time (release profile) may

extend over a period of at least 6 hours, preferably over at least 8 hours, and more preferably

over at about 12 hours. The compositions may provide in single dose a mean maximum

plasma concentration of aminopyridine in the patient of from about 15 to about 180 ng/ml; a

mean Tmaxfrom about 1 to about 6 hours, more preferably about 2 to about 5.2 hours after

administration of the composition to the patient.

[0046] In one embodiment, aminopyridine is administered to a subject at a dose and

for a period sufficient to allow said subject to tolerate said dose without showing any adverse

effects and thereafter increasing the dose at selected intervals of time until a therapeutic dose

is achieved. In one embodiment, the medicament is administered to a subject at a dose and for a period sufficient to allow said subject to tolerate said dose without showing any adverse effects and thereafter increasing the dose of aminopyridine at selected intervals of time until a therapeutic dose is achieved. For example, at the commencement of treatment aminopyridine is preferably administered at a dose less than 15 mg/day until a tolerable state is reached.

Suitably when said tolerable state is reached, the dose administered may be increased by

amounts of at least 5-15 mg/day until said therapeutic dose is reached.

[0047] Preferably, aminopyridine is administered at a dose of about 10-15 mg twice

daily (20-30 mg/day) depending upon the condition or symptoms being treated. The method

can include scheduling administration of doses of the pharmaceutical so that the

concentration of the aminopyridine in the patient is at about the minimum therapeutically

effective level to ameliorate the neurological condition, yet relatively lower compared to the

maximum concentration in order to enhance restful periods for the patient during the day or

night, depending on desired results or dosage schedule. Preferably the method provides for

the treatment of neurological diseases characterized by a degradation of nerve impulse

transmission comprising the step of administering to a patient a composition of the present

invention.

[0048] The formulations and compositions of the present invention exhibit a

specific, desired release profile that maximizes the therapeutic effect while minimizing

adverse side effects. The desired release profile may be described in terms of the maximum

plasma concentration of the drug or active agent (Cax) and the plasma concentration of the

drug or active agent at a specific dosing interval (Ct). A ratio of Cmax to Cr (Cmax:CT)may

be calculated from the observed Cmax and C. A dosing interval (t) is the time since the last

administration of the drug or active agent. In the present application, the dosing interval (t) is

twelve (12) hours, therefore C, is the concentration of the drug or active agent at twelve (12)

hours from the last administration.

[0049] Additionally, the formulations and compositions of the present invention

exhibit a desired release profile that may be described in terms of the maximum plasma

concentration of the drug or active agent at steady state (Cmaxss) and the minimum plasma

concentration of the drug or active agent at steady state (Cmjnss). Steady state is observed

when the rate of administration (absorption) is equal to the rate of elimination of the drug or

active agent. A ratio of Cmaxss to Cinss (Cmaxss:Cminss) may be calculated from the observed

CmaxSs and Cminss. In addition, the formulations and compositions of the present invention

exhibit a desired release profile that may be described in terms of the average maximum

plasma concentration of the drug or active agent at steady state (Cavss).

[0050] Another embodiment is a sustained release tablet of a sustained release

matrix and an aminopyridine, said tablet exhibits a release profile to obtain a Cmax:C, ratio in

vivo of 1.0 to 3.5, and more preferably a Cmax:C, ratio of about 1.5 to about 3.0. In another

preferred embodiment, the Cmax:C, ratio is about 2.0 to about 3.0. The aminopyridine may

comprise 4-aminopyridine. The sustained release matrix may include for example,

hydroxypropylmethylcellulose, or other rate controlling matrices that are suitable for

controlling the release rate of an aminopyridine for use in the pharmaceutical compositions of

the present invention.

[0051] Another embodiment is a sustained release tablet of a sustained release

matrix and an aminopyridine, said tablet exhibits a release profile to obtain aCmax:C, ratio in

vivo of 1.0 to 3.5 and a Cavss of about 15 ng/ml to about 35 ng/ml, and more preferably a

Cmax:C, ratio of about 1.5 to about 3.0. In another preferred embodiment, the Cmax:C, ratio is

about 2.0 to about 3.0.

[0052] A further aspect is a sustained release composition comprising a sustained

release matrix and an aminopyridine, wherein said composition provides aCavss of about 15

ng/ml to about 35 ng/ml. In a further aspect, a sustained release tablet comprising a sustained release matrix and an aminopyridine, said tablet exhibiting a Cmaxss of about 20 ng/ml to about 35 ng/ml is provided. The pharmacokinetic characteristics of sustained release aminopyridine compositions and methods of treating various neurological disorders are described in co-pending PCT/US2004/008101 entitled "Stable Formulations of

Aminopyrdines and Uses Thereof' filed April 17, 2004 and U.S. Application No. 11/010,828

entitled "Sustained Release Aminopyridine Composition" filed December 13, 2004, the

contents of which are incorporated herein by reference in their entireties.

[0053] The amount of a pharmaceutically acceptable quality aminopyridine, salt,

solvated, or prodrug thereof included in the pharmaceutical composition of the present

invention will vary, depending upon a variety of factors, including, for example, the specific

potassium channel blocker used, the desired dosage level, the type and amount of rate

controlling polymer matrix used, and the presence, types and amounts of additional materials

included in the composition. Preferably, the aminopyridine comprises from about 0.1 to

about 13%w/w, more preferably from about 0.5 to about 6.25 %w/w. In an even more

preferable embodiment of the present invention the aminopyridine is present from about 0.5

to 4.75 %w/w of the pharmaceutical composition. Accordingly, a weight percentage less

than about 4.75% is desired. The amount of aminopyridine, or a derivative thereof, in the

formulation varies depending on the desired dose for efficient drug delivery, the molecular

weight, and the activity of the compound. The actual amount of the used drug can depend on

the patient's age, weight, sex, medical condition, disease or any other medical criteria. The

actual drug amount is determined according to intended medical use by techniques known in

the art. The pharmaceutical dosage formulated according to the invention may be

administered once or more times per day, preferably two or fewer times per day as

determined by the attending physician.

[0054] Suitable formulations and methods of manufacture are further described in

co-pending PCT/US2004/008101 entitled "Stable Formulations of Aminopyrdines and Uses

Thereof' filed April 17, 2004 and U.S. Application No. 11/010,828 entitled "Sustained

Release Aminopyridine Composition" filed December 13, 2004, the contents of which are

incorporated herein by reference in their entireties.

[0055] The release matrix aminopyridine formulation is preferably fabricated into

tablets, capsules or granules for oral use. The rate of aminopyridine release from the tablets

may be controlled by the erosion mechanism of the release matrix from which aminopyridine

is released. In general, for producing a tablet on an industrial scale, the drug and polymer are

granulated alone or in combination. Preferably the release of the aminopyridine from the

matrix of the pharmaceutical composition is relatively linear over time. Preferably the matrix

provides a release profile that gives a therapeutically effective concentration of the

aminopyridine in the plasma of the patient permitting a once per day or twice per day dosing.

Preferably the sustained release aminopyridine formulation for oral administration to patients

includes from about 0.0001 mole to about 0.0013 mole aminopyridine that provides a mean

maximum plasma concentration of aminopyridine from about 15 to about 180 ng/ml, a mean

Tmax of about 2 to about 5 hours after administration, and a mean minimum plasma

concentration of from about 10 to 60 ng/ml at about 8-24 hours after administration.

[0056] The formulations of the invention are prepared by procedures known in the

art, such as, for example, by the dry or wet method. The method selected for manufacturing

affects the release characteristics of the finished tablet. In one method, for example, the

tablet is prepared by wet granulation in the presence of either water or an aqueous solution of

the hydrophilic polymer or using other binder as a granulating fluid. In alternative, organic

solvent, such as isopropyl alcohol, ethanol and the like, may be employed with or without

water. The drug and polymer may be granulated alone or in combination. Another method in for preparation of the tablet which may be used requires using a drug-polymer dispersion organic solvents in the presence or absence of water. Where the aminopyridine or its derivative has very low solubility in water it may be advantageous to reduce the particle size, for example, by milling it into fine powder and in this way to control the release kinetics of the drug and enhance its solubility.

[0057] The hardness of the tablets of the present invention may vary, depending on

a variety of factors, including, for example, the relative amounts and specific types of

ingredients used,, the tableting equipment employed, and the selected processing parameters.

