MODIFIED RELEASE FORMULATIONS COMPRISING S1 P

RECEPTOR MODULATORS

Field of the Invention

The present invention relates to formulations of immunosuppressant compounds, and particularly to formulations of S1P receptor modulators. More particularly the invention relates to formulations of selective S1P1/S1P5 modulators, particularly 1-{4-[1-(4-cyclohexyl- 3-trifluoromethyl-benzyloxyimino)-ethyl]-2-ethyl-benzyl}-aze tidine-3-carboxylic acid, pharmaceutically acceptable salts, and related compounds.

Background to the Invention

S1 P receptors belong to a family of closely related, lipid activated G- protein coupled receptors. S1P1, S1P3, S1 P2, S1P4, and S1 P5 (also respectively termed EDG-1 , EDG-3, EDG-5, EDG-6, and EDG-8) are identified as receptors specific for sphingosine-1 -phosphate (S1P). Certain S1P receptors are associated with diseases mediated by lymphocyte interactions, for example, in transplantation rejection, autoimmune disease, inflammatory diseases, infectious diseases and cancer.

WO2004/103306 discloses immunosuppressant compounds that are useful in the treatment

non-limiting example paragraphs [0041]-[0054], inclusive, of US 2009/0036423 and each example thereof.

WO2010/020610 discloses use of S1 P receptor agonists, for example 1-{4-[1-(4-cyclohexyl- 3-trifluoromethyl-benzyloxyimino)-ethyl]-2-ethyl-benzyl}-aze tidine-3-carboxylic acid, in the treatment of peripheral neurapathies, such as Guillain-Barre syndrome (GBS), chronic inflammatory demyelinating polyradiculoneuropathy (CI DP), multifocal motor neuropathy with conduction block (MMN), and paraproteinaemic demyelinating peripheral neuropathy (PDN).

WO2007/021666 discloses a concentrate for dilution comprising a S1 P receptor agonist, propylene glycol and optionally glycerin, which concentrate is described as being stable for extended periods of time. One compound mentioned as an S1 P receptor agonist is 1-{4-[1- (4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethyl]-2-eth yl-benzyl}-azetidine-3-carboxylic acid. The dilution disclosed in WO2007/021666 is provided in a liquid form and is therefore particularly suitable for patients who struggle to swallow solid medications.

WO2009/048993 discloses compositions comprising S1 P receptor modulators, such as 2- substituted 2-amino-propane-1 ,3-diol or 2-amino-propanol derivatives, which are suitable for use as an oral dosage form. The composition is disclosed to comprise the active ingredient and one or more of various specified excipients. Example 10 mentions glyceryl behenate as a non-feasible excipient, seemingly because of degradation of the active (FTY720). One S1 P modulator metioned is 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethy l]-2- ethyl-benzyl}-azetidine-3-carboxylic acid.

WO2010/072703 teaches that SIP receptor modulators or agonists may produce a negative chronotropic effect, i.e. reduce the heart rate. There is disclosed a dosage regimen for minimising the negative chronotropic effect of an S1 P receptor modulator, e.g. when used for the treatment of patients suffering from an autoimmune disease, for example multiple sclerosis. The dosage regimen comprises administering a lower dosage of an S1 P receptor modulator during the initial days of treatment compared to the standard daily dosage. The dosage is then increased stepwise up to the standard daily dosage of the S1 P receptor modulator. One S1 P modulator mentioned is 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl- benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azetidine-3-carboxyli c acid. Summary of the Invention

The present invention provides novel oral compositions comprising SIP modulators or agonists. In an embodiment, it addresses the problem of reducing the negative chronotropic effect of such compounds. Embodiments of the invention are predicated at least in part on the provision of such a compound in a modified release formulation, thereby to reduce or eliminate the negative chronotropic effect.

The compounds to which the application relates are therefore SIP receptor modulators or agonists. In an embodiment, the compounds are as disclosed in WO 04/103306 and and US

wherein

A is COORs, OPO(OR ₅ ) ₂ , PO(OR ₅ ) ₂ , S0 ₂ OR ₅ , POR ₅ OR ₅ or 1 H-tetrazol-5-yl, R ₅ being H or an ester-forming group, e.g. C^alkyl, and in one implementation of the invention being H;

W is a bond, C _1-3 alkylene or C _2-3 alkenylene;

Y is G ₆ .i ₀ aryl or C _3-9 heteroaryl, optionally substituted by 1 to 3 radicals selected from halogen, N0 ₂ , Ci _-6 alkyl, C _1-6 alkoxy; halo-substituted Ci. ₆ alkyl and halo-substituted

Ci. ₆ alkoxy;

Z is chosen from:

wherein the asterisks of Z indicate the point of attachment between -C(R ₃ )(R ₄ )- and A of Formula la or lb, respectively; R ₆ is chosen from hydrogen and C _-6 alkyl; and Ji and J ₂ are independently methylene or a heteroatom chosen from S, O and NR ₅ ; wherein R ₅ is chosen from hydrogen and C _h alky!; and any alkylene of Z can be further substituted by one to three radicals chosen from halo, hydroxy, C _1-6 alkyl; or R ₆ can be attached to a carbon atom of Y to form a 5-7 member ring;

Ri is C ₆ -ioaryl or C ₃ . ₉ heteroaryl, optionally substituted by Ci _-6 alkyl, Ce-ioaryl, C ₆ . ₁₀ arylCi. alkyl, C _3-9 heteroaryl, C ₃ . ₉ heteroarylCi. alkyl, C ₃ . ₈ cycloalkyl, C ₃ . ₈ cycloalkylC _1-4 alkyl,

C _3-8 heterocycloalkyl or Cs-eheterocycloalkylCMalkyl; wherein any aryl, heteroaryl, cycloalkyi or -heterocycloalkyl of Ri may be substituted by 1 to 5 groups selected from halogen, d. ₆ alkyl, d-ealkoxy and halo substituted-C _1-6 alkyl or -d- ₆ alkoxy;

R ₂ is H, C _1-6 alkyl, halo substituted C _-6 alkyl, C ₂ . ₆ alkenyl or C ₂ - ₆ alkynyl; and each of R ₃ or R ₄ , independently, is H, halogen, OH, C _1-6 alkyl, C _1-6 alkoxy or halo substituted C _1-6 alkyl or d-ealkoxy; or a pharmacologically acceptable salt, solvate or hydrate thereof.

In the above formula, the designation "Ci. ₆ " means "having 1 , 2, 3, 4, 5 or 6 carbon atoms" and the designation "CW means "having 3, 4, 5, 6, 7 or 8 carbon atoms". The designation "C _1-4 " means "having 1 , 2, 3 or 4 carbon atoms". The designation "C ₃ - ₉ " means "having 3, 4, 5, 6, 7, 8 or 9 carbon atoms".

The invention particularly, but not exclusively, involves compounds of formula A1 or A2 in which R ₅ is H, e.g. moiety A is -COOH, in its acid form (not therefore as a salt thereof).

An exemplary compound of formula A1 or A2 is 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl- benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azetidine-3-carboxyli c acid and pharmaceutically acceptable salts, e.g. hemifumarate salts.

In one aspect, the invention provides a pharmaceutical composition for oral administration which comprises an active pharmaceutical ingredient ("API") selected from SIP receptor modulators and agonists, for example the compounds of Formulae A1 and A2, wherein the composition is adapted to provide sustained release of the API. The composition may increase Tmax or reduce Cmax, or both increase Tmax and reduce Cmax, as compared to an immediate release composition. A sustained release composition as compared to an immediate release composition comprises one or more agents which act to prolong release of the API; for example, the API may be embedded in a matrix and/or surrounded by a_ membrane which, in either case, controls (reduces) the rate of diffusion of the API into the Gl tract. One version of membrane-control is osmotic pump systems in which a core comprises e.g. solid drug or highly concentrated drug in combination with an osmotic agent which acts to imbibe water from the Gl tract through a semi-permeable membrane, the API being driven out of an orifice in the device by the osmotic pressure generated in the device, such that release of the API is controlled by water influx across the semipermeable membrane. In an alternative embodiment, the controlled release may be achieved using a multiparticulate system comprising a number of independent sub-units to release the active.

