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Title:
SALTS OF OXADIAZOLE DERIVATIVES AS DGAT INHIBITORS
Document Type and Number:
WIPO Patent Application WO/2008/129319
Kind Code:
A1
Abstract:
Sodium, magnesium, tert -butylammonium salt, tris( hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium and nicotinamide salts of a compound of formula (I) are provided, which compounds inhibit acetyl CoA(acetyl coenzyme A):diacylglycerol acyltransferase (DGAT1) activity: Formula (I) wherein R 1 is phenyl optionally substituted with 1, 2 or 3 substituents independently selected from fluoro, chloro, bromo, trifluoromethyl, methoxy, difluoromethoxy, trifluoromethoxy, cyano, methyl and ethyl; R A is hydrogen or methyl; p is 0 or 1; with the proviso that the salt of formula (I) is not sodium ( trans -4-{4-[({5-[(2,4,5- trifluorophenyl) amino]-1,3,4-oxadiazol-2-yl} carbonyl)amino]phenyl}c yclohexyl)acetate; together with processes for preparing such compounds, their utility in treating clinical conditions associated with obesity, methods for their therapeutic use and pharmaceutical compositions containing them.

Inventors:
BACK KEVIN RICHARD (GB)
WHALLEY DAVID PAUL (GB)
Application Number:
PCT/GB2008/050270
Publication Date:
October 30, 2008
Filing Date:
April 18, 2008
Export Citation:
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Assignee:
ASTRAZENECA AB (SE)
ASTRAZENECA UK LTD (GB)
BACK KEVIN RICHARD (GB)
WHALLEY DAVID PAUL (GB)
International Classes:
C07D271/113; A61K31/4245; A61P3/10
Domestic Patent References:
WO2006064189A12006-06-22
Attorney, Agent or Firm:
ASTRAZENECA INTELLECTUAL PROPERTY (Södertälje, SE)
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Claims:

Claims

1. A sodium, magnesium, tert-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium or nicotinamide salt of a compound of formula (I):

(I) wherein R 1 is phenyl optionally substituted with 1, 2 or 3 substituents independently selected from fiuoro, chloro, bromo, trifiuoromethyl, methoxy, difluoromethoxy, trifluoromethoxy, cyano, methyl and ethyl;

R A is hydrogen or methyl; p is 0 or 1 ; with the proviso that the salt of formula (I) is not sodium (trans -4- {4-[({5-[(2,4,5- trifluorophenyl) amino]- 1 ,3 ,4-oxadiazol-2-yl} carbonyl)amino]phenyl} cyclohexyl)acetate.

2. A salt of a compound of formula (I), as claimed in claim 1, being a sodium or magnesium salt.

3. A salt of a compound of formula (I), as claimed in claim 1, being a tert- butylammonium, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium or nicotinamide salt.

4. A salt of a compound of formula (I), as claimed in any one of claims 1 to 3, wherein the compound of formula (I) is selected from the following:

(trans-4- {4-[( {5-[(4-ethylphenyl)amino]- 1 ,3 ,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4- {4-[( {5-[(4-methoxyphenyl)amino]- 1 ,3 ,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid; [trans-4-(4-{[(5-{[3-(trifluoromethyl)phenyl]amino}-l,3,4-oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid;

(trans-4- {4-[({5-[(3-cyanophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4- {4-[({5-[(3-methylphenyl)amino]-l,3,4-oxadiazol-2- yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid;

[trans-4-(4-{[(5-{[4-(trifluoromethyl)phenyl]amino}-l,3,4-oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid;

[trans-4-(4-{[(5-{[4-(trifluoromethoxy)phenyl]amino}-l,3,4-oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid; (trans-4-{4-[({5-[(4-fluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4-{4-[({5-[(2,4-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4- {4-[({5-[(2,6-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid;

(trans-4- {4-[({5-[(3-fluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4- {4-[({5-[(2,5-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid; (trαfts-4-{4-[({5-[(2,4,5-trifluorophenyl) amino]- 1,3, 4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

2-(trans-4-{4-[({5-[(2,4,5-trifluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)propanoic acid; trαn5-2-(4-{4-[({5-[(3,4-difluorophenyl)amino]-l,3,4-oxadiazol-2-yl}carbonyl)amino]- phenyl} cyclohexyl)propanoic acid; and

(trans-4- {4-[({5-[(3,4-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid.

5. A pharmaceutical composition which comprises a salt of a compound of formula (I), as claimed in any one of claims 1 to 4, in association with a pharmaceutically-acceptable excipient or carrier.

6. A salt of a compound of formula (I), as claimed in any one of claims 1 to 4 for use in the treatment of diabetes mellitus and/or obesity in a warm-blooded animal such as a human being.

7. A method of treating diabetes mellitus and/or obesity 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 a salt of a compound of formula (I), as claimed in any one of claims 1 to 4.

A. CLASSIRCATION OF SUBJECT MATTER ,

INV. C07D271/113 A61K31/4245 A61P3/10

According to International Patent Classification (IPC) or to both national classification and IPC

B. FIELDS SEARCHED

Minimum documentation searched (classification system followed by classification symbols)

C07D A61K A61P

Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched

Electronic data base consulted during the international search (name of data base and, where practical, search terms used)

EPO-I nternal

C. DOCUMENTS CONSIDERED TO BE RELEVANT

Category * Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No.

WO 2006/064189 A (ASTRAZENECA AB [SE]; 1-6 ASTRAZENECA UK LTD [GB]; BIRCH ALAN MARTIN [GB];) 22 June 2006 (2006-06-22) cited in the application page 38, lines 12-25

D Further documents are listed in the continuation of Box C. See patent family annex

* Special categories of cited documents :

'T' later document published after the international filing date or priority date and not in conflict with the application but

'A* document defining the general state of the art which is not cited to understand the principle or theory underlying the considered to be of particular relevance invention 'E' earlier document but published on or after the International "X" document of particular relevance; the claimed invention . filing date cannot be considered novel or cannot be considered to 1L" document which may throw doubts on priority claιm(s) or involve an inventive step when the document is taken alone which is cited to establish the publication date of another 'Y' document of particular relevance; the claimed invention citation or other special reason (as specified) cannot be considered to involve an inventive step when the

1 O" document referring to an oral disclosure, use, exhibition or document is combined with one or more other such docuother means ments, such combination being obvious to a person skilled

'P' document published pnor to the international filing date but in the art. later than the priority date claimed '&' document member of the same patent family

Date of the actual completion of the international search Date of mailing of the international search report

18 June 2008 24/06/2008

Name and mailing address of the ISA/ Authorized officer

European Patent Office, P B. 5818 Patentlaan 2 NL - 2280 HV Rijswijk TeI (+31-70) 340-2040, Tx. 31 651 epo nl, Fax: (+31-70) 340-3016 Lauro, Paol a

Form PCT/ISA/210 (second sheβtl (April 2005)

Description:

SALTS OF OXADIAZOLE DERIVATIVES AS DGAT INHIBITORS

The present invention relates to certain novel salts of compounds which inhibit acetyl CoA(acetyl coenzyme A):diacylglycerol acyltransferase (DGATl) activity, to processes for preparing such compounds, to their utility in treating clinical conditions associated with obesity, to methods for their therapeutic use and to pharmaceutical compositions containing them.

Acyl CoA:diacylglycerol acyltransferase (DGAT) is found in the microsomal fraction of cells. It catalyzes the final reaction in the glycerol phosphate pathway, considered to be the main pathway of triglyceride synthesis in cells by facilitating the joining of a diacylglycerol with a fatty acyl CoA, resulting in the formation of triglyceride. Although it is unclear whether DGAT is rate-limiting for triglyceride synthesis, it catalyzes the only step in the pathway that is committed to producing this type of molecule [Lehner & Kuksis (1996) Biosynthesis of triacylglycerols. Prog. Lipid Res. 35: 169-201]. Two DGAT genes have been cloned and characterised. Both of the encoded proteins catalyse the same reaction although they share no sequence homology. The DGATl gene was identified from sequence database searches because of its similarity to acyl CoAxholesterol acyltransferase (ACAT) genes. [Cases et al (1998) Identification of a gene encoding an acyl CoA: diacylglycerol acyltransferase, a key enzyme in triacylglycerol synthesis. Proc. Natl. Acad. Sci. USA 95: 13018-13023]. DGATl activity has been found in many mammalian tissues, including adipocytes.

