ZA200201437B

ZA200201437B - Substituted N-phenyl 2-hydroxy 2-methyl-3,3,3-trifluoropropanamide derivates which elevate pyruvate dehydrogenase activity.

Info

Publication number: ZA200201437B
Application number: ZA200201437A
Authority: ZA
Inventors: Roger John Butlin; Janet Elizabeth Pease; Michael Howard Block; Thorsten Nowak; Jeremy Nicholas Burrows
Original assignee: Astrazeneca Ab
Priority date: 1999-09-04
Filing date: 2002-02-20
Publication date: 2003-05-20
Also published as: UA73319C2; GB9920814D0

Description

SUBSTITUTED N-PHENYL 2-HYDROXY-2-METHYL-3, 3, 3-TRIFLUOROPROPANAMIDE DERIVATIVES

WHICH ELEVATE PYRUVATE DEHYDROGENASE ACTIVITY

The present invention relates to compounds which elevate pyruvate dehydrogenase (PDH) activity, processes for their preparation, pharmaceutical compositions containing them as active ingredient, methods for the treatment of disease states associated with reduced PDH activity, to their use as medicaments and to their use in the manufacture of medicaments for use in the elevation of PDH activity in warm-blooded animals such as humans, in particular the treatment of diabetes mellitus, peripheral vascular disease and myocardial ischaemia in : warm-blooded animals such as humans, more particularly to their use in the manufacture of : medicaments for use in the treatment of diabetes mellitus in warm-blooded animals such as humans.

Within tissues adenosine triphosphate (ATP) provides the energy for synthesis of complex molecules and, in muscle, for contraction. ATP is generated from the breakdown of energy-rich substrates such as glucose or long chain free fatty acids. In oxidative tissues such as muscle the majority of the ATP is generated from acetyl CoA which enters the citric acid cycle, thus the supply of acetyl CoA is a critical determinant of ATP production in oxidative tissues. Acetyl CoA is produced either by B-oxidation of fatty acids or as a result of glucose . metabolism by the glycolytic pathway. The key regulatory enzyme in controlling the rate of acetyl CoA formation from glucose is PDH which catalyses the oxidation of pyruvate to acetyl .

CoA and carbon dioxide with concomitant reduction of nicotinamide adenine dinucleotide (NAD) to NADH.

In disease states such as both non-insulin dependent (NIDDM) and insulin-dependent diabetes mellitus (IDDM), oxidation of lipids is increased with a concomitant reduction in © utilisation of glucose, which contributes to the hyperglycaemia. Reduced glucose utilisation in both IDDM and NIDDM is associated with a reduction in PDH activity. In addition, a further consequence of reduced PDH activity may be that an increase in pyruvate concentration results in increased availability of lactate as a substrate for hepatic gluconeogenesis. It is , reasonable to expect that increasing the activity of PDH could increase the rate of glucose oxidation and hence overall glucose utilisation, in addition to reducing hepatic glucose output.

Another factor contributing to diabetes mellitus is impaired insulin secretion, which has been shown to be associated with reduced PDH activity in pancreatic B-cells (in a rodent genetic model of diabetes mellitus Zhou et al. (1996) Diabetes 45: 580-586).

Oxidation of glucose is capable of yielding more molecules of ATP per mole of oxygen than is oxidation of fatty acids. In conditions where energy demand may exceed energy supply, such as myocardial ischaemia, intermittent claudication, cerebral ischaemia and reperfusion, (Zaidan et al., 1998; J. Neurochem. 70: 233-241), shifting the balance of substrate utilisation in favour of glucose metabolism by elevating PDH activity may be expected to improve the ability to maintain ATP levels and hence function.

An agent which is capable of elevating PDH activity may also be expected to be of benefit in treating conditions where an excess of circulating lactic acid is manifest such as in certain cases of sepsis.

The agent dichloroacetic acid (DCA) which increases the activity of PDH after acute administration in animals, (Vary et al., 1988; Circ. Shock, 24: 3-18), has been shown to have the predicted effects in reducing glycaemia, (Stacpoole et al., 1978; N. Engl. J. Med. 298: 526-530), and as a therapy for myocardial ischaemia (Bersin and Stacpoole 1997; American

Heart Journal, 134: 841-855) and lactic acidaemia, (Stacpoole etal, 1983; N. Engl. J. Med. 309: 390-396).

