CA2653550A1

CA2653550A1 - Crystalline form of (trans-4- [( { 5-[(3,4-difluorophenyl) amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl) acetic acid

Info

Publication number: CA2653550A1
Application number: CA002653550A
Authority: CA
Inventors: Andrew Hornby Dobson; Walter Grundy
Original assignee: Individual
Current assignee: AstraZeneca AB
Priority date: 2006-06-12
Filing date: 2007-06-08
Publication date: 2007-12-21
Also published as: CN101466690A; MX2008015762A; NO20084963L; TW200815378A; JP2009539954A; IL195348A0; AR061332A1; EP2041101A1; BRPI0712354A2; KR20090015980A; AU2007259031A1; WO2007144571A1; GB0611552D0; UY30404A1; CL2007001700A1

Abstract

Novel crystalline forms of the compound (trans-4- {4-[({5-[(3,4-difluorophenyl)amino]- 1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid are provided, together with processes for the manufacture of such forms, pharmaceutical compositions comprising them and the use of such forms in medical treatment. Formula (I)

Description

CRYSTALLINE FORM OF
(TRANS-4-[({5-[(3,4-DIFLUOROPHENYL)AMINO]-1,3,4-OXADIA7,OL-2-YL}CARBONYL)AMINO]PH
ENYL}CYCLOHEXYL)ACETIC
ACID

The present invention relates to a novel crystalline chemical compound and more particularly to novel crystalline forms of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid illustrated in Formula (I) hereinafter, which compound is an inhibitor of acetyl CoA(acetyl coenzyme A):diacylglycerol acyltransferase (DGAT1) activity. The invention also relates to processes for the manufacture of the crystalline forms, pharmaceutical compositions comprising the crystalline forms and the use of the crystalline forms in medical treatment.

O N_ ~ F
HO N~-( N
~ , ~ O/T O N F
H
(I) Our co-pending International Application, PCT/GB2005/004726 describes oxadiazole containing compounds which inhibit DGAT1. The compound of formula (I) is exemplified in that application (Example 541 of International Application, PCT/GB2005/004726; WO 2006/064189), and was obtained as a crystalline solid after recrystallisation from acetic acid, hereinafter known as Form 1. An X-ray powder diffraction pattern of Form 1 is shown in Figure 2A and thermal data for the acetic acid solvate of Form 1 is shown in Figure 2B.
We have now surprisingly and unexpectedly discovered that the Agent of formula (I) can be prepared in other crystalline forms, hereinafter referred to as Form 2 and Form 3.
Such polymorphic forms may have different solubilities and/or stabilities and/or bioavailabilities and/or different impurity profiles (minor impurities which arise for example because of the process of manufacture and/or isolation) and/or crystal forms which are easier to handle, micronise and/or form into tablets.
According to the present invention there is provided a crystalline form of (I), Form 2, having an X-ray powder diffraction pattern with peaks at 2-theta (20) =
21.4 and 22.7 .
According to the present invention there is provided a crystalline form of (I), Form 2, having an X-ray powder diffraction pattern with peaks at 2-theta (20) =
16.8, 21.4 and 22.7 .

