GB2498968A

GB2498968A - Pharmaceutical combination of a GPR119 agonist and a GPR40 agonist

Info

Publication number: GB2498968A
Application number: GB1201720.8A
Authority: GB
Inventors: John Keily; Colin Peter Sambrook-Smith; Donald Smyth; Sonal Patel; Andreas Scheel; Kirsty Louise Smith; Jonathan Richard White
Original assignee: Prosidion Ltd
Current assignee: Prosidion Ltd
Priority date: 2012-02-01
Filing date: 2012-02-01
Publication date: 2013-08-07
Also published as: GB201201720D0

Abstract

A pharmaceutical combination comprising at least one GPR119 agonist and at least one GPR40 agonist. Provided is a pharmaceutical composition comprising at least one GPR119 agonist and at least one GPR40 agonist. The combination may alternatively comprise a single agent that has both GPR119 agonist and GPR40 agonist activity. Further provided are methods of treating metabolic disorders, including type 2 diabetes and diseases associated with type 2 diabetes in a mammal, said methods comprising providing the mammal with a therapeutically effective combination of at least one GPR119 agonist and at least one GPR40 agonist. The said combination has been found to be additive, more than additive and synergistic in nature in the treatment of the aforementioned disease states.

Description

NOVEL COMBINATIONS

BACKGROUND OF THE: INVENTION

5 The present invention is directed to a novel combination of biological mechanisms -useful for the treatment of metabolic disorders including- type II diabetes. The invention provides novel compositions comprising a GPR119 agonist and a GPR40 agonist and uses thereof. These compositions are useful for the treatment of metabolic disorders including type. II diabetes.

10 Type 2 diabetes mellitus :(T2DM) is one of the most common chronic diseases affecting over 200 million people wordwide. The disease is characterized by abnormally high blood glucose levels (hyperglycemia) which is believed to cause long-term microvascular and.microvascular complications, such as heart disease, stroke, high blood pressure, hyperiipidemia, kidney failure, blindness, limb 15 amputation, nerve pain and other neurologic problems. (Xu J. et al, IDrngs vol? no3 p249-256, 2004),. Insulin plays a central role in controlling blood glucose levels, and secretion of this hormone is abnormal in pati ents with T2DM, for example being at levels that are insufficient to normalise blood glucose levels.

Obesity Is characterized by an excessive adipose tissue mass relative to body 20 size.. Clinically, body fat mass is estimated by the body mass index (BMI;

wei^it(kg)/height(m)2), or waist circumference. Individuals are considered obese when the BMI is greater than 3D and there are established medical consequences of being overweight. It has been an accepted medical view for some time that an increased body weight, especially as a result of abdominal body fat, is associated with 25 an increased risk for diabetes,, hypertension, heart disease, and numerous other health complications, such as arthritis, stroke, gallbladder disease, muscular and respiratory problems, back pain and even certain cancers

A range of drug therapies have been tested or are in current use for the treatment of T2D.M and related metabolic conditions. These include enhancement of 30 insulin secretion by pancreatic islet beta cells using insulin secretagogues such as sutphonylurea drugs or -glucagon-like peptide-1- (GLP-l)-based therapies, or direct adnainistration of insulin.

1

There is a continuing need for novel antidiabetic- agents, particularly ones that are more efficacious than ciutent standards of care or are better tolerated with fewer adverse effects. There is a particular need for agents which could potentially ameliorate multiple risk factors,

5 However treatment with drugs modulating a singl e molecular target is frequently unable to produce the desired level of clinical efficacy, such as defined by reducing blood glucose levels or HbAlc, a marker of long tern glucose control.

As a solution to inadequate efficacy achieved by agents modulating single targets, combination therapy is a strategy, delivered either by co-administration of 10 single agents, or by creation of fixed-dose combination products. It has been hypothesized that modulation of complex biological responses to achieve therapeutics with improved efficacy and/or safety may be achievable by targeting multiple biological targets or pathways.

Thus we have sought to identify novel combinations of biological mechanisms 15 which, when modulated simultaneously, could give desirable anti-diabetic biological responses which are additi ve or synergistic when compared to modulation of either mechanism alone, In addition, we have sought to combine these dual or multiple biological activities into single small molecule drag candidates, called designed multiple ligands or poly pharmacology compounds. Single compounds with multiple 20 biological actions which display additive, better than additive or synergistic biological responses over single agents alone could have certain advantages over fixed-dose combination therapies, and over 'co-administration of existing single agents. For example, a fixed-dose combination pill delivers two or more separate entities which have individual drug disposition and pharmacokinetic properties, and the safety of the 25 combination has to be ascertained independently from the safety of the two separate entities alone. In the case of co-administration of multiple drugs, there is a greater risk of drug-drug interactions and different dosing regimens for the individual drugs can cause poor patient compliance, and thus compromise patient benefit.

Therefore, we .have sought to identify novel combinations of biological 30 mechanisms which, when modulated simultaneously, could give desirable additive or synergistic biological responses and which are tractable to- single small molecule modulation in designed multiple ligand.suitable..for oral administration.