PDH is an intramitochondrial multienzyme complex consisting of multiple copies of . several subunits including three enzyme activities El, E2 and E3, required for the.completion of the conversion of pyruvate to acetyl CoA (Patel and Roche 1990; FASEB J., 4: 3224-3233). ) El catalyses the non-reversible removal of CO, from pyruvate; E2 forms acetyl CoA and E3 reduces NAD to NADH. Two additional enzyme activities are associated with the complex: a specific kinase which is capable of phosphorylating E1 at three serine residues and a loosely-associated specific phosphatase which reverses the phosphorylation. Phosphorylation of a single one of the three serine residues renders the E1 inactive. The proportion of the PDH in its active (dephosphorylated) state is determined by a balance between the activity of the kinase and phosphatase. The activity of the kinase may be regulated in vivo by the relative concentrations of metabolic substrates such as NAD/NADH, CoA/acetylCoA and adenine diphosphate (ADP)/ATP as well as by the availability of pyruvate itself.

European Patent Publication Nos. 617010 and 524781 describes compounds which are capable of relaxing bladder smooth muscle and which may be used in the treatment of urge incontinence. We have found that the compounds of the present invention are very good at elevating PDH activity, a property nowhere disclosed in EP 0617010 and EP 524781.

The present invention is based on the surprising discovery that certain compounds elevate PDH activity, a property of value in the treatment of disease states associated with

. -3. . o disorders of glucose utilisation such as diabetes mellitus, obesity, (Curto et al., 1997; Int. J.

Obes. 21: 1137-1142), and lactic acidaemia. Additionally the compounds may be expected to have utility in diseases where supply of energy-rich substrates to tissues is limiting such as peripheral vascular disease, (including intermittent claudication), cardiac failure and certain cardiac myopathies, muscle weakness, hyperlipidaemias and atherosclerosis (Stacpoole et al., 1978; N. Engl. J. Med. 298: 526-530). A compound that activates PDH may also be useful in treating Alzheimer’s disease (AD) (J Neural Transm (1998) 105, 855-870).

Accordingly the present invention provides a compound of formula (I):

R4

R33 je! o

AL

N

ROT om 1) wherein: nislor2; . R! is chloro, fluoro, bromo, methyl or methoxy; .

R? is selected from one of the following three groups: i) halo, nitro, hydroxy, amino or cyano; ’ . ii) -X'-R® wherein X' is a direct bond, -O-, -S-, -SO-, -SO»-, -NR®-, -CO-, -CONR®-, -NR®CO-, -NR®SO;- or NR’ CONR’-; wherein R® and R’ are independently hydrogen or

C.salkyl optionally substituted with one or more A; and R® is selected from Cj_salkyl optionally substituted with one or more A, Cj.;cycloalkyl optionally substituted with one or more A, Cs.scycloalkylC, alkyl optionally substituted with one or more A, C; ¢alkenyl optionally substituted with one or more A, C;.alkynyl optionally substituted with one or more

A, phenyl optionally substituted with one or more D, phenylC,.calkyl optionally substituted with one or more D, heteroaryl ring optionally substituted on a ring carbon by one or more D ‘ or (heteroaryl ring)C;.salkyl optionally substituted on a ring carbon with one or more D; wherein said heteroaryl ring is a carbon linked 6-membered ring containing 1-2 nitrogen atoms or a carbon linked 5-membered ring containing 1-3 heteroatoms selected independently from O, N and S; and wherein if said S-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

iii) a nitrogen-linked 4-8 membered heterocyclic group optionally substituted on a ring carbon by one or more D and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

R} is C,.¢alky] optionally substituted with one or more A, Cs.scycloalkyl optionally substituted with one or more A, phenyl optionally substituted with one or more D, a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D, or a carbon linked 5-membered heteroaryl ring containing 1-3 heteroatoms selected independently from O, N and S optionally substituted on aring carbon by one or more D and wherein if said 5-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

A is selected from hydroxy, amino, halo, carboxy, N-(C;.4alkyl)amino,

N,N-di-(C,.4alkyl)amino, carbamoyl and C;_¢alkoxy;

D is selected from: i) -X°-R° wherein X* is a direct bond, -O-, -S-, -SO-, -SO,-, -CO-, -NR®SO,-, -NRCO-, -NR®CONR®-, -NR- or -CONR’-; wherein R? and R°® are independently hydrogen or C; 4alkyl optionally substituted with one or more hydroxy or C.4alkoxy; and R°® is selected from . hydrogen or C, alkyl optionally substituted with one or more hydroxy or C;.salkoxy; ii) a 4-8 membered Het which is optionally substituted on a ring carbon with one or more , groups selected from hydroxy, halo, C,4alkoxy, C;.salkyl or cyano and wherein if said 4-8 membered Het contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G; iii) -X?-C).¢alkyl-X°-R® wherein X® and R°® are as defined hereinbefore and X" is -S-, -SO- or -SO,-; iv) cyano or halo; and v) -X°-R wherein X® is -C(O)- or -SO,- and R'is a nitrogen-linked 4-8 membered heterocyclic group optionally substituted on a ring carbon by one or more groups selected from hydroxy, halo, C;.salkoxy, C;.salkyl or cyano and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