According to the present invention there is provided a crystalline form of (I), Form 2, having an X-ray powder diffraction pattern with peaks at 2-theta (20). =
4.7, 9.3, 16.8, 21.4 and 22.7 .
According to the present invention there is provided a crystalline form of (I), Form 2, having an X-ray powder diffraction pattern with peaks at 2-theta (20) =
4.7, 9.3, 16.8, 21.4, 22.7, 23.3 and 25.9 .
According to the present invention there is provided a crystalline form of (I), Form 2, having an X-ray powder diffraction pattern with peaks at 2-theta (20) =
4.7, 9.3, 16.2, 16.8, 17.7, 18.1, 18.6, 21.4, 22.7, 23.3, 25.9 and 29.2 0 .
io The X-Ray powder diffraction pattern for Form 2 is shown in Figure 1. A
comparison between Forms 1 and 2 is shown in Figure 3 and thermal data for Form 2 is shown in Figure 4 (Form 2 in anliydrous form).
According to the present invention there is provided a crystalline form of (I), Form 2, having an X-ray powder diffraction pattern substantially as shown in Figure 1.
is According to the present invention there is also provided a crystalline form of (I), Form 3, having an X-ray powder diffraction pattern with peaks at 2-theta (20) = 8.4, 13.8 and 16.7 .
According to the present invention there is also provided a crystalline form of (I), Form 3, having an X-ray powder diffraction pattern with peaks at 2-theta (20) = 7.0, 8.4, 20 13.8, 16.7, 21.6 and 24.3 .
According to the present invention there is also provided a crystalline form of (I), Form 3, having an X-ray powder diffraction pattern with peaks at 2-theta (20) = 4.8, 5.6, 7.0, 8.4, 13.8, 16.7, 19.5, 20.0, 21.6 and 24.3 .
The X-Ray powder diffraction pattern for Form 3 is shown in Figure 5 and thermal 25 data for Form 3 is shown in Figure 6 (Form 3 in anhydrous form).
According to the present invention there is also provided a crystalline form of (I), Form 3, having an X-ray powder diffraction pattern substantially as shown in Figure 5.
Tables 1, 2 and 3 show, respectively, major peaks for Forms 2, 1 and 3.
Forms 2 and 3 obtained according to the present invention are respectively 30 substantially free from other crystal and non-crystal forms of compound (I). The term "substantially free from other crystal and non-crystal forms" shall be understood to mean that the desired crystal form contains less than 50%, preferably less than 20%, more preferably less than 10%, more preferably less than 5% of any other forms of compound (I).
The X-ray powder diffraction patterns were determined by mounting a sample of the crystalline material on Siemens single silicon crystal (SSC) wafer mounts and spreading out the sample into a thin layer with the aid of a microscope slide.
The sample was spun at 30 revolutions per minute (to improve counting statistics) and irradiated with X-rays generated by a copper long-fine focus tube operated at 40 kV and 40 mA
with a wavelength of 1.5406 Angstroms using a Bruker D5000 powder X-ray diffractometer (Bruker AXS, Banner Lane Coventry CV4 9GH). The collimated X-ray source was passed through an automatic variable divergence slit set at V20 and the reflected radiation directed through a 2 mm antiscatter slit and a 0.2 mm detector slit. The sample was exposed for 1 second per 0.02 degree 2-theta increment (continuous scan mode) over the range 2 degrees to 40 degrees 2-theta in theta-theta mode. The instrument was equipped with a scintillation counter as detector. Control and data capture was by means of a Dell Optiplex 686 NT 4.0 is Workstation operating with Diffrac+ software.
The skilled person is aware that an X-ray powder diffraction pattern may be obtained which has one or more measurement errors depending on measurement conditions (such as equipment, sample preparation or machine used). In particular, it is generally known that intensities in an X-ray powder diffraction pattern may fluctuate depending on measurement conditions and sample preparation. For example, the skilled person will realize that the relative intensity of peaks can be affected by, for example, grains above 30 microns in size and non-unitary aspect ratios, which may affect analysis of samples. The skilled person will also realize that the position of reflections can be affected by the precise height at which the sample sits in the diffractometer and the zero calibration of the diffractometer. The surface planarity of the sample may also have a small effect.
Hence a person skilled in the art will appreciate that the diffraction pattern data presented herein is not to be construed as absolute (for further information see Jenkins, R & Snyder, R.L. `Introduction to X-Ray Powder Diffractometry' John Wiley & Sons, 1996).
Therefore, it shall be understood that the crystalline forms of compound (I) are not limited to the crystals that provide X-ray powder diffraction patterns identical to the X-ray powder diffraction patterns respectively shown in Figures 1 and 5 and any crystals providing X-ray powder diffraction patterns substantially the same as that shown respectively in Figures 1 and 5 fall within the scope of the present invention.
Forms 2 and 3 may also be characterised by other analytical techniques known in the art such as Differential scanning calorimetry (DSC) and Thermogravimetric analysis (TGA) according to standard methods, such as, for example those described in Hohne, G.
W. H. et al (1996), Differential Scanning Calorimetry, Springer, Berlin.
It will be understood that the onset/peak teinperature values of the DSC and the weight loss values for TGA may vary slightly from one machine to another or from one sample to another, and so the values quoted in the thermal trace figures are not to be construed as absolute. The techniques used are described in more detail below.
Differential Scanning Calorimetry was performed using analytical instrument Mettler DSC820e. Typically less than 5mg of material contained in a 40 l aluminium pan fitted with a pierced lid was heated over the temperature range 25 C to 325 C
at a constant heating rate of 10 C per minute. A purge gas using nitrogen was used - flow rate 100m1 per minute.
Thermogravimetric Analysis was performed using analytical instrument: Mettler TG85 1. Typically between 3 and 12 mg of material contained in a 70 1 alox (aluminium oxide) crucible was heated over the temperature range 25 C to 325 C at a constant heating rate of 10 C per minute. A purge gas using helium was used - flow rate 50m1 per minute.
Form 2 and Form 3 may be crystallised as illustrated in the accompanying examples.
Therefore in a further aspect of the present invention is provided a process for the manufacture of crystalline Form 2 of a compound of formula (I) which comprises preparation and isolation of compound (I) from water and methanol, then stirring a suspension of the obtained solid in water (for example for 1-5 days, such as 3 days), and isolation of the obtained solid (for example by filtration, optionally washing with water, and drying).

In a further aspect of the invention there is provided a process for the manufacture of crystalline Form 2 of a compound of formula (I) which comprises stirring a suspension of compound (I) in water.
In a further aspect of the present invention is provided a process for the manufacture of crystalline Form 3 of a compound of formula (I) which comprises stirring a mixture in acetonitrile of compound (I) Forms 1 and 2 (for example stirred for 1-5 days, such as 3 days) at elevated temeprature (for example, 30 C to 70 C, particularly at, or about, 50 C), and isolation of the obtained solid (for example by filtration, optionally washing with a suitable solvent, and drying).
According to a further feature of the invention there is provided a novel crystalline form of a compound of formula (I) obtainable by any of the methods or Examples disclosed herein.
According to a further aspect of the invention there is provided a pharmaceutical coinposition which comprises Form 2 or Form 3, as defined hereinbefore or hereinafter, in io association with a pharmaceutically-acceptable excipient or carrier.
The compositions of the invention may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular or intramuscular dosing or as a suppository for rectal dosing).
The compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and/or preservative agents.
Suitable pharmaceutically acceptable excipients for a tablet formulation include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and disintegrating agents such as corn starch or algenic acid; binding agents such as starch; lubricating agents such as magnesium stearate, stearic acid or talc; preservative agents such as ethyl or propyl p-hydroxybenzoate, and anti-oxidants, such as ascorbic acid. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case, using conventional coating agents and procedures well known in the art.

Compositions for oral use may be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin, or olive oil.
Aqueous suspensions generally contain the active ingredient in finely powdered form together with one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents such as lecithin or condensation products of an alkylene oxide with fatty acids (for example polyoxethylene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation is products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid), colouring agents, flavouring agents, and/or sweetening agents (such as sucrose, saccharine or aspartame).
Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil (such as arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin). The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set out above, and flavouring agents may be added to provide a palatable oral preparation.
These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water generally contain the active ingredient together with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.