Pancreatic islet cells, which include beta cells, express at least 20 types of G protein-coupled receptors (GPCR), which through ligand interactions may modulate secretion of key hormones of glucose control such as insulin (Ahren- B, Nature Reviews Drug Discovery vol8 p369~385, 2009], Since enhancement of insulin 5 secretion is a known strategy for the treatment of T2DM, the identification of certain novel combinations of GPCR agonists that lead: to additive or synergistic levels of insulin secretion relative to the modulation of either GPCR alone, are a promising avenue to achieve superior efficacy,

GPRl19 {previously referred to as GPRl 16) is a GPCR identified as 10 S.NORF25 in WQ00/5Q562 which discloses: both: the human and rat receptors, US 6,468,756 also discloses the mouse receptor (accession numbers; AAN95194 (human), AAN95195 (rat) and ANN95196 (mouse)), .In humans, GPRl. 19 is expressed in the pancreas, small intestine, large intestine and stomach. The expression profile of the human GPRl19 receptor indicates its potential utility as a 15 target for the treatment of diabetes. GPRl19 agonists have been shown to stimulate the release of GLP-1 from the GI tract. In doing so, GPRl 19 agonists (1) enhance glucose-dependent insulin, release from the pancreas leading to improvements in oral glucose tolerance; (2) attenuate disease progression by increasingJJ-eell eAMP concentrations; and (3) induce weight loss possibly through GLP-1's ability to reduce 20 food intake.

GPR40 is a GPCR and has been shown to be highly expressed in the pancreatic islets -of humans (and rodents) and insulin secreting cell lines. Saturated and. unsaturated medium and long chain fatty acids have been identified as ligands for GPR40 (ItoliY et.al, Nature 422: 173-176, Briscoe et al J Biol ChemY1%\ 1 Hu 25 11311), The use of selective GPR40 agonists have confirmed the role of GPK40 in insulin secretion (Briscoe1 et al Br J Pharmacol 148 619-628 and Tan et al Diabetes 57 2211-2219). A selective GPR40 agonist has been shown to improve both fasting and postprandial glyeaemic control in type 2 diabetic patients, (Leifke et al ADA 2011 414PP),

30 The inventors have identified novel combinations of biological mechanisms which, when modulated simultaneously, give desirable anti-diabetic biological, responses which are more than additive or synergistic when compared to modulation of either mechanism alone. The invention provides novel combinations of GPRl 19

3

agonism and GPR40 agonism which, when modulated simultaneously, displays greater than additive insulin release in HIT-T15 islet-derived cells, and thus may be a useful approach to the treatment of metabolic disorders and type 2 diabetes.

The inventors have identified novel compositions comprising a GPRl 19 5 agonist and a GPR40 agonist that are useful for the treatment of metabolic disorders including type II diabetes.: The invention provides composition, dosage forms comprising the compositions and methods of treating or preventing metabolic disorders, including type 2 diabetes and diseases associated with type 2 diabetes in a mammal, said methods comprising providing the mammal with a therapeutically 10 effecti ve amount of the compositio n or dosage form,

Combining GPCR agonists may elicit a response level that is either less than (sub-additive combination), equal to (additive combination), or greater than .(synergistic combination) the simple arithmetic addition of response levels obtained with the same agents when tested separately. Alternatively the combined response i 5 may be less than that of just the single agents alone (antagonistic combination). In the context of combinations of drugs ultimately used for treatment of disease, synergistic or additive combinations are desirable. In this sense it is not possible to predict how two agonists of different GPR targets will -interact,- and it is therefore not possible to determine suitable therapeutic combinations based solely on their 20 individual activity -at their target receptor or downstream effector,

The inventors have now surprisingly discovered that when -a GPRl 19 agonist and a GPR4Q agonist are combined in a composition the response elicited is either additive, more than additive or synergistic,

25 SUMMARY OF THE INVENTION

The invention provides compositions comprising a GPRl 19 agonist and a GPR40 agonist wherein the agonists are separate agents or a single agent that is-:both a GPRl 19 agonist and a GPR4Q agonist.

30 DETAILED DECRYPTION OF THE INVENTION

The invention will now be described in detail referring to the following figures:

4

FIGURE 1 shows the more than additive effect on insulin secretion of coadministration of Compound A and Compound C at ECjo concentrations.

FIGURE 2 shows the more than additive effect on insulin secretion of co-5 admini stration of Compound A and Compound C at optimum concentrations.

FIGURE 3 shows the more than additive effect on insulin secretion of coadministration of Compound A and Compound D at EC50 concentrations.

10 FIGURE .4 shows the more than additive effect 011 insulin secretion of coadministration of Compound A and Compound D at optimum concentrations,

FIGURE 5 shows the more than additive effect on insulin secretion of coadministration of Compound B and Compound D at ECso concentrations.-

15

FIGURE 6 shows the more than additive effect on insulin secretion of coadministration of Compound B and Compound D at optimum concentrations,

FIGURE 7 shows the less than additive effect on insulin secretion of co-20 administration of GLP-1 and Compound A at optimum concentrations.