Because of the previous lack of molecular probes, little is known about the regulation of DGATl. DGATl is known to be significantly up-regulated during adipocyte differentiation. Studies in gene knockout mice has indicated that modulators of the activity of

DGATl would be of value in the treatment of type II diabetes and obesity. DGATl knockout (Dgatl '1' ) mice, are viable and capable of synthesizing triglycerides, as evidenced by normal fasting serum triglyceride levels and normal adipose tissue composition. Dgatl '1' mice have less adipose tissue than wild-type mice at baseline and are resistant to diet-induced obesity. Metabolic rate is -20% higher in Dgatl '1' mice than in wild-type mice on both regular and high-fat diets [Smith et al (2000) Obesity resistance and multiple mechanisms of triglyceride synthesis in mice lacking DGAT. Nature Genetics 25: 87-90].

Increased physical activity in Dgatl '1' mice partially accounts for their increased energy expenditure. The Dgatl '1' mice also exhibit increased insulin sensitivity and a 20% increase in glucose disposal rate. Leptin levels are 50% decreased in the Dgatl '1' mice in line with the 50% decrease in fat mass. When Dgatl '1' mice are crossed with ob/ob mice, these mice exhibit the ob/ob phenotype [Chen et al (2002) Increased insulin and leptin sensitivity in mice lacking acyl CoA:diacylglycerol acyltransferase J. Clin. Invest. 109:1049-1055] indicating that the Dgatl '1' phenotype requires an intact leptin pathway. When Dgatl '1' mice are crossed with Agouti mice a decrease in body weight is seen with normal glucose levels and 70% reduced insulin levels compared to wild type, agouti or ob/ob/ Dgatl '1' mice.

Transplantation of adipose tissue from Dgatl '1' mice to wild type mice confers resistance to diet-induced obesity and improved glucose metabolism in these mice [Chen et al (2003) Obesity resistance and enhanced glucose metabolism in mice transplanted with white adipose tissue lacking acyl CoA:diacylglycerol acyltransferase J. Clin. Invest. I l l: 1715-1722].

Our International Application, PCT/GB2005/004726 describes, inter alia, compounds of formula (I):

(I) wherein R 1 is phenyl optionally substituted with 1, 2 or 3 substituents independently selected from fiuoro, chloro, bromo, trifluoromethyl, methoxy, difluoromethoxy, trifluoromethoxy, cyano, methyl and ethyl; R A is hydrogen or methyl; p is 0 or 1. Specific pharmaceutically-acceptable salts of compounds of the formula (I) are not generally disclosed in PCT/GB2005/004726. Compounds of formula (I) exemplified in PCT/GB2005/004726 are generally isolated as the free acids, except for sodium (trans-4-

{4-[({5-[(2,4,5-trifluorophenyl) amino]-l,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl} cyclohexyl)acetate.

However, although some of these compounds may be crystalline, they generally have poor aqueous solubility which is not ideal for use in pharmaceutical formulations. Therefore there exists a need for a solid form of these compounds that has physical and chemical properties suitable for use in pharmaceutical formulations. Salts with suitable properties for pharmaceutical formulation have now been found.

In the formulation of drug compositions, it is important for the drug substance to be in a form in which it can be conveniently handled and processed. This is of importance, not only from the point of view of obtaining a commercially viable manufacturing process, but also from the point of view of subsequent manufacture of pharmaceutical formulations comprising the active compound.

Further, in the manufacture of drug compositions, it is important that a reliable, reproducible and constant plasma concentration profile of drug is provided following administration to a patient.

Chemical stability, solid state stability, and "shelf life" of the active ingredients are also very important factors. The drug substance, and compositions containing it, should preferably be capable of being effectively stored over appreciable periods of time, without exhibiting a significant change in the active component's physico-chemical characteristics (e.g. its chemical composition, density, hygroscopicity and solubility).

Moreover, it is also important to be able to provide the drug in a form which is as chemically pure as possible.

The skilled person will appreciate that, typically, if a drug can be readily obtained in a stable form, such as a stable crystalline form, advantages may be provided, in terms of ease of handling, ease of preparation of suitable pharmaceutical formulations, and a more reliable solubility profile.

The present invention provides a sodium, magnesium, tert-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of a compound of formula (I):

(I) wherein R 1 is phenyl optionally substituted with 1, 2 or 3 substituents independently selected from fiuoro, chloro, bromo, trifiuoromethyl, methoxy, difluoromethoxy, trifluoromethoxy, cyano, methyl and ethyl; R A is hydrogen or methyl; p is 0 or 1 ; with the proviso that the salt of formula (I) is not sodium {trans -4- {4-[({5-[(2,4,5- trifluorophenyl) amino]- l,3,4-oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetat e.

In one embodiment, R 1 is phenyl, optionally substituted with 1 , 2 or 3 substituents independently selected from fiuoro, chloro, trifiuoromethyl, trifluoromethoxy, cyano, methyl and ethyl.

In another embodiment, R 1 is phenyl substituted with 1, 2 or 3 fiuoro. In another embodiment, R 1 is phenyl substituted with 1 or 2 fiuoro.

In another embodiment, R 1 is phenyl substituted with 1 fiuoro. In another embodiment, R 1 is phenyl substituted with 2 fiuoro. In another embodiment, R 1 is phenyl substituted with 3 fiuoro. In another embodiment, R 1 is phenyl substituted with 1 methyl group. In another embodiment, R 1 is phenyl substituted with 1 ethyl group.

In another embodiment, R 1 is phenyl substituted with 1 methoxy group. In another embodiment, R 1 is phenyl substituted with 1 cyano group. In another embodiment, R 1 is phenyl substituted with 1 trifiuoromethyl group. In another embodiment, R 1 is phenyl substituted with 1 trifluoromethoxy group. In one embodiment, R A is methyl. In another embodiment, R A is hydrogen.

In one embodiment p is 0. In another embodiment p is 1. In one embodiment p is 1 and R A is hydrogen.

In one embodiment, there is provided the sodium salt of a compound of formula (I). In another embodiment there is provided the magnesium salt of a compound of formula (I). In another embodiment there is provided the tert-butylammonium salt of a compound of formula (I). In another embodiment there is provided the tris(hydroxymethyl)methylammonium salt of a compound of formula (I). In another embodiment there is provided the triethanolammonium salt of a compound of formula (I). In another embodiment there is provided the diethanolammonium salt of a compound of formula (I). In another embodiment there is provided the ethanolammonium, salt of a compound of formula (I). In another embodiment there is provided the methylethanolammonium salt of a compound of formula (I). In another embodiment there is provided the nicotinamide salt of a compound of formula (I).

Specific compounds of formula (I) are one or more of the following:

(trans-4- {4-[( {5-[(4-ethylphenyl)amino]- 1 ,3 ,4-oxadiazol-2- yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid;

(trans-4- {4-[( {5-[(4-methoxyphenyl)amino]- 1 ,3 ,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

[trans-4-(4-{[(5-{[3-(trifluoromethyl)phenyl]amino}-l,3,4 -oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid; (trans-4- {4-[({5-[(3-cyanophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4- {4-[({5-[(3-methylphenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

[trans-4-(4-{[(5-{[4-(trifluoromethyl)phenyl]amino}-l,3,4 -oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid;

[trans-4-(4-{[(5-{[4-(trifluoromethoxy)phenyl]amino}-l,3, 4-oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid;

(trans-4- {4-[({5-[(4-fluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid; (trans-4-{4-[({5-[(2,4-difiuorophenyl)amino]-l,3,4-oxadiazol -2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4-{4-[({5-[(2,6-difluorophenyl)amino]-l,3,4-oxadia zol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trans-4- {4-[({5-[(3-fluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid; (trans-4- {4-[({5-[(2,5-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

(trαfts-4-{4-[({5-[(2,4,5-trifluorophenyl) amino]- 1,3, 4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid;

2-(trans-4-{4-[({5-[(2,4,5-trifluorophenyl)amino]-l,3,4-o xadiazol-2- yl}carbonyl)amino]phenyl}cyclohexyl)propanoic acid; trαn5-2-(4-{4-[({5-[(3,4-difluorophenyl)amino]-l,3,4-oxadia zol-2-yl}carbonyl)amino]- phenyl} cyclohexyl)propanoic acid; and

(trans-4- {4-[({5-[(3,4-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid. In one aspect there is provided the sodium, magnesium, fe/t-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of(trans-4-{4-[({5-[(4-ethylphenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid. In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of

(trans-4- {4-[( {5-[(4-methoxyphenyl)amino]- 1 ,3 ,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid. In another aspect there is provided the sodium, magnesium, tert-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of

[trans-4-(4-{[(5-{[3-(trifluoromethyl)phenyl]amino}-l,3,4 -oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid. In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt

of(trans-4-{4-[({5-[(3-cyanophenyl)amino]-l,3,4-oxadiazol -2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of(trans-4-{4-[({5-[(3-methylphenyl)amino]-l,3,4-oxadiazol-2 - yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid.