In an embodiment, the composition comprises a hydrogel-former, for example a hydrogel- forming polymer. The composition may comprise hydrogel-former in admixture with the API. The hydrogel-forming polymer may be, or comprise, a cellulosic polymer, e.g. hypromellose.

In embodiments, the compositions of the invention further have one, two or three of the following features which serve to enhance stability of the API when it is of formula A1 or A2:

• the API is not exposed to a basic compound (other than the API itself when in basic form)

• the API is in the form of particles having a crystallinity of 80% or more • the API is in the form of particles having an X90 diameter of at least 8pm, e.g. at least 10pm.

A particular embodiment resides in a tablet having a compressed composition comprising a hydrogel-former in admixture with a compound mentioned herein, e.g. 1-{4-[1-(4-cyclohexyl- 3-trifluoromethyl-benzyloxyimino)-ethyl]-2-ethyl-benzyl}-aze tidine-3-carboxylic acid and its pharmaceutically acceptable salts e.g. its hemifumarate salt as the API. The hydrogel- former may be non-basic and in particular is hypromellose, e.g. a mixture of two or more hypromellose materials of different grades. The compressed composition may consist of non-basic materials; the matrix may for example contain, in addition to the API and a hydrogel-former, a non-basic filler, e.g. lactose (particularly as the monohydrate), a non- basic lubricant, e.g. glyceryl behenate, and a non-basic glidant, e.g. colloidal silicon dioxide. The 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethy l]-2-ethyl-benzyl}-azetidine- 3-carboxylic acid and its pharmaceutically acceptable salts e.g. its hemifumarate salt (i.e. the API) may be in the form of particles which have an X90 diameter of from 10 pm to 200 pm and/or which are at least 80% crystalline. The compressed mixture may include a desiccant, as which colloidal silicon dioxide also functions. In particular embodiments, the compressed composition consists of non-basic materials in the form of particles which have an X90 diameter of from 10 pm to 200 pm and/or which are at least 80% crystalline.

The pharmaceutical compositions of the present invention may be used to treat autoimmune diseases, e.g. rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis and multiple sclerosis, amongst many others, e.g. as disclosed in ^' WO2004/103306 and US 2009/0036423 for example in paragraphs [0041]-[0042] of US 2009/0036423. The pharmaceutical compositions of the present invention may be used to treat peripheral neurapathies, for example motor neuron disease, Guillain-Barre syndrome (GBS), chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), multifocal motor neuropathy with conduction block (M N), or paraproteinaemic demyelinating peripheral neuropathy (PDN). In particular embodiments, the compositions of the invention are used to treat multiple sclerosis. The invention therefore includes a method for treating a subject in need thereof, e.g. having or suspected of having one of the aforesaid diseases, e.g. multiple sclerosis, comprising administering to the patient a composition of the invention. The composition is desirably administered in an effective amount. Also disclosed are compositions of the invention for use in treating at least one of the aforesaid diseases, e.g. multiple sclerosis. Brief description of the Figures

Figure 1: shows simulated plasma levels of compound NVS-A following single oral administration of a 4 mg immediate release ("IR") and modified release (F10 and F16) tablet.

Figure 2: is a plot of heart rate reduction following administration of a 4 mg immediate release and expected reduction following administration of modified release tablets compared to placebo (PBO). Profiles are shown for two modified release tablets, namely a tablet with a 10 hour release profile (designated F10). and a tablet with a 16 hour release profile (designated F16).

Figure 3 is a box plot which demonstrates the effect of drug particle size on the content uniformity of a tablet comprising 5 mg (equivalent base weight) of the hemifumarate salt of 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl- benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azetidine-3- carboxylic acid.

Figure 4 is a box plot which demonstrates the effect of drug particle size on the content uniformity of a tablet comprising 0.25 mg (equivalent base weight) of the hemifumarate salt of 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl- benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azetidine-3- carboxylic acid.

Figure 5 is a line graph which compares the sum of degradation products of four different compositions, each comprising 0.25 mg (equivalent base weight) of the hemifumarate salt of 1 -{4-[1 -(4-cyclohexyl-3-trif luoromethyl- benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azetidine-3- carboxylic acid mixed with a lubricant.

Detailed description of the invention

For the avoidance of doubt, it is hereby stated that the information disclosed earlier in this specification under the heading "Background to the Invention" is relevant to the invention and is to be read as part of the disclosure of the invention.

Throughout the description and claims of this specification, the words "comprise" and "contain" and variations of them mean "including but not limited to", and they are not intended to (and do not) exclude other moieties, additives, components, integers or steps. Throughout the description and claims of this specification, the singular encompasses the plural unless the context otherwise requires. In particular, where the indefinite article is used, the specification (which term encompasses both the description and the claims) is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and/or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and/or steps are mutually exclusive. The invention is not restricted to the details of any foregoing embodiments. The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel

combination, of the steps of any method or process so disclosed.

The term "treat" includes: (1) preventing or delaying the appearance of clinical symptoms of the state, disorder or condition developing in an animal, particularly a mammal and especially a human, that may be afflicted with or predisposed to the state, disorder or condition but does not yet experience or display clinical or subclinical symptoms of the state, disorder or condition; (2) inhibiting the state, disorder or condition (e.g., arresting, reducing or delaying the development of the disease, or a relapse thereof in case of maintenance treatment, of at least one clinical or subclinical symptom thereof); and/or (3) relieving the condition (i.e., causing regression of the state, disorder or condition or at least one of its clinical or subclinical symptoms). The benefit to a patient to be treated is either statistically significant or at least perceptible to the patient or to the physician. However, it will be appreciated that when a medicament is administered to a patient to treat a disease, the outcome may not always be effective treatment.

"Effective amount" means an amount of an API or pharmaceutical composition sufficient to result in the desired therapeutic response. The therapeutic response can be any response that a user (e.g., a clinician or patient) will recognize as an effective response to the therapy.

"Cmax" means the the peak concentration of the drug in the plasma. Tmax" means the time from administration to reach Cmax.

The invention relates to oral pharmaceutical compositions comprising as an API an SIP receptor modulator or agonist, e.g. a compound of formula A1 or A2 as defined above. The disclosure describes solid phase dosage units, which may be a tablet or capsule, particularly a tablet. Exemplary compositions, e.g. tablets, contain 10mg (equivalent base weight) or less of the API, e.g. 7.5mg or less of the API, for example 5mg or less of the API. Some solid phase dosage units, e.g. tablets, contain 0.1 mg (equivalent base weight) or more API, e.g. 0.2mg or more of the API, for example 0.25mg or more API. There are therefore included in the invention dosage units, e.g. tablets, which contain from 0.1 mg to 10mg of the API (equivalent base weight), e.g. 0.2mg to 7.5mg of the API; particular dosage units, e.g. tablets, contain from 0.25mg to 5mg of the API, for example 2.5mg to 5mg of the API.

In one implementation of the invention, the dosage units (e.g. tablets) contain no more than 4mg of the API (equivalent base weight), e.g. from 0.2mg or 0.25mg to 4mg of the API. Some dosage units contain from 2mg or 2.5mg to 4mg of the API. In one embodiment, the dosage units (e.g. tablets) contain 4mg of the API (equivalent base weight).

In one embodiment, the compound is of formula A1. It will be understood that this embodiment, like all embodiments mentioned herein, is applicable across the entire scope of the disclosure, including to ail other embodiments disclosed herein, including those disclosed in the following paragraphs.

In one embodiment, R ₅ is H.

In one embodiment, A is COOR ₅ , and in particular is COOH. In one embodiment, Z is

In one embodiment, A-Z is

In one embodiment, Y is phenyl optionally substituted by 1 , 2 or 3 radicals selected from halogen, N0 ₂ , C _1-6 alkyl, C _1-6 alkoxy; halo-substituted Ci _-6 alkyl and halo-substituted

d-ealkoxy, and in particular Y is phenyl substituted by a single said radical, e.g. by a single C _1-6 alkyl radical. The designation "C _1-6 " means "having 1 , 2, 3, 4, 5 or 6 carbon atoms" and an exemplary d-β alkyl radical is ethyl.