G is selected from Cj_salkyl optionally substituted with one or more A, C,_salkanoyl optionally substituted with one or more A, C, salkylsulphonyl optionally substituted with one or more A, Cj.salkoxycarbonyl optionally substituted with one or more A, carbamoyl,

N-(C,.salkyl)carbamoy! optionally substituted with one or more A, N-(C;.¢alkyl),carbamoyl

-5. R © optionally substituted with one or more A and benzoyl optionally substituted with one or more

A; and

Ris hydrogen or fluoro; : or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

In this specification the term “alkyl” includes both straight and branched chain alkyl groups but references to individual alkyl groups such as “propyl” are specific for the straight chain version only. For example, “C,alkyl” includes C,.4alkyl, C5 4alkyl, propyl, isopropyl and ¢-butyl. However, references to individual alkyl! groups such as ‘propyl’ are specific for the straight chained version only and references to individual branched chain alkyl groups such as ‘isopropyl’ are specific for the branched chain version only. The term “halo” refers to fluoro, chloro, bromo and iodo.

Suitable values for “a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms” include pyridyl, pyrimidyl, pyrazinyl and pyridazinyl. Preferably “a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms” is pyridyl. In another aspect of the invention preferably “a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms” is pyridazinyl.

Suitable values for “a carbon-linked 5-membered heteroaryl ring containing 1-3 . heteroatoms” include pyrrolyl, furyl, thienyl, pyrazolyl, oxazolyl, oxadiazolyl, imidazolyl and triazolyl. : .

A “nitrogen-linked 4-8 membered heterocyclic group” is a saturated, partially saturated or unsaturated, monocyclic ring containing 4-8 atoms of which at least one is a nitrogen atom with optionally 1-3 further heteroatoms selected independently from O, N and S wherein a -CH,- group can optionally be replaced by a -C(O)- and a ring nitrogen and/or sulphur atom may be optionally oxidised to form the N-oxide and or the S-oxides. It will be appreciated that in forming this nitrogen link, the nitrogen atom is not quaternised, i.e. a neutral compound is formed. Suitable values for “nitrogen-linked 4-8 membered heterocyclic group” include morpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, ‘ homopiperazinyl, pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazolidinyl and triazolyl. Further suitable values for “nitrogen-linked 4-8 membered heterocyclic group” include azetidinyl, morpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, homopiperazinyl, pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazolidinyl and triazolyl. Preferably a “nitrogen-linked 4-8 membered heterocyclic group” is morpholino, piperidyl, piperazinyl, pyrrolidinyl,

RX WO 01/17956 PCT/GB00/03314 thiomorpholino, pyrrolinyl or homopiperazinyl. More preferably a “nitrogen-linked 4-8 membered heterocyclic group” is azetidinyl, morpholino, piperidyl, piperazinyl, pyrrolidiny], thiomorpholino, pyrrolinyl or homopiperazinyl. Additional suitable values for “nitrogen-linked 4-8 membered heterocyclic group” include azetidinyl, morpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, pyrrolinyl, homopiperazinyl, pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazolidinyl and triazolyl. Preferably a “nitrogen-linked 4-8 membered heterocyclic group” is morpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl or homopiperazinyl. More preferably a “nitrogen-linked 4-8 membered heterocyclic group” is azetidinyl, morpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, pyrrolinyl or homopiperazinyl. Particularly R' as a “nitrogen-linked 4-8 membered heterocyclic group” is azetidinyl, morpholino or pyrrolidinyl. Particularly when R’isa “nitrogen-linked 4-8 membered heterocyclic group” it is thiomorpholino. In another aspect of the invention, particularly when R? is a “nitrogen-linked 4-8 membered heterocyclic group” it is thiomorpholino, piperazinyl, 1-oxothiomorpholino, 1,1-dioxothiomorpholino or \ morpholino.