Additional excipients such as sweetening, flavouring and colouring agents, may also be present.
The pharmaceutical compositions of the invention may also be in the fonn of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as for example liquid paraffin or a mixture of any of these.
Suitable emulsifying agents may be, for example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soya bean, lecithin, an esters or partial esters derived from fatty acids and hexitol anhydrides (for example sorbitan monooleate) and condensation products of the said partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavouring and preservative agents.
Syrups and elixirs may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, and may also contain a demulcent, preservative, flavouring and/or colouring agent.
The pharmaceutical compositions may also be in the form of a sterile injectable aqueous or oily suspension, which may be formulated according to known procedures using one or more of the appropriate dispersing or wetting agents and suspending agents, which have been mentioned above. A sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example a solution in 1,3-butanediol.
Compositions for administration by inhalation may be in the form of a conventional pressurised aerosol arranged to dispense the active ingredient either as an aerosol containing finely divided solid or liquid droplets. Conventional aerosol propellants such as volatile fluorinated hydrocarbons or hydrocarbons may be used and the aerosol device is conveniently arranged to dispense a metered quantity of active ingredient.

For further information on formulation the reader is referred to Chapter 25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990 and Particle size reduction for improvement of oral bioavailability of hydrophobic drugs: G.G. Liversidge, K.C. Cundy, International J.
Pharmaceutics, 125 (1995), 91-97 (relevant sections of which are hereby incorporated by reference).

The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans will generally contain, for example, from 0.5 mg to 2 g of active agent compounded with an appropriate and convenient amount of excipients which may vary from about 5 to about 98 percent by weight of the total composition.
Dosage unit forms will generally contain about 1 mg to about 500 mg of an active ingredient. For further information on Routes of Adininistration and Dosage Regimes the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch;
io Chairman of Editorial Board), Pergamon Press 1990.
According to a further aspect of the present invention there is provided Form 2 or Form 3 as defined hereinbefore or hereinafter for use in a method of treatment of the liuman or animal body by therapy.
We have found that compounds of the present invention inhibit DGATl activity and are therefore of interest for their effects on triglyceride synthesis, and/or weight loss and/or blood-glucose lowering. Information on the role of DGAT1 is contained in our International Application W02005/044250 and references therein.
A further feature of the present invention is Form 2 or Form 3 for use as a medicament. Conveniently this is Form 2 or Form 3 for use as a medicament for producing an inhibition of DGATI activity in a warm-blooded animal such as a human being.
Particularly this is Form 2 or Form 3 for use as a medicarnent for treating diabetes mellitus and/or obesity in a warm-blooded animal such as a human being.
Thus according to a further aspect of the invention there is provided the use of Form 2 or Form 3 in the manufacture of a medicament for use in the production of an inhibition of DGATl activity in a warm-blooded animal such as a human being.
Thus according to a further aspect of the invention there is provided the use of Form 2 or Form 3 in the manufacture of a medicament for use in the treatment of diabetes mellitus and/or obesity in a warm-blooded animal such as a human being.
According to a further aspect of the invention there is provided a pharmaceutical composition which comprises Form 2 or Form 3 in association with a pharmaceutically-acceptable excipient or carrier for use in producing an inhibition of DGATl activity in an warm-blooded animal, such as a human being.

According to a further aspect of the invention there is provided a pharmaceutical composition which comprises Form 2 or Form 3 in association.with a pharmaceutically-acceptable excipient or carrier for use in the treatment of diabetes mellitus and/or obesity in an warm-blooded animal, such as a human being.
According to a further feature of the invention there is provided a method for producing an inhibition of DGAT1 activity in a warm-blooded animal, such as a human being, in need of such treatment which comprises administering to said animal an effective amount of Form 2 or Form 3 defined hereinbefore or hereinafter.
According to a further feature of the invention there is provided a method of io treating diabetes mellitus and/or obesity in a warm-blooded animal, such as a human being, in need of such treatinent which comprises administering to said animal an effective amount of Form 2 or Form 3 as defined hereinbefore or hereinafter.
As stated above the size of the dose required for the therapeutic or prophylactic treatment of a particular disease state will necessarily be varied depending on the host treated, the route of administration and the severity of the illness being treated. Preferably a daily dose in the range of 1-50 mg/kg is employed. However the daily dose will necessarily be varied depending upon the host treated, the particular route of administration, and the severity of the illness being treated. Accordingly the optimum dosage may be determined by the practitioner who is treating any particular patient. As stated above compounds defined in the present invention are of interest for their ability to inhibit the activity of DGAT1. A compound of the invention may therefore be useful for the prevention, delay or treatment of a range of disease states including diabetes mellitus, more specifically type 2 diabetes mellitus (T2DM) and complications arising there from (for example retinopathy, neuropathy and nephropathy), impaired glucose tolerance (IGT), conditions of impaired fasting glucose, metabolic acidosis, ketosis, dysmetabolic syndrome, arthritis, osteoporosis, obesity and obesity related disorders, (which include peripheral vascular disease, (including intermittent claudication), cardiac failure and certain cardiac myopathies, myocardial ischaemia, cerebral ischaemia and reperfusion, hyperlipidaemias, atherosclerosis, infertility and polycystic ovaiy syndrome); the compounds of the invention may also be useful for muscle weakness, diseases of the skin such as acne, Alzheimer's disease, various immunomodulatory diseases (such as psoriasis), HIV infection, inflarmnatory bowel syndrome and inflammatory bowel disease such as Crohn's disease and ulcerative colitis.
In particular, the compounds of the present invention are of interest for the prevention, delay or treatment of diabetes mellitus and/or obesity and/or obesity related disorders. In one aspect, the compounds of the invention are used for prevention, delay or treatment of diabetes mellitus. In another aspect, the compounds of the invention are used for prevention, delay or treatment of obesity. In a further aspect, the compounds of the invention are used for prevention, delay or treatment of obesity related disorders.
The inhibition of DGAT1 activity described herein may be applied as a sole therapy or in combination with one or inore other substances and/or treatments for the indication being treated. Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate administration of the individual components of the treatment. Simultaneous treatment may be in a single tablet or in separate tablets. For example such conjoint treatment may be beneficial in the treatment of metabolic syndrome [defined as abdominal obesity (as measured by waist circumference against ethnic and is gender specific cut-points) plus any two of the following:
hypertriglyceridemia (> 150 mg/dl; 1.7mmo1/1); low HDLc (<40 mg/dl or <1.03mmo1/1 for men and <50 mg/dl or 1.29 mmol/1 for women) or on treatment for low HDL (high density lipoprotein);
hypertension (SBP > 130 mmHg DBP > 85 mmHg) or on treatment for hypertension; and hyperglycemia (fasting plasma glucose > 100 mg/dl or 5.6 mmol/1 or impaired glucose tolerance or pre-existing diabetes mellitus) - International Diabetes Federation & input from IAS/NCEP].
Such conjoint treatments may include the following main categories:
1) Anti-obesity therapies such as those that cause weight loss by effects on food intake, nutrient absorption or energy expenditure, such as orlistat, sibutramine and the like.
2) Insulin secretagogues including sulphonylureas (for example glibenclamide, glipizide), prandial glucose regulators (for example repaglinide, nateglinide);
3) Agents that improve incretin action (for example dipeptidyl peptidase IV
inhibitors, and GLP-1 agonists);