In one embodiment, A-Z-C( R3)-Y- is

where R ₇ is H, halogen, Nd ₂ , C _-6 alkyl, d-ealkoxy; halo-substituted d-ealkyl and halo- substituted d-ealkoxy, and in particular is d-ealkyl e.g. ethyl.

In one embodiment, W is d, d or d alkylene, particularly methylene. Since every embodiment disclosed herein is applicable to the entire disclosure of the invention, it will be understood that in this case (where W is d, d or C ₃ alkylene, particularly methylene), the compound may be of the formula A1 and/or Y may be an optionally substituted phenyl group as previously described herein and in articular where A-Z-C(R ₄ )(R ₃ )-Y- may be

In one embodiment (for example applicable to the embodiments mentioned in the immediately preceding paragraph), Ri is phenyl substituted by phenyl or by C ₃ -C ₈ cycloalkyl, e.g. by cyclohexyl, wherein each phenyl and cyclohexyl are each independently optionally substituted by 1 or 2 substituents selected from halogen, d. ₆ alkyl, d _-6 alkoxy and halo substituted-d _-6 alkyl or -d. ₆ alkoxy. Ri may therefore be phenyl substituted by -d cycloalkyl, each optionally substituted as just mentioned; in one sub-embodiment, phenyl of RT is substituted by a single substituent, e.g. fluoro or in particular trifluoromethyl, and cycloalkyl of Ri is unsubstituted cyclohexyl. In an embodiment where the compound is of formula A1 , phenyl is 1 ,4-substituted by W and cycloalkyl (e.g. unsubstituted cyclohexyl). The designation "C ₃ -C ₈ " means having 3, 4, 5, 6, 7 or 8 carbon atoms, e.g. 5 or 6 carbon atoms.

In one embodiment, the compound is of formula A1 ; W is C _1t C ₂ or C ₃ alkylene; Y is a phenyl group optionally substituted by 1 , 2 or 3 radicals selected from halogen, N0 ₂ , C _1-6 alkyl, ₆ alkoxy; halo-substituted d _-6 alkyl and halo-substituted Ci. ₆ alkoxy; and is phenyl substituted by C ₃ -C ₈ cycloalkyi, wherein phenyl and cyclohexyl are each independently optionally substituted by 1 or 2 substituents selected from halogen, C _h alky!, Ci. ₆ alkoxy and halo substituted-C _1-6 alkyl or -C ealkoxy.

Pharmaceutical compositions

WO 20 0/072703 teaches that SIP receptor agonists have a negative chronotropic side effect. The invention is predicated at least in part by an insight that the negative

chronotropic effect may be ameliorated without rendering the drug therapeutically useless by administering the drug in sustained release form.

In one aspect, therefore, the invention provides a pharmaceutical composition for oral administration which comprises an active pharmaceutical ingredient ("API") selected from SIP receptor modulators and agonists, wherein the composition is adapted to provide sustained release of the API. In embodiments, the API is selected from the compounds of Formulae A1 and A2.

The invention in some embodiments is in part predicated on a finding that an active compound as disclosed above, namely the hemifumarate salt of 1-{4-[1-(4-cyclohexyl-3- trifluoromethyl-benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azeti dine-3-carboxylic acid, has poor stability in certain settings and that stability of the compound in a pharmaceutical composition may be improved by adopting one, two or three of the following measures:

• not exposing the active compound to a basic compound, e.g. excluding basic

compounds from the composition (other than the active compound itself when in basic form)

• having the active compound in the form of particles having a crystallinity of above 80%

• having the active compound in the form of particles having an X90 diameter of at least

8μηη, e.g. at least 10pm. Advantageously, the composition is substantially dry, e.g. is made of substantially moisture- free constitutents, and is provided with a moisture-protective layer as a barrier to ingress of water and/or includes a desiccant.

In one implementation of the invention, the composition, for example in the form of a tablet, comprises a desiccant. In another implementation of the invention, the composition, for example in the form of a tablet, is provided with a moisture-protective layer. In a particular implementation, the composition, for example in the form of a tablet, comprises a desiccant and is provided with a moisture-protective layer.

It will be appreciated that 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethy l]-2- ethyl-benzyl}-azetidine-3-carboxylic acid and its pharmaeutically acceptable salts e.g.

hemifumarate salt is a representative of the compounds of formula A1 or A2 as described above and that the just-mentioned measures may therefore be beneficially applied to any such compound as an active pharmaceutical ingredient (API) of a composition.

Where the API is formula A1 or A2, it is advantageous to combine two or more of the above measures for imparting stability, as well as to formulate the API for sustained release. For example embodiments of the invention lie in sustained release compositions, for example capsules or tablets, particularly tablets, having one the following characteristics (1) to (6):

(1) the API is not exposed to basic compounds, e.g. basic compounds are excluded from the composition, and the API is in the form of particles having a crystallinity of 80% or more;

(2) the API is not exposed to basic compounds, e.g. basic compounds are excluded from the composition, and the API is in the form of particles having an X90 diameter of at least 8μηι e.g. at least 10pm;

(3) the API is in the form of particles having an X90 diameter of at least 8pm e.g. at least 10pm and a crystallinity of 80% or more;

(4) the API is not exposed to basic compounds, e.g. basic compounds are excluded from the composition, and the API is in the form of particles having an X90 diameter of at least 8μηι e.g. at least 10pm and a crystallinity of 80% or more; (5) any of (1), (2), (3) and (4) in combination with inclusion of a desiccant in the composition;

(6) any of (1), (2), (3), (4) and (5) in combination with provision of a moisture-protective layer or coating.

In one embodiment, therefore, the compositions of the invention comprise an API selected from the compounds of Formulae A1 and A2, wherein the API is not exposed to a basic compound.

In another embodiment, the compositions of the invention comprise an API selected from the compounds of Formulae A1 and A2, wherein the API is in the form of particles having an X90 diameter of at least 8pm and optionally of at least 1ΰμπι.

Also provided is an embodiment in which the compositions of the invention comprise an API selected from the compounds of Formulae A1 and A2, wherein the API is in the form of particles having a crystallinity of 80% or more.

Further included are embodiments in which the compositions comprise an API selected from the compounds of Formulae Aland A2, wherein the API is in the form of particles having an X90 diameter of at least 8pm and optionally of at least 10pm and a crystallinity of 80% or more.

A further embodiment resides in compositions of the invention which comprise an API selected from the compounds of Formulae A1 and A2, wherein the API is in the form of particles which have an X90 diameter of at least 8pm and optionally of at least 10pm and which are not exposed to a basic compound. In an embodiment, the composition is free of basic compounds.

Certain implementations of the invention reside in compositions of the invention which comprise an API selected from the compounds of Formulae A1 and A2, wherein the API is in the form of particles which have a crystallinity of 80% or more and which are not exposed to a basic compound.

Additionally to be mentioned are implementations in which the compositions of the invention comprise an active pharmaceutical ingredient ("API") selected from the compounds of Formulae A1 and A2, and one or more pharmaceutically acceptable excipients, wherein the API is in the form of particles which have have an X90 diameter of at least 10pm and a crystallinity of 80% or more and which are not exposed to a basic compound.

In practice, the compositions of the invention comprise one or more pharmaceutically acceptable excipients, as well as an API. The excipient(s) will include a sustained release component, e.g. polymer.

The invention is not limited as to the manner in which sustained release is achieved.

In one embodiment, the sustained release is pH-independent. Thus, a polymer may be included in the formulation which acts to control the release of the API, e.g. in a pH- independent manner. In embodiments, a polymer, for example a hydrogel-forming polymer, is in admixture with the API to form a matrix, or it is comprised in a coating, or it both forms a matrix and is comprised in a coating.