The pressure used to prepare the tablets can influence the release profile of the aminopyridine

into the patient. The pressure used to prepare the tablets of the present invention may vary

depending upon their surface area and the amount and particle size of aminopyridine,

additive, excipients, or binders included in the tablet. The degree of hydration and solvation

of the components in the composition will also be important in determining the hardness of

the tablets. Preferably the formed tablets have a hardness in the range of from 80-400 N, and

more preferably from 150 to 300 N.

[0058] The effects of various matrices, concentrations of aminopyridine, as well as

various excipients and additives to the composition on the concentration of the channel

blocker on the dissolution rate may be monitored for example using a type H dissolution

apparatus according to U.S. Pharmacopoeia XXII, or USP Apparatus II (Paddle Method).

Clinical evaluations may be used to study the effects on plasma levels of various release

matrices, concentrations of aminopyridine, as well as various excipients and additives.

Plasma aminopyridine concentrations may be used to calculate pharmacokinetic data (release

profiles) including apparent absorption and elimination rates, area-under-the curve (AUC),

maximum plasma concentration (Cmax), time to maximum plasma concentration (Tmax),

1 2(abs)), and elimination half-life (T 2 (elim)). absorption half-life (Tu Pharmacodynamic effects may be assessed based upon response tests, such as muscle strength improvement or reduction in spasticity for patients with multiple sclerosis or spinal cord injury or other tests as would be known to those skilled in the art. Plasma aminopyridine concentration in blood plasma or cerebral spinal fluid may be monitored using liquid chromatography/MS/MS assay methods.

[0059] The drug delivery of the invention can utilize any suitable dosage unit form.

Specific examples of the delivery system of the invention are tablets, tablets that disintegrate

into granules, capsules, sustained release microcapsules, spheroids, or any other means that

allow for oral administration. These forms may optionally be coated with pharmaceutically

acceptable coating which allows the tablet or capsule to disintegrates in various portions of

the digestive system. For example a tablet may have an enteric coating that prevents it from

dissolving until it reaches the more basic environment of the small intestine.

[0060] The dispersion of the aminopyridine throughout the release matrix imparts

enhanced stability characteristics in the dosage formulation. This enhanced stability is

achieved without loss of the desired sustained-release profile. Preferably the release profile,

which may be measured by dissolution rate is linear or approximately linear, preferably the

release profile is measured by the concentration of the aminopyridine in the plasma in the

patient and is such to permit twice daily (BID) dosing.

[0061] The pharmaceutical composition of the present invention can include also

auxiliary agents or excipients, for example, glidants, dissolution agents, surfactants, diluents,

binders including low temperature melting binders, disintegrants, solubilizing agents and/or

lubricants as described in co-pending PCT/US2004/008101 entitled "Stable Formulations of

Aminopyrdines and Uses Thereof' filed April 17, 2004 and U.S. Application No. 11/010,828

entitled "Sustained Release Aminopyridine Composition" filed December 13, 2004, the

contents of which are incorporated herein by reference in their entireties.

[0062] The active ingredient of the present invention may be mixed with excipients

which are pharmaceutically acceptable and compatible with the active ingredient and in

amounts suitable for use in the therapeutic methods described herein. Various excipients may

be homogeneously mixed with the aminopyridines of the present invention as would be

known to those skilled in the art. For example, aminopyridines may be mixed or combined

with excipients such as but not limited to microcrystalline cellulose, colloidal silicon dioxide,

lactose, starch, sorbitol, cyclodextrin and combinations of these.

[0063] To further improve the stability of the aminopyridine in the sustained release

composition, an antioxidant compound can be included. Suitable antioxidants include, for

example: sodium metabisulfite; tocopherols such as x, @, 8-tocopherol esters and

ax.-tocopherol acetate; ascorbic acid or a pharmaceutically acceptable salt thereof; ascorbyl

palmitate; alkyl gallates such as propyl gallate, Tenox PG, Tenox s-1; sulfites or a

pharmaceutically acceptable salt thereof; BHA; BHT; and monothioglycerol.

[0064] In another embodiment, the pharmaceutical composition of the present

invention comprises a rate-controlling polymeric matrix comprising of a hydrogel matrix.

For instance, an aminopyridine may be compressed into a dosage formulation containing a

rate-controlling polymer, such as HPMC, or mixture of polymers which, when wet, will swell

to form a hydrogel. The rate of release of the aminopyridine from this dosage formulation is

sustained both by diffusion from the swollen tablet mass and by erosion of the tablet surface

over time. The rate of release of the aminopyridine may be sustained both by the amount of

polymer per tablet and by the inherent viscosities of the polymers used.

[0065] According to another aspect of the invention, there is provided a stable,

sustained-release oral dosage formulation which includes an effective amount a

aminopyridine dispersed in a release matrix, and which, upon administration to a patient or as

part of a therapy regiment, provides a release profile (of therapeutically effective blood plasma level of the aminopyridine) extending for a period of at least 6 hours, preferably at least 12 hours. In another embodiment, the stable, controlled-release oral dosage form of provides, upon administration to a patient, a therapeutically effective blood plasma level the aminopyridine for a period of at least 6 hours, preferably at least 12 hours, and more preferably at least 24 hours.

[0066] The dosage formulation may assume any form capable of delivering orally to

a patient a therapeutically effective amount of an aminopyridine dispersed in a rate

controlling polymer. Preferably, the dosage formulation comprises a monolithic tablet.

[0067] Tablet weight will also vary in accordance with, among other things, the

aminopyridine dosage, the type and amount of rate-controlling polymer used, and the

presence, types and amounts of additional materials. Assuming 4-aminopyridine dosages of

from about 2 mg to about 120 mg; tablet weights can range from about 50 mg to about 1200

mg per tablet, and preferably from 250 to 500 mg, and more preferably about 400 mg.

[0068] The dosage formulation of the present invention may comprise also one or

more pharmaceutically acceptable excipients as mentioned above. In preferred embodiments,

the dosage formulation will comprise diluents and a lubricant in addition to the

aminopyridine unit dose and the rate-controlling polymer. Particularly preferred diluents is

microcrystalline cellulose sold under the name Avicel PH101, and a particularly preferred

lubricant is magnesium stearate. When these materials are used, the magnesium stearate

component preferably comprises from about 0.2 to about 0.75 %w/w of the dosage

formulation, and the microcrystalline cellulose along with the rate controlling polymer and

aminopyridine comprises the balance of the formulation. For example, a tablet formulation

including a aminopyridine x % w/w, a rate-controlling polymer y % w/w, and

microcrystalline cellulose z %, the magnesium stearate amount would be (100-(x+y+z))

where 0.2%:5 (100-(x+y+z)) < 0.75% w/w. As would be known to those skilled in the art, the amount of an additives such as magnesium stearate may vary depending upon the shear rate used to perform the mixing and the amount of such an additive may be changed without limitation to obtain a satisfactory dissolution rate or plasma level of the aminopyridine.

[0069] As used herein, the term "sustained-release" as it relates to the

aminopyridine compositions includes the release of a aminopyridine from the dosage

formulation at a sustained rate such that a therapeutically beneficial blood level below toxic

levels of the aminopyridine is maintained over a period of at least about 12 hours, preferably

about 24 hours or more. Preferably, the amount of the aminopyridine in the oral dosage

formulations according to embodiments of the present invention establish a therapeutically

useful plasma concentration through BID administration of the pharmaceutical composition.

[0070] If desired, the dosage formulations of this invention may be coated with a

sustained-release polymer layer so as to provide additionalsustained-release properties.

Suitable polymers that can be used to form this sustained release layer include, for example,

the release matrices listed above. As desired, the dosage formulation of the invention can be

provided also with a light-protective and/or cosmetic film coating, for example, film-formers,

pigments, anti-adhesive agents and politicizes. Such a film-former may consist of

fast-dissolving constituents, such as low-viscosity hydroxypropylmethylcelluose, for

example, Methocel E5 or D14, or Pharmacoat 606 (Shin-Etsu). The film coating may also

contain excipients or enteric coatings customary in film-coating procedures, such as, for

example, light-protective pigments, for example, iron oxide, or titanium dioxide,

anti-adhesive agents, for example, talc, and also suitable plasticizers such as, for example,

PEG 400, PEG 6000, diethyl phthalate or triethyl citrate.