4) Insulin sensitising agents including PPARgamma agonists (for example pioglitazone and rosiglitazone), and agents with combined PPARalpha and gamma activity;

-Il-5) Agents that modulate hepatic glucose balance (for example metformin, fructose 1, 6 bisphosphatase inhibitors, glycogen phopsphorylase inhibitors, glycogen synthase kinase inhibitors, glucokinase activators);
6) Agents designed to reduce the absorption of glucose from the intestine (for example acarbose);
7) Agents that prevent the reabsorption of glucose by the kidney (SGLT
inhibitors);
8) Agents designed to treat the complications of prolonged hyperglycaemia (for example aldose reductase inhibitors);
9) Anti- dyslipidaemia agents such as, HMG-CoA reductase inhibitors (eg statins);
PPAR alpha-agonists (fibrates, eg gemfibrozil); bile acid sequestrants (cholestyramine);
cholesterol absorption inhibitors (plant stanols, synthetic inhibitors); bile acid absorption inhibitors (IBATi) and nicotinic acid and analogues (niacin and slow release formulations);
10) Antihypertensive agents such as beta-blockers (eg atenolol, inderal); ACE
inhibitors (eg lisinopril); Calcium antagonists (eg. nifedipine); Angiotensin receptor antagonists (eg candesartan), alpha-antagonists and diuretic agents (eg.
furosemide, benzthiazide);

11) Haemostasis modulators such as, antithrombotics, activators of fibrinolysis and antiplatelet agents; thrombin antagonists; factor Xa inhibitors; factor VIIa inhibitors);
antiplatelet agents (eg. aspirin, clopidogrel); anticoagulants (heparin and Low molecular weight analogues, hirudin) and warfarin;

12) Agents which antagonise the actions of glucagon; and 13) Anti-inflammatory agents, such as non-steroidal anti-inflammatory drugs (eg.
aspirin) and steroidal anti-inflammatory agents (eg. cortisone).
The utility of the compound forms of the invention may be demonstrated by the following;
Human Enzyme Assay The in vitf=o assay to identify DGAT1 inhibitors uses human DGAT1 expressed in insect cell membranes as the enzyme source (Proc. Natl. Acad. Sci. 1998, 95, 13018-13023). Briefly, sf9 cells were infected with recombinant baculovirus containing human DGAT1 coding sequences and harvested after 48 h. Cells were lysed by sonication and membranes isolated by centrifuging at 28000 rpm for 1 h at 4 C on a 41%
sucrose gradient. The membrane fraction at the interphase was collected, washed, and stored in liquid nitrogen.
DGAT1 activity was assayed by a modification of the method described by Coleman (Methods in Enzymology 1992, 209, 98-102). Compound at 1-10 M was incubated with 0.4 g membrane protein, 5 mM MgC12, and 10 0 M 1,2 dioleoyl-sn-glycerol in a total assay volume of 200 l in plastic tubes. The reaction was started by adding 14C oleoyl coenzyme A(30 M final concentration) and incubated at room temperature for 30 minutes. The reaction was stopped by adding 1.5 mL
2-propanol:heptane:water (80:20:2). Radioactive triolein product was separated into the organic phase by adding 1mL heptane and 0.5 mL 0.1 M carbonate buffer pH 9.5.