In another aspect there is provided the sodium, magnesium, tert-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of[trans-4-(4-{[(5-{[4-(trifluoromethyl)phenyl]amino}-l,3,4- oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of[trans-4-(4-{[(5-{[4-(trifluoromethoxy)phenyl]amino}-l,3,4 -oxadiazol-2- yl)carbonyl]amino}phenyl)cyclohexyl]acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of (trans-4- {4-[({5-[(4-fluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of (trans-4- {4-[({5-[(2,4-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of (trans-4- {4-[({5-[(2,6-difluorophenyl)amino]-l,3,4-oxadiazol-2- yl} carbonyl)amino]phenyl} cyclohexyl)acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of(trans-4-{4-[({5-[(3-fluorophenyl)amino]-l,3,4-oxadiazol-2 - yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of (trans-4-{4-[({5-[(2,5-difluorophenyl)amino]-l,3,4-oxadiazol -2- yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid.

In another aspect there is provided the magnesium, tert-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of (trαfts-4-{4-[({5-[(2,4,5-trifluorophenyl) amino]- 1,3, 4-oxadiazol-2- yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of2- (trans-4-{4-[({5-[(2,4,5-trifluorophenyl)amino]-l,3,4-oxadia zol-2- yl}carbonyl)amino]phenyl}cyclohexyl)propanoic acid.

In another aspect there is provided the sodium, magnesium, tert-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of trαn5-2-(4-{4-[({5-[(3,4-difluorophenyl)amino]-l,3,4-oxadia zol-2-yl}carbonyl)amino]- phenyl} cyclohexyl)propanoic acid.

In another aspect there is provided the sodium, magnesium, tøt-butylammonium salt, tris(hydroxymethyl)methylammonium, triethanolammonium, diethanolammonium, ethanolammonium, methylethanolammonium, diethylammonium, or nicotinamide salt of(trans-4-{4-[({5-[(3,4-difluorophenyl)amino]-l,3,4-oxadiaz ol-2- yl}carbonyl)amino]phenyl}cyclohexyl)acetic acid.

In one aspect of the invention, the salts of the invention are crystalline solids.

Compounds of the present invention exist as cis/trans isomers across the cycloalkyl ring. It is to be understood that the present invention encompasses all such isomers. It is also to be understood that certain compounds of the formula (I) and salts thereof can exist in solvated as well as unsolvated forms such as, for example, hydrated forms. It is to be understood that the invention encompasses all such solvated forms which inhibit DGATl activity.

The compound of the invention can be prepared as outlined below. However, the invention is not limited to these methods.

Compounds of the formula (I) may be prepared by a process a) to b) as follows (wherein all variables are as hereinbefore defined for a compound of formula (I) unless otherwise stated): a) reaction of an amine of formula (2) with a carboxylate salt of formula (3) wherein P is a protecting group;

(2) (3) b) cyclisation of a compound of formula (4) (where X is S or O) wherein P is a protecting group;

(4)

and thereafter if necessary, removing any protecting groups.

Suitable protecting groups P are (l-6C)alkyl groups, which may be removed by any suitable known process known, such as by hydrolysis under acidic or basic conditions. Process a)

Certain compounds of formula (2) may also have chiral centres or can exist in different isomeric forms such as cis/trans isomers, and may be prepared as individual isomers, as illustrated below in Schemes 1 and 2 and in the accompanying examples.

(4)

Scheme 1 It will be appreciated that for compounds of formula (I) wherein R A is methyl, there is a chiral centre in the molecule as well as cis/trans isomers across the cyclohexyl ring. The chiral centre may be resolved either at an early stage in the synthesis so that is it resolved in the compound of formula (2) (by standard chromatographic separation for example of diastereomeric amide derivatives, or by chiral chromatography on a suitable chiral stationary phase) or may be resolved at a late stage in the synthesis by, for example chiral chromatography of the compound of formula (I) itself, as illustrated in the accompanying examples.

Compounds of formula (I) in which R A is methyl may be prepared from intermediate (5) by the analogous method to that described in Scheme 1. Intermediate (5) may be prepared by palladium catalysed amination of intermediate (6), for example using benzophenone imine in the presence of palladium acetate, followed by removal of the protecting group, for example by hydrogeno lysis in the presence of palladium on carbon.

Intermediate (6) may be prepared by reaction of intermediate (7) with a trifluoromethanesulfonylating agent such as N-phenyltrifluoromethanesufonimide. Intermediate (7) may be prepared by reaction of intermediate (8) with ethyl 2- (diethoxyphosphoryl)propanoate, followed by hydrogenation of the double bond, for example using palladium on carbon catalyst. The pure trans isomer may be obtained by recrystallisation, for example from a mixture of isohexane and ethyl acetate.

Compounds of formula (3) may be made by alkaline hydrolysis of ester (9a) as prepared using a published procedure (J. Het. Chem. 1977, 14, 1385-1388). Ester (9a) may be made by cyclisation of a compound of formula (9b) (where X is O or S) in a similar manner as described in process b) for compounds of formula (4).

(9a) (9b)

An alternative method for making compounds of formula (9a) is illustrated below:

Compounds of formula (2) may be coupled with compounds of formula (3) under standard conditions for formation of amide bonds. For example using an appropriate coupling reaction, such as a carbodiimide coupling reaction performed with EDAC, optionally in the presence of DMAP, in a suitable solvent such as DCM, chloroform or DMF at room temperature. Process b)

Compounds of formula (4) and (9b) where X is S may be made by reaction of an aminocarbonyl acylhydrazine or ethoxycarbonyl acylhydrazine with a thioisocyanate or thioisocyanate equivalent such as aminothiocarbonylimidazole in a suitable solvent such as DMF or MeCN at a temperature between 0 and 100 0 C. The preparation of aminocarbonyl acylhydrazines from anilines and of ethoxycarbonyl acylhydrazines is well known in the art. For example reaction of an aniline with methyl chlorooxoacetate in the presence of pyridine in a suitable solvent such as DCM followed by reaction with hydrazine in a suitable solvent such as ethanol at a temperature between 0 and 100 0 C . The compound of formula (4) may then be cyclised using, for example agents such as carbonyldiimidazole, or tosyl chloride and a suitable base (such as triethylamine), under conditions known in the art.

Iso(thio)cyanates R 1 - NCX (where X is O or S) are commercially available or may be made by reaction of the acid chlorides R^-NH 2 with for example (thio)phosgene or a (thio)phosgene equivalent followed by a suitable base (such as triethylamine).

Compounds of formula (4) may be made from compounds of formula (2) as illustrated in Scheme 1.

The amine salts of compounds of formula (I) may be prepared by reacting an acid of formula (I) with the appropriate amine in an inert solvent, for example ethanol, methanol, propan-2-ol, ethyl acetate, dichloromethane, toluene or mixtures thereof or a mixture of ethanol or methanol or propan-2-ol and water, at a temperature in the range of 0-100 0 C and isolating the solid salt. The salt may be isolated by cooling the reaction solution and optionally seeding the solution with the desired product and/or concentrating the solution. Optionally the product may be isolated by adding an antisolvent to a solution of the product in an inert solvent. The solid may be collected by methods known to those skilled in the art for example filtration or centrifugation.

The alkali metal salts of formula (I), such as the sodium salts, may be prepared by reacting an acid of formula (I) with an alkali metal alkoxide, for example sodium methoxide, or hydroxide, for example sodium hydroxide, in an appropriate alcohol solvent such as methanol, or water at a temperature in the range of 0-100 0 C and isolating the solid salt. The salt may be isolated by cooling the reaction solution and optionally seeding the solution with the desired product and/or concentrating the solution. Optionally the product may be isolated by adding an antisolvent to a solution of the product in an inert solvent. The solid may be collected by methods known to those skilled in the art for example filtration or centrifugation. The mono- or di- alkali salts may be prepared by this method, requiring the addition of one or two equivalents of alkali metal alkoxide or hydroxide as appropriate.

The magnesium salts of formula (I) may be prepared by an analogous method to the sodium salts above.

In one embodiment the acid was suspended in methanol and 1 equivalent of tert- butylamine in methanol was added at room temperature. A transient solution was observed followed by precipitation of the product tøt-butylammonium salt, which was collected by filtration and dried under vacuum at room temperature.

In a second embodiment the acid was suspended in methanol and 1 equivalent of a solution of sodium methoxide in methanol was added. The solvents were evaporated at room temperature and the residual mono- sodium salt was collected by filtration and dried under vacuum at room temperature.