[0071] The compositions of the present invention may be used for the treatment of

neurological diseases characterized by a degradation of nerve impulse transmission by

administering to a patient the oral dosage formulation of the present invention. Preferably, the administration is twice daily dosage of a therapeutically effective amount of an aminopyridine, even more preferably, 4-AP dispersed in HPMC. The administration can also include scheduling administration of doses of the pharmaceutical so that the concentration of the aminopyridine in the patient is at about the minimum therapeutically effective level to ameliorate the neurological condition, yet relatively low compared to the maximum concentration in order to minimize side effects. The compositions may be administered to a subject at a dose and for a period sufficient to allow said subject to tolerate said dose without showing any adverse effects and thereafter increasing the dose of said active agent in the tablets at selected intervals of time until a therapeutic dose is achieved in the subject. For example, at the commencement of treatment the active agent is preferably administered at a dose less than about 15 mg/day until a tolerable state is reached. The dose administered may then be increased by amounts of at least 5-10 mg/day until a therapeutic dose is reached, preferably less than about 30 mg/day. For other diseases the amount of the aminopyridine required to reach a therapeutically effective amount for treatment is described in U.S. Pat.

No. 5, 952,357 the contents of which are incorporated herein by reference in their entirety.

[0072] Compositions of the present invention where the potassium channel blocker

is a mono- or di-aminopyridine active agent are particularly suitable for use in the treatment

of a neurological disease that is characterized by demyelination of the central nervous system,

more especially multiple sclerosis.

[0073] In one embodiment of the present invention, a method of treating multiple

sclerosis is provided. Compositions of the present invention containing a therapeutically

effective amount of mono- or di-aminopyridine active agent may be administered to a patient

in need thereof. In particular, sustained release compositions comprising at least about 5

milligrams of an aminopyridine, preferably 4-aminopyridine may be administered at least

once daily. In a preferred embodiment, a sustained release composition containing from about 10 to about 15 milligrams of 4-aminopyridine is administered twice daily. Treatment of multiple sclerosis may include increased walking speed, improved lower extremity muscle strength or improved lower extremity muscle tone. The sustained release aminopyridine composition is preferably administered twice daily. In certain embodiments, the composition may be administered about every 12 hours.

[0074] A further embodiment is a method of increasing walking speed in patients

with multiple sclerosis comprising administering to a patient at least about 5 milligrams of a

sustained release aminopyridine composition, preferably at least about 10 to about 15

milligrams of a sustained release aminopyridine composition.

[0075] A further embodiment is a method of increasing muscle tone or muscle

strength in patients with multiple sclerosis comprising administering to a patient at least

about 5 milligrams of a sustained release aminopyridine composition, preferably at least

about 10 to about 15 milligrams of a sustained release aminopyridine composition.

[0076] Fampridine is a potential therapy for MS with a unique mechanism of action.

At concentrations of 1-2 sM or less, fampridine appears to be a specific blocker of voltage

dependent, neuronal potassium channels that affect conduction in demyelinated axons.

Fampridine has been shown to restore action potential conduction in damaged, poorly

myelinated nerve fibers, and it may also directly enhance synaptic transmission. In previous

clinical trials, treatment with fampridine has been associated with a variety of neurological

benefits in people with MS including faster walking and increased strength, as measured by

standard neurological assessments.

[0077] Another aspect of the present invention provides for a method of selecting

individuals based on responsiveness to a treatment. In one embodiment, the method

comprises identifying a plurality of individuals; administering a test to each individual prior

to a treatment period; administering a treatment, including, but not limited to administering a therapeutic agent or drug, to one or more of the individuals during the treatment period; administering the test a plurality of times to each individual during the treatment period; and selecting one or more individuals, wherein the selected individuals exhibit an improved performance during a majority of the tests administered during the treatment period as compared to the test administered prior to the treatment period. In certain embodiments, the method may further comprise administering the test to each individual after the treatment period, wherein the selected individuals further exhibit an improved performance during a majority of the tests administered during the treatment period as compared to the test administered after the treatment period.

[0078] It is important to note that this embodiment selects subjects who show a

pattern of change that is consistent with a treatment response, but does not define the full

characteristics of that response. The criterion itself does not specify the amount of

improvement nor does it specify that the improvement must be stable over time. For

example, a progressive decline in effect during the course of the study period, even one

resulting in speeds slower than the maximum non-treatment value, would not be excluded by

the criterion; as a specific example, changes from the maximum non-treatment value of,

respectively, +20%, +5%, + 1% and -30% during the double blind treatment period would

qualify as a response under the criterion, but would actually show a net negative average

change for the entire period, poor stability and a negative endpoint. Post-hoc analyses of

studies discussed in greater detail below indicate that we may expect responders defined by

consistency of effect also to demonstrate increased magnitude and stability of benefit.

[0079] We have found this embodiment particularly applicable in our analysis of

fampridine in patients suffering from multiple sclerosis. Clinicians who regularly prescribe

compounded fampridine for MS have reported that only a proportion of their patients appear

to respond with clear clinical benefits, and that, in their judgment, this proportion may be around one third. This extent of responsiveness may be related to the proposed mechanism of action, which is the restoration of conduction in demyelinated axons via the blockade of voltage-dependent potassium channels. Only a proportion of MS patients would be expected to possess axons of appropriate functional relevance that are susceptible to these drug effects, given the highly variable pathology of the disease. Currently, there is insufficient understanding of the disease to allow for pre-trial selection of potentially responsive patients.

However, the existence of a subset of patients who respond consistently to the drug can be

supported by quantitative observations in our own clinical studies discussed below.

[0080] Before treatment, the subjects in these two trials exhibited average walking

speeds on the TW25 measure of approximately 2 feet per second (ft/sec). This is a

significant deficit, since the expected walking speed for an unaffected individual is 5-6 ft/sec.

Subjects in MS-F202 were selected for TW-25 walking time at screening of 8-60, which is

equivalent to a range in speed of 0.42-3.1 ft/sec. Variability of functional status is an

inherent characteristic of MS, and this can be seen in repeated measurement of walking speed

over the course of weeks or months. At any of the three visits during the stable treatment

period, 15-20% of placebo-treated subjects showed >20% improvement from baseline

walking speed, a threshold chosen as one that indicates a true change in walking speed over

background fluctuations. A larger proportion of the Fampridine-SR treated subjects showed

such improvements, but this difference was not statistically significant, given the sample size

and placebo response rate.

[0081] Given the often large variations in function experienced by people with MS,

it is difficult for the subject or a trained observer to separate a treatment-related improvement

from a disease-related improvement without the element of consistency over time.

Consistency of benefit might therefore be expected to be a more selective measure of true

treatment effect than magnitude of change. Based on this rationale, the responses of the individual subjects in the MS-F202 trial were examined for the degree to which their walking speed showed improvement during the double-blind treatment period and returned towards pre-treatment values after they were taken off drug, at follow-up. This subject-by-subject examination yielded a subgroup of subjects whose pattern of walking speed over time appeared to be consistent with a drug response. This led to the analysis illustrated in Figure

1. This compares the placebo and Fampridine-SR treated groups with respect to the number

of visits during the double-blind treatment period in which walking speed on the TW25 was

faster than the maximum speed out of all five of the non-treatment visits (four visits prior to

randomization and one follow-up visit after the drug treatment period).

[0082] The placebo-treated group showed a clear pattern of exponential decline in

numbers of subjects with higher numbers of "positive" visits. This is what would be

expected from a random process of variability. In contrast, the pattern of response in the

Fampridine-SR treated group strongly diverged from this distribution; much larger numbers

of Fampridine-SR treated subjects showed three or four visits with higher walking speeds

than the maximum speed of all five non-treatment visits and less than half of the expected

proportion had no visits with higher speeds. These results indicate that there was a sub

population of subjects in the Fampridine-SR treated group that experienced a consistent

increase in walking speed related to treatment.

[0083] This analysis suggests that a relatively highly selective criterion for a likely

treatment responder would be: a subject with a faster walking speed for at least three (i.e.,

three or four) of the four visits during the double blind treatment period compared to the

maximum value for all five of the non-treatment visits. The four visits before initiation of

double-blind treatment provide an initial baseline against which to measure the consistency of

response during the four treatment visits. The inclusion of the follow-up visit as an additional

component of the comparison was found valuable primarily in excluding those subjects who did not show the expected loss of improvement after coming off the drug. These are likely to be subjects who happened by chance to have improved in their MS symptoms around the time of treatment initiation, but whose improvement did not reverse on drug discontinuation because it was actually unrelated to drug. Thus, incorporating the follow-up visit as part of the criterion may help to exclude false positives, if the TW25 speed remains high at follow up.