activity was quantified by counting aliquots of the upper heptane layer by liquid scintillography. In this test the compound of formula (I) has an ICso of 2.5 nM.
The ability of the compound of formula (I), and corresponding pharmaceutically-acceptable acid salts, to inhibit DGAT1 may further be demonstrated is employing the following whole cell assays 1) and 2):
1) Measurement of Triglyceride Synthesis in 3T3 Cells Mouse adipocyte 3T3 cells were cultured to confluency in 6 well plates in new born calf serum containing media. Differentiation of the cells was induced by incubating in medium containing 10% foetal calf serum, 1 p.g/mL insulin, 0.25 M dexamethasone and 0.5 mM
isobutylmethyl xanthine. After 48 h the cells were maintained in medium containing 10%
foetal calf serum and 1 g/mL insulin for a further 4-6 days. For the experiment, the medium was changed to serum-free medium and the cells pre-incubated with compound solubilised in DMSO (final concentration 0.1%) for 30 minutes. De novo lipogenesis was measured by the addition of 0.25 mM sodium acetate plus 1 Ci/mL 14C-sodium acetate to each well for a further 2 h (J. Biol. Chem., 1976, 251, 6462-6464). The cells were washed in phosphate buffered saline and solubilised in 1% sodium dodecyl sulfate. An aliquot was removed for protein determination using a protein estimation kit (Perbio) based on the method of Lowry (J. Biol. Chem., 1951, 193, 265-275). The lipids were extracted into the organic phase using a heptane:propan-2-ol:water (80:20:2) mixture followed by aliquots of water and heptane according to the method of Coleman (Methods in Enzymology, 1992, 209, 98-104). The organic phase was collected and the solvent evaporated under a stream of nitrogen. The extracts solubilised in iso-hexane:acetic acid (99:1) and lipids separated via normal phase high performance liquid chromatography (HPLC) using a Lichrospher diol-5, 4 x 250 mm column and a gradient solvent system of iso-hexane:acetic acid (99:1) and iso-hexane:propan-2-ol:acetic acid (85:15:1), flow rate of 1 mL/minute according to the method of Silversand and Haux (1997). Incorporation of radiolabel into the triglyceride fraction was analysed using a Radiomatic Flo-one Detector (Packard) connected to the HPLC machine.
2) Measurement of Triglyceride Synthesis in MCF7 Cells Human mammary epithelial (MCF7) cells were cultured to confluency in 6 well plates in foetal calf serum containing media. For the experiment, the medium was changed to serum-free medium and the cells pre-incubated with compound solubilised in DMSO (final concentration 0.1%) for 30 minutes. De novo lipogenesis was measured by the addition of 50 M sodium acetate plus 3 Ci/mL 14C-sodium acetate to each well for a further 3 h (J.
Biol. Chem., 1976, 251, 6462-6464). The cells were washed in phosphate buffered saline is and solubilised in 1% sodium dodecyl sulfate. An aliquot was removed for protein determination using a protein estimation kit (Perbio) based on the method of Lowry (J.
Biol. Chem., 1951, 193, 265-275). The lipids were extracted into the organic phase using a heptane:propan-2-ol:water (80:20:2) mixture followed by aliquots of water and heptane according to the method of Coleman (Methods in Enzymology, 1992, 209, 98-104).
The organic phase was collected and the solvent evaporated under a stream of nitrogen. The extracts solubilised in iso-hexane:acetic acid (99:1) and lipids separated via normal phase high performance liquid chromatography (HPLC) using a Lichrospher diol-5, 4x 250 mm column and a gradient solvent system of iso-hexane:acetic acid (99:1) and iso-hexane:propan-2-ol:acetic acid (85:15:1), flow rate of 1 mL/minute according to the method of Silversand and Haux (J. Chromat. B, 1997, 703, 7-14). Incorporation of radiolabel into the triglyceride fraction was analysed using a Radiomatic Flo-one Detector (Packard) connected to the HPLC machine.
The invention will now be illustrated by the following Examples.
Examples The invention will now be illustrated by the following Examples in which, unless stated otherwise:
(i) temperatures are given in degrees Celsius ( C); operations were carried out at room or ambient temperature, that is, at a temperature in the range of 18-25 C and under an atmosphere of an inert gas such as argon;
(ii) organic solutions were dried over anhydrous magnesium sulfate;
evaporation of solvent was carried out using a rotary evaporator under reduced pressure (600-4000 Pa;
4.5-30 mmHg) with a bath temperature of up to 60 C;
(iii) chromatography means flash chromatography on silica gel; where a Biotage cartridge is referred to this means a cartridge containing KP-SILTM silica, 60A, particle size 32-63 mM, supplied by Biotage, a division of Dyax Corp., 1500 Avon Street Extended, Charlottesville, VA 22902, USA;
(iv) in general, the course of reactions was followed by TLC and reaction times are given for illustration only;
(v) yields are given for illustration only and are not necessarily those which can be obtained by diligent process development; preparations were repeated if more material was required;
is (vi) wliere given, NMR data (H) is in the form of delta values for major diagnostic protons, given in parts per million (ppm) relative to tetramethylsilane (TMS), determined at 300 or 400 MHz (unless otherwise stated) using perdeuterio dimethyl sulfoxide (DMSO-d6) as solvent, unless otherwise stated; peak multiplicities are shown thus: s, singlet; d, doublet; dd, doublet of doublets; dt, doublet of triplets; dm, doublet of multiplets; t, triplet, q, quartet; m, multiplet; br, broad;
(vii) chemical symbols have their usual meanings; SI units and symbols are used;
(viii) solvent ratios are given in volume : volume (v/v) terms;
(ix) mass spectra (MS) (loop) were recorded on a Micromass Platform LC
equipped with HP 1100 detector; unless otherwise stated the mass ion quoted is (MH+);
(x) LCMS (liquid chromatography-mass spectrometry) were recorded on a system comprising Waters 2790 LC equipped with a Waters 996 Photodiode array detector and Micromass ZMD MS, using a Phenomenex Gemini 5u C 18 110A 50x2 mm column and eluting with a flow rate of 1.1 ml/min with 5% (Water/Acetonitrile (1:1) + 1%
formic acid) and a gradient increasing from 0-95% of acetonitrile over the first 4 minutes, the balance (95-0%) being water and where HPLC Retention Times are reported these are in minutes in this system unless otherwise stated; unless otherwise stated the mass ion quoted is (MH);