In a third embodiment, the acid was suspended in methanol and 1 equivalent of a solution of 2-amino-2-(hydroxymethyl)propane-l,3-diol in methanol was added. A transient solution was observed followed by precipitation of the product tris(hydroxymethyl)methylammonium (that is, l,3-dihydroxy-2-(hydroxymethyl)prop-2- ylammonium salt), which was collected by filtration and dried under vacuum at room temperature.

The expression "inert solvent" refers to a liquid in which the acid and the amine are both soluble that does not react with either of these starting materials. In one aspect of the invention there is provided a salt of a compound of formula (I) obtainable by a process as described above.

The compounds of the invention are conveniently presented as a pharmaceutical composition.

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 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 form 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. 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 Administration and Dosage Regimes the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990. According to a further aspect of the present invention there is provided a compound of formula (I) as defined hereinbefore for use in a method of treatment of the human or animal body by therapy.

A further feature of the present invention is a compound of formula (I) for use as a medicament.

Conveniently this is a compound of formula (I) for use as a medicament for producing an inhibition of DGATl activity in a warm-blooded animal such as a human being.

Particularly this is a compound of formula (I) for use as a medicament 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 a compound of formula (I) 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 a compound of formula (I) 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 a compound of formula (I) as defined hereinbefore 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 a compound of formula (I) as defined hereinbefore, 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 DGATl 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 a compound of formula (I) as defined hereinbefore.

According to a further feature of the invention there is provided a method of treating diabetes mellitus and/or obesity 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 a compound of formula (I) as defined hereinbefore. 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 DGATl. 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 ovary 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, inflammatory 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. In another aspect, compounds of the invention are used for treatment of

Alzheimer's disease and/or dementia.

The inhibition of DGATl activity described herein may be applied as a sole therapy or in combination with one or more 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 gender specific cut-points) plus any two of the following: hypertriglyceridemia (> 150 mg/dl; 1.7mmol/l); low HDLc (<40 mg/dl or <1.03mmol/l 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-I agonists);

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

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); PPARα-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, β-blockers (eg atenolol, inderal); ACE inhibitors (eg lisinopril); Calcium antagonists (eg. nifedipine); Angiotensin receptor antagonists (eg candesartan), α 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 Vila 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).

In addition to their use in therapeutic medicine, compounds of formula (I) are also useful as pharmacological tools in the development and standardisation of in vitro and in vivo test systems for the evaluation of the effects of inhibitors of DGATl activity in laboratory animals such as cats, dogs, rabbits, monkeys, rats and mice, as part of the search for new therapeutic agents.

As indicated above, all of the compounds, are useful in inhibiting DGATl. The ability of the compounds of formula (I) to inhibit DGATl may be demonstrated employing the following enzyme assay: Human Enzyme Assay The in vitro assay to identify DGATl inhibitors uses human DGATl 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 DGATl 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 0 C on a 41% sucrose gradient. The membrane fraction at the interphase was collected, washed, and stored in liquid nitrogen.

DGATl activity was assayed by a modification of the method described by Coleman (Methods in Enzymology 1992, 209, 98-102). Compound at 1-10 μM was incubated with 0.4 μg membrane protein, 5 mM MgCl 2 , and 10 OμM 1,2 dioleoyl-sft-glycerol in a total assay volume of 200 μl in plastic tubes. The reaction was started by adding 14 C oleoyl coenzyme A (30μM 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 ImL heptane and 0.5 mL 0.1 M carbonate buffer pH 9.5. DGATl activity was quantified by counting aliquots of the upper heptane layer by liquid scintillography.

Using this assay the compounds generally show activity with IC 50 <10μM, particularly< 1 μM.

The ability of the compounds of formula (I), and their corresponding pharmaceutically-acceptable acid salts, to inhibit DGATl may further be demonstrated 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 μg/mL insulin, 0.25 μM dexamethasone and 0.5 mM isobutylmethyl xanthine. After 48 h the cells were maintained in medium containing 10% foetal calf serum and 1 μg/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 μCi/mL 14 C-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 μM sodium acetate plus 3 μCi/mL 14 C-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 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 (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.

Examples

The invention will now be illustrated by the following Examples in which, unless stated otherwise:

(i) temperatures are given in degrees Celsius ( 0 C); operations were carried out at room or ambient temperature, that is, at a temperature in the range of 18-25 0 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 0 C;

(iii) chromatography means flash chromatography on silica gel; where a Biotage cartridge is referred to this means a cartridge containing KP-SIL™ silica, 6OA, 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;

(vi) where given, NMR data ( 1 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-dβ) 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 C18 HOA 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 70ml 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; (xiϋ) where an Isco Companion is referred to then a Combiflash companion chromatography instrument, supplied by ISOC Inc. Address Teledyne ISOC Inc, 4700 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 2Oi 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 μm 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, IPl 5AP, UK; (xvii) where chiral chromatography is referred to this is carried generally out using a 20μm Merck 50mm Chiralpak AD column, (Chiral Stationary Phase supplied by Chiral Technologies Europe, Pare 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 (200ml heads); (xviii) melting points were determined using a Buchi 530 apparatus and are uncon n ected; (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:

Et 2 O or ether diethyl ether

DMF dimethylformamide DCM dichloromethane

DME 1 ,2-dimethoxyethane

MeOH methanol

EtOH ethanol

H 2 O water

TFA trifluoroacetic acid

THF tetrahydrofuran

DMSO dimethylsulfoxide

HOBt 1 -hydroxybenzotriazole

EDCI (EDAC) l-ethyl-3-(3-dimethylaminopropyl)carbodi-imide hydrochloride

DIPEA diisopropylethylamine

DEAD diethyl azodicarboxylate

EtOAc ethyl acetate

NaHCO 3 sodium bicarbonate / sodium hydrogencarbonate

K 3 PO 4 potassium phosphate

PS polymer supported

BINAP 2,2 ' -bis(diphenylphosphino)- 1 , 1 'binaphthyl

Dppf 1 , 1 ' -bis(diphenylphosphino)ferrocene dba dib enzy lidineacetone

PS-CDI polymer supported carbonyldiimidazole

CH 3 CN 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

MgSO 4 magnesium sulfate

Na 2 SO 4 sodium sulfate

CDCl 3 deutero chloroform

CD 3 OD per-deuterated methanol

Boc tert-butoxycarbonyl

AIl compound names were derived using ACD NAME computer package.

Example 1: (?mns-4-{4-[({5-[(4-Ethylphenyr)aminol-l,3.,4-oxadiazol-2- vUcarbonvDaminol phenyl} cyclohexyDacetic acid

A solution of lithium hydroxide monohydrate (110 mg, 2.64 mmol) in water (1.4mL) was added to a suspension of methyl (trαfts-4-{4-[({5-[(4-ethylphenyl)amino]-l,3,4-oxadiazol- 2-yl}carbonyl)amino]phenyl}cyclohexyl)acetate (Intermediate 2, 152 mg, 0.33mmol) in THF (1.6mL) and MeOH (2.6 mL). After 24 h, IM aqueous HCl (5mL) was added followed by water (1OmL). The solid was filtered off and washed with water (2 x 5mL), Et 2 O (2 x 5mL) then hexane (2 x 5mL) to give the title compound as a solid (105 mg, 71%); 1 R NMR δ 1.04-1.19 (5H, m), 1.37-1.51 (2H, m), 1.65-1.85 (5H, m), 2.11-2.15 (2H, m), 2.39-2.47 (IH, m), 2.57 (2H, q), 7.21 (4H, m), 7.49 (2H, d), 7.68 (2H, d), 10.84 (IH, s), 10.91 (IH, s), 11.99 (IH, s); MS m/e MH + 449.

Examples 2-7

The following examples were prepared by the general procedure of Example 1 using the appropriate ester as starting material selected from Intermediates 3-8.