[0084] As described in Example 5, below, this responder criterion was met by 8.5%,

35.3%, 36.0%, and 38.6% of the subjects in the placebo, 10 mg, 15 mg, and 20 mg b.i.d.

treatment groups, respectively, showing a highly significant and consistent difference

between placebo and drug treatment groups. Given that there was little difference in

responsiveness between the three doses examined, more detailed analyses were performed

comparing the pooled Fampridine-SR treated groups against the placebo-treated group. The

full results of this analysis for study are described in the following sections. These show that

the responder group so identified experienced a >25% average increase in walking speed over

the treatment period and that this increase did not diminish across the treatment period. The

responder group also showed an increase in Subject Global Impression score and an

improvement in score on the MSWS-12.

[0085] Additional features and embodiments of the present invention are illustrated

by the following non-limiting examples.

EXAMPLE 1

[0086] This example illustrates preparation of compositions of the present invention

and their release of an aminopyridine. Tablets in accordance with the present invention

having dosages of 5 mg, 7.5 mg and 12.5 mg respectively were manufactured at 5Kg scale.

Materials were used in the amounts shown in Table 1.

TABLE % w/w % w/w % w/w Milled 4-AP (#50 mesh) 1.25 1.875 3.125 Methocel K100LV 60 60 60 Avicel PH101 38.15 37.525 36.275 Magnesium stearate 0.2 0.2 0.2 Aerosil 200 0.4 0.4 0.4 Equipment Tablet Press Horn Noak equipped with 13 x 8mm oval tooling pre s speed 42,000 tablets / hr Tablet Weight Range (mg) 386-404 388-410 388-406 (96.5-101.0%) (97.0-102.5%) (97.0-101.5%) Tablet Hardness Range (N) 200-262 179- 292 150-268

Tablet Potency - mg/tab. (%LC) 97.1 99.1 100.2

Mean CU (mg/tab.)/%CV 5.0mg / 1.0% 7.4mg / 0.7% 12.4mg / 1.1% CU Discrete Samples 5.0mg /1.2% 7.5mg / 1.8% 12.3/1.1% (mg/tab.)/%CV Dissolution (%/hr) Mean (SD) Mean (SD) Mean (SD) 1 28.9 1.1 29.2 1.8 25.9 1.1 2 42.7 1.8 42.1 1.6 40.2 2.5 3 52.8 1.4 53.0 1.0 49.8 2.1 4 61.4 2.2 61.8 1.5 60.1 2.4 6 75.7 3.1 75.2 1.6 74.8 2.7 10 95.5 3.3 98.7 1.4 93.2 0.9

[0087] Prior to blending, 4-AP was milled through #50 mesh screen using a

Fitzmill* comminutor. The materials were added into a Gral 25 bowl in the following order:

half Methocel K100LV, Avicel PH101, Aerosil 200, milled 4-AP and the remaining

Methocel K100LV. The mix was blended for 15 minutes at 175 rpm, then the magnesium

stearate was added and was further blended for 5 minutes at 100 rpm. Samples were taken

from top and bottom positions for blend potency analysis. Weight and hardness checks were

performed every 15 minutes by the check-master E3049. Discrete tablet samples were taken

during the compression process to evaluate intra batch content uniformity.

EXAMPLE2

[0088] This example illustrates that the pharmacokinetic profile of fampridine in

compositions of the present invention is altered by administration in a sustained release tablet

matrix compared to immediate release and controlled release formulations.

[0089] There is a delay in absorption manifested by a lower peak concentration,

without any effect on the extent of absorption. When given as a single 12.5 mg dose, the

peak concentration is approximately two-thirds lower as compared to peak values following

administration of the IR formulation; the time to reach peak plasma levels was delayed by

about 2 hours. As with the IR formulation, food delayed the absorption of Fampridine-SR.

The absorption of fampridine was approximately 50% slower following ingestion of a fatty

meal, although due to the flatness of the absorption curve, this may be exaggerated value.

Extent of absorption did not differ, as values for Cmax and AUC were comparable as

summarized in Table 2.

Table 2 Pharmacokinetic Parameter Values (Mean ±SD) in Studies Using Fampridine SR, CR, and IR Formulations: Single Dose Studies in Healthy Adult Male Volunteers

Study Number Dose (mg) Fed/Fasted CMx tvx (hours) AUC (0-oo) (ng/mL) (ng hr/mL) 0494006 12.5 SR Fed 28.7 ±4.3 5.3 ±0.8 257.0 ±62.7 N=12 (PD12265) Fasted 25.6 ±3.8 2.8 ±1.3 269.9 ±44.4 12.5 IR Fasted 79.3 16.3 0.9 ±0.4 294.2 ±55.6 (PD12266) 1194002 12.5 SR Fasted 28.5 4.3 2.9 ±2.4 285.9 ±37.8 N= 12 (PD12907) 12.5 CR Fasted 37.7 9.9 3.6 ±0.9 300.0 ±53.6 (4n806) 12.5 IR Fasted 83.5 23.5 0.79 ±0.3 274.0 ±59.2 (PS644) I I

EXAMPLE3

[0090] This example details the pharmacokinetic properties of Fampridine-SR in

tablets of the present invention administered to patients with multiple sclerosis. Plasma

samples were analyzed for fampridine using a validated LC/MS/MS assay with a sensitivity

of 2 ng/mL. Noncompartmental pharmacokinetic parameter values were calculated using

standard methodology.

[0091] This was an open-label, multi-center, dose proportionality study of orally

administered fainpridine in patients with multiple sclerosis. Single doses of fampridine were

to be given in escalating doses (5 mg, 10 mg, 15 mg, and 20 mg) with at least a four-day

interval between administration of each dose of drug. Safety evaluations were to be

performed during the 24 hour period following administration of fampridine and blood

samples were to be taken at the following times to determine pharmacokinetic parameters:

hour 0 (pre-dose), hours 1-8, and hours 10, 12, 14, 18, and 24.

[0092] Twenty-three subjects received all 4 treatments, and one subject received

only 3 treatments; data from all treatments were analyzed. Dose-dependent parameters (e.g.,

peak plasma concentration and areas-under-the curve) were normalized to a 10 mg dose for

among-dose comparisons. Overall observed time of the peak plasma concentration (mean

and its 95% confidence interval) was 3.75 (3.52, 3.98) h, observed peak plasma fampridine

concentration (normalized to a 10 mg dose) was 24.12 (23.8, 26.6) ng/ml, area-under-the

concentration-time curve (normalized to a 10 mg dose) was estimated to be 254 (238, 270)

ng-h/ml, extrapolated area-under-the-concentration-time curve (normalized to a 10 mg dose)

was 284 (266, 302) ng-h/ml, terminal rate constant equaled 0.14 (0.13, 0.15) h-, terminal

half-life was 5.47 (5.05, 5.89) h and clearance divided by bioavailability (CL/F) was equal to

637 (600, 674) ml/ min.

[0093] Dizziness was the most common treatment-related adverse event. Other

treatment related adverse events included amblyopia, asthenia, headache, and ataxia. There

were no clinically significant changes in clinical laboratory values, ECG parameters, vital

signs, physical examination findings, or neurological examination findings noted over the

course of this study.

[0094] When the plasma concentrations of fampridine were normalized to the 10.0

mg dose levels, there were no significant differences between any pharmacokinetic parameter

(AUC, Cmx, t 2) in the 5-20 mg dose range. Fampridine was well tolerated at the doses used

in this study. Dose-normalized (to a 10 mg dose) pharmacokinetic parameter values are

summarized in Table3.

Table 3. Dose-Normalized (at 10 mg) Pharmacokinetic Parameter Values (Mean ±SEM) Following Single Oral Administration of Fampridine-SR to Patients with MS.

Dose CmAx-norm tMx AUC-norm tv2 Cl/F (Mg) (ng/mL) (hours) (ng hr/mL) (hours) (mL/min) 5 26.2 ±0.6 3.9 ±0.2 244.2 ±9.4 5.8 ±0.5 619.8 ±36.2 (n=24) 10 25.2 ±0.7 3.9 ±0.3 252.2 ±7.8 5.6 ±0.4 641.4 ±39.1 (n=24) 15 24.6 ±0.7 3.6 ±0.3 263.0 ±7.4 5.5 ±0.4 632.4 ±39.0 (n=24) 20 24.6 ±0.8 3.6 ±0.3 255.6 ±6.9 5.1 ±0.3 653.9 ±37.1 (n=23) II

EXAMPLE4

[0095] This example describes the results of an open-label study to assess the steady

state pharmacokinetics of orally administered fampridine (4-aminopyridine) compositions of

the present invention in subjects with Multiple Sclerosis. This study was an open-label

multiple dose study of Fampridine-SR intended to assess steady state pharmacokinetics in 20

patients with MS who previously completed the study summarized in Table 4. Fampridine

SR (40 mg/day) was administered as two 20 mg doses, given as one morning and one

evening dose for 13 consecutive days, with a single administration of 20 mg on Day 14.