In one embodiment, the compositions of the invention comprise a hydrogel-former, for example a single polymer or a combination of two or more polymers. The hydrogel-former may be in admixture with the API, or in a coating, or both in admixture with the API and in a coating. In a particular embodiment, the hydrogel former comprises, or is, at least one hypromellose. It is feature of embodiments of the invention that the API is a compound of formula A1 or A2 and that sustained release agent is non-basic, for example a non-basic hydrogel-former such as, e.g., hypromellose. Some compositions of the invention comprise a combination of two or more different hypromellose materials, for example having different viscosities, whereby a desired balance of properties may be achieved.

Additional or alternative, e.g. alternative, materials which may be included in the composition to provide sustained release are hydrophobic polymers, for example ethyl cellulose or a methacrylic acid polymer, or a combination thereof. Such polymers, whether used singly or in combination, may be comprised in a coating or may be included in admixture with the API (i.e. may be used as a matrix-former), or may be present both in a coating and in admixture with the API.

Further additional or alternative, e.g. alternative, materials which may be included in the composition to provide sustained release are insoluble erodible materials, for example a wax or a hydrogenated vegetable oil, or a combination thereof. Such materials, whether used singly or in combination, may be comprised in a coating or may be included in admixture with the API (i.e. may be used as a matrix-former), or may be present both in a coating and in admixture with the API.

Other additional or alternative, e.g. alternative, materials which may be included in the composition to provide sustained release are hydrophilic polymers other than hypromellose as just mentioned, for example a non-basic poly(ethylene oxide) or sodium alginate, or a combination thereof. Such materials, whether used singly or in combination, may be comprised in a coating or may be included in admixture with the API (i.e. may be used as a matrix-former), or may be present both in a coating and in admixture with the API. Where a basic material is used, e.g. sodium alginate, it is in the case of APIs of formula A1 or A2 preferably confined to a coating and not used in admixture with the API.

As mentioned previously herein, an osmotic pump system may be used to obtain sustained release.

It will be recalled that, for all compositions comprising an API of Formula A1 or A2, it is desirable that the API should not be in contact with an basic sustained release agent. Thus, basic matrix materials are desirably avoided in relation to such APIs.

In one embodiment, the sustained release composition provides a zero order (linear) release profile, for example for a period of at least 5 hours, e.g. at least 10 hours and optionally for 16 hours or more.

Without wishing to be bound by theory, it is believed that a zero order (linear) release profile provides the optimal profile to minimise undesirable cardiac side effects e.g. the negative chronotropic effect and or AV blocks sometimes associated with administration of some S1P receptor modulators.

Suitable compositions may be determined empirically. Linear release compositions may conveniently be made by using a hydrogel-former as a matrix, for example a single grade of hypromellose or a combination of different grades of hypromellose may be used to achieve linear release, e.g. in the compositions described in the examples of this specification. In one embodiment, there is used as a matrix hypromellose having an apparent viscosity of 80- 120 mPa.s. In another embodiment, there is used as a matrix equal parts by weight of hypromellose having an apparent viscosity of 80-120 mPa.s and of hypromellose having an apparent viscosity of 75000-140000 mPa.s. In a further embodiment, the invention includes compositions comprising an insoluble, erodible matirx material such as a wax or cellulosic polymer.

The invention includes embodiments which have the effect of increasing Tmax. The invention includes embodiments which have the effect of reducing Cmax. The invention includes embodiments which have the effect of increasing Tmax and reducing Cmax. It is contemplated that the aforementioned compositions comprising a hypromellose matrix (e.g. a single hypromellose material or a combination thereof) will both increase Tniax and reduce Cmax but the invention is not limited to such hypromellose matrices. Sustained release agents which reduce Cmax, increase Tmax or both may be chosen empirically.

APIs of formula A1 or A2 have been found to have poor stability in admixture with the following basic compounds, which should therefore be avoided from being combined with such APIs in the tableting composition: sodium starch glycolate (sold under the trade mark . Primojel®), sodium lauryl sulfate, magnesium stearate, calcium stearate, calcium carbonate, calcium sulfate, sodium stearyl fumarate and sodium bicarbonate. The above-mentioned APIs also have poor stability in admixture with gelatine, and this is preferably not included with these APIs.

Not exposing the active compounds of formula A1 and A2 to a basic compound may be achieved by not having the active compound in admixture with a basic compound. In an embodiment, therefore, any such API is in a mixture of materials which is free of basic compounds. Conveniently, therefore, the composition is free of basic compounds (provided that the compound of formula A1 or A2 may have its moiety A (e.g. -COOH) in acid or salt form). However, the invention includes, for _. example, the provision of two- or multi-part compositions of which one part incorporates a compound of formula A1 or A2 but is free of any basic compound (provided that the compound of formula A1 or A2 may have its moiety A (e.g. -COOH) in acid or salt form) and a second part does not incorporate a compound of formula A1 or A2 but does incorporate an ionic compound. It will be understood that the expression "free of any basic compound" and similar expressions do not mean that no base may be present in the composition (apart from the API if it should be in basic form) but does allow very low amounts of base to be present, such an amount therefore being at a concentration which does not for practical purposes promote degradation of the API. For example, a basic compound may be an impurity which it is not possible or practicable to remove entirely or to an undetectable level. In embodiments, moiety A is not in the form of a salt but ₅ is present as H or an ester- forming group. R ₅ particularly is present as H (i.e. moiety A is in its acid form and not as a salt).

In one embodiment the pharmaceutical composition comprises a tablet comprising a compressed composition which consists of a compound of formula A1 or A2 in admixture with a non-basic sustained release agent (e.g. a hydrogel former) and one or more other non-basic compounds. The one or more other non-basic compounds may comprise or consist of non-basic excipients, e.g. selected from: binders, glidants, lubricants, fillers, diluents, and sorbents. The tablet may have one or more coating layers and/or may have an basic component separated from said solid phase mixture by a barrier. Said solid phase mixture may comprise one or more non-basic APIs in addition to one or more APIs of formula A1 or A2. The tablet may have a coating layer which is a moisture barrier, for example as sold under the trade mark Opadry® amb. A moisture barrier film composition is described in W01996/001874, included herein by reference for all purposes, and comprises polyvinyl alcohol, soya lecithin, and optionally a flow aid, a colorant, and/or a suspending agent. Conveniently, the composition includes a desiccant, e.g. colloidal silica.

In one embodiment the pharmaceutical composition comprises a tablet comprising a compressed composition which consists of a compound of formula A1 or A2 in admixture with hypromellose and further excipients selected from lactose (e.g. as lactose

monohydrate), microcrystalline cellulose, stearic acid, hydrogenated vegetable oil, mineral oil, polyethylene glycol (e.g. polyethylene glycol 4000-6000), glyceryl palmitostearate, colloidal silicon dioxide, and glyceryl behenate. In such a formulation, hypromellose is considered to act as a sustained release agent, lactose as a filler, microcrystalline cellulose as a binder, colloidal silicon dioxide as a glidant and the remaining materials as lubricants; in embodiments, only a single such lubricant is included in the composition, e.g. glyceryl behenate. Colloidal silicon dioxide acts also as a desiccant.

An embodiment of the invention therefore comprises a tablet which comprises a compressed phase which consists of a compound of formula A1 or A2 in admixture with lactose (e.g. as lactose monohydrate), microcrystalline cellulose, colloidal silicon dioxide, hypromellose and a lubricant selected from stearic acid, hydrogenated vegetable oil, mineral oil, polyethylene glycol (e.g. polyethylene glycol 4000-6000), glyceryl palmitostearate and glyceryl behenate, and combinations of the aforesaid lubricant compounds. For example, such an embodiment may comprise a tablet which comprises a compressed phase which consists of a compound of formula A1 or A2 in admixture with lactose (e.g. as lactose monohydrate), microcrystalline cellulose, colloidal silicon dioxide, hypromellose and a lubricant selected from hydrogenated vegetable oil, mineral oil, polyethylene glycol (e.g. polyethylene glycol 4000-6000), glyceryl palmitostearate and glyceryl behenate, and combinations of the aforesaid lubricant compounds. In embodiments, a single lubricant is present in the solid phase mixture, particularly glyceryl behenate.