1.14-1.18 (2H, m), 1.42-1.51 (2H, m), 1.71-1.85 489 (5H, m), 2.15 (2H, d), 2.39-2.47 (IH, m), 7.23 (2H, d), 7.42 (IH, d), 7.63-7.71 (3H, m), 7.85- 7.87 (IH, m), 8.01 (IH, s), 10.94 (IH, s), 11.35 (IH, s), 11.97 (IH, s)

1.05-1.20 (2H, m), 1.40-1.55 (2H, m), 1.70-1.95 446 (5H, m), 2.14-2.16 (2H, m), 2.46-2.55 (IH, m), 7.23 (2H, d), 7.53 (IH, d), 7.63 (IH, t), 7.70 (2H, d), 7.87 (IH, d), 8.02 (IH, s), 10.97 (IH, s), 11.43 (IH, s), 11.97 (IH, s)

1.08-1.18 (2H, m), 1.42-1.50 (2H, m), 1.71-1.85 435 (5H, m), 2.15 (2H, d), 2.33 (2H, s), 2.46-2.55 (IH, m), 6.89 (IH, d), 7.22-7.29 (3H, m), 7.39 (IH, s), 7.44 (IH, d), 7.68-7.71 (2H, m), 10.90 (IH, s), 11.97 (IH, s)

1.11-1.18 (2H, m), 1.42-1.51 (2H, m), 1.73-1.85 489 (5H, m), 2.14-2.16 (2H, m), 2.42-2.53 (IH, m), 7.23 (2H, d), 7.69-7.72 (2H, m), 7.76-7..82 (4H, m), 10.96 (IH, s), 11.49 (IH, s) 12.00 (IH, s)

1.11-1.18 (2H, m), 1.42-1.50 (2H, m), 1.71-1.85 505 (5H, m), 2.14-2.16 (2H, m), 2.42-2.54 (IH, m), 7.23 (2H, d), 7.42 (2H, d), 7.68-7.73 (4H, m), , 10.94 (IH, s), 11.18 (IH, s), 11.97 (IH, s)

The following examples were made by the method of Example 1, starting from Intermediates 9-13, but the final compounds were recrystallised from acetic acid.

Example R 1 H NMR δ MS m/e MH +

1.11 (q, 2H), 1.45 (q, 2H), 1.64-1.88 (m, 5H), 439 2.15 (d, 2H), 2.44 (t, IH), 7.17-7.31 (m, 4H), 7.58-7.66 (m, 2H), 7.66-7.77 (m, 2H), 10.98 (s, IH), 11.04 (s, IH), 12.03 (s, IH)

1.11 (q, 2H), 1.45 (q, 2H), 1.66-1.89 (m, 5H), 457 2.15 (d, 2H), 2.37-2.45 (m, IH), 7.10-7.29 (m, 3H), 7.36-7.48 (m, IH), 7.69 (d, 2H), 7.95- 8.11 (m, IH), 10.78 (s, IH), 10.98 (s, IH), 12.03 (s, IH)

10 1.11 (q, 2H), 1.45 (q, 2H), 1.64-1.88 (m, 5H), 457 2.14 (d, 2H), 2.36-2.48 (m, IH), 7.22 (d, 2H), 7.28 (t, 2H), 7.37-7.49 (m, IH), 7.67 (d, 2H), 10.56 (s, IH), 10.93 (s, IH), 12.03 (s, IH)

11 1.12 (q, 2H), 1.46 (q, 2H), 1.67-1.88 (m, 5H), 453 2.15 (d, 2H), 2.44 (t, IH), 6.82-6.98 (m, IH), 7.23 (d, 2H), 7.31-7.39 (m, IH), 7.39-7.50 (m, IH), 7.49-7.59 (m, IH), 7.70 (d, 2H), 11.01 (s, IH), 11.28 (s, IH), 12.03 (s, IH)

12 1.12 (q, 2H), 1.46 (q, 2H), 1.66-1.89 (m, 5H), 457 2.15 (d, 2H), 2.44 (t, IH), 6.92-7.03 (m, IH), 7.23 (d, 2H), 7.32-7.45 (m, IH), 7.70 (d, 2H), 7.95-8.09 (m, IH), 11.03 (s, IH), 11.14 (s, IH), 12.03 (s, IH)

Example 13: (?mns-4-{4-[U5-[d,4,5-Trifluorophenyl)aminol-l,3.,4-oxadiazo l-2- vUcarbonvDaminol phenyl} cyclohexyDacetic acid

To a stirred suspension of methyl (trαfts-4-{4-[({5-[(2,4,5-trifluorophenyl)amino]-l,3,4- oxadiazol-2-yl}carbonyl)amino]phenyl}cyclohexyl)acetate (Intermediate 14; 923 mg) in methanol (26 mL) and THF (13 mL) was added a 1 M aqueous solution of lithium hydroxide (3.8 mL; 2 equivalents). The resulting solution was allowed to stir at ambient temperature for -41 h. A further 0.2 equivalents of IM LiOH solution (380 μL) were added and stirring continued for a further 24h. A further 0.2 equivalents of IM LiOH solution (380 μL) were added and stirring continued for another -15 h. The organic solvents were evaporated. NaOH (2 M, 13 mL) was added to the aqueous residue and it was washed with ether (4 x 20 mL). The aqueous phase was acidified to pH 2-3 with 2 M HCl and the resulting cream suspension filtered, washed with a little water and dried at 60 0 C under high vacuum overnight, leaving an amorphous powder (737 mg). 1 H NMR 1.11 (q, 2H), 1.45 (q, 2H), 1.66 - 1.75 (m, IH), 1.79 (t, 4H), 2.15 (d, 2H), 2.38 - 2.48 (m, 43H), 7.23 (d, 2H), 7.69 (d, 2H), 7.72 - 7.78 (m, IH), 8.10 - 8.25 (m, IH), 11.02 (s, IH), 11.10 (s, IH), 12.01 (s, IH). MS m/e (M+H) + 487

Example 14: ^m«5-4-{4-[({5-[(3,4-Difluorophenyl)aminol-l,3,4-oxadiazol- 2- vUcarbonvDaminol phenyl} cyclohexyDacetic acid

This example was made by the method of Examples 8-12, starting from Intermediate 15 as starting material. 1 U NMR δ 1.12 (q, 2H), 1.46 (q, 2H), 1.65-1.75 (m, IH), 1.79 (t, 4H), 2.15 (d, 2H), 2.37-2.48 (m, IH), 7.23 (d, 2H), 7.31-7.39 (m, IH), 7.49 (q, IH), 7.70 (d, 2H), 7.71-7.75 (m, IH); MS m/e MH + 457. This example was also made by the followng method.

Lithium hydroxide monohydrate (10 equivalents) in water (2.23 L per mole of Intermediate 15) was added to a stirred suspension of methyl (trans-4- {4-[({5-[(3, 4- difluorophenyl)amino]-l,3,4-oxadiazol-2-yl}carbonyl)amino]ph enyl}cyclohexyl)acetate (Intermediate 15; 1 equivalent) in methanol (9.38 L per mole of Intermediate 15). The reaction mixture was stirred at 3O 0 C for 2 hours then cooled to O 0 C and acidified to pH2 with concentrated hydrochloric acid (keeping the temperature below 1O 0 C). The resulting white precipitate was filtered, washed with water and methanol and then dried under vacuum at 5O 0 C to give the title compound as a solid (82% yield). This solid was then suspended in water (approximately 28ml 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 0 C in a high vacuum drying oven.

Example 15 : 2-(trans-4- {4- [({5- [(2,4.,5-trifluor ophenvDaminol - 1 ,3,4-oxadiazol-2- vUcarbonvDaminolphenvUcyclohexyDpropanoic acid

A solution of lithium hydroxide (17 mg, 0.39 mmol) in H 2 O (1.0 mL) was added in one portion to a solution of trans-ethyl 2-(4-{4-[({5-[(2,4,5-trifluorophenyl)amino]-l,3,4- oxadiazol-2-yl}carbonyl)-amino]phenyl}cyclohexyl)propanoate (Intermediate 16, 25 mg, 0.05 mmol) in a mixture of THF (1.5 mL) and MeOH (1.5 mL) and the reaction mixture was stirred at room temperature for 24 h. A IM aqueous solution of citric acid (10 mL) was added and the mixture was filtered to leave a solid. The solid was washed with H 2 O (2 mL) and purified by reverse phase preparative HPLC, eluting with a gradient of CH 3 CN and H 2 O containing 0.2 % TFA to give the title compound (10 mg, 42 %) as a solid. The individual isomers were separated by chiral chromatography under standard conditions and purified by recrystallisation from EtOH to give:

(2i?)-2-(trαn5-4-(4-r((5-r(2.4.5-Trifluorophenyl)amino1- 1.3.4-oxadiazol-2- yl} carbonyDaminolphenyU cyclohexyD-propanoic acid

1 H NMR O 1.01-1.1 (5H, m), 1.1-1.28 (2H, m), 1.37-1.5 (2H, m), 1.51-1.63 (IH, m), 1.71- 1.9 (4H, m), 2.14-2.22 (IH, m), 2.4-2.5 (IH, m), 3.45 (2H, dq), 4.32 (IH, t), 7.21 (2H, d), 7.62-7.75 (m, IH), 7.7 (2H, d), 8.1-8.22 (m, IH), 10.95 (IH, s), 11.03 (IH, s), 12.0 (IH, s); MS nVe MH + 489.