Blood samples for pharmacokinetic analysis were collected on Days 1, 7/8, and 14/15 at the

following intervals: immediately prior to drug administration (baseline), hourly for the first 8

hours, and 10, 12, and 24 hours post-dose. Additional blood samples were collected 14, 18,

and 20 hours post-dose on Day 14, and 30 and 36 hours post-dose on Day 15.

[0096] Pharmacokinetic parameter estimates following the first dose in these

patients in this study on Day1 were comparable to those determined when they participated

in the study summarized in Table 4. No significant difference in Tmax was detected among the four means (Single dose = 3.76 h; Day 1 = 3.78 h; Day 8 = 3.33 h; Day 15 = 3.25 h). Cmax and Cmax/C, on Days 8 (Cmax = 66.7 ng/ml) and 15 (Cmax = 62.6 ng/ml) were significantly greater than those of the single dose treatment and of Day 1 (Cmax = 48.6 ng/ml), reflecting accumulation of the drug with multiple dosing.

[0097] There was no significant difference among the four occasions with regard to

either T or C and no difference in Cmax, Cmax/ C , CL/F or AUCo, between Days 8 and 15.

Further AUC on Days 8 and 15 did not differ significantly from total AUC with single dose

treatment. Likewise, the estimates of CL/F on Days 8 and 15 and of k and T 1/2 on Day 15

did not differ significantly from those with single dose.

[0098] Steady-state was attained by Day 7/8 as evidence by the lack of differences

in Cmax or AUC between Days 7/8 and 14/15; there was no apparent unexpected

accumulation. Likewise, the estimates of C/F on Days 7/8 and 14/15 of and of T1 /2 on Day

14/15 did not differ significantly from those given a single dose. On the final day of dosing,

mean Cmaxwas 62.6 ng/mL, occurring 3.3 hours post-dose. The T1 /2 was 5.8 hours. These

values are similar to those observed in patients with chronic SCI receiving similar doses of

this formulation. These results are summarized in Table 4.

Table 4. Pharmacokinetic Parameter Values (Mean and 95% CI) Following Multiple Oral Doses of Fampridine-SR (40 mg/day) to 20 Patients with MS.

Parameter Day CMx tWx AUC (o-12) tI/2 Cl/F (ng/mL) (hours) (ng hr/mL) (hours) (L/mmin) Day 1 48.6 3.8 NE NE NE (42.0, 55.3) (3.2,4.3) Day 7/8 66.7 3.3 531 NE 700 (57.5, 76.0) (2.8, 3.9) (452, 610) (557,884) Day 14/15 62.6 3.3 499 5.8 703 (55.7, 69.4) (2.6, 3.9) (446, 552) (5.0, 6.6) (621, 786)

[0099] Dizziness was the most common treatment-related adverse event. Other

treatment-related adverse events that occurred included nausea, ataxia, insomnia, and tremor.

There were no clinically significant changes in mean clinical laboratory values, vital signs, or

physical examination findings from baseline to last visit. There were no apparent clinically

significant changes in corrected QT intervals or QRS amplitudes after administration of

fampridine.

[00100] Fampridine was well tolerated in subjects with multiple sclerosis who

receive twice daily doses (20 mg/dose) of fampridine for two weeks. A significant increase

was observed in Cmax, and Cmax/ C, on Days 8 and 15 relative to those on Day 1 and with

single dose treatment, reflecting accumulation of fampridine with multiple dosing. A lack of

significant differences in Cmax, Cmax/ C, CLF or AUCo, between Days 8 and 15 suggest that

near steady-state is reached by Day 8. There was no evidence of significant changes in

pharmacokinetics during a two-week period of multiple dosing with fampridine.

EXAMPLE5

[00101] This example provides an embodiment of a method of treating subjects with

a sustained release fampridine formulation and a responder analysis of the present invention.

This was a Phase 2, double-blind, placebo-controlled, parallel group, 20-week treatment

study in 206 subjects diagnosed with Multiple Sclerosis. This study was designed to

investigate the safety and efficacy of three dose levels of Fampridine-SR, 10 mg b.i.d., 15 mg

b.i.d., and 20 mg b.i.d. in subjects with clinically definite MS. The primary efficacy endpoint

was an increase, relative to baseline, in walking speed, on the Timed 25 Foot Walk.

Secondary efficacy measurements included lower extremity manual muscle testing in four

groups of lower extremity muscles (hip flexors, knee flexors, knee extensors, and ankle

dorsiflexors); the 9-Hole Peg Test and Paced Auditory Serial Addition Test (PASAT 3"); the

Ashworth score for spasticity; Spasm Frequency/Severity scores; as well as a Clinician's

(CGI) and Subject's (SGI) Global Impressions, a Subject's Global Impression (SGI), the

Multiple Sclerosis Quality of Life Inventory (MSQLI) and the 12-Item MS Walking Scale

(MSWS-12).

[00102] At the first visit (Visit 0) subjects were to enter into a two-week single-blind 2 placebo run-in period for the purpose of establishing baseline levels of function. At Visit

subjects were to be randomized to one of four treatment groups (Placebo or Fampridine-SR

10 mg, 15 mg, 20 mg) and begin two weeks of double-blind dose-escalation in the active

drug treatment groups (B, C and D). Group A were to receive placebo throughout the study.

Subjects in the 10 mg (Group B) arm of the study took a dose of 10 mg approximately every

12 hours during both weeks of the escalation phase. The 15 mg (Group C) and 20 mg (Group

D) dose subjects took a dose of 10 mg approximately every 12 hours during the first week of

the escalation phase and titrated up to 15 mg b.i.d. in the second week. Subjects were to be

instructed to adhere to an "every 12 hour" dosing schedule. Each subject was advised to take

the medication at approximately the same time each day throughout the study; however,

different subjects were on differing medication schedules (e.g., 7 AM and 7 PM; or 9 AM

and 9 PM). After two weeks, the subjects were to return to the clinic at Visit 3 for the start of

the stable dose treatment period. The first dose of the double-blind treatment phase at the

final target dose (placebo b.i.d. for the Group A, 10 mg b.i.d. for Group B, 15 mg b.i.d. for

Group C, and 20 mg b.i.d. for Group D) was taken in the evening following Study Visit 4.

Subjects were to be assessed five times during the 12-week treatment period. Following the

12-week treatment phase there was to be a one-week down titration starting at Visit 9.

During this down-titration period, group B was to remain stable at 10 mg b.i.d. and Group C

was to be titrated to 10 mg b.i.d., while group D was to have a change in the level of dose

during the week (15 mg b.i.d. for the first three days and 10 mg b.i.d. for the last four days).

At the end of the down titration period at Visit 10, subjects were to enter a two-week washout

period where they did not receive any study medication. The last visit (Visit 11) was to be scheduled two weeks after the last dosing day (end of the downward titration). Plasma samples were collected at each study site visit other than Study Visit 0.

[00103] The primary measure of efficacy was improvement in average walking from speed, relative to the baseline period (placebo run-in), using the Timed 25 Foot Walk

the Multiple Sclerosis Functional Composite Score (MSFC). This is a quantitative measure

of lower extremity function. Subjects were instructed to use whatever ambulation aids they

normally use and to walk as quickly as they could from one end to the other end of a clearly

marked 25-foot course. Other efficacy measures included the LEMMT, to estimate muscle

strength bilaterally in four groups of muscles: hip flexors, knee flexors, knee extensors, and

ankle dorsiflexors. The test was performed at the Screening Visit and at Study Visits 1, 2, 4,

7, 8, 9 and 11. The strength of each muscle group was rated on the modified BMRC scale: 5

= Normal muscle strength; 4.5= Voluntary movement against major resistance applied by the

examiner, but not normal; 4= Voluntary movement against moderate resistance applied by

the examiner; 3.5= Voluntary movement against mild resistance applied by the examiner; 3=

Voluntary movement against gravity but not resistance; 2= Voluntary movement present but

not able to overcome gravity; 1= Visible or palpable contraction of muscle but without limb

movement; and 0= Absence of any voluntary contraction. Spasticity in each subject was

assessed using the Ashworth Spasticity Score. The Ashworth Spasticity Exam was

performed and recorded at the Screening Visit and at Study Visits 1, 2, 4, 7, 8, 9 and 11.

[00104] Protocol Specified Responder Analysis. To supplement the primary

analysis, a categorical "responder" analysis was also conducted. Successful response was

defined for each subject as improvement in walking speed (percent change from baseline) of

at least 20%. Subjects who dropped out prior to the stable dose period were considered non

responders. The proportions of protocol specified responders were compared among

treatment groups using the Cochran-Mantel-Haenszel test, controlling for center.