A “nitrogen-linked 4-6 membered heterocyclic group” is a saturated, partially } saturated or unsaturated, monocyclic ring containing 4-6 atoms of which at least one is a nitrogen atom with optionally 1-3 further heteroatoms selected independently from O, N and S wherein a -CH,- group can optionally be replaced by a -C(O)- and a ring nitrogen and/or sulphur atom may be optionally oxidised to form the N-oxide and or the S-oxides. It will be appreciated that in forming this nitrogen link, the nitrogen atom is not quaternised, i.e. a neutral compound is formed. Suitable values for a “nitrogen-linked 4-6 membered heterocyclic group” include azetidinyl, morpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazolidinyl and triazolyl. Preferably a “nitrogen-linked 4-6 membered heterocyclic group” is azetidinyl, morpholino or pyrrolidinyl.

A “nitrogen-linked 5 or 6 membered heterocyclic group” is a saturated, partially saturated or unsaturated, monocyclic ring containing 5 or 6 atoms of which at least one is a nitrogen atom with optionally 1-3 further heteroatoms selected independently from O, N and S wherein a -CH,- group can optionally be replaced by a -C(O)- and a ring nitrogen and/or sulphur atom may be optionally oxidised to form the N-oxide and or the S-oxides. It will be

-7- R « appreciated that in forming this nitrogen link, the nitrogen atom is not quaternised, i.e. a neutral compound is formed. Suitable values for a “nitrogen-linked 5 or 6 membered heterocyclic group” include morpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazolidinyl and triazolyl. Preferably a “nitrogen-linked 5 or 6 membered heterocyclic group” is morpholino, piperidyl, piperazinyl, pyrrolidinyl, thiomorpholino or pyrrolinyl.

A “nitrogen-linked 6 membered heterocyclic group” is a saturated, partially saturated or unsaturated, monocyclic ring containing 6 atoms of which at least one is nitrogen atom with optionally 1-3 further heteroatoms selected independently from O, N and S wherein a -CH,- group can optionally be replaced by a -C(O)- and a ring nitrogen and/or sulphur atom may be optionally oxidised to form the N-oxide and or the S-oxides. It will be appreciated that in forming this nitrogen link, the nitrogen atom is not quaternised, i.e. a neutral compound is formed. Suitable values for a “nitrogen-linked 6 membered heterocyclic group” include morpholino, piperidyl, piperazinyl, thiomorpholino.

A “4-8 membered Het” is a saturated, partially saturated or unsaturated monocyclic ring containing 4-8 atoms including 1-4 heteroatoms selected independently from O, N and S, which may be carbon or nitrogen linked, wherein a -CH,- group can optionally be replaced by . a -C(0)- and a ring nitrogen and/or sulphur atom may be optionally oxidised to form the

N-oxide and or the S-oxides. Suitable values for “4-8 membered Het” are morpholino, . piperidyl, pyridyl, pyranyl, pyrrolyl, isothiazolyl, thienyl, thiadiazolyl, piperazinyl, thiazolidinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, homopiperazinyl, tetrahydropyranyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl, isoxazolyl, 4-pyridone, 2-pyrrolidone, 4-thiazolidone, pyridine-N-oxide and quinoline-N-oxide.

A “5 or 6 membered Het” is a saturated, partially saturated or unsaturated monocyclic ring containing 4-8 atoms including 1-4 heteroatoms selected independently from O, N and S, which may be carbon or nitrogen linked, wherein a -CH,- group can optionally be replaced by a -C(O)- and a ring nitrogen and/or sulphur atom may be optionally oxidised to form the

N-oxide and or the S-oxides. Suitable values for “5 or 6 membered Het” are morpholino, piperidyl, pyridyl, pyranyl, pyrrolyl, isothiazolyl, thienyl, thiadiazolyl, piperazinyl, thiazolidinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, tetrahydropyranyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl, isoxazolyl, 4-pyridone, 2-pyrrolidone, 4-thiazolidone, pyridine-N-oxide and quinoline-N-oxide.

» . N 8 =

Examples of “C; alkoxycarbonyl” include methoxycarbonyl, ethoxycarbonyl, n- and t-butoxycarbonyl. Examples of “C,;.salkoxy” include C,.4alkoxy, methoxy, ethoxy and propoxy. Examples of “Cj4alkylsulphiny}l” include methylsulphinyl and ethylsulphinyl.

Examples of “Cj.salkylsulphonyl” include C,_4alkylsulphonyl, mesyl and ethylsulphonyl.

Examples of “C,.salkanoyl” include Ci .salkanoyl, propionyl and acetyl. Examples of “Cs.7cycloalkyl” are cyclopropyl and cyclohexyl. Examples of “Cj. salkenyl” are vinyl, allyl and 1-propenyl. Examples of “Ca salkynyl” are ethynyl, 1-propynyl and 2-propynyl. Examples of “N-(C.salkyl)carbamoy]” are methylaminocarbonyl and ethylaminocarbonyl. Examples of “N-(C).salkyl),carbamoyl” are dimethylaminocarbonyl and methylethylaminocarbonyl.