(2y)-2-(trαn5-4-(4-r((5-r(2.4.5-Trifluorophenvπamino1-1 .3.4-oxadiazol-2- yl} carbonyDaminolphenyU cyclohexyDpropanoic acid

1 H NMR O 1.01-1.1 (5H, m), 1.1-1.28 (2H, m), 1.37-1.5 (2H, m), 1.51-1.63 (IH, m), 1.71- 1.9 (4H, m), 2.14-2.22 (IH, m), 2.4-2.5 (IH, m), 3.45 (2H, dq), 4.32 (IH, t), 7.21 (2H, d), 7.62-7.75 (m, IH), 7.7 (2H, d), 8.1-8.22 (m, IH), 10.95 (IH, s), 11.03 (IH, s), 12.0 (IH, s); MS nVe MH + 489.

Example 16 : trans-2-(4- {4- [({5- [(3,4-Difluorophenyl)aminol - 1 ,3,4-oxadiazol-2- yl} carbonvDaminol -phenyl} cvclohexyDpr opanoic acid

The following example was prepared using the method described for Example 15 but using

Intermediate 18 as starting material to give the title compound (9.5 mg, 40 %) as a solid. The individual isomers were separated by chiral chromatography under standard conditions and purified by recrystallisation from EtOH to give:

(2i?)-2-(trαn5-4-(4-r((5-r(3.4-difluorophenvπamino1-1.3 .4-oxadiazol-2- yl} carbonyDaminolphenyU cyclohexyDpropanoic acid

1 H NMR O 1.05 (3H, d), 1.1-1.26 (2H, m), 1.35-1.5 (2H, m), 1.51-1.62 (IH, m), 1.7-1.9 (4H, m), 2.12-2.23 (IH, m), 2.38-2.5 (IH, m), 7.21 (2H, d), 7.29-7.38 (IH, m), 7.41-7.51

(IH, m), 7.64-7.76 (IH, m), 7.69 (2H, d), 11.0 (IH, s); MS m/e MH + 471.

(2y)-2-(trαn5-4-(4-r((5-r(3.4-difluorophenvDaminol-1.3.4 -oxadiazol-2- yl} carbonvDaminol -phenyl} cyclohexyDpropanoic acid

1 H NMR O IH NMR δ: 1.05 (3H, d), 1.1-1.26 (2H, m), 1.35-1.5 (2H, m), 1.51-1.62 (IH, m), 1.7-1.9 (4H, m), 2.12-2.23 (IH, m), 2.38-2.5 (IH, m), 7.21 (2H, d), 7.29-7.38 (IH, m),

7.41-7.51 (IH, m), 7.64-7.76 (IH, m), 7.69 (2H, d), 11.0 (IH, s), 11.7 (IH, s); MS m/e MH + 471.

Starting materials Intermediate 1: Methyl \tmns-4-(4-

{ [hydr azino(oxo)acetyll aminolphenyPcyclohexyll acetate

i) Methyl (?mns-4-phenylcvclohexyl)acetate

10% Pd/C (4520 mg) was added to a solution of methyl [trans-4-(4-

{[(trifluoromethyl)sulfonyl]oxy}phenyl)cyclohexyl]acetate (prepared as described in Patent Application WO2004/047755) (8100 mg) in MeOH (150 mL). The resulting suspension was stirred for 16 h under an atmosphere of hydrogen. The suspension was filtered through diatomaceous earth and concentrated in vacuo to give a slurry. This was extracted into EtOAc (300 mL). The organic extract was washed with saturated aqueous sodium hydrogen carbonate solution (75 mL) and then brine (75 mL). The organic layer was dried and concentrated in vacuo to give the title compound (4720 mg) as an oil; 1 H NMR δ 7.28-7.11 (5H, m), 3.58 (3H, s), 2.43 (1H+DMSO, m), 2.22 (2H, d), 1.83-1.67 (5H, m), 1.44 (2H, m), 1.13 (2H, m); MS m/e MH + 233.

ii) Methyl [?mns-4-(4-aminophenyl)cyclohexyll acetate

A mixture of 65% nitric acid (3.95 mL) and 95% sulphuric acid (4.97 mL) was added dropwise to a stirred solution of methyl (trαfts-4-phenylcyclohexyl)acetate (4710 mg) in carbon tetrachloride (20 mL) at 5 0 C and the solution was allowed to warm to ambient temperature and stirred for 16 hours. Ice/water (50 mL) was added and the mixture was extracted with DCM (2 x 40 mL). The organic extracts were combined, washed with brine

(50 mL), dried, and concentrated in vacuo to give an oil. This oil was purified by flash chromatography on a 80 g Biotage™ silica column, using a gradient of 0-20% EtOAc in hexane as the eluent to give crude methyl [trαn5-4-(4-nitrophenyl)cyclohexyl]acetate which was dissolved in ethyl acetate (30 mL). 10% palladium on carbon (0.400 g) was added and the resulting suspension was stirred at ambient temperature for 16 hours under an atmosphere of hydrogen. The suspension was filtered through diatomaceous earth and concentrated in vacuo to give a solid. This was purified by flash chromatography on a 40 g Biotage™ silica column using a gradient of 20-45% EtOAc / hexane as eluent to give the title compound (1740 mg) as a solid; 1 H NMR δ 6.83 (2H, d), 6.46 (2H, d), 4.72 (2H, s), 3.59 (3H, s), 2.23 (3H, m), 1.72 (5H, m), 1.35 (2H, m), 1.09 (2H, m); MS m/e MH + 248.

Alternatively, methyl [/rαfts-4-(4-aminophenyl)cyclohexyl] acetate can be made by the following method: a) Methyl 2-r4-(4-hvdroxyphenyl)cvclohexylidenelacetate

Trimethyl phosphonoacetate (170 mL, 1.05 mol) 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 completion of addition, the reaction mixture was allowed to warm to ambient temperature and stirred for I 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 0 C and the guanidine solution added slowly, controlling the temperature between 8 and 12 0 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 (MgSO 4 ) 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%). 1 R NMR δ 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 (IH, m), 6.66 (2H, d), 6.99 (2H, d), 9.10 (IH, s)

b) trans -M ethyl 2- r4-(4-hvdroxyphenyl)cvclohexyll acetate

10% Palladium on carbon (50% water wet, 6.9 mmol) was added to methyl 2-[4-(4- hydroxyphenyl)cyclohexylidene]acetate (100 g, 0.41 mol) in dry THF (400 mL). The reaction mixture was heated at 3O 0 C under a hydrogen atmosphere (2 bar). The mixture was filtered over Celite to leave a solid, which was washed 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 with ethyl acetate (50 mL) to give the title compound as a solid (42 g, 42%). 1 H NMR δ 1.02 - 1.17 (2H, m), 1.31 - 1.46 (2H, m), 1.66 - 1.82 (5H, m), 2.23 (2H, d), 2.28 - 2.38 (IH, m), 3.63 (3H, s), 6.66 (2H, d), 6.99 (2H, d), 9.10 (IH, s).

c) trans -M ethyl 2- | " 4-(4-aminophenyl)cyclohexyl] acetate

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 0 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 (MgSO 4 ) and concentrated in vacuo to leave the intermediate trifiate as a yellow solid (4.59 g, 106%), which was used with no further purification.

The intermediate trifiate (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 mL) 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 (MgSO 4 ) 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 I 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). 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 (MgSO 4 ) 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). 1 H NMR (CDCl 3 ) δ 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 (IH, m), 3.61 (3H, s), 6.68 (2H, d), 6.96 (2H, d).

iii) Methyl f{4-[frfl«5-4-(2-methoxy-2-oxoethyl)cvclohexyllphenyl}amino )(oxo)acetate

Methyl chloro(oxo)acetate (0.842 mL) was added to a stirred solution of methyl [trans -4- (4-aminophenyl)cyclohexyl]acetate (1740 mg) and pyridine (0.689 mL) in DCM (50 mL) at 0 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; 1 U NMR δ 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), 1.12 (2H, m); MS m/e MH " 332.

iv) Hydrazine hydrate (0.361 mL) was added to a stirred solution of methyl ({4-[trans-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 Et 2 O, and dried under vacuum overnight to give the title compound (Intermediate 1, 1845 mg) as a solid; 1 U NMR δ 10.44 (IH, s), 10.20 (IH, s), 7.70 (2H, d), 7.21 (2H, d), 4.60 (2H, s), 3.60 (3H, s), 2.42 (IH, m), 1.79 (5H, m), 1.45 (2H, m), 1.11 (2H, m); MS m/e MH + 334.

Intermediates 2-8

The following Intermediates were prepared by the following general procedure, using Intermediate 1 and commercially available isothiocyanates.