[00105] Post hoc analysis of this study suggested that a relatively highly selective

criterion for a likely treatment responder would be a subject with a faster walking speed for at

least three visits during the double blind treatment period as compared to the maximum value

among a set of five non-treatment visits (four before treatment and one after discontinuation

of treatment). The four visits before initiation of double-blind treatment provided an initial

baseline against which to measure the consistency of response during the four double-blind

treatment visits. The inclusion of the follow-up visit as an additional component of the

comparison was useful primarily in excluding those subjects who may be false positives, i.e.,

did not show the expected loss of improvement after coming off the drug. Treatment

differences in the proportion of theses post hoc responders were analyzed using the Cochran

Mantel-Haenszel (CMH) test, controlling for center.

[00106] To validate the clinical meaningfulness of the post hoc responder variable,

(post hoc) responders were compared against the (post hoc) non-responders, on the subjective

variables: (i) Change from baseline in MSWS-12 over the double-blind; (ii) SGI over the

double-blind; and (iii) Change from baseline in the CGI over the double-blind; to determine if

subjects with consistently improved walking speeds during the double-blind could perceive

improvement relative to those subjects who did not have consistently improved walking

speeds. For the subjective variables, differences between responder status classification

(responder or non-responder) were compared using an ANOVA model with effects for

responder status and center.

[00107] Results. A total of 206 subjects were randomized into the study: 47 were

assigned to placebo, 52 to 10 mg bid Fampridine-SR (10 mg bid), 50 to 15 mg bid

Fampridine-SR (15 mg bid), and 57 to 20 mg bid Fampridine-SR (20 mg bid). The

disposition of subjects is presented in Table 5 below.

Table 5 Summary of subject disposition (all randomized population)

Treatment Group: N (%) Placebo 10 mg bid 15mg bid 20 mg bid Total

SubjectsRandomized __47 52 50 57 206- - Took at Least One Dose 47(100%) 52(100%) 50(100%) 57(100%) 206(100%) (Included inSafe!Analysis) . .. .. ITT Population__ 47(100%) 51(98.1%) 50(100%) 57(100%) 205(99.5%) ._....... . ...... _. .. .. ....... .. .. . ....... . ---- - - - ...

Discontinued Subjects . 6 (10.5%) Note: Percentages are based on the number of randomized subjects.

and

[00108] All 206 randomized subjects took at least one dose of study medication

were included in the safety population. One subject (subject# 010/07 10 mg bid group) was

excluded from the ITT population (lost to follow-up after 8 days of placebo run-in). A total

of 11 subjects discontinued from the study.

[00109] The population consisted of 63.6% females and 36.4% males. The majority

of the subjects were Caucasian (92.2%), followed by Black (4.9%), Hispanic (1.5%), those

classified as 'Other' (1.0%), and Asian/Pacific Islander (0.5%). The mean age, weight, and

height of the subjects were 49.8 years (range: 28-69 years), 74.44 kilograms (range: 41.4

145.5 kilograms), and 168.84 centimeters (range: 137.2-200.7 centimeters), respectively.

Most of the subjects (52.4%) had a diagnosis type of secondary progressive with about equal

amounts of relapsing remitting (22.8%) and primary progressive (24.8%) subjects. The mean

duration of disease was 12.00 years (range: 0.1-37.5 years) while the mean Expanded

Disability Status Scale (EDSS) at screening was 5.77 units (range: 2.5-6.5 units). The

treatment groups were comparable with respect to all baseline demographic and disease

characteristic variables.

[00110] Results for the key efficacy variables at baseline for the ITT population are

further summarized in Table 6 below.

Table 6 Sunmar ofkey efficacy variables at baseline (ITT population) Treatment Group: Mean (SD) placebo 10 mg bid 15mg bid 20 mg bid Treatment. Parameter N=47 N=51 N=50 N=57 p-value

Walking(Sped(ft/sec) 1.87 (0.902) 1.94 (0.874) 1.99 (0.877) 2.04 (0.811) 0.752 4.00 (0.737) 3.98 (0.634) 0.964 LEMMT . . .-- ......... 4.05 (0.690) ....... ........ . ..... ... . . . . .... _..-.......... ~ (0.661) 3.98 ........-........... ..

SGI 4.38(0.795) 4.32(0.999)* 4.56(1.110) 4.25(0.969) 0.413 MSWS-12 731(16.186) (16.566) 74.60 (17.671) 76.83 (18.124) 0.923 *:One subject did not have a baseline value.

[00111] With respect to the 205 subjects in the ITT population, mean values for

baseline walking speed, LEEMT, SGI, and MSWS-12 were approximately 2 feet per second,

4 units, 4.5 units,and76units, respectively. The treatment groups were comparable with

respect to these variables as well as all the other efficacy variables at baseline.

[00112] Descriptive statistics for the average walking speed (ft/sec) by study day

based on the Timed 25-Foot Walk are presented in Table 7 and Figure 2. The timed 25 foot

walk showed a trend toward increased speed during the stable dose period for all three dose

groups, though the average improvement declined during the treatment period.

Table 7 Average walking speeds (ft/sec) by study day (observed cases, ITT population) Summary Statistics Over Time Study day Treatment base titration 1st stbl 2nd stbl 3rd stbl follow-up placebo Mean 1.87 1.89 1.90 1.89 1.89 1.86 (SD) (0.902) (0.876) (0.908) (0.891) (0.914) _ (0.933) N# 47 47 46 46 45 45 mg id Mean 1.94 2.20 2.09 2.12 2.00 1.88 (SD) (0.874) (0.979) (0.955) (1.043) (1.016) (0.970) N 51 51 51 51 50 48 15mg bid Mean 1.99 2.25 2.16 2.14 2.18 1.83 (SD) (0.877) (0.995) (0.986) (0.957) (0.932) (0.952) N 50 49 49 48 48 47 20mg bid Mean 2.04 2.26 2.22 2.19 2.04 1.83 (SD) (0.811) (0.936) (0.893) (0.936) (0.996) (0.822) N 57 55 52 51 49 55

#: The treatment sample sizes presented in the figure legend represent the number of ITT subjects. Sample sizes at individual time points may be smaller than those in the ITT population due to dropouts or missed assessments.

[00113] During double-blind treatment, all the Fampridine-SR groups exhibited mean

walking speeds between 2.00 and 2.26 feet per second, while the mean value in the placebo

group was consistently about 1.90 feet per second. It should be noted that, at the third stable dose visit, both the 10 mg bid and 20 mg bid group means dropped-off from what would be expected under the assumption that treatment benefit is consistent over time. This may or may not have been due to chance; further studies should provide additional evidence for either case. After double-blind medication was discontinued, all the treatment groups converged to approximately the same mean value at follow-up.

[00114] Results for the primary efficacy variable (percent change in average walking

speed during the 12-week -stable dose period relative to baseline based on the 25-foot walk)

are summarized in Figure 3. The timed 25 foot walk showed a trend toward increased speed

during the stable dose period for all three dose groups, though the average improvement

declined during the treatment period, as shown in Figure 3. The mean percent changes in

average walking speed during the 12-week stable dose period (based on adjusted geometric

mean change of the log-transformed walking speeds) were 2.5%, 5.5%, 8.4%, and 5.8% for

the placebo, 10 mg bid, 15 mg bid, and 20 mg bid groups, respectively. There were no

statistical differences between any Fampridine-SR groups and the placebo group.

[00115] Results for the protocol specified responder analysis (subjects with average

changes in walking speed during the 12 weeks of stable double-blind treatment of at least

20%) are summarized in Figure 4. The percentages of subjects with average changes in

walking speed during the 12-week stable dose period of at least 20% (pre-defined responders)

were 12.8%, 23.5%, 26.5%, and 16.1% for the placebo, 10 mg bid, 15 mg bid, and 20 mg bid

groups, respectively. There were no statistically significant differences between any of the

Fampridine-SR groups and the placebo group.

[00116] Descriptive statistics for the average overall Lower Extremity Manual

Muscle Testing (LEMMT) by study day are presented in Table 8 and in Figure 5.