Particle Size

It will be recalled that an embodiment of the invention is in part predicated on a finding that the stability in a solid phase composition, particularly a tablet, of an active compound as disclosed above, namely 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethy l]-2- ethyl-benzyl}-azetidine-3-carboxylic acid, and pharmaceutically acceptable salts, e.g.

hemifumarate salt, may be improved by the active compound being in a relatively large particle size but, in most instances, a particle size not so large that the composition fails to comply with the the USP, EP and JP harmonised content uniformity requirement.

Content uniformity is influenced by the particle size of the drug substances. Median particle size is denoted by X _m , where m is a percentage of the particle size distribution.

It will be appreciated that 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethy l]-2- ethyl-benzyl}-azetidine-3-carboxylic acid and pharmaceutically acceptable salts, e.g.

hemifumarate salt, is representative of the compounds of formula A1 or A2 as described above and that the just-mentioned measures relating to particle size may therefore be beneficially applied to any such compound as an active pharmaceutical ingredient (API) of a composition. The invention therefore provides a solid phase sustained release formulation, particularly a tablet, comprising a compound of formula A1 or A2 in a relatively large particle size, e.g. an X90 diameter of at least 8pm and often of 10pm or more, for example 20pm or 25pm or more, optionally of 100pm or more and further optionally of 150pm or more. The particle size (X90 diameter ) may be up to 300pm, e.g. up to 250pm, and optionally up to 200pm (e.g. up to 195pm). In some embodiments, the X90 diameter is of from 10pm to 300pm, e.g. 10pm to 250pm or 10pm to 200pm. Also included are embodiments in which the particle size (X90 diameter) is from 25pm to 300pm, e.g. 25pm to 250pm or 25pm to 200pm. Further included are embodiments in which the X90 diameter is from 100pm to 300pm, e.g. 100pm to 250pm or 100pm to 200pm. In a particular embodiment, the X90 particle diameter is at least 10 pm and is less than 191 pm, e.g. less than 180pm, less than 170 pm, less than 160 pm, less than 150 pm, less than 140 pm or less than 130 pm, e.g. no more than 125 pm, such as no more than 121 pm; in a sub-embodiment, the X90 particle diameter is at least 25 pm and is less than 191 pm, e.g. less than 180pm, less than 170 pm, less than 160 pm, less than 150 pm, less than 140 pm or less than 130 pm, e.g. no more than 125 pm, such as no more than 121 pm. In some instances the X90 particle diameter is at least 10 pm or 25 pm but no more than 100pm, e.g. is no more than 90pm, no more than 80pm, no more than 70pm, no more than 60pm, no more than 50pm or no more than 40pm, for example is no more than 30pm. In other embodiments, the X90 particle diameter is from 121 pm to 191 m. In another embodiment, the X90 particle diameter is from 1 1 pm to 29pm.

An embodiment of the invention is in part predicated on a finding that, in the case of an API of formula A1 or A2, API particles of the sizes mentioned herein of at least 8pm may be formulated into tablets which have a tablet content uniformity which fulfills the requirement of the USP, EP and JP harmonised content uniformity requirement, in particular as in force on 1 January 2011. The sustained release pharmaceutical compositions of the present invention may fulfil, and desirably do fulfill, the USP, EP and JP harmonised content uniformity requirement (refer to chapter 2.9.40. Uniformity of dosage unit in the EP). A maximum particle size which enables the requirement to be met cannot be stipulated because the maximum possible particles size varies with the drug content of the dosage unit. For example, in the case of a sustained release tablet containing 0.25mg of an API of formula A1 or A2, API particles having an X90 diameter of 121 pm complied with the harmonised content uniformity requirement whereas API particles having an X90 diameter of 191pm failed to comply. In contrast, in the case of a tablet containing 5mg of an API, API particles having an X90 diameter of up to 191 pm complied with, the harmonised content uniformity requirement. A maximum particle size which enables any particular tablet composition to meet the harmonised content uniformity requirement may be determined empirircally.

The particle size distribution (by volume) may be measured using a laser diffraction sizing instrument, for example the Sympatec Helos device (available from Sympactec GmbH, Germany) using the Cuvette dispersion device. The X ₉₀ diameter is the spherical diameter corresponding to the X ₉₀ volume.

To make the measurement, a stock dispersion may be prepared by mixing the drug substance with a dispersing aid (e.g Octastat 5000 (Octel corp)) using a vortex until a smooth and homogeneous paste is formed. The paste may then be diluted and mixed to a final volume of 3 to 6 ml using white spirit. The optical concentration of the final solution should remain below 5%. The percent values are calculated from the mean cumulative volume size curve by the software of the Sympatec instrument. X90 mean 90% of the particle size population is below the specified value in volume.

Additional information on particle size may be obtained using SEM (scanning electron microscopy).

The invention includes the following embodiments (optionally in combination with the features of other embodiments disclosed herein):

1) a solid phase sustained release pharmaceutical composition in unit dosage form, particularly a tablet, which comprises one or more pharmaceutically acceptable excipients, in particular non-basic excipients, and an API as described herein and which complies with the US Pharmacopeia, European Pharmacopeia and Japanese Pharmacopeia harmonised content uniformity requirements as in force on 1 January 2011 ;

2) a solid phase sustained release pharmaceutical composition in unit dosage form, particularly a tablet, which comprises one or more pharmaceutically acceptable excipients, in particular non-basic excipients, and an API as described herein in an amount of from 4 mg to 6mg API, the API being as particles have an X90 diameter of at least 100pm but no more than 300pm, e.g. from 100pm to 250μιη, from 100pm to 200pm, from 100μηι to 190μητ, from 100pm to 180pm, from 100pm to 170pm; from 100μιη to 160pm or from 100pm to 150pm;

3) a solid phase sustained release pharmaceutical composition in unit dosage form, particularly a tablet, which comprises one or more pharmaceutically acceptable and preferably non-basic excipients, including a sustained release agent, and an API as described herein in an amount of from from 0.2 mg to 1 mg API, the API being as particles have an X90 diameter of at least 10μιη but no more than 100pm, e.g. 10pm to 90μιη, 10pm to 80pm, 10μι ^η to 70pm, 10pm to 60μιτι, 10pm to 50pm, 10pm to 40pm or 10μιη to 30pm.

An aspect of the invention resides in compounds of formula A1 or A2 when in particulate form having an X90 diameter as disclosed herein, as well as products or compositions of matter comprising such particulate compounds. CrystalHnity

The invention is in part also predicated on a finding that the stability in a solid phase formulation, particularly a tablet, of an active compound as disclosed above, namely 1-{4-[1- (4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethyl]-2-eth yl-benzyl}-azetidine-3-carboxylic acid, may be enhanced by the active compound being in particles of relatively high crystallinity, e.g. a crystallinity of 80% or more, for example 85% or more and optionally of 90% or more.

It will be appreciated that 1-{4-[1-(4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethy l]-2- ethyl-benzyl}-azetidine-3-carboxylic acid is a representative of the compounds of formula A1 or A2 as described above and that the just-mentioned measures may therefore be beneficially applied to any such compound as an active pharmaceutical ingredient (API) of a sustained release composition. The invention therefore provides a sustained release solid phase formulation, particularly a tablet, comprising a compound of formula A1 or A2 in a crystallinity of 80% or more, for example 85% or more and optionally of 90% or more.

A aspect of the invention resides in compounds of formula A1 or A2 when having a crystallinity of 80% or more, for example 85% or more and optionally of 90% or more, as well as products or compositions of matter comprising compounds having such crystallinity.

Conveniently, a crystallinity of 80% or more may be attained by a compound of formula A1 or A2 being as particles having an X90 diameter of 10pm or more and particularly 11 pm or more, for example 20pm or 25pm or more, optionally of 100pm or more and further optionally of 150pm or more. The X90 diameter may be as previously described herein Under the heading "Particle Size".