Examples of “N-(C)4alkyl)amino” include methylamino and ethylamino. Examples of “N-(C,4alkyl);amino” include di-N-methylamino, di-(N-ethyl)amino and

N-ethyl-N-methylamino. Examples of *phenylC, alkyl” include phenylC,.salkyl, benzyl and phenethyl. Examples of “Cj.scycloalkylCy alkyl” include cyclopropylmethyl, cyclopentylethyl and 2-cyclohexylpropyl. Examples of ‘(heteroaryl ring)C; salkyl” include pyridylmethyl, pyrazinylethyl and imidazolylpropyl.

According to a further aspect of the present invention there is provided a compound of formula (I) (as depicted above) wherein: nislor2;

R!is chloro, fluoro, bromo, methyl or methoxy;

R? is selected from one of the following three groups: i) halo, nitro, hydroxy, amino or cyano; ii) -X'-R® wherein X' is a direct bond, -O-, -S-, -SO-, -SO,-, -NR-, -CO-, -CONR®-, -NR®CO-, -NR°SO;- or NR®CONR’-; wherein R® and R’ are independently hydrogen or

C,.4alkyl optionally substituted with one or more A; and R® is selected from C;galkyl optionally substituted with one or more A, Cs.ocycloalkyl optionally substituted with one or more A, Ca.¢alkenyl optionally substituted with one or more A, Ca salkynyl optionally substituted with one or more A, phenyl optionally substituted with one or more D, a carbon ) linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D or a carbon linked 5-membered heteroaryl ring containing 1-3 heteroatoms selected independently from O, N and S optionally substituted on a ring carbon by one or more D and wherein if said 5-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

R3 is Cy 6alkyl optionally substituted with one or more A, Cs.scycloalkyl optionally substituted with one or more A, phenyl optionally substituted with one or more D, a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D, or a carbon linked 5-membered heteroaryl ring containing 1-3 heteroatoms selected independently from O, N and S optionally substituted on : a ring carbon by one or more D and wherein if said 5-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

A is selected from hydroxy, amino, halo, carboxy and C,_salkoxy;

D is selected from: i) -X*-R° wherein X® is a direct bond, -O-, -S-, -SO-, -80;-, -CO-, -NR‘SO,-, -NR’CO-, “NRYCONR®-, -NR’- or -CONRY-; wherein R? and R° are independently hydrogen or Cy.alkyl optionally substituted with one or more hydroxy or Cj_salkoxy; and R® is selected from hydrogen or C;.salkyl optionally substituted with one or more hydroxy or C, salkoxy; ii) a 4-8 membered Het which is optionally substituted on a ring carbon with one or more . groups selected from hydroxy, halo, Ci.salkoxy, C;.4alkyl or cyano and wherein if said 4-8 membered Het contains an -NH- moiety that nitrogen may be optionally substituted with a i group selected from G; iii) -X*-C.salkyl-X"-R® wherein X® and R® are as defined hereinbefore and X® is -S-, -SO- or -SO,-; and iv) cyano or halo;

G is selected from Cj_galkyl optionally substituted with one or more A, Cj.salkanoyl optionally substituted with one or more A, C,_¢alkylsulphonyl optionally substituted with one or more A, C;galkoxycarbonyl optionally substituted with one or more A, carbamoyl,

N-(C,.¢alkyl)carbamoyl optionally substituted with one or more A, N-(C,.salkyl);carbamoyl optionally substituted with one or more A and benzoyl optionally substituted with one or more

A; and

R* is hydrogen or fluoro; or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

Accordingly to an additional aspect of the present invention there is provided a compound of formula (I) (as depicted above) wherein: : nislor2;

R! is chloro, fluoro, bromo, methyl or methoxy;

R? is selected from one of the following three groups: 1) halo, nitro, hydroxy, amino or cyano; ii) -X'-R’ wherein X' is a direct bond, -O-, -S-, -SO-, -SO;-, -NR®-, -CO-, -CONR-,

NR®CO-, -NR®SO,- or NR®CONR’-; wherein RS and R’ are independently hydrogen or

Cialkyl optionally substituted with one or more A; and RS is selected from C,salkyl optionally substituted with one or more A, Cs.scycloalkyl optionally substituted with one or more A, C, ¢alkenyl optionally substituted with one or more A, C; alkynyl optionally substituted with one or more A, phenyl! optionally substituted with one or more D, a carbon linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D or a carbon linked 5-membered heteroaryl ring containing 1-3 heteroatoms selected independently from O, N and S optionally substituted on a ring carbon by one or more D and wherein if said 5-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G; iii) a nitrogen-linked 4-8 membered heterocyclic group optionally substituted on a ring carbon by one or more D and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