Table 8. Average overall LEMMT by Study Day Summary Statistics Oer Time Study day Treatment base titration 1st stbI 2nd stbl 3rd stb1 follow-up placebo Mean 4.05 4.00 4.02 4.03 4.00 4.02 (SD) (0.690) (0.75) (0.687) (0.69) (0.679) (0.73) N# 47 46 46 46 45 45 bid Mean 3.98 4.08 4.06 4.09 4.07 3.89 (SD) (0.631) (0.641) (0.650) (0.685) (0.642) (0.631) N 51 50 51 51 50 49 15-ibid_ _Mean 4.00 4.16 4.11 4.09 4.17_ 4.08 (__SD)_ (0.737) (.653) (065) (0.659) _(0.618)_ _(0K.674) N 50 49 49 49 49 46 2mgbid_ Mean 3.98 4.08 4.03 3.98 4.07 _3.92 (SD) (0.634) (0.639) (0.659) (0.714) (0.649) (0.650) N- 57 54 52 52 48 55

#: The treatment sample sizes presented at individual time points may be smaller than those in the ITT population due to dropouts or missed assessments.

[00117] During double-blind treatment, all the Fampridine-SR groups exhibited a

numerical pattern of larger mean LEMMT scores than placebo (except the 20 mg bid group at

the 2" stable dose visit). After double-blind medication was discontinued, with the exception

of the 15 mg bid group, all the group means were lower than they were at baseline.

[00118] Results for the average change in LEMMT during the 12-week stable dose

period relative to baseline are summarized in Figure 6. The mean changes in overall

LEMMT during the 12-week stable dose period were -0.05 units, 0.10 units, 0.13 units, and

0.05 units for the placebo, 10 mg bid, 15 mg bid, and 20 mg bid groups, respectively.

Improvements in LEMMT were significantly greater in the 10 mg bid and 15 mg bid groups

compared to the placebo group; there was no significant difference between the 20 mg bid

group and the placebo group.

[00119] No statistically significant differences were detected among treatment group

based on any of the other secondary efficacy variables, as shown in Table 9.

Table 9 Changes in secondary efficacy variables from baseline during the 12-week stable dose period Treatment Group placebo 10 mg bid 15mg bid 20 mg bid Parameter N=47 N=51 N=50 N=57 Ashworth Score N 46 51 49 53 Mean (SD) -0.11 (0.377) -0.04 (0.449) -0.06 (0.375) 0.02 (0.466) p-valueeahdsepaceb0.802 0.826 0.275

CGI N 45 50 49 52 Mean (SD) 0.0(0.66) -0.2(0.72) -0.1(0.85) 0.0(0.78) _p-value(eachdosevsplacebo) 0.772 0.997 0.996

SGI N 46 50 49 53 Mean (SD) -0.2(0.96) 0.0(1.27) -0.1(1.11) -0.1(0.86) _ 0.704 0.953 0.968

PASAT N 46 51 49 53 Mean (SD) 2.17 (4.016) 2.13 (3.394) 0.90 (3.274) 0.65 (4.590) p-value>eachdosevsplace >0.999 0.306 0.218

MSFC N 46 51 49 52 Mean (SD) 0.08 (0.205) 0.10 (0.310) 0.90 (0.224) 0.06 (0.194) p-vatSleachdo~se vs.-placebo) ---------------- 0.977 >0.999 0.96.8----

MSWS-12 N 46 51 49 52 Mean (SD) -3.56 (14.548) -5.53 (16.154) -7.32 (16.295) -5.76(15.296) p-value (each dose vs. placebo) 0.718 0.445 0.617 Note: The treatment sample sizes presented in the treatment heading represent the number of ITT subjects. Sample sizes for individual variables may be smaller due to dropouts or missed assessments. Note: For each variable, the p-values (versus placebo) are Dunnett-adjusted.

[00120] While pre-planned analyses of the primary efficacy endpoint provided

insufficient evidence of treatment benefits for any of the Fampridine-SR doses, subsequent

analysis revealed the existence of a subset of subjects who responded to the drug with clinical

meaningfulness. These subjects exhibited walking speeds while on drug that were

consistently better than the fastest walking speeds measured when the subjects were not

taking active drug.

[00121] The post hoc responder rates based on consistency of improved walking

speeds were significantly higher in all three active dose groups (35, 36 and 39%) compared to placebo (9%; p<0.00 6 for each dose group, adjusting for multiple comparisons) as shown in

Figure 7.

[00122] Given that there was little difference in responsiveness between the three

doses examined, more detailed analyses were performed comparing the pooled Fampridine

SR treated groups against the placebo-treated group. Figure 8 summarizes, for the placebo

and the pooled Fampridine-SR group, the percentage of post hoc responders. The number of

subjects who met he post hoc responder criterion in the pooled Fampridine-SR treated group

was 58 (36.7%) compared to 4 (8.5%) in the placebo-treated group, and this difference was

statistically significant (p<0.001).

[00123] To validate the clinical meaningfulness of the post hoc responder variable,

the 62 responders (58 fampridine and 4 placebo) were compared against the 143 non

responders (100 fampridine and 43 placebo) on the subjective variables to determine if

subjects with consistently improved walking speeds during the double-blind could perceived

benefit relative to those subjects who did not have consistently improved walking speeds.

The results are summarized in Figure 9 and indicate that consistency in walking speed had

clinical meaningfulness for the subjects in this study since the responders had (over the

double-blind period) significantly better changes from baseline in MSWS-12 and

significantly better subjective global scores. In addition, the responders were rated

marginally better than the non-responders by the clinicians during the double-blind. Thus,

responders experienced clinically meaningful improvements in their MS symptoms, and

treatment with fampridine significantly increased the chances of such a response.

[00124] To establish baseline comparability among the responder analysis groups,

analyses were performed on the baseline demographic variables, key neurological

characteristics and the relevant efficacy variables at baseline. In general, the responder

analysis groups were comparable for all demographic and baseline characteristics variables.

[00125] Having demonstrated the clinical meaningfulness of consistently improved

walking speeds during the double-blind as a criterion for responsiveness, the question of the

magnitude of benefit becomes of interest. The fampridine non-responders, although

providing no relevant efficacy information, do provide safety information regarding those

individuals who are treated with fampridine but show no apparent clinical benefit. As such,

responder analyses of these groups were performed.

[00126] With respect to magnitude of benefit, Figure 10 and Table 12 below

summarizes the percent changes in walking speed at each double-blind visit by responder

analysis grouping. The mean improvement for the fainpridine responders during the double

blind across 14 weeks of treatment ranged from 24.6% to 29.0% compared to 1.7% to 3.7%

for the placebo group; this was highly significant (p<0.001) at every visit. Although

providing no relevant efficacy information, results for the fampridine non-responders are also

illustrated and show that there was, and could be, some worsening in walking speeds after 12

weeks when a non-responder is treated with fampridine. The improvement was stable (± 3%)

across 14 weeks of treatment, and was associated with improvement in two global measures

(Subject Global Impression and Multiple Sclerosis Walking Scale-12). The four placebo

responders showed a 19% improvement in walking speed but there were too few subjects in

this group for meaningful statistical comparison. Response status was not significantly

related to baseline demographics, including type or severity of MS. Adverse events and

safety measures were consistent with previous experience for this drug.

Table 12. Summary of percent change in Walking Speed at each double-blind visit by responder analysis grouping.

Summary Statistics Oer Time Study day Treatment titration 1st stbl 2nd stbl 3rd stbl Placebo Mean 1.7 2.6 1.8 3.7 (SEM) (2.21) (3.23) (3.11) (3.38) N# 47 46 46 45 Fampridine Mean 8.3 3.5 -0.2 -6.5 Non-responders (SEM) (2.05) (1.90) (1.76) (2.49) N 97 94 93 89 ampridine Mean 27.4 24.6 29.0 27.3 Responders (SEM) (2.43) (2.44) (4.31) (3.52) N 58 58 57 58 FRvs.Placebo p-value^ <0.001 <0.001 <0.001 <0.001 FR vs. FNR p-value^ <0.001 <0.001 <0.001 <0.001 FNR vs. PBO p-value^ 0.080 0.884 0.497 0.022

ABBREVIATIONS: FR=Fampridine Responders; FNR=Fampridine Non-responders. #: The treatment sample sizes presented at individual time points may be smaller than those in the ITT population due to dropouts or missed assessments. #: The treatment sample sizes presented in the figure legend represent the number of ITT subjects. Sample sizes at individual time points may be smaller due to dropouts or missed assessments. ^: P-values from t-tests of the least-squares means using the mean square error via an ANOVA model with effects for responder analysis grouping and center.