Crystallinity may be measured using X-ray powder diffraction (XRPD), for example using a Bruker D8 device.

It will be appreciated that solid phase compositions, e.g. tablets, may advantageiously combine the features bf particle size and/or crystallinity disclosed herein with the feature of not exposing the API to an basic compound.

Methods for making the pharmaceutical compositions W 201

22

Compositions of the invention may.suitably be made by combining the components as dry powders, for example tablets may be made by dry granulating the components of the tablet mix and optionally applying a film coating, for example a sustained release film, to the compressed tablet.

API particles can be prepared by suitable milling techniques, e.g. those well known in the art such as, for example, pin-milling, wet-jet milling and wet-ball milling.

Where the API particles are derived from coarse API particle crystals, the coarse crystals may be obtained using any suitable methodolgy. For example any of the methodologies set . out in WO2010/071794, WO2010/080455 or WO2010/080409.

Of interest in the invention are the following numbered paragraphs.

1. A pharmaceutical composition for oral administration which comprises an active pharmaceutical ingredient ("API") selected from SIP receptor modulators and agonists, wherein the composition is adapted to provide sustained release of the API.

2. A composition of numbered paragraph 1 which increases Tmax as compared to an immediate release composition.

3. A composition of numbered paragraph 1 or 2 which reduces Cmax as compared to an immediate release composition.

4. A composition of any preceding numbered paragraph which has a zero order release profile for a period of at least 5 hours.

5. A composition of numbered paragraphs which has a zero order release profile for a period of at least 10 hours and optionally for 16 hours or more.

6. A composition of any previous numbered paragraph which comprises a polymer which controls the release of the API in a pH-independent manner.

7. A composition of any previous numbered paragraph which comprises a hydrogel- former. W

23

8. A composition of numbered paragraph 7 wherein the hydrogel-former is in admixture with the API.

9. A composition of numbered paragraph 7 or 8 wherein the hydrogel-former comprises, or is, at least one hypromeliose.

10. A composition of numbered paragraph 9 wherein the hydrogel-former is hypromeliose having an apparent viscosity of 80-120 mPa.s or a mixture of equal parts by weight of hypromeliose having an apparent viscosity of 80-120 mPa.s and of hypromeliose having an apparent viscosity of 75000-140000 mPa.s.

11. A composition of any preceding numbered paragraph which is a tablet. . 2. A composition of any preceding numbered paragraph wherein the API is a compound ereof, and is

wherein

A is COORs, OPO(OR ₅ ) ₂ , PO(OR ₅ ) ₂ , S0 ₂ OR _5l POR ₅ OR ₅ or 1 /-tetrazol-5-yl, R ₅ being H or an ester-forming group;

W is a bond, Ci- ₃ alkylene or C ₂ - ₃ alkenylene;

Y is C ₆ .i ₀ aryl or C ₃ - ₉ heteroaryl, optionally substituted by 1 to 3 radicals selected from halogen, N0 ₂ , C^alkyl, C _1-6 alkoxy; halo-substituted C _-6 alkyl and halo-substituted

Ci _-6 alkoxy;

Z is chosen from:

wherein the asterisks of Z indicate the point of attachment between -C(R ₃ )(R ₄ )- and A of Formula la or lb, respectively; R ₆ is chosen from hydrogen and C _h alky!; and and J ₂ are independently methylene or a heteroatom chosen from S, O and NR ₅ ; wherein R ₅ is chosen from hydrogen and C^alkyl; and any alkylene of Z can be further substituted by one to three radicals chosen from halo, hydroxy, C _1-6 alkyl; or R ₆ can be attached to a carbon atom of Y to form a 5-7 member rin rylC ₁ . ₄ alkyl, , cycloalkyl

or heterocycloalkyl of may be substituted by 1 to 5 groups selected from halogen, d. ₆ alkyl, C^alkoxy and halo substituted-C^alkyl or -d-ealkoxy;

R ₂ is H, d-ealkyl, Halo substituted C _h alky!, C ₂ . ₆ alkenyl or C _2-6 alkynyl; and each of R ₃ or R ₄ , independently, is H, halogen, OH, C _1-6 alkyl, d-ealkoxy or halo substituted C _1-6 alkyl or C _1-6 alkoxy.

13. A composition of numbered paragraph 12 wherein A is COOH.

14. A composition of numbered paragraph 2 or 13 wherein the compound is of formula A1. 15. A composition of numbered paragraph 14 wherein:

W is CL C ₂ or C ₃ alkylene;

Y is a phenyl group optionally substituted by 1 , 2 or 3 radicals selected from halogen, N0 ₂ , C^alkyl, Ci. ₆ alkoxy; halo-substituted C _-6 alkyl and halo-substituted C _-6 alkoxy; and

Ri is phenyl substituted by C ₃ -C ₈ cycloalkyl, wherein phenyl and yclohexyl are each independently optionally substituted by 1 or 2 substituents selected from halogen, d. ₆ alkyl, C _1-6 alkoxy and halo substituted-Ci. ₆ alkyl or -C _1-6 alkoxy.

16. A composition of numbered paragraph 12 wherein the API is 1-{4-[1-(4-cyclohexyl-3- t fluoromethyl-benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azetid ^' ine-3-carboxylic acid or a pharmaceutically acceptable salt thereof.

17. A composition of numbered paragraph 16 wherein the API is 1-{4-[1-(4-cyclohexyl-3- tr ^' ifluoromethyl-benzyloxyimino)-ethyl]-2-ethyl-benzyl}-a zetidine-3-carboxylic acid or a hemifumarate salt thereof.

18. A composition of any of numbered paragraph 12 to 17 wherein the API is in a mixture of materials which is free of basic compounds.

19. A composition of numbered paragraph 18 wherein the mixture is comprised in a tablet and the mixture consists of the API in admixture with lactose, microcrystalline cellulose, a hypromeilose, colloidal silicon dioxide and a lubricant selected from hydrogenated vegetable oil, mineral oil, polyethylene glycol, gyceryl palmitostearate and glyceryl behenate.

20. A composition of any preceding numbered paragraph wherein the API is selected from the compounds as defined in any of numbered paragraphs 12 to 17 and is in the form of particles having an X90 diameter of at least 8pm.

21. A composition of numbered paragraph 20 wherein the API particles have an X90 diameter of from 1 Opm to 300pm.

22. A composition of numbered paragraph 21 wherein the particles have an X90 diamater of at least 10pm but no more than 100pm. 23. A composition of numbered paragraph 21 wherein the particles have an X90 diameter of at least 100pm but no more than 250pm.

24. A composition of any of numbered paragraphs 20 to 23 which is in unit dosage form and complies with the US Pharmacopeia, European Pharmacopeia and Japanese

Pharmacopeia harmonised content uniformity requirements as in force on 1 January 2011.

25. A composition of numbered paragraph 24 which is in the form of a tablet containing from 4 mg to _. 6mg API (equivalent free base weight) and particles have an X90 diameter of at least 100pm but no more than 200pm.

26. A composition of numbered paragraph 24 which is in the form of a tablet containing . from 0.2 mg to 1 mg API (equivalent free base weight) and particles have an X90 diameter of at least 10pm but no more than 50pm.

27. A composition of any preceding numbered paragraph wherein the API is selected from the compounds as defined in any of claims 12 to 17 and has a crystallinity of 80% or more.

28. A composition of numbered paragraph 27 wherein the API has a crystallinity of 85% or more.

29. A tablet comprising a compressed mixture consisting of 1-{4-[1-(4-cyclohexyl-3- trifluoromethyl-benzyloxyimino)-ethyl]-2-ethyl-benzyl}-azeti dine-3-carboxylic acid or its hemifumarate salt and one or more non-basic excipients including a hydrogel-former, the 1- {4-[1-(4-cyclohexyl-3-trifluoromethyl-benzyloxyimino)-ethyl] -2-ethyl-benzyl}-azetidine-3- carboxylic acid or hemifumarate salt being in the form of particles having an X90 diameter of from 10 pm to 200 pm.