R® is C;.¢alkyl optionally substituted with one or more A, Cj.scycloalkyl optionally substituted with one or more A, phenyl optionally substituted with one or more D, a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D, or a carbon linked 5-membered heteroaryl ring containing 1-3 heteroatoms selected independently from O, N and S optionally substituted on a ring carbon by one or more D and wherein if said S-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

A is selected from hydroxy, amino, halo, carboxy, N-(C;4alkyl)amino,

N,N-di-(C;.4alkyl)amino and C,_salkoxy;

D is selected from: i) -X*-R® wherein X® is a direct bond, -O-, -S-, -SO-, -SO,-, -CO-, -NR?SO,-, -NRCO-, -NRUCONR®-, -NR?- or -CONR"-; wherein R® and R°® are independently hydrogen or C,_salkyl optionally substituted with one or more hydroxy or C;4alkoxy; and R® is selected from hydrogen or C,alkyl optionally substituted with one or more hydroxy or C,_salkoxy; ii) a 4-8 membered Het which is optionally substituted on a ring carbon with one or more groups selected from hydroxy, halo, C;.4alkoxy, C;4alkyl or cyano and wherein if said 4-8 membered Het contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G; iii) -X®-C,_¢alkyl-X"-R°® wherein X* and R° are as defined hereinbefore and X® is -S-, -SO- or -SO,-; iv) cyano or halo; and v)-X°-R! wherein X° is -C(0)- or -SO,- and R' is a nitrogen-linked 4-8 membered heterocyclic group optionally substituted on a ring carbon by one or more groups selected from hydroxy, halo, C;_4alkoxy, C «alkyl or cyano and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G;

G is selected from C,.galkyl optionally substituted with one or more A, C.¢alkanoyl optionally substituted with one or more A, Cy.salkylsulphonyl optionally substituted with one or more A, C;.salkoxycarbonyl optionally substituted with one or more A, carbamoyl, .

N-(C,¢alkyl)carbamoyl optionally substituted with one or more A, N-(C,.galkyl).carbamoyl optionally substituted with one or more A and benzoyl optionally substituted with one or more )

A; and

Preferred values of R!, R?, R® and R* are as follows. Such values may be used where appropriate with any of the definitions, claims or embodiments defined hereinbefore or hereinafter.

In one aspect of the invention preferably n is 1.

In another aspect of the invention preferably n is 2. :

Preferably R'is chloro, fluoro or bromo.

More preferably R! is chloro or fluoro.

Particularly R' is chloro.

In another aspect of the invention, preferably R' is methyl, chloro, fluoro or bromo.

In another aspect of the invention, more preferably R! is methyl, chloro or fluoro.

In another aspect of the invention, particularly Rlis methyl or chloro.

Where R? is selected from group i):

Preferably group i) is halo or hydroxy.

More preferably group i) is halo.

Particularly group i) is chloro or fluoro.

More particularly group i) is chloro.

In another aspect of the invention, where R? is selected from group i):

Preferably group i) is nitro, halo, amino or hydroxy.

More preferably group i) is nitro, amino or halo.

Particularly group i) is nitro, bromo, iodo, amino, chloro or fluoro.

Where R? is selected from group ii):

Preferably in group ii) X! is -S-, -SO-, -SO,-, -NR"- or -NR°CO-; preferably RS is hydrogen; and preferably R3 is selected from C;_galkyl optionally substituted with one or more

A, phenyl! optionally substituted with one or more D or a carbon linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D.

More preferably in group ii) X' is -S-, -SO-, -SO;- or -NR®CO-; more preferably R® is hydrogen; and more preferably R? is selected from C,4alkyl optionally substituted with one or more A, phenyl optionally substituted with one or more D or a carbon linked pyridyl! optionally substituted on a ring carbon by one or more D.

Particularly in group ii) X' is -S- or -NR®CO-; particularly R® is hydrogen; and particularly R® is selected from methyl optionally substituted with one or more A, ethyl optionally substituted with one or more A, pheny! optionally substituted with one or more D or a carbon linked pyridyl optionally substituted on a ring carbon by one or more D.