The invention provides compositions comprising a GPRl 19 agonist and a GPR40 agonist wherein the agonists are separate agents or a single agent that is both a GPRl 19 agonist and a: GPR40 agonist. In ■■certain embodiments the compositions 25 comprise a single agent that is both a GPRl 19 agonist and a GPR40 agonist, In further embodiments the compositions comprise at least one agent that is a GPRl 19 agonist and at least one further agent that is a GPR40 agonist. Compositions comprising two or more agents may be suitable for simultaneous or sequential administration of the agonists.

30 When used herein the term GPRl 19 agonist 1 to any agent that binds to a

GPRl 19 receptor and triggers afunctional response in the GPRl 19 receptor,

GPRl 19 agonists suitable for use in the present invention may be identified by methods known in the art and disclosed in, For example US7Q83933, Preferably the:

5

GPRl 19 agonist is a selective GPRl 19 agonist. For the purpose of the present invention, the following protocol may be followed to determine, whether a compound is a GPRl 19 agonist:.

A stable cell tine expressing recombinant human GPRl 19 is established by 5 any method known in the art. This cell line is used to investigate the: effect of a candidate compound on intracellular levels of cyclic AMP (cAMP), The cell monolayers are washed with phosphate buffered saline and stimulated, at 37°C for 30 min. with various concentrations of compound in stimulation buffer plus 1% DMSO, Cells are then iysed and cAMP content determined using, the Perkin Elmer 10 AlpliaScreen1 m (Ampii fled Luminescent ProximityHomogeneous Assay) cAMP kit Buffers and assay conditions are as described in the manufacturer's protocol. Compounds found to display a concentration-dependent increase in intraeellu] ar cAMP levels with an ECso of <20 pM.are considered to be GPRl 19 agonists. International Patent Applications WO 200.5/061489, WO 2006/070208, 1.5 WG 2006/067.532, WO 2006/067531, WO 2007/003960, WO 2007/003961, WO 2007/003962, WO 2007/003964, WO 2007/116229, WO 2007/116230, WO 2008/081204, WO 2008/081205, WO 2008/081206, WO.2008/081207, WO 2008/081208, WO 2009/050522, WO 2009/050523, WfO 2010/001166, WO 2010/004343, WO 2010/004344, WO 2010/004345, WO 2010/004346, 20 WO 2010/004347 and WO 2010/004348 disclose GPRl 19 receptor agonists suitable for use in the invention.

Preferred GPRl 19 agonists include:

5 - [ 1-(3-isoprop yl~ [ 1. ,2,4] oxadi azol-5-yl)-pip eridin-4-ylmethoxy ] - 2-{4-ni:ethanesul.fonylphenyl)-pyridine,

25 5-ethyl-2~ {4-[4-(4-pentazol-l -ylplienoxymefhyl)-thiazol-2-yl]-pipendin-1 -yl} -

pyrimidine,

4-[I-(2-fluoro-4-methanesulfenylphenyl)-lFl-pyra2oloj;3,4-d]pyrimidin-4 -yIoxy3-pi.peridine~l -carboxylic acid isopropyl ester,

4-[5-methyl-6-(2-methylpyridin-3-yloxy)-pyrimidin-4-yloxy]-piperidine-1 -30 carboxylic acid isopropyl .ester,

4-[6-(6-methanesuifoiiyI-2-methylpyridin-3-yiamino)-5-methyipyrimidin-4-yloxy]-piperidine-l-carboxylic acid isopropyl ester,

6

4-[6-(Cv-metlianesii!fbnyl-2-methylpyridin-3-y1a]iiino)-5-metIioxypyi'imidin-4-yloxyl piperidine-1 -carboxylic acid isopropyl ester and 4-[5-methox.y-6-(2-inethyl-6-[ 1,2,4]triazol-1 -yl-pyridiii-3-y!affiino)-pyrimidin-4-yloxy]-piperidine~l-carboxylic aeid isopropyl ester.

Further GPRl 19 agonists that may be used in the invention include a compound of formula (I), or a phamiaceutically acceptable salt thereof:

wherein one of X and Y is O and the other is N;:

R1 is -GONHR5;

R2 is hydrogen,, halo or methyl;

R3 is hydrogen or methyl;

R4 is C2-5 alkyl; and

R5 is hydrogen, Cl-3alfcyi, or C2-3 alkyl substituted by hydroxy. Examples of such agonists include the foilowing:

HO

OH

and pharmaceutically acceptable salts thereof.

Still further GPRl 19 agonists that may be used in the invention include a 10 compound of formula (I), or a pharmaceutical!}' acceptable salt thereof:

wherein Q is CH or N;

15 one of W, X and Y is N or CH and the others ai i ( H where the H may be replaced by R5 when present;

R1 is -SG2Me or -CQNHR6;

8

R2, :R3 and R4 are independently selected from hydrogen, and methyl;

n is 0, 1 or 2;

R5 is independently Cl-4 alkyl, Cl-4 alkoxy, tluoro, chloro, Cl-3 ftuoroalkyl or benzyl;

R6 is hydrogen, 3-azetidinyl, 3-pyrrolidinyi, 3-piperidinyI or 4-piperidinyl, wherein the azetidihyl, pyrrolidinyl and piperidiiiyl rings may be optionally substituted with OH, CH20H or C3H3; Cl-3 alkyl, C2-4 alkyl substituted by -N(R7.)2 and/or one or two hydroxy groups, or Cl-4 alkyl substituted by a 4- to 6-membered nitrogen-containing heterocyclic ring; and R.7 Is independently hydrogen or methyl.