(xi) where phase separation cartridges are stated then ISOLUTE Phase Separator 70m1 columns, supplied by Argonaut Technologies, New Road, Hengoed, Mid Glamorgan, CF82 8AU, United Kingdom, were used;
(xii) where a SiliCycle cartridge is referred to this means a cartridge containing Ultra Pure Silica Gel particle size 230-400 mesh, 40 -63 um pore size, supplied by SiliCycle Chemical Division, 1200 Ave St-Jean-Baptiste, Suite 114, Quebec City, Quebec, G2E 5E8, CANADA;
(xiii) where an Isco Companion is referred to then a Combiflash companion chromatography instrument, supplied by ISOC Inc. Address Teledyne ISOC Inc, io Superior Street, Lincoln, NE 68504, USA, was used;
(xiv) where a microwave is referred to this means a Biotage Initiator sixty or Smith Creator microwave, supplied by Biotage, a division of Dyax Corp., 1500 Avon Street Extended, Charlottesville, VA 22902, USA;
(xv) where GCMS is referred to then a Gas Chromatography -Mass Spectrometry analysis was carried out on a QP-2010 GC-MS system fitted with an AOC 20i autosampler and controlled by `GCMS solutions' software, version 2.0, supplied by Shimadzu, Milton Keynes, MK12 5RE, UK; the GC column was a DB-5MS of length 25 m, 0.32 mm i.d.
with a film thickness of 0.52 m supplied by J & W Scientific, Folsom, CA, USA;
(xvi) where a centrifuge is referred to this means a Genevac EZ-2plus, supplied by Genevac Limited, The Soveriegn Centre, Farthing Road, Ipswich, IP1 5AP, UK;
(xvii) where chiral chromatography is referred to this is carried generally out using a 20 m Merck 50mm Chiralpak AD column, (Chiral Stationary Phase supplied by Chiral Technologies Europe, Parc d'Innovation, Bd. Gonthier d'Andernach, 67404 Illkirch Cedex, France), using MeCN/2-propanol/AcOH (90/10/0.1) as eluent, flow rate 80 mL/min, wavelength 300nm, using a Gilson prep HPLC instrument (200m1 heads);
(xviii) melting points were determined using a Buchi 530 apparatus and are uncorrected;
(xix) where equivalents (equiv) are referred to, they are intended to mean molar equivalents.
(xx) The following abbreviations may be used below or in the process section hereinbefore or hereinafter:
Eta0 or ether diethyl ether DMF dimethylformamide DCM dichloromethane DME 1,2-dimethoxyethane MeOH methanol EtOH ethanol Ha0 water TFA trifluoroacetic acid THF tetrahydrofuran DMSO dimethylsulfoxide HOBt 1-hydroxybenzotriazole EDCI (EDAC) 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide hydrochloride DIPEA diisopropylethylamine DEAD diethyl azodicarboxylate EtOAc ethyl acetate is NaHCO3 sodium bicarbonate / sodium hydrogencarbonate K3P04 potassium phosphate PS polymer supported BINAP 2,2'-bis(diphenylphosphino)-1,1'binaphthyl Dppf 1,1' -bis(diphenylphosphino)ferrocene dba dibenzylidineacetone PS-CDI polymer supported carbonyldiimidazole CH3CN or MeCN acetonitrile h hour min minute HATU O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexofluorophosphate NaOH sodium hydroxide AcOH acetic acid DMA dimethyl acetamide nBuLi n-butyl lithium MgSO4 magnesium sulfate NaZSO4 sodium sulfate CDC13 deutero chloroform CD3OD per-deuterated methanol Boc tert-butoxycarbonyl Example 1: Form 2(trans-4-{4-(({5-f(3,4-Difluoronhenyl)aminol-1,3,4-oxadiazol-yl}carbonyl)aminolphenyl}cyclohexyl)acetic acid ~~ NN
HO O N
~ O O N F
H
Lithium hydroxide monohydrate (10 equivalents) in water (2.23 L per mole of Intermediate 1) was added to a stirred suspension of methyl (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetate (Intermediate 1; 1 equivalent) in methanol (9.3 8 L per mole of Intermediate 1). The reaction mixture was stirred at 30 C for 2 hours then cooled to 0 C and acidified to pH2 with concentrated hydrochloric acid (keeping the temperature below 10 C). The resulting white precipitate was filtered, washed with water and methanol and then dried under vacuum at 50 C to give the title compound as a solid (82% yield). This solid was then suspended in water (approximately 28m1 per g of compound) with stirring, the suspension was then stirred for 3 days. The solid was then filtered off and washed with fresh water (slow filtration). The resulting white solid was dried to constant weight at 50 C in a high vacuum drying oven and analysed (see Figures 1, 3 & 4).

Intermediate 1: methyl (trafas-4-{4- f ({5-[(3,4-difluorouhenyl)aminol-1,3,4-oxadiazol-2-yl}carbonyl)aminolphenyl} cyclohexyl)acetate 3,4-Difluoroisothiocyanate (1.2 equivalent), was added to a stirred suspension of methyl [trans-4-(4-{[hydrazino(oxo)acetyl]amino}phenyl)cyclohexyl]acetate (Intermediate 2, 1 equivalent) in DMA (approximately 5.3 litres per mole of Intermediate 2) and the mixture was heated to 45 C and stirred for 2 hours. EDAC (1.2 equivalent) was added and the resulting mixture was heated to 85 C and stirred for 3 hours. Water (approximately 4.3 litres per mole of Intermediate 2) was added. The precipitate was filtered off and washed with water, then dried under vacuum to give the title compound as a yellow solid.

Intermediate 2: Methyl f trans-4-(4-{ f hydrazino(oxo)acetyll amino}phenyl)cvclohexyll acetate C~~ N N-NH2 MeO2C ~ l ~ ,, ~\
O O
i) Methyl 2- f 4-(4-hydroxyphenyl)cyclohexylidenel acetate OMe ~ 0 HO C/