The relevant substituted phenylisothiocyanate (1.25 equivalents) was added to a stirred suspension of methyl [£rαfts-4-(4-{[hydrazino(oxo)acetyl] amino} phenyl) cyclohexyl] acetate (Intermediate 1, 1 equivalent) in DMF (approx 16 mL per mmol) and stirred at 50 0 C until a clear solution was obtained. PS-Carbodiimide (2 equivalents) was added and the reaction was heated at 80 0 C for a further 4 h. The reaction was filtered and the resin washed with DMF (5 mL). The combined DMF solutions were concentrated in vacuo and the residue triturated with diethyl ether to give the title compound.

Intermediates 9-13

The following Intermediates were prepared by the following general procedure:

The appropriate commercially available arylisothiocyanates, R-NCS, (1.2 equivalent) was added to a stirred suspension of methyl |/r<my-4-(4-{[hydrazino(oxo)acetyl] amino} phenyl)cyclohexyl] acetate (Intermediate 1) (1 equivalent) in DMA (approximately 3L per mole of Intermediate 1) and the mixture was heated to 40 0 C and stirred for 60 minutes. EDAC (1.2 equivalents) was added and the resulting mixture was heated to 85°C and stirred for 16 hours. Water (approximately 3L per mole of Intermediate 1) was added. The

precipitate was filtered off and washed with water, then diethylether, then dried under vacuum overnight to give the title compound.

Intermediate R 1 U NMR δ MS m/e MH +

1.01-1.22 (m, 2H), 1.35-1.56 (m, 2H), 1.66- 453 1.88 (m, 5H), 2.25 (d, 2H), 2.39-2.47 (m, IH), 3.60 (s, 3H), 7.17-7.31 (m, 4H), 7.56- 7.66 (m, 2H), 7.66-7.75 (m, 2H), 10.98 (s, IH), 11.04 (s, IH)

10 1.00-1.23 (m, 2H), 1.34-1.59 (m, 2H), 1.66- 471 1.89 (m, 5H), 2.25 (d, 2H), 2.37-2.46 (m, IH), 3.60 (s, 3H), 7.11-7.31 (m, 3H), 7.30- 7.49 (m, IH), 7.68 (d, 2H), 7.95-8.13 (m, IH), 10.77 (s, IH), 10.98 (s, IH)

11 1.01-1.21 (m, 2H), 1.36-1.56 (m, 2H), 1.69- 471 1.86 (m, 5H), 2.25 (d, 2H), 2.38-2.47 (m, IH), 3.60 (s, 3H),7.21 (d, 2H), 7.28 (t, 2H),7.36-7.50 (m, IH), 7.66 (d, 2H), 10.55 (s, IH), 10.93 (s, IH)

12 1.01-1.23 (m, 2H), 1.36-1.56 (m, 2H), 1.66- 453 1.87 (m, 5H), 2.25 (d, 2H), 2.37-2.46 (m, IH), 3.60 (s, 3H), 6.89 (t, IH), 7.21 (d, 2H), 7.36 (d, IH), 7.43 (q, IH), 7.51-7.57 (m, IH), 7.70 (d, 2H), 11.01 (s, IH), 11.28 (s, IH)

Intermediate 14: Methyl (?mns-4-{4-[({5-[(2.,4.,5-trifluorophenyr) aminol-1,3.,4- oxadiazol-2-yl} carbonvDaminol phenyl} cyclohexyDacetate

To a suspension of methyl [7rαfts-4-(4-{[hydrazino(oxo)acetyl] amino} phenyl) cyclohexyl] acetate (Intermediate 1; 667 mg) in anhydrous DMA (10 mL) was added 2,4,5- trifluorophenylisothiocyanate (454 mg) and the solution was heated to 45 0 C for 1 h. EDAC (461 mg) was added and the suspension heated at 85 0 C for 3 hours. The reaction mixture was cooled and water (-10 mL) was added. The precipitated solid was filtered off and dried to leave an off-white powder (964 mg). 1 H NMR δ 1.03 - 1.23 (m, 2H), 1.37 - 1.55 (m, 2H), 1.68 - 1.86 (m, 5H), 2.25 (d, IH), 3.61 (s, 3H), 7.23 (d, 2H), 7.63 - 7.79 (m, 3H), 8.17 (q, IH), 10.97 (s, IH), 11.06 (s, IH). MS m/e (M-H) " 487

Intermediate 15: methyl (?mns-4-{4-[({5-[(3.,4-difluorophenyr)aminol-l.,3.,4-oxadiaz ol- 2-yl}carbonyl)aminol phenyl} cyclohexyDacetate

This intermediate was made by the method used for Intermediates 9-13 using 3,4- difluoroisothiocyanate as starting material.

1 H NMR S 1.04-1.21 (m, 2H), 1.37-1.54 (m, 2H), 1.69-1.87 (m, 5H), 2.25 (d, 2H), 2.38- 2.48 (m, IH), 3.61 (s, 3H), 7.22 (d, 2H), 7.31-7.40 (m, IH), 7.48 (q, IH), 7.65-7.76 (m,

3H), 10.96 (s, IH), 11.24 (s, IH); MS m/e (M-H) " 469.

This intermediate was also made by a slight variation of this method:

3,4-Difiuoroisothiocyanate (1.2 equivalent), was added to a stirred suspension of methyl

[trαfts-4-(4-{[hydrazino(oxo)acetyl]amino}phenyl)cyclohe xyl]acetate (Intermediate 1, 1 equivalent) in DMA (approximately 5.3 litres per mole of Intermediate 1) 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 1) 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 16: me- trans-εthγ\ 2-(4-{4-[({5-[(2,4,5-trifluorophenyl)aminol-l,3,4- oxadiazol-2-yl} carbonvD-aminol phenyl} cyclohexyDpropanoate

2,4,5-Trifluorophenyl isothiocyanate (252 mg, 1.33 mmol) was added in one portion to a suspension of trans-ethyl 2-[4-(4-{[hydrazino(oxo)acetyl]amino}phenyl)cyclo- hexyl]propanoate (Intermediate 17, 400 mg, 1.11 mmol) in DMA (6 mL) and the reaction mixture was stirred at room temperature for 2 h. EDCI (319 mg, 1.66 mmol) was added in one portion and the reaction mixture was heated to 90 0 C for 10 mins in the microwave. The mixture was then cooled to room temperature and H 2 O (20 mL) was added. The mixture was filtered to leave a solid, which was washed with H 2 O (10 mL) to give the title compound (550 mg, 95%) as a solid; 1 H NMR δ 1.1 (3H, d), 1.12-1.28 (2H, m), 1.2 (3H, t), 1.35-1.51 (2H, m), 1.52-1.64 (IH, m), 1.65-1.75 (IH, m), 1.77-1.9 (3H, m), 2.22-2.32 (IH, m), 2.38-2.51 (IH, m), 4.01-4.16 (2H, m), 7.2 (2H, d), 7.65-7.78 (IH, m), 7.69 (2H, d), 8.1-8.25 (IH, m), 10.95 (IH, s), 11.05 (IH, s); MS m/e MH + 517.

Intermediate 17: trans-εthyl 2-[4-(4-Uhydrazino(oxo)acetyllamino}phenyl) cyclohexyllpropanoate

i) Ethyl 2-[4-(4-hydroxyphenyl)cyclohexylidenelpropanoate

A solution of ethyl 2-(diethoxyphosphoryl)propanoate (12.5 g, 52.6 mmol) in THF (10 mL) was added in one portion to a stirred mixture of sodium hydride (60 % in mineral oil, 2.3 g, 57.6 mmol) in THF (75 mL) and the reaction mixture was stirred at 10 0 C under an argon atmosphere for I h. In a separate flask tetramethylguanidine (6.61 g, 57.4 mmol) was added in one portion to a solution of 4-(4-hydroxyphenyl)cyclohexanone (9.1 g, 47.8 mmol) in THF (40 mL) at 10 0 C under an argon atmosphere and the stirred mixture was warmed to room temperature over 1 h. This mixture was added in one portion to the ethyl 2-(diethoxyphosphoryl)propanoate mixture at 10 0 C and the combined reaction mixture was warmed to ambient temperature and stirred for 18 h under an argon atmosphere. A saturated aqueous solution of ammonium chloride (75 mL) and EtOAc (100 mL) were added. The layers were separated and the aqueous layer was extracted with EtOAc (100 mL). The combined organic layers were dried and concentrated in vacuo to leave a residue. The residue was triturated with a mixture of ether and isohexane (2:1), filtered and the filtrate was concentrated in vacuo to give the title compound (6.9 g, 53 %) as a solid; 1 R NMR δ 1.21 (3H, t), 1.33-1.5 (2H, m), 1.78-2.1 (4H, m), 1.84 (3H, s), 2.6-2.78 (2H, m), 2.99-3.09 (IH, m), 4.12 (2H, q), 6.67 (2H, d), 7.02 92H, d), 9.08 (IH, s); MS m/e MH + 275.