When used herein the term GPR40 agonist .refers to any molecule that binds to a GPR40 receptor and triggers a functional response in the GPR40 receptor. QPR40 agonists suitable for use in the present invention may be identified by methods known in the art, Preferably, the GPR.4Q agonist is a selective GPR40 agonist. For example, the following protocol may be followed to determine whether a compound is a GP.R4Q agonist:

A stable HEK293 cell line expressing recombinant human GPR4Q receptors is established using methods known in the art. This cell line is used to investigate the effect of candidate compounds on mobilisation of intracellular calcium through the GPR40 receptor. Cells of the HEK293 cell line are cultured in growth medium overnight in black, clear bottom, poly-D-lysine-coated:, 96-well plates at 37CC with 5% C02. Growth medium is then removed from each well and replaced immediately with Fluo-4 Direct™ reagent (Invitrdgen). Cell plates are subsequently incubated for 60 min at 37°C + 5% CO2. Cell plates are then transferred to a FLIPR tetra® plate reader where test compounds are added and levels , of intracellular fluorescence measured (Ex 47Q-49.5nmj Em 515-575nm), Compounds that stimulate a calcium response in a concentration-dependent manner with an EG50 value <20jlM may be considered to be GPR40 agonists.

GPR.40 agonists are well known in the ait. For example, GPR40 agonists are disclosed in the following references, the contents of which are incorporated by reference: Shawn D. Walker, et at, Development of a Scalable iSynthesis of a GPR40 Receptor Agonist, Organic Process Research and Development, especially AMG.B37,

9

Stephen C. McReown, et at, Solid phase synthesis and SAR of small molecule agonists for the OPR4Q receptor, Bioorganic & Medicinal Chemistry Letters 17 (2007) 1584—1589, especially compounds 1- 45. Elisabeth Christiansen etal, Structure-Activity Study of Dihydroeimiamic Acids and Discovery of the Potent.

5 FFA1 (OPR4Q) Agonist TUG-469, ACS Med. Chem. Lett., 2010,1 (7), pp 345-349, especially any of the compounds in Figure 1, and any of compounds 1-31. Shawn P. Walsh et al, 3-Substituted 3-(4-Aryloxyaryl)-Propanoie Acids as GPR40 Agonists; Bioorg Med Chem Left 2011 Jun 1;21(11):3390-4, especially compounds 1-25. Gelia P. Briscoe et al, Pharmacological regulation of insulin secretion in M.1.N6 cells 10 through the fatty acid receptor GPR40: identification, of agonist and antagonist small molecules, British Journal of Pharmacology (2006) 148, 619-628, especially GW9508 of figure 1. Dulce M. Garrido etal, Synthesis and activity of small molecule GPR40 agonists; Bioorganic & Medicinal Chemistry Letters 16 (2006) 1840-1845, especially compounds 2A-14A, 2B-7B, 10B-12B and 14B-15B. 15 Changyoii Zhou et al\ Discovery of 5-aryloxy~2,4-thiazolidinediot3es as potent GPR40 agonists; Bioorganie & Medicinal Chemistry Letters 20 (2010) 1298-1301, especially the compounds in Tables 1-5. Fengbin Song, et at, Synthesis and Biological Evaluation of 3-Aryl-3-(4-phenoxy)-pTopionic Acid as a Novel Series of G Protein-Coupled Receptor 40 Agonists; J Med Chem. 2007 Jim 14;SO(12):2807-i7, especially 20 compounds .1-61, Nobuyuki Negoro et at, Discovery of TAK-875: A Potent,

Selective* and Orally Bioavailable GPR40 Agonist; ACS; Med. Chem. Lett., 2010,1 (6), pp 290-294, especially compounds 1-9.

Preferred GPR40 agonist include:

{(S)-6-[4<-(3-methanesulfonylpropoxy)-2,,6'-dimethylbiphenyl-3-y:imethoxy]-25 2,3 -dihydrob enzof Uf an-3 -yl} -acetic aci d,

(lS,laS,6:aR)-4-(2-difluorome.thoxy-4-trifIuoromethylphenoxy)-l ,1 a,6,6a-tetrahydro cyelopropa[a]indene-l -carboxylic acid,

(S)-3-[4-(4,-trifluorO:methylbiphenyl-3-ylmethoxy)-phenyl]-hex-4-3moic acid. 3-[4~(3-phenoxybenzylamino)-phenyl]-propionie acid, 30 and 5-[4-(3-Chloro-5-methyl-pyndin-2-yIoxy)-2-methyl-beBzyl]-thiaz:oIidine-

2,4-dione.

10

When used -herein the term agent refers to the active compound or to a substance that provides or delivers the active compound, e,g. a carrier or matrix comprising the active compound.

The- compositions may suitably comprise further therapeutic ingredients or 5 adjuvants.