Trimethyl phosphonoacetate (170 mL, 1.05 inol) was added dropwise to a stirred suspension of sodium hydride (60 % in mineral oil, 27.5 g, 1.14 mol) in THF
(3.5 L) cooled to 12 C. After coinpletion of addition, the reaction mixture was allowed to warm to io ambient temperature and stirred for 1 h. In a separate vessel, N,N-tetramethyl guanidine (144 mL, 1.14 mol) was added to a suspension of 4-(4-hydroxyphenyl)cyclohexan-l-one (235 g, 0.95 mol) in THF (1.2 L) and the reaction mixture was stirred for 1 h at ambient temperature. The phosphonoacetate mixture was cooled to 10 C and the guanidine solution added slowly, controlling the temperature between 8 and 12 C until no residual exotherm was observed. The temperature was allowed to rise to ambient temperature and the reaction mixture was stirred for 16 h. The mixture was partitioned between a dilute aqueous solution of ammonium chloride (2.4 L) and ethyl acetate (2.4 L). The aqueous phase was separated and extracted with ethyl acetate (1.2 L). The organic phases were combined and washed with brine (2.4 L), dried (MgSO4) and concentrated in vacuo to leave an off-white solid. The solid was slurried in a mixture of ether and hexane (2:1; 470 mL), filtered and washed with a mixture of ether and isohexane (2:1; 240 mL) to give the product as a white solid (285 g, 94%). 'H NMR S 1.35 - 1.55 (2H, m), 1.85 -2.05 (4H, m), 2.25-2.40 (2H, m), 2.65 - 2.75 (IH, m), 3.60 (3H, s), 3.80 (1H, m), 6.66 (2H, d), 6.99 (2H, d), 9.10 (1 H, s) ii) traits-Methyl2-[4-(4-hydroxyphenyllcyclohexyllacetate ,,,=~OMe ~ O

HO ~ /

10% Palladium on carbon (50% water wet, 6.9 mmol) was added to methyl2-[4-(4-hydroxyphenyl)cyclohexylidene]acetate (100 g, 0.41 mol) in dry THF (400 mL).
The reaction mixture was heated at 30 C under a hydrogen atmosphere (2 bar). The mixture was filtered over Celite to leave a solid, which was waslzed with THF (50 mL).
The THF
solution was concentrated in vacuo to leave a residue, which was washed with ethyl acetate. The crude mixture was dissolved in hot ethyl acetate (100 mL) and then cooled to ambient temperature. After chilling with ice water, the precipitate was filtered and washed io with ethyl acetate (50 mL) to give the title compound as a solid (42 g, 42%). 'H NMR S
1.02 - 1.17 (2H, in), 1.31 - 1.46 (2H, m), 1.66 - 1.82 (5H, m), 2.23 (2H, d), 2.28 - 2.38 (1H, m), 3.63 (3H, s), 6.66 (2H, d), 6.99 (2H, d), 9.10 (1H, s).

iii) trafzs-Methyl2-f4-(4-aminophenyl)cyclohexyllacetate ,.%--~OMe ~ O

1~
HZN
A solution of trans-methyl 2-[4-(4-hydroxyphenyl)cyclohexyl] acetate (2.82 g, 11.4 mmol) and diisopropylethylamine (2.32 mL, 13.3 mmol) in DCM (40 mL) was cooled to 4 C and trifluoromethanesulfonyl chloride (1.42 mL, 13.3 mmol) was added over 30 mins, maintaining the temperature below 6 C. The reaction mixture was stirred at 4 C
for 45 mins and then warmed to 15 C. Stirring was stopped and the reaction mixture was left for 16 h. The mixture was poured into ice water (18 mL), the layers separated and the aqueous layer extracted with DCM (7 mL). The combined organic phases were washed with a 2N
aqueous solution of sodium hydroxide (2 mL) and then brine (9 mL), dried (MgSO4) and concentrated in vacuo to leave the intermediate triflate as a yellow solid (4.59 g, 106%), which was used with no further purification.
The intermediate triflate (12 g, 32 mmol) was added to a mixture of cesium carbonate (14.4 g, 44 mmol), palladium acetate (0.43 g, 1.9 mmol), BINAP (1.2 g, 1.9 mmol), and benzophenone imine (7.9 mL, 47 mmol) in THF (200 mL). Stirring was started and the vessel was evacuated and purged with nitrogen 5 times. The stirred mixture was heated to reflux for 16 h. The reaction mixture was cooled to ambient temperature and concentrated in vacuo to leave a residue. The residue was partitioned between ether (360 s inL) and water (210 mL) and the layers were separated. The aqueous layer was extracted with ether (3 x 360 mL) and the combined organic layers were dried (MgSO4) and concentrated in vacuo to leave a crude yellow oil which was used with no further purification.
The crude imine (21 g, 51 mmol) was dissolved in methanol (300 mL) and the solution cooled to 4 C. A 1 M solution of hydrochloric acid (100 mL) was added slowly, maintaining the temperature below 7 C. The suspension was warmed to ambient temperature over 16 h. The methanol was removed in vacuo and the resulting mixture diluted with water (100 mL). The aqueous mixture was washed with ether (2 x 30 mL) and the combined organic layer washed with a 1 M solution of hydrochloric acid (2 x 30 mL).
is The combined aqueous layers were basified to pH9 with a 10% aqueous solution of sodium carbonate to give a precipitate. Ethyl acetate (3 x 200 mL) was added and the layers were separated. The combined organic layers were dried (MgSO4) and concentrated in vacuo until a precipitate formed. The mixture was cooled, filtered and washed with hexane (20 mL) to give the product as a pale yellow solid. The filtrates were concentrated in vacuo to give additional product, which were combined, concentrated in vacuo and purified by column chromatography, using a gradient of 10 - 50 % EtOAc and isohexane as eluent to give the product as a yellow solid (5.1 g, combined yield 65% over 2 steps).
'H NMR
(CDC13) 6 0.98 - 1.06 (2H, m), 1.33 - 1.42 (2H, m), 1.72 - 1.81 (5H, m), 2.16 -2.18 (2H, m), 2.28 - 2.34 (1H, m), 3.61 (3H, s), 6.68 (2H, d), 6.96 (2H, d).

iv) Methyl ({4-ftraizs-4-(2-methoxy-2-oxoethyl)cyclohexyllphenyl}aminol(oxo)-acetate H OMe - N
MeOZC ~~
// O