30. A tablet of numbered paragraph 29 wherein said particles are at least 80% crystalline.

31. A tablet of numbered paragraph 29 or 30 wherein the hydrogel-former is a

hypromellose or a mixture of hypromelloses. Examples

The hemifumarate salt form of compound 1-{4-[1-(4-cyclohexyl-3-trif|uoromethyl- benzyloxyimino)-ethy|]-2-ethyl-benzyl}-azetidine-3-carboxyli c acid is referred to as "NVS-A" in the following examples.

Example 1

A clinical ascending dose study is undertaken with compound NVS-A. A decrease in heart rate was demonstrated at single doses > 0.3 mg . The maximum bradycardia effect approximately occured 3 hours post dose, the heart rate then returned to its initial rhythm 10 hours post dose. During multidose treatment, the maximum decrease in heart rate was observed on the first day of treatment; the bradycardia then tended to disappear after 4 days which seems to indicate a "tolerance build up".

Example 2

Clinical results highlighted that Cmax and Tmax were most likely the two critical

pharmacokinetic parameters impacting the cardiac adverse effect. A Gastroplus™ (PK/PD modeling software) simulation was thus performed in order to first assess the potential for absorption along the G I tract based on physicochemical properties. Secondly, the

Gastroplus™ data were used to optimise the pharmacokinetic profile so that a reduction in heart rate could be achieved.

In Figure 1 , the PK profile obtained with the 4 mg immediate release tablet is presented. Typically, the plasma concentration peaked between 3 to 6 hours post dose. From the Gastroplus™ simulation, it was recommended to generate a release profile shifting the Tmax to around 10 hours for a first modified release prototype and at least 16 hours for a second one.

The expected benefit on the heart rate was also simulated and is shown in Figure 2. Example 3

Hydrogel matrix tablets having a linear release profile over, respectively, 10 hours and at least 16 hours were made by direct compression of the dry blend in accordance with the compositions given in Table 1 : Table 1 : Composition of the NVS-A modified release tablets

Ingredient Amount per film-coated tablet Function Reference to

(mg) standards

4 mg (F16) 4 mg (F10)

Tablet core

NVS-A 4.448 ¹ 4.448 ¹ Active Novartis

ingredient monograph

Lactose monohydrate 45:277 40.177 Filler Ph.Eur./NF

Hypromellose/ Hypromellose 10.200 . Controlled Ph.Eur./USP (Type 2208) ² . Release

agent

Hypromellose/ Hypromellose 10.200 25.500 Controlled Ph.Eur./USP (Type 2208) ³ Release

agent

Cellulose, microcrystalline/ 8.500 8.500 Filler/ Ph.Eur./NF Microcrystalline cellulose Binder

Glycerol dibehenate/ Glyceryl 5.950 5.950 Lubricant Ph.Eur./NF behenate

Silica, colloidal anhydrous/ 0.425 0.425 Glidant Ph.Eur./NF Colloidal silicon dioxide

Core tablet weight 85.000 85.000

Coating

Basic coating premix white ⁴ 4.600 4.600 Coating agent Novartis

monograph

Water, purified ⁵ — ... Solvent Ph.Eur./USP Total film-coated tablet 89.600 ⁶ 89.600 ⁶

weight

Corresponds to 4 mg NVS-A base respectively

² Apparent viscosity 75000 - 140000 mPa.s

³ Apparent viscosity 80 - 120 mPa.s

⁴ The qualitative composition of the coating premix is provided in Table 2

⁵ Removed during processing

⁶ Tablet target weight of 89.6 mg corresponding to a 4.6 mg coating after dry weight adjustment

Table 2: qualitative composition of coating premix white

Coating premix ingredient Reference to standards

Hypromellose/Hydroxypropylmethy!-cellulose (HPMC 2910), (E464) Ph.Eur./USP

Titanium Dioxide (E171 , C.I. 77891 ) Ph.Eur./USP

Talc Ph.Eur./USP

Macrogol / Polyethylene glycol / PEG 4000 Ph.Eur./NF

Example 4

The following Example discloses the stability of compound NVS-A in the presence of various compounds. Mixtures of NVS-A with various compounds were exposed in a sealed vial to conditions of 50°C temperature and 75% relative humidity for 4 weeks. After 4 weeks, the degradation of of NVS-A and sum of degradation products was assayed.

Assay method: HPLC using agilent 11000 and a Phenomenex Gemini C18 column. UV detection at 260 nm was used. The mobile phase used was a gradient water- acetonitrile containing 0.2% (v/v) formic acid and 10% (v/v) Triethylamine.

The results are displayed in Table 3.

Table 3

Mixture Sum of unknown peaks (%)* Assay (%)*

NVS-A alone ^: Ϊ86 100.2

NVS-A + Lactose (1 :1 ) 1.74 96.5

NVS-A + Maize Starch (1 :1 ) 1.82 99.4 _.

NVS-A + . Microcrystalline Cellulose 1.80 99.0 (1 :1)

NVS-A + Mannitol (1 :1 ) 1.80 99.2

NVS-A + HPMC (1 :1) 2.45 97.7

NVS-A + PVP K30 (1 :1 ) 2.07 99.6

NVS-A + Ac-Di-Sol (1 :1 ) 1.89 99.7

NVS-A + Primojel (1:1) 4.08 100.9

NVS-A + Crospovidone (1 :1 ) 1.89 99.7

NVS-A + Mgstearate (1 :0.85) 0.31 98.2

NVS-A + Mg Stearate (1:0.85)* 3.68 90.0

NVS-A + Sodium LaurylSulfate (SLS) + 8.54 - 88.3 - Microcrystalline Cellulose (2:1 :1)

NVS-A + Gelatin (1 :1) 3.86 ^s 96.1

NVS-A + Aerosil + Microcrystalline 1.89 99.9 Cellulose (2:1:1)

NVS-A +. Magnesium Stearate (Mg St) 6.13 89.2 + Microcrystalline Cellulose (2:1 :1)

NVS-A + Magnesium stearate + 10.53 80.7 Microcrystalline Cellulose (2:1 :1

NVS-A + Magnesium stearate + lactose 9.31 82.2 (2:1:1)

NVS-A + Calcium stearate + 4.55 89.8 Microcrystalline Cellulose (2:1 :1)

NVS-A + sodium stearyl fumarate + 7.52 83.1 Microcrystalline Cellulose (2:1 :1) Mixture Sum of unknown peaks (%)* Assay (%)*

NVS-A + Sodium bicarbonate + . 848 618

Microcrystalline Cellulose (2 :1 :1)

NVS-A + Tween 80 + Microcrystalline . 2.07 100.1

Cellulose (2:1:1)

* at 50°C/75%RH opened vial/ 4 weeks ^{— ~~}

Example 5

The following Example demonstrates the detrimental effect of magnesium stearate on NVS- A.

An NVS-A film coated tablet was developed comprising micronised drug substance, lactose monohydrate, microcrystalline cellulose, crospovidone, colloidal silicon dioxide, and magnesium stearate. A moisture protective Opadry® AMB film coating was applied. Tablet, strengths from 0.25 mg to 5 mg (equivalent free base weight) were produced (see Table 4 below). ^]

Table 4 Composition of the immediate release 0.25 and 5 mg tablet containing magnesium stearate as lubricant.

Ingredient Amount (mg) per Amount (mg) per Function Reference to

0.25 mg tablet 5 mg tablet standards

Tablet core

NVS A 0.278 ¹ 5.560 ² Drug substance Novartis

monograph

Lactose . 63.897 58.615 Diluent Ph. Eur. /NF monohydrate

Ingredient Amount (mg) per Amount (mg) per Function Reference to 0.25 mg tablet 5 mg tablet standards

Tablet core

Coating premix 3.400 3.400 Coating agent

Opadry A B

white**

Water, purified* 13.600 13.600 Solvent Ph.Eur. /USP

Total film-coated 88.400 88.400

tablet weight

Corresponds to 0.25 mg (e.g. 0.294% w/w) NVS-A base respectively

Corresponds to 5 mg (e.g 5.88% w/w) NVS-A base respectively

* Removed during processing

** The qualitative composition of the coating premix is as follows:

Coating premix ingredient Reference to standards

Polyvinyl alcohol - part hydrolised Ph. Eur. / USP

Titanium dioxide Ph.Eur. / USP

talc Ph.Eur. / USP

Lecithin (soya) NF

Xanthan gum Ph.Eur. / NF

The tablets were only stable for prolonged periods at 2-8°C. At 25°C, the 0.25 mg strength tablet was stable for only 6 months and the 5 mg tablet was stable for only 12 months. Beyond 6 months and 12 months at 25°C respectively for the 0.25 and 5 mg tablet.the stability data did not fulfill the stability guidance from the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH).