The compositions may be conveniently presented in unit dosage form. The compositions may be prepared by any of the methods well known in the art of pharmacy. The agonists may be combined either together or separately as the active ingredient(s) in intimate admixture with a pharmaceutical carrier according to J .0 conventional pharrnaeeutieal compounding"techniques. The earner may take a wide variety of forms depending on the form of preparation desired for administration. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions may be prepared by uniformly and intimately admixing the active 15 ingredient with liquid carriers or finely divided solid earners or both, The product can then be conveniently shaped into the desired presentation.

The invention also provides dosage forms comprising a composition as described herein and a pharmaceutical^ acceptable carrier. In certain embodiments the dosage forms comprise a single agent that is both a GPRl 19 agonist and a CP 1140 20 agonist. In further embodiments the dosage forms comprise at least one agent that is a GPRTT9 agonist and at least one further agent that is a GPR40 agonist. In certain embodiments the dosage forms are in a fixed-dose combination. The dosage form maybe solid liquid or semi-solid depending on the intended route of administration. Suitable dosage forms include oral dosage forms and , parenteral injectionssnch as 25 pills, tablets, capsules, liquids, suspensions, powders, granules, solid crystals and pastes. Further dosage forms include controlled release means and delivery devices.

The dosage forms are provided for use in the treatment of a disorder selected from the group comprising diabetes, obesity, metabolic syndrome (syndrome X), impaired glucose tolerance, insulin resistance, pancreatic beta-cell insufficiency, 30 hyperglycemia, enteroendocrine cell insufficiency, glueosuria, metabolic acidosis,

cateracts, diabetic nephropathy, diabetic retinopathy, diabetic coronary artery disease, diabetic cerebrovascular disease, diabetic peripheral vascular disease, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, dyslipidemia, low HDL levels,

11

hypertension, myocardial infarction, atherosclerosis and stroke, preferably for use in the treatment, or prophylaxis of the disorder is diabetes.

The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas, Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples -of liquid carriers are sugar syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

In preparing compositions for oral dosage form any convenient pharmaceutical media may be employed. For example water, glycols, oils, alcohols, flavoring agents, preservatives, coloring .agents, and the like may be used to form oral liquid preparations such as suspensions, elixirs and solutions;: while carriers Sueh as starches, sugars, mierocrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like may be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and eapsul.es are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets may be coated by standard aqueous or nonaqueous techniques,

A tablet containing the composition may be prepared by compression or moulding, optionally with one or more accessory ingredients or adjuvants.

Compressed tablets may be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant* inert diluent,. surface active or dispersing agent. Moulded tablets may be made by moulding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent Each tablet may suitably contain from about 100 mg to about .2 g of one or more active ingredient and. each cachet or capsule may suitably contain from about 100 mg to about 2 g of one or more active ingredient

For example, a formulation intended for the oral administration to humans may contain from about 100 mg: to about 2 g of one or more of the: active agent(s), compounded with an appropriate and convenient amount of earner material which may vary from about 5 to about 95 percent of the total compositi on. Unit dosage forms will generally contain between from about 250 mg to about 2 g of the active

12

ingredient, typically 250 mg, 300 mg, 350 mgs 4()0 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 800 rag or 1000 mg.

Pharmaceutical compositions for use by parenteral administration may be prepared as solutions or suspensions of the active compounds in water, A suitable 5 surfactant can be included such as, for example, hydroxypropylceliulose, Dispersions can also be prepaid in glycerol, liquid polyethylene .glycols, and mixtures thereof m oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.

The pharmaceutical compositions must be stable: under the conditions of 10 manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as; bacteria and fungi . The carrier can be a solvent or dispersion medium containing, for example, water, efhanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.

15 In addition to the aforementioned, carrieringredients, the pharmaceutical formulations described above may include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like, Furthermore, other adjuvants can be included to render the foniiulation 20 isotonic with the blood of the intended recipient. Compositions containing a compound or pharmaceutically acceptable salts thereof for use in accordance with the present invention may also be prepared in powder or liquid concentrate form.

The present invention also provides the use of a GPRl 19 agonist or a GPR40 agonist in the preparation of a composition or dosage form as described herein. 25 The present invention also provides methods of preparing a pharmaceutical composition comprising admixing a GPRl 19 agonist and a GPR40 agonist with, at least one pharmaceutically acceptable Carrier. Included are methods that comprise admixing a single agent that is both a GPRl 19 agonist and a GPR4Q agonist with at least one pharmaceutically acceptable carrier. Also included are methods that 30 comprise admixing at least one agent that is a GPRl 19 agonist and at least one further agent that is a. GPR4Q agonist with at least one pharmaceutically acceptable carrier,

The present invention.also provides a composition as described herein for use in a method of treatment of the human or animal body.

13

The present invention also provides the use of a composition as described herein in the manufacture of a medicament for the treatment or prevention of diabetes or a condition related thereto.