In another aspect of the invention, where R? is selected from group ii):

Preferably in group ii) X' is -S-, -SO-, -SO;-, -NR®- or -NR®CO-; preferably R® is hydrogen; and preferably RS is selected from C,.salkyl optionally substituted with one or more

A or phenyl optionally substituted with one or more D.

More preferably in group ii) X' is -S-, -SO-, -SO;- or -NR®CO-; more preferably R® is hydrogen; and more preferably R’ is selected from C)_salkyl optionally substituted with one or more A or phenyl optionally substituted with one or more D.

Particularly in group ii) X' is -S- or -NR°CO-; particularly R® is hydrogen; and particularly R’ is selected from methyl optionally substituted with one or more A, ethyl

-13- ' a optionally substituted with one or more A or phenyl optionally substituted with one or more

D.

In another aspect of the invention, where R? is selected from group ii): : Preferably in group ii) X' is -O-, -S-, -SO-, -SO,-, -NR®- or -NR®CO-; preferably R® is hydrogen; and preferably R’ is selected from C 1-salkyl optionally substituted with one or more oe A, phenyl optionally substituted with one or more D or phenylC,_ salkyl optionally substituted with one or more D. - More preferably in group ii) X' is -O-, -S-, -SO-, -SO;- or -NR®CO-; more preferably

R® is hydrogen; and more preferably R? is selected from Ci.salkyl optionally substituted with one or more A, phenyl optionally substituted with one or more D or phenylC; 4alkyl optionally substituted with one or more D. :

Where R? is selected from group iii):

Preferably group iii) is a nitrogen-linked 5 or 6 membered heterocyclic group “ optionally substituted on a ring carbon by one or more D and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G.

More preferably group iii) is a nitrogen-linked 6 membered heterocyclic group ; optionally substituted on a ring carbon by one or more D and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group . selected from G.

Particularly group iii) is morpholino optionally substituted on a ring carbon by one or more D, piperidin-1-yl optionally substituted on a ring carbon by one or more D or piperazin-1-y] optionally substituted on a ring carbon by one or more D and optionally substituted on the -NH- moiety by a group selected from G.

In another aspect of the invention, where R? is selected from group iii):

Particularly group iii) is morpholino optionally substituted on a ring carbon by one or more D, thiomorpholino optionally substituted on a ring carbon by one or more D, piperidin-1-yl optionally substituted on a ring carbon by one or more D or piperazin-1-yl optionally substituted on a ring carbon by one or more D and optionally substituted on the -NH- moiety by a group selected from G.

More particularly group iii) is thiomorpholino.

In another aspect of the invention, where R? is selected from group iii):

0 . -14-

Particularly group iii) is morpholino, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino or piperazin-1-yl optionally substituted on the -NH- moiety by a group selected from G.

Preferably A is hydroxy, amino, halo, carboxy and methoxy. 3 More preferably A is hydroxy.

In another aspect of the invention, preferably A is hydroxy, amino, dimethylamino, halo, carboxy and methoxy.

In another aspect of the invention, more preferably A is hydroxy, methoxy and dimethylamino.

In another aspect of the invention, preferably A is hydroxy, amino, dimethylamino, halo, carboxy, methoxy and carbamoyl.

Where D is selected from group i):

Preferably X* in group i) is -S-, -SO-, -SO-, -NR%-, -NR'CONR"- or -CONR’-; preferably R? is hydrogen or Cj_alkyl optionally substituted with one or more hydroxy; and preferably RC is selected from hydrogen or C;.¢alkyl optionally substituted with one or more hydroxy. } More preferably X* in group i) is -S-, -SO-, -SO,-, -NR%- or -CONR-; more preferably

RY is hydrogen, methyl or ethyl optionally substituted with hydroxy; and more preferably R°® is selected from hydrogen or C,4alkyl optionally substituted with one or more hydroxy.

More preferably X? in group i) is -SO-, -SO;-, -NR* or -CONR’-; more preferably R® is hydrogen, methyl or ethyl optionally substituted with hydroxy; and more preferably R® is selected from hydrogen, methyl or ethyl optionally substituted with hydroxy.

In another aspect of the invention, where D is selected from group i):

Preferably X® in group i) is -SO-, -SO;-, -NR% or -CONR-; more preferably Ris hydrogen, methyl or ethyl optionally substituted with hydroxy; and more preferably R® is selected from hydrogen, methyl, ethyl or butyl optionally substituted with hydroxy.

Where D is selected from group ii):

Preferably group ii) isa 5 or 6 membered Het which is optionally substituted on a ring carbon with one or more groups selected from hydroxy, halo, C,4alkoxy, Cj4alkyl or cyano and wherein if said 5 or 6 membered Het contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G.