[00127] Figure 11 and Table 13 summarize the changes in LEMMT at each double

blind visit by responder analysis grouping. The mean improvement for the fampridine

responders during the double-blind ranged from 0.09 to 0.18 units compared to -0.04 units at

each visit for the placebo group; this was significant at every visit except the second stable

dose visit (p=0.106). Although providing no relevant efficacy information, results for the

fampridine non-responders are also illustrated and show that there was, and could be, some

significant improvement in leg strength when non-responder is treated with fampridine. This

suggests that although a clinically meaningful response can be linked to about 37% of

subjects treated with Fampridine-SR, additional subjects may have functional improvements

on variables other than walking speed.

Table 13. Summary of percent change in LEMMT at each double-blind visit by responder analysis grouping.

Summary Statistics Oer Time I Study day Treatment Ititration 1st stbl 2nd stbl 3rd stbl Placebo Mean -0.04 -0.04 -0.04 -0.04 (SEM) (0.035) (0.042) (0.039) (0.042) N# 46 46 46 45 Fampidne Mean 0.12 0.10 0.09 0.10 Non--responders (SEM) (0.028) (0.033) (0.036) (0.038) N 95 94 94 89 Fampridine Mean 0.18 0.09 0.09 0.17 Responders (SEM) (0.029) (0.032) (0.043) (0.045) N 58 58 58 58 FR vs. Placebo p-valueA <0.001 0.023 0.106 0.004 FRvs. FNR p-valueA 0.178 0.627 0.739 0.311 FNRvs.PBO p-value^ <0.001 0.003 0.038 0.032

ABBREVIATIONS: FR=Fampridine Responders; FNR=Fampridine Non-responders. #: The treatment sample sizes presented at individual time points may be smaller than those in the ITT population due to dropouts or missed assessments. Treatment sample sizes presented in the figure legend represent the number of ITT subjects. Sample sizes at individual time points may be smaller due to dropouts or missed assessments. A: P-values from t-tests of the least-squares means using the mean square error via an ANOVA model

with effects for responder analysis grouping and center.

[00128] Figure 12 and Table 14, below, summarize the changes in Overall Ashworth

Score at each double-blind visit by responder analysis grouping. The mean reduction from

baseline (indicative of improvement) for the fampridine responders during the double-blind

ranged from -0.18 to -0.11 units compared to -0.11 to -0.06 for the placebo group. The

fampridine responders were numerically superior to placebo but there was insufficient

evidence to detect significant differences. Although appearing to provide little relevant

efficacy information, results for the fampridine non-responders are also illustrated.

Table 14. Summary of change in overall Ashworth score at each double-blind visit by responder analysis grouping. ~_Summary Statistics Over Time

Treatment I_ titration 1st stbl Study day 2nd stbl 3rd stbl Placebo Mean -0.06 -0.11 -0.06 -0.13 (SEM) (0.069) (0.073) (0.070) (0.073) N# 46 46 46 45 Fampridine Mean -0.16 -0.08 -0.07 0.00 Non-responders (SEM) (0.044) (0.053) (0.054) (0.056) N 95 94 94 89 Fampridine Mean -0.14 -0.18 -0.11 -0.18 Responders (SEM) (0.058) (0.066) (0.060) (0.055) N 58 58 58 58 FR vs. Placebo p-value^ 0.343 0.374 0.717 0.680 FRvs.FNR p-value^ 0.675 0.210 0.911 0.064 FNR vs. PBO p-value^ 0.151 0.823 0.772 0.189

ABBREVIATIONS: FR=Fampridine Responders; FNR=Fampridine Non-responders. #: The treatment sample sizes presented at individual time points may be smaller than those in the ITT population due to dropouts or missed assessments. A: P-values from t-tests of the least-squares means using the mean square error via an ANOVA model

with effects for responder analysis grouping and center.

[00129] Adverse events most commonly reported prior to treatment were accidental

injury, reported by 12 (5.8%) subjects, nausea, reported by 9 (4.4%) subjects, and asthenia,

diarrhea, and paresthesia, each reported by 8 (3.9%) subjects. Six (2.9%) subjects also

reported headache, anxiety, dizziness, diarrhea, and peripheral edema. These adverse events

are indicative of the medical conditions affecting people with MS.

[00130] Conclusions. The data does not appear to support either a number of

anecdotal reports or expectations from preclinical pharmacology that doses higher than about

10 to 15 mg b.i.d., and even about 10 mg b.i.d., should be associated with greater efficacy.

The data presented below in Table 15 support this, based on the new responder analysis

methodology.

Table 15. Cormparison of 10_mg vs. 15 mg among Responders 10 mg 15 mg (N=51) (N=50) Responders N (%) 18(35.3) 18(36.0) Average %CFB in Walk Speed: Mean (SD) 27.6% (18.39) 29.6% (22.43) %Change in Walk Speed by Visit: minimum - maximum 26%- 32% 27%-31% Average SGI 4.8(1.09) 4.7(1.09) Average Change in MSWS-12* -11.1 (21.9) -7.8(19.6) * For the average change in the MSWS-12, a negative score is indicative subjective of improvement.

[00131] A responder analysis based on consistency of improvement provides a

sensitive, meaningful approach to measuring effects on the timed 25 foot walk and may be

used as a primary endpoint for future trials. This data suggest that for responsive subjects

(approximately 37%), treatment with fampridine at doses of 10-20 mg bid produces

substantial and persistent improvement in walking.

[00132] Efficacy. There are no notable differences between 10 mg bid and 15 mg bid

among subjects who respond to drug. In fact, the largest difference, favors the 10 mg bid

group (see MSWS-12 result).

[00133] Safety. With respect to safety, there are three considerations: There was an

apparent decline below baseline walking speed at the last visit on drug in the fampridine non

responders in the 10 mg bid and 20 mg bid groups, but not the 15 mg bid group. This may or

may not be significant, but is not clearly dose related. There was an apparent rebound effect,

with walking speed dropping below baseline, among fampridine treated subjects at the two

week follow-up visit; this occurred in the 15 and 20 mg but not the 10 mg bid group. Serious

AE's were more frequent in the 15 mg and 20 mg bid groups 10% and 12% rates vs. 0% rate

in 10 mg bid and 4% in placebo groups. This may or may not be significant, but the risk of

potentially related SAEs, particularly seizures appears to be dose-related from all available

data and based on mechanism of action. Based on this data, it would appear that a 10 mg bid dose is preferred because of its favorable risk to benefit ratio compared with the 15 and 20 mg doses.

[00134] Although the present invention has been described in considerable detail

with reference to certain preferred embodiments thereof, other versions are possible.

Therefore the spirit and scope of the appended claims should not be limited to the description

and the preferred versions contain within this specification.

Claims (16)

1. We claim: 1. A method of increasing walking speed comprising administering to a patient with multiple sclerosis an effective amount of a sustained release aminopyridine composition twice daily, wherein said effective amount is less than about 15 milligrams of aminopyridine.
2. 2. The method of claim 1, wherein said effective amount is about 10 to about 15 milligrams of aminopyridine.
3. 3. The method of claim 1, wherein said effective amount is about 10 milligrams of aminopyridine.
4. 4. The method of claim 1, wherein said aminopyridine is 4-aminopyridine.
5. 5. The method of claim 4, wherein said sustained release 4-aminopyridine composition exhibits a Cmax:Cr ratio from about 1.0 to 3.5.
6. 6. The method of claim 1, wherein said twice daily administration comprises about every 12 hours.
7. 7. A method of improving lower extremity muscle tone comprising administering to a patient with multiple sclerosis an effective amount of a sustained-release aminopyridine composition twice daily, wherein said effective amount is less than about 15 milligrams of aminopyridine.
8. 8. The method of claim 7, wherein said effective amount is about 10 to about 15 milligrams of aminopyridine.
9. 9. The method of claim 7, wherein said effective amount is about 10 milligrams of aminopyridine.
10. 10. The method of claim 7, wherein said aminopyridine is 4-aminopyridine.
11. 11. The method of claim 10, wherein said sustained release 4-aminopyridine composition exhibits a Cmax:Cr ratio from about 1.0 to 3.5.
12. 12. A method of improving lower extremity muscle strength comprising administering to a patient with multiple sclerosis an effective amount of a sustained-release aminopyridine composition twice daily, wherein said effective amount is less than about 15 milligrams of aminopyridine.
13. 13. The method of claim 12, wherein said effective amount is about 10 to about 15 milligrams of aminopyridine.
14. 14. The method of claim 12, wherein said effective amount is about 10 milligrams of aminopyridine.
15. 15. The method of claim 12, wherein said aminopyridine is 4-aminopyridine.
16. 16. The method of claim 15, wherein said sustained release 4-aminopyridine composition exhibits a Cmax:Cr ratio from about 1.0 to 3.5.