5 EXAMPLES

As a model system to investigate the effects of combining islet GPCR modulators, insulin secretion is measured, using the.HIT-T.13 islet-derived cell line (Santerre RF et al,PNAS vbl'78 no? p4339-4343 1981). H1T-T15 is derived from hamster pancreatic beta cells and allows the precise measurement of insulin levels in 10 response to test, agents or combinations thereof. Functional expression of several islet GPCR genes is known to be retained in this cell line compared to primary islet cells, as: well as the effects of specifi c agonist: compounds on the rate of insulin secretion

METHODOLOGY

15 FIIT-Tl 5 cells are seeded in 96 well cell culture microti tre plates at a density of 80,000 cells/ well in 0.2ml ../ well RPMI-1640 medium supplemented with 10%

fetal calf serum. Enhanced reproducibility is obtained by avoiding edge wells and by allowing cells to settle at room temperature on the bench for 45 min.immediately following seeding, Culture plates are incubated at 37°C in a humidified .5% C02 20 atmosphere for 24 hr. All assay buffers and diluted compounds are pre-warmed to 37°C prior to addition. Plates: are maintained on. a 37°C hotplate during manipulations. Medium is discarded from .assay plate wells. Cells are washed twice with 0.2ml and pre-incubated. for 30 min in 0.1 ml ICrebs-Ringer-HEPES assay buffer (K.RHB; Poitout ¥ et al, Diabetes vo!44 p3 06-313, 1995) supplemented with 0,1% 25 BSA which is then discarded. Test compounds in single or combination doses are prepared in KRHB-BSA buffer supplemented with 5,6 mM glucose, at a final DMSO concentration of 0.5% and added to wells of the 96-well plate, which is incubated at 37GC for 30 min. This glucose concentration in the;HtT-Tl5 cell line represents a stimulatory concentration, in effect mimicking hyperglycemia, Samples containing 30 secreted insulin are collected, clarified by centrifugation at 2500 rpm for 5 minutes, and,measured for insulin .concentration using a commercial ELISA kit which includes standards of known insulin concentration (Rat insulin ELISA kit; Mercodia). Absorbarscs data from test samples is interpolated against the ELISA standard curve

14

to generate nM insulin concentrations.. Experiments are designed with duplicate wells for each test combination... Data is subtracted from adjacent vehicle control wells containing 0.5% DMSO alone, generating an increment above control insulin secretion value.

Data is provided sho wing effects of co-dosing agonist compounds of GPRl 19 and GPR40, both of which are individually functionally acti ve in HIT-T15 cells in causing an increase in insulin secretion levels.. It is: shown that the combinations of GPR40 and GPRl 19 agonists elicit, a greater' than additive interaction. This.occurs irrespective of the selection of alternative agonist .compounds for each GPCR, and irrespective of whether maximal or EC50 concentrations are used. In contrast, combining agonists to GPR119 with GLP-1 receptor or GIF receptor, generates a total response that is sub-additive. This occurs irrespective of whether maximal or EG50 concentrations are used. Although agonists of all GPCRs tested are active on their own, the outcome of combining.them reveals an unexpected beneficial response for the combination of GPRl 19 and GPR40 agonism,

The following compounds are used as tool GPCR agonists:

Compound A- GPRl 19 Agonist (GPRl I 9agA)

4-[3-(3-Fluoro-4-methanesulfonylpheri.oxy)propyl] piperidine-1 -carboxylic acid tert-hutyl ester. The synthesis of this compound is disclosed inPCT/GB2Q06/050178.

Compound B- GPRl 19. Agonist fGPRH9agB)

N -(2~Hydroxyethyl)-4 - {3- [ 1 -(3 -isopropyl-[1,2,4] oxadiazol -3 -yl)piperidin-4-■yl]propoxy}-2 methylbenisamide. The synthesis of this compound is disclosed in PCT/GB20Q8 ^050011.

o ©

/

HO'

15

Compound C- GPR40 Agonist fGPR4QagC)

identified as Cpd B in Tan CP et al, Diabetes vo!57 p2211-2219, 2008. 5 Comoonnd D- GPR40 Agonist (G.PR40agD)

OH

identified as TAK-Si75 inNegoro N et al, ACS Med, Chem. Lett. Vol.! No6 p290-294. Example 1

10 Compound A and Compound C were added to HiT-Tl 5 ceils at concentrations equal to their individual ECjo concentrations for enhancement of insulin Secretion as determined Froin a foil dose-response test in the same experiment (Compound A; .1 OOnM; Compound C: 4000 nM). Responses for the two compounds at this concentration either alone or in combination delivered simultaneously are shown -graphically in Figure 1. Increase in insulin 15 secretion in response to combined agents (0,84 nM insulin) exceeds that of simple addition of the levels achieved with respective single agents (0.55+0.22 nM insulin).

Example 2

Compound A and Compound C were added to HIT-T l 5 cells at concentrations equal 20 to their most effective concentrations for enhancement of insulin secretion as determined from a full dose-response test in the same experiment (Compound A; IDDOnM, Compound C : 15000nM). Responses for the two compounds at ibis concentration either alone or m combination delivered simultaneously are shown graphically in Figure 2, Increase in insulin secretion in response to combined agents (1.57 nM insulin) exceeds that Of simple addition of 25 the levels achieved with respective single agents (0.88+0.51 nM insulin):.

Example 3

Compound A and Compound D were added to HiT-T l 5 cells at concentrations approximately equal to their individual ECje concentrations for enhancement of insulin 30 secretion as determined from a fill! dose-response test in the same experiment (Compound A: 30nM, Compound I): 1 5QQ0nM). Responses for the two compounds at this concentration either alone or ii combination delivered simultaneously are shown graphically in Figure.3.