Example 6

The effect of drug particle size on the content uniformity of a tablet comprising 5 mg of NVS- A was studied. The results are presented in Figure 3.

The X ₉₀ particle size distribution (by volume) was measured using a laser diffraction sizing instrument [the Sympatec Helos device (available from Sympactec GmbH, Germany) using the Cuvette dispersion device. To make the measurement, a stock dispersion was prepared by mixing the drug substance with a dispersing aid (Octastat 5000 (Octel corp)) using a vortex until a smooth and homogeneous paste was formed. The paste was then diluted and mixed to a final volume of 3 to 6 ml using white spirit. The optical concentration of the final solution were kept below 5%. The percentage values were calculated from the mean cumulative volume size curve by the software of the Sympatec instrument. Additional confirmatory information on particle size was obtained using SEM (scanning electron microscopy).

The finer API material (X ₉₀ < 10pm (4 and 7 pm) was obtained by micronisation using a jet mill [ Hosokawa Alpine AFG100] using a pressure in the range from 2 to 5 bars. The material with X ₉₀ at 1 1 , 29 pm and 121 μηη was obtained by milling in this case with a pin mill with the relative tin speed being adjusted from 1 15, to 75 to 40 m/sec respectively. Finally, the coarser material with X ₉₀ pm of 191 urn was obtained by sieving on a 457um sieve.

The drug substance particle sizes considered showed reduced impact on the mean content uniformity value and its variability for the 5 mg tablet:

• where X90 = 191 μιτι, the content uniformity ranged from about 97.3% to about

104.0%

• where X90 = 121 pm, the content uniformity ranged from about 92.8% to about 99.7%

• where X90 = 29 pm, the content uniformity ranged from about 97.2% to about 102.9%

• where X90 = 11 μητι, the content uniformity ranged from about 97.3% to 00.1 %

• where X90 = 7 pm, the content uniformity ranged from about 94.1% to 98.9%

• where X90 = 4 μηι, the content uniformity ranged from about 93.3% to 99.1%.

This study demonstrated that despite the low tablet strength, micronised drug substance (where X90 = 4 pm to 7 μηη) was not essential to fulfil the USP, EP and JP harmonised content uniformity requirement. This requirement could be achieved when using milled drug substance (where X90 = 11 pm to 29 μιη) and unexpectedly also using the coarse drug substance (where X90 = 121 μιτι to 191 pm).

Example 7

The effect of drug particle size on the content uniformity of a tablet comprising 0.25 mg (equivalent free base weight) of NVS-A was studied. API drug particles were obtained and measured as in Example 6.

The results are presented in Figure 4.

Coarse drug substance with an X90 diameter of 121 pm and. 191 pm led to a significant increase in content uniformity variability:

• where X90 = 191 pm, the content uniformity ranged from about 81.2% to about

113.9% ,

• where X90 = 121 pm, the content uniformity ranged from about 87.6% to about

106.5%.

Milled drug substance with an X90 of 11 pm or 29 pm led to a better centered mean content uniformity and a slightly lower content variability than the one observed with the micronised drug substance (X90 = 4 and 7 μηι):

• where X90 = 29 pm, the content uniformity ranged from about 98.5% to about 101.6%

• where X90 = 11 μιη, the content uniformity ranged from about 98.9% to about 101.8%

• where X90 = 7 pm, the content uniformity ranged from about 95.9% to about 102.2%

• where X90 = 4 pm, the content uniformity ranged from about 94.0% to about 101.3%.

This example demonstrates that despite the low tablet strength, micronised drug substance (where X90 = 4 pm to 7 pm) is not essential to fulfil the USP, EP and JP harmonised content uniformity requirement. This requirement can unexpectedly be achieved when using milled drug substance (where X90 = 11 pm to 29 μιη). The upper limit that achieved the USP, EP and JP harmonised content uniformity requirement was a 0.25 mg tablet with a particle size characterised by a X90 = 121 pm.

Example 8

The following example compares the sum of degradation products of four different compositions, each comprising 0.25 mg (equivalent free base weight) of NVS-A mixed with a lubricant. The lubricants and the particle sizes of each composition are as follows: The table below summarises the difference in composition between the tablets made with magnesium stearate and glyceryl behenate as lubricant. It should be noted that the tablet made with glyceryl behenate and with the different drug substance particle size were not coated with the moisture protective film coated opadry AMB in order to better demonstrate the stability benefit. API drug particles were obtained and measured as in Example 6.

Table 5: Composition of the immediate release 0.25 mg tablet (equivalent free base weight) containing magnesium stearate and glyceryl behenate as lubricant.

Ingredient Amount (mg) per Amount (mg) per Function Reference to

0.25 mg tablet 0.25 mg tablet ⁽ standards with magnesium with glyceryl

sterate as behenate as

lubricant lubricant

Tablet core

NVS-A 0.278 ¹ 0.278 ¹ Drug substance Novartis

monograph

Lactose 63.897 58.797 Diluent Ph. Eur. /NF monohydrate

Microcrystalline 17.000 17.000 Diluent . Ph. Eur. /NF cellulose/

Cellulose

microcrystalline

Crospovidone 2.550 2.550 Disintegrant Ph. Eur. /NF

Silica, colloidal 0.425 0.425 Gliding agent Ph. Eur. /NF anhydrous /

Colloidal silicon

dioxide

Magnesium 0.850 0 Lubricant Ph. Eur. /NF stearate

Glyceryl behenate 0 5.950 Lubricant

Core tablet 85.000 85.000

weight

Coating

Coating premix 3.400 0 Coating agent

Opadry AMB

white**

Water, purified* 13.600 0 Solvent Ph.Eur. /USP

Total film-coated 88.400 Not applicable

tablet weight

Corresponds to 0.25 mg (e.g. 0.294% w/w) NVS-A base respectively

* Removed during processing

** The qualitative composition of the coating premix is as follows:

Coating premix ingredient Reference to standards W

35

Ingredient Amount (mg) per Amount (mg) per Function Reference to

0.25 mg tablet 0.25 mg tablet standards with magnesium with glyceryl

sterate as behenate as

lubricant lubricant

Tablet core

Polyvinyl alcohol - part hydrolised Ph.Eur. / USP

Titanium dioxide Ph.Eur. / USP

Talc Ph.Eur. / USP

Lecithin (soya) NF

Xanthan gum Ph.Eur. / ^' NF

1. Magnesium stearate, micronised drug substance X90 < 8 μιτι.

2. Glyceryl behenate, micronised drug substance X90 < 8 μιτι.

3. Glyceryl behenate, milled drug substance X90 = 1 1 μηη.

4. Glyceryl behenate, milled drug substance X90 = 29 pm.

Each of the 4 compositions were stored at 40°C in 75% relative humidity. The evolution of degradation product for each composition was followed over a period of six months. The results are shown in Figure 5.

• Composition 1 exhibited around 16% degradation product after 3 months of storage.

• Composition 2 exhibited around 4% degradation product after 3 months of storage.

• Composition 3 exhibited around 2% degradation product after 3 months of storage.

• Composition 4 exhibited around 1 % degradation product after 3 months of storage.

These results demonstrate that replacing magnesium stearate with glyceryl behenate reduces the sum of degradation product. Furthermore, changing the drug substance quality from micronized (less than 80% crystallinity) to milled (greater than 80% crystallinity) also reduces the sum of degradation product.