ii) trans-εthyl 2-[4-f4-hvdroxyphenyl)cvclohexyllpropanoate

10% Palladium on carbon (250 mg) was added in one portion to a stirred solution of ethyl

2-[4-(4-hydroxyphenyl)cyclohexylidene]propanoate (6.9 g, 25.3 mmol) in EtOAc (50 mL) and the reaction mixture was stirred under a hydrogen atmosphere for 24 h. The reaction mixture was filtered and concentrated in vacuo to give a solid that was recrystallised from a mixture of isohexane and EtOAc (10:1) to give the title compound (2.2 g, 32 %) as a solid; 1 U NMR δ (CDCl 3 ) 1.0-1.3 (2H, m), 1.08 (3H, d), 1.2 (3H, t), 1.3-1.45 (2H, m), 1.45-1.64 (2H, m), 1.65-1.75 (IH, m), 1.77-1.9 (3H, m), 2.15-2.26 (IH, m), 2.28-2.39 (IH, m), 4.1 (2H, q), 6.68 (2H, d), 7.0 (2H, d).

iii) trans-εthyl 2-[4-f4-{[ftrifluoromethyl)sulfonylloxy}phenyl)cvclohexyllpr opanoate

Ethyldiisopropylamine (1.6 mL) and N-phenyltrifluoromethanesulfonimide (4.12 g, 11.5 mmol) was added sequentially, each in portions over 1 minute, to a stirred solution of trans-ethyl 2-[4-(4-hydroxyphenyl)cyclohexyl]propanoate (2.6 g, 9.23 mmol) in DCM (25 mL) at 5 0 C. The reaction mixture was warmed to room temperature and stirred for 18 h and then an aqueous solution of sodium hydroxide (2M, 30 mL) was added. The layers were separated and the organic layer was washed with H 2 O (30 mL), dried and concentrated in vacuo to leave a residue. The residue was purified by column chromatography, eluting with 0-20% EtOAc in isohexane as eluent, to give the title compound (3.6 g, 96 %) as an oil; 1 H NMR δ (CDCl 3 ) 1.0-1.25 (2H, m), 1.1 (3H, d), 1.2 (3H, t), 1.3-1.45 (2H, m), 1.5-1.64 (2H, m), 1.7-1.79 (IH, m), 1.8-1.95 (3H, m), 2.17-2.28 (IH, m), 2.38-2.48 (IH, m), 4.1 (2H, q), 7.1 (2H, d), 7.2 (2H, d).

iv) trans-Ethyl 2-(4- {4- [(diphenylmethylene)aminol phenyl} cyclohexyDpropanoate

Caesium carbonate (4.0 g, 12.4 mmol), R -BINAP (330 mg, 0.53 mmol), palladium acetate (119 mg, 0.53 mmol) and benzophenone imine (2.2 mL, 13.2 mmol) were added sequentially, each in one portion, to a stirred solution of trans-ethyl 2-[4-(4- {[(trifluoromethyl)-sulfonyl]oxy}phenyl)cyclohexyl]propanoat e in THF (50 mL) and the reaction mixture was heated at 65 0 C for 70 h under an argon atmosphere. The reaction mixture was cooled to room temperature and concentrated in vacuo to leave a residue. The residue was taken up in EtOAc (75 mL) and H 2 O (25 mL) was added. The layers were separated and the organic layer was extracted with brine (25 mL), dried and concentrated in vacuo to leave a residue. The residue was purified by chromatography, eluting with 10- 50% EtOAc in isohexane, to give the title compound (2.3g, 59 %) as an oil; 1 H NMR δ (CDCl 3 ) 1.07-1.25 (2H, m), 1.15 (3H, d), 1.3 (3H, t), 1.35-1.5 (2H, m), 1.55-1.7 (2H, m), 1.72-1.8 (IH, m), 1.83-1.95 (2H, m), 2.24-2.43 (2H, m), 4.15 (2H, q), 6.7 (2H, d), 7.0 (2H,

d), 7.15 (2H, d), 7.23-7.35 (IH, m), 7.4 (2H, dd), 7.47-7.55 (2H, m), 7.78 (2H, d), 7.84 (IH, d); MS nVe MH + 440.

v) trans-εthyl 2-[4-(4-aminophenyl)cvclohexyllpropanoate

10% Palladium on carbon (500 mg) was added in one portion to a stirred solution of trans- ethyl 2-(4-{4-[(diphenylmethylene)amino]phenyl}cyclohexyl)propanoa te (2.3 g, 5.23 mmol) in ethanol (50 mL) and the reaction mixture was stirred under a hydrogen atmosphere for 6 h. The reaction mixture was filtered and concentrated in vacuo to leave an oil. The oil was purified by chromatography, using 20-50% EtOAc in isohexane as eluent, to give the title compound (1.0 g, 71 %) as an oil; 1 H NMR δ (CDCl 3 ) 0.9-1.1 (2H, m), 0.95 (3H, d), 1.1 (3H, t), 1.18-1.32 (2H, m), 1.38-1.5 (IH, m), 1.54-1.64 (IH, m), 1.65- 1.8 (3H, m), 2.04-2.15 (IH, m), 2.18-2.3 (IH, m), 3.98 (2H, q), 6.8 (2H, d), 6.92 (2H, d); MS m/e MH + 276.

vi) trans-Ethyl 2-[4-(4-{[methoxy(oxo)acetyllamino}phenyl)cvclohexyllpropano ate

A solution of methyl chlorooxoacetate (670 mg, 5.45 mmol) in DCM (3 mL) was added in one portion to a solution of trans-ethyl 2-[4-(4-aminophenyl)cyclohexyl]propanoate (1.0 g, 3.63 mmol) and pyridine (0.59 mL, 7.26 mmol) in DCM (22 mL) at 5 0 C and the mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was concentrated in vacuo and EtOAc (30 mL) was added. The layers were separated and the organic layer was washed with water (25 mL) and brine (25 mL). The organic layer was dried and concentrated in vacuo to give the title compound (1.3 g, 99 %) as a solid; 1 H NMR δ (CDCl 3 ) 1.0-1.1.25 (2H, m), 1.1 (3H, d), 1.2 (3H, t), 1.3-1.47 (2H, m), 1.48-1.55 (2H, m), 1.68-1.79 (IH, m), 1.8-1.95 (2H, m), 2.18-2.28 (IH, m), 2.35-2.5 (IH, m), 3.9 (3H, t), 4.08 (2H, q), 7.12 (2H, d), 7.47 (2H, d); MS m/e (M-H) " 360.

vii) Hydrazine hydrate (229 mg, 4.57 mmol) was added in one portion to a stirred solution of trans-ethyl 2-[4-(4-{[methoxy(oxo)acetyl]amino}phenyl)cyclohexyl]propano ate (1.3 g, 3.6 mmol) in ethanol (25 mL) and the reaction mixture was stirred at room temperature for 18 h. The mixture was filtered to leave a solid, which was washed with ethanol (10 mL) to give the title compound (Intermediate 99) (1.2 g, 92 %) as a solid; 1 H NMR δ 1.07 (3H, d), 1.1-1.3 (2H, m), 1.2 (3H, t), 1.36-1.5 (2H, m), 1.5-1.63 (IH, m), 1.65-1.72 (IH, m), 1.75- 1.88 (3H, m), 2.2-2.32 (IH, m), 2.35-2.49 (IH, m), 4.0-4.17 (2H, m), 4.6 (2H, s), 7.19 (2H, d), 7.7 (2H, d), 10.44 (IH, s); MS m/e MH + 362.

Intermediate 18 : trans-εthyl 2-(4- {4- [({5- [(3,4-difluor ophen v Darni no I -1 ,3,4-oxadiazol- l-vUcarbonvD-aminolphenvUcyclohexyDpropanoate

This intermediate was made by an analogous method to Intermediate 16, using Intermediate 17 and appropriate commercially available isothiocyanate. 1 H NMR O 1.1 (3H, d), 1.12-1.28 (2H, m), 1.2 (3H, t), 1.37-1.51 (2H, m), 1.52-1.64 (IH, m), 1.65-1.75 (IH, m), 1.77-1.9 (3H, m), 2.22-2.34 (IH, m), 2.39-2.5 (IH, m), 4.03-4.17 (2H, m), 7.22 (2H, d), 7.32-7.4 (IH, m), 7.44-7.55 (IH, m), 7.65-7.75 (IH, m), 7.7 (2H, d), 10.95 (IH, s), 11.25 (IH, s); MS m/e (M+H) + 499