More preferably group ii) is a 5 or 6 membered Het which is optionally substituted on a ring carbon with one or more groups selected from hydroxy, halo, methyl, ethyl, methoxy,

-15- . x ethoxy or cyano and wherein if said 5 or 6 membered Het contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G.

Particularly group ii) is morpholino, morpholinyl, piperidinyl or piperazinyl optionally substituted on the -NH- moiety by a group selected from G.

Where D is selected from group iii):

Preferably group iii) is -X*-C4alkyl-X-R® wherein X® and R° are as defined hereinbefore and X" is preferably -SO- or -SO»-.

Where D is selected from group iv):

Preferably group iv) is cyano, fluoro or chloro.

More preferably group iv) is fluoro or chloro.

In another aspect of the invention, where D is selected from group iv):

Preferably group iv) is fluoro.

Where D is selected from group v):

Preferably X° is -C(O)- and R'is a nitrogen-linked 4-6 membered heterocyclic group optionally substituted by hydroxy.

More preferably X° is -C(O)- and R'is azetidinyl, morpholino or pyrrolidinyl (optionally substituted by hydroxy). .

Particularly X° is -C(O)- and Rf is azetidinyl, morpholino or 3-hydroxypyrrolidinyl.

Preferably G is Cj.¢alkanoyl optionally substituted with one or more A or C, alkyl } optionally substituted by one or more A.

More preferably G is C)4alkanoyl optionally substituted with one or more A or

C.4alkyl optionally substituted by one or more A.

Particularly G is acetyl or C, 4alkyl substituted by one or more A.

More particularly G is acetyl.

Preferably R?is chloro, fluoro, methylthio, acetylamino, hydroxy, C,.salkylsulphiny! (optionally substituted with hydroxy), Cj4alkylsulphonyl (optionally substituted with hydroxy), phenylsulphonyl [optionally substituted with halo, amino, N-(C,4alkyl),carbamoyl (optionally substituted with hydroxy), N-(C,4alkyl)carbamoy! (optionally substituted with hydroxy), N-(Cj.salkyl)amino (optionally substituted with hydroxy), N-(C;.4alkyl);amino (optionally substituted with hydroxy), C;4alkylsulphinyl (optionally substituted with hydroxy), C.salkylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)-piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl}, pyridylsulphony! [optionally substituted with halo,

amino, N-(C,4alkyl),amino (optionally substituted with hydroxy), N-(C;.4alkyl),carbamoyl (optionally substituted with hydroxy), N-(C;4alkyl)carbamoyl (optionally substituted with hydroxy), N-(C;.salkyl)amino (optionally substituted with hydroxy), C;.salkylsulphinyl (optionally substituted with hydroxy), C.4alkylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethy!)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl}, N-(C;salkyl)amino (optionally substituted with hydroxy), morpholino, 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropy!)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yL

More preferably R? is chloro, fluoro, methylthio, acetylamino, hydroxy, methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), phenylsulphonyl [optionally substituted with halo, amino, N,N-dimethylcarbamoyl, N,N-diethylcarbamoy! (optionally substituted with hydroxy),

N-methyl-N-ethylcarbamoyl (optionally substituted with hydroxy), N-methylcarbamoyl,

N-ethylcarbamoyl (optionally substituted with hydroxy), methylamino, ethylamino (optionally substituted with hydroxy), N,N-dimethylamino, N,N-diethylamino (optionally substituted with hydroxy), N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)-piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], pyridylsulphonyl [optionally substituted with halo, amino, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl (optionally substituted with hydroxy),

N-ethylcarbamoyl (optionally substituted with hydroxy), N,N-dimethylamino,

N,N-diethylamino (optionally substituted with hydroxy), N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), methylamino, ethylaminio (optionally substituted with hydroxy), mesyl, } ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl}, methylamino, ethylamino (optionally substituted with hydroxy), morpholino, 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl.

Particularly R? is chloro, fluoro, methylthio, acetylamino or hydroxy.

In another aspect of the invention, preferably R? is chloro, fluoro, bromo, iodo, nitro, amino, methylthio, acetylamino, hydroxy, C;alkylsulphanyl (optionally substituted with hydroxy), C;.salkylsulphinyl (optionally substituted with hydroxy), C;_alkylsulphonyl (optionally substituted with hydroxy), N-(C,4alkyl)amino (optionally substituted with hydroxy, methoxy or dimethylamino), phenylsulphonyl [optionally substituted with halo, amino, N-(C,4alkyl),carbamoyl (optionally substituted with hydroxy), N-