16

Increase in insulin secretion in response to combined agents (3 .89 nM: insulin) exceeds that of simple addition of the levels achieved- with respective single agents (0.79+1.61 -nM insulin).

Example 4

5 Compound A and Compound D were added, to: I-IIT-Tl 5 cells at concentrations equal to their most effecti ve concentrations for -enhancement of insulin secretion as determined from: a full dose-response test in the same experiment (Compound A: lOOOnM, Compound C: 30QGQnM).. Responses for the two compounds at this concentration either alone or in •combination delivered simultaneously are shown graphically in Figure 4, Increase in insulin 10 secretion in response to combined agents (6.16 nM insulin) exceeds that of simple addition of the levels achieved with respective single agents (1.28+3.13 nM insulin).

Example 5

Compound B and Compound D were added to HIT-T15 cells at concentrations 15 approximately equal'to their individual EC50 concentrations for enhancement of insulin secretion as determined from a full dose-response test in the same, experiment (Compound B: lOQnM, Compound D: .15000nM). Responses for the two compounds at this concentration either alone or in combination delivered simultaneously are shown graphically in Figure 5. Increase in insulin secretion in response to combined agents (1.78 nM insulin) exceeds that of 20 simple addition of the levels achieved with respective single agents (0.03+0.7.1 nM insulin).

Example 6

Compound B and Compound D were added to HIT-T15 cells at concentrations equal to their most effective concentrations for enhancement of insulin secretion as determined 25 from a full dose-response test in the same experiment (Compound B: lOOOnM, Compound D: BOGOOnM), .Responses for the two compounds at this concentration either alone or m combination delivered simultaneously are shown graphically in Figured, Increase in insulin secretion in response to combined agents (3.37 nM insulin) exceeds that of simple addition of the levels achieved with respective single agents (0.33+1.77 nM insulin).

30

Comparative Example 7 GLP-1 + GFK119agA- Comparative Example

GLP-1 and Compound A were added to HIT-T15 cells at 'concentrations equal to their most effective concentrations for enhancement of insulin secretion as determined from a full dose-response test in the same experiment (GLP-1; 300nM* Compound A: 1000 riM). 3 5 Responses for the two compounds at this concentration either alone or in combination delivered simultaneously are shown graphically in Figure 7, Increase in insulin secretion in

17

response to combined agents (0,55 nM insulin) is less than that of simple addition of the levels achieved with respective single agents (0.51+0.17 nM insulin).

18

Claims

CLAIMS:

1. A composition comprising; a GPR l 19 agonist and a GPR40 agonist wherein the agonists are separate agents or a single agent that is both a GPRl 19 agonist and a GPR40 agonist.

2, A composition as claimed in Claim X comprising a single agent that is both a GPRl 19 agonist and a GPR40 agonist,

3. A composition as claimed in Claim 1 comprising at least one agent that is a GPRl 19 agonist and at least one farther agent that is a GPR40 agonist,

4, A composition as claimed in any one of Claims; 1 to 3 wherein the GPRl 19 agonist is a selective GPRl 19 agonist.

5:, A composition as claimed in any one of claims 1 to 4 wherein the GPRl 19 agonist is selected from:

5-[ 1 -(3 -isopropyl-[ 1,2,4] oxadiazol- 5-yl)~pi peridin-4-ylmethoxy] ~2~ {4methattesulfonylphenyl)-pyridine)

5-ethyl-2- (4- [4-(4 -pentazol- 3 .-ylph;enoxymethyl)4hiazo]-2-yl J -piperidin-1 -y!} -pyrimidine,

4-[l-(2-fluoro-4-methanesirifonylphenyl)-l.H-pyrazo1o[3,4d]p}frimidin4-yloxy]-pipe.ridine-l -carboxylic acid isopropyl ester,

4-[5-metliyl-6-(2--metliylpyridm-3-yloxy)-pyEimidin4-yloxy'j-piperidine-l-carboxylic acid isopropyl ester,

4-[6-(6-metlianesuifonyI-2-m:ethylp>tidin-3-ylamino)-5--methylpyrimidin-4-yloxy]-piperidine-l-carboxylic acid isopropyl ester,

4-[CH6-methanesuifonyl-2-methylpyridin-3-ylamino)-5-methoxypyrimidin-4-yloxyl piperidinc-1-carboxylic acid isopropyl ester,

4-[5-metlioxy-6-{2-methyl~6-[I,2,4]triazol-l-yl-pyridin-3-ylamij]o)-pyri;midin-4-ylGxy]-piperidine-l-earboxylic acid isopropyl ester and pharmaceutically acceptable salts thereof.

19

6. A composition as claimed in any one of claims 1 to 4 wherein ....,, agonist is a. compound of formula (I), or a pharmaceutically acceptable salt thereof:

XtJ

Xo

■R4

wherein one of X and Y is: O and the other is N;

R1 is -CONHR5;

R2 is hydrogen, halo or methyl;

R3 is hydrogen or methyl ;

R4 is C2-5 alkyl; and

R5 is hydrogen, Cl~3atkyl, or G2-3alkyl substituted by hydroxy,

7. A. composition as claimed in Claim 6, wherein the compound is seleeted from

20

Q

bo-^N-

iHO

.NH

(id)

OH

(tf)

and pharmaceutically acceptable salts thereof.