Methyl chloro(oxo)acetate (0.842 niL) was added to a stirred solution of trans-methyl2-[4-(4-aminophenyl)cyclohexyl]acetate (1740 mg) and pyridine (0.689 mL) in DCM
(50 mL) at 0 C. After the addition was complete the mixture was allowed to warm to ambient temperature and stirred for 64 hours. The solution was diluted with DCM (100 mL), washed with water (50 mL) and brine (50 mL), then dried and concentrated in vacuo to give the title compound (2267 mg) as a solid; 'H NMR 8 7.60 (2H, d), 7.18 (2H, d), 3.83 (3H, s), 3.58 (3H, s), 2.58-35 (1H+DMSO, m), 2.21 (2H, d), 1.75 (5H, m), 1.43 (2H, m), s 1.12 (2H, m); MS mle (M-H)" 332.

iv) Hydrazine hydrate (0.361 mL) was added to a stirred solution of methyl ({4-[tf=ans-4-(2-methoxy-2-oxoethyl)cyclohexyl]phenyl}amino)(oxo)acetate (2260 mg) in EtOH
(50 mL). The mixture was stirred for 1 hour. The precipitate was filtered off, washed with Et20, and dried under vacuum overnight to give the title compound (Intermediate 2, 1845 mg) as a solid; 1H NMR 8 10.44 (1H, s), 10.20 (1H, s), 7.70 (2H, d), 7.21 (2H, d), 4.60 (2H, s), 3.60 (3H, s), 2.42 (1H, m), 1.79 (5H, m), 1.45 (2H, m), 1.11 (2H, m);
MS m/e MH+ 334.

Example 2 : Form 3(trans-4-14-1({5-f(3,4-Difluoronhenyl)aminol-1,3,4-oxadiazol-yl}carbonyl)aminolphenyl}cyclohexyl)acetic acid Form 2 (as prepared in Example 1; approximately 20mg) was placed in a small vial containing a magnetic stirrer. A small amount of Form 1 was added as seed and acetonitrile (approximately lml) added. The vial was sealed with a lid, and placed on a hotplate stirrer at 50 C for 3 days, stirring throughout. After 3 days, the vial was removed from the stirrer, the lid taken off and any remaining solvent allowed to evaporate. The resulting Form 3 solid was analysed (see Figures 5 & 6 and Table 3).

The Form 1 material was prepared as follows...
Methyl 2-[4-[4-[[5-[(3,4-difluorophenyl)amino] 1,3,4-oxadiazole-2-carbonyl]amino]-phenyl]cyclohexyl]acetate (see Example 1; leq) was suspended, with stirring under nitrogen, in methanol/tetrahydrofuran (14 vol/7vol) and a solution of lithium liydroxide (l0eq) in water (5 vol) added. A yellow solution was formed which was heated to 30 C
(complete by (lc/ms) after 2 hours). The mixture was then cooled to 0 C and acidified to pH2 with concentrated hydrochloric acid, keeping the temperature below 10 C. A
white precipitate formed which was filtered off and washed with water and methanol.
The product obtained was dried at 50 C under high vacuum to give a white solid.
The solid was slurried in water (50 vol) and stirred for 24 hours at ambient temperature, then heated to 60 C for 2 hours and allowed to cool overnight. The solid was filtered off and vacuum dried to give the product (THF 0.57w/w present).

Table 1: X-Ray Diffraction Pattern for Form 2 Peak 20 d-Spacing Relative Intensity Number (>20%) 1 4.7 19.0 54 2 9.3 9.5 32 3 16.2 5.5 40 4 16.8 5.3 99 5 17.7 5.0 40 6 18.1 4.9 41 7 18.6 4.8 44 8 21.4 4.1 92 9 22.7 3.9 100 23.3 3.8 53 11 25.9 3.4 73 12 29.2 3.1 41 Table 2: X-Ray Diffraction Pattern for Form 1 Peak 20 d-Spacing Relative Intensity Number (>20%) 1 5.1 17.4 19 2 7.8 11.4 100 3 14.8 6.0 36 4 16.5 5.4 30 5 22.1 4.0 52 Table 3: X-Ray Diffraction Pattern for Form 3 Peak Number 20 1 4.8 2 5.6 3 7.0 4 8.4 13.8 6 16.7 7 19.5 8 20.0 9 21.6 24.3

Claims

1. A crystalline form of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid having an X-ray powder diffraction pattern with peaks at 2-theta (2.theta.) = 16.8, 21.4 and 22.7°.

2. A crystalline form of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid, as claimed in claim 1, having an X-ray powder diffraction pattern with peaks at 2-theta (2.theta.) =
4.7, 9.3, 16.8, 21.4 and 22.7°.

3. A crystalline form of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid, as claimed in claim 1 or 2 having an X-ray powder diffraction pattern substantially as shown in Figure 1.

4. A crystalline form of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid having an X-ray powder diffraction pattern with peaks at 2-theta (2.theta.) = 8.4, 13.8 and 16.7°.

5. A crystalline form of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid, as claimed in claim 4, having an X-ray powder diffraction pattern with peaks at 2-theta (2.theta.) =
7.0, 8.4, 13.8, 16.7, 21.6 and 24.3°.

6. A crystalline form of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid, as claimed in claim 4 or 5, having an X-ray powder diffraction pattern with peaks at 2-theta (2.theta.) =
4.8, 5.6, 7.0, 8.4, 13.8, 16.7, 19.5, 20.0, 21.6 and 24.3°.

7. A crystalline form of (trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-1,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid, as claimed in claim 4, 5 or 6, having an X-ray powder diffraction pattern substantially as shown in Figure 5.

8. A pharmaceutical formulation comprising a compound as defined in any one of claims 1 to 7 in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

9. A compound as claimed in any one of claims 1 to 7 for use as a pharmaceutical.

10. The use of compound as defined in any one of claims 1 to 7 for the manufacture of a medicament for the treatment of a condition where inhibition of DGAT1 is required or desired.

11. A method of treatment of a condition where inhibition of DGAT1 is required or desired which method comprises administering a therapeutically effective amount of a compound according to any one of claims 1 to 7 to a patient in need of such treatment.