8, A composition as claimed in any one of claims 1 to 4 wherein the GPRl 19 agonistis a compound of formula (I), or a pharmaceutically-acceptable-salt thereof:

wherein Q is CH or N;

one of W, X and Y is N or CH and the others are CH where the H may he replaced by

R5 when present;

Rt is -S02.Me or -CONHM;

R2, R3 and R4 are independently seleeted from hydrogen and methyl;

n is 0, 1 or 2;

R5 is independently Cl-4 alkyl, Cl-4 alkoxy, fluaro, chloro, Cl-3 fluoroalkyl or benzyl;

21

R.6 is hydrogen, 3-azetidinyl, 3-pyrrolidmyl, 3-piperidinyl or 4-pipe i i.vj.ixijf t ^ v) i.ivi v'1-LJL the azetidinyl, pyrmlidinyl and piperidinyl rings may be optionally Substituted with OB, CH20H or CH3; CI-3 alkyl, C2-4 alkyl substituted by -K(R7)2 and/or one or two hydroxy groups, or Gl-4 alkyl substituted by a 4- to 6-membered nitrogen-containing heterocyclic ring; and R7 is independently hydrogen or methyl,

9. A composition as claimed in arty one of Claims 1 to 8 wherein the GPR40 agonist is; a selective GPR40 agonist.

10. A composition as claimed in any one of Claims 1 to 9 wherein the GPR40 agonist is selected from:

{(S.)-6-[4'-(3-methan.esulfonyIpropoxy)-2'16'-dimethyl:biphetiyl-3"ylmethoxy]-2,3 -dibydrob enzofiiran- 3 -y 1}- acetic acid,

(IS. 1 aS .oaR)-4-(2-di:fluoromethoxy-4-trifluoromethylphenoxy)-l, I a,6,6a-tetrahydro cyclopropa[a]indene-1 -carboxylic acid,

(S J-3 - [4-(4,4rifluo romethylbiphenyl-3 -ylmethoxy)-phenyl] -liex-4-ynoie acid, 3~[4-(3-phenoxybenzylamino)-piienyI]-pmpionic acid, 5-[4-(3-Chloro-5~methyl-pyridin-2-yloxy)-2-methyl-benzyljduazolidir!e-2,4-

di.one and pharmaceutically acceptable salts thereof.

11. A dosage form comprising a composition as claimed in any one of Claims 1 to 10 and a pharmaceutically acceptable carrier.

12. A dosage form as claimed in Claim 11 comprising a single; agent that is both a GPRl 19 agonist and a. GPR40 agonist

13. A dosage form as claimed in Claim 11 comprising at least one agent that is a GPRl 19 agonist and at least one further agent that is a GPR40 agonist.

14. A dosage form as claimed in Claim 13 wherein the agents are in a fixed-dose combination.

15. A dosage form as: claimed in any one of Claims 11 to 14 wh— ~

dosage form.

16. A dosage form as claimed in Claim 1S which is a tablet, capsule, granule or pellet.

17. A dosage form as claimed in any one of Claims 11 to 16 for use in the treatment or prophylaxis of a disorder selected from the group comprising diabetes, obesity, metabolic syndrome (syndrome X), impaired glucose tolerance, insulin resistance, pancreatic beta-eell insufficiency, hyperglycemia, enteroendocrine cell insufficiency, glucosuria, metabolic acidosis, cateracts. diabetic nephropathy, diabetic retinopathy, diabetic coronary artery disease, diabetic cerebrovascular disease, diabetic peripheral vascular disease, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, dislypidaemia, low HDL levels, hypertension, myocardial infarction, atherosclerosis and stroke.

18. A dosage form as claimed in Claim 17 wherein the disorder is diabetes.

19. Use of a GPRl 19 agonist in the preparation of a composition as claimed in any one of Claims 1 to 10 or in the preparation of a dosage form as claimed in any one of Claims 11 to 16.

20. Use of a GPR40 agonist in the preparation of a composition as claimed in any one of Claims 1 to 10 or in the preparation of a dosage form as claimed in any one of Claims 11 to 16.

21. A method of preparing a pharmaceutical composition compri sing admixing a GPRl 19 agonist and a GPR40 agonist with at least one pharmaceutically acceptable carrier.

22. A method as claimed in Claim 21 comprising admixing a single agent that is both a GPRl 19 agonist and a GPR40 agonist with at least one pharmaceutically acceptable carrier.

23

23, A method as claimed in Claim 21 comprising admixing at least one agent that is a GPRl 19 agonist and at least one further agent that is a GPR40 agonist with at least one pharmaceutically acceptable carrier.

24; A. composition as elaimed in any one of Claims 1 to 10 or a dosage form as claimed in 'any one of Claims 11 to 16 for use in a method of treatment of the human or animal body.

25, Use of a composition as Claimed in any one of Claims 1 to 10 in the manufacture of a medicament for the treatment or prevention of diabetes or a condition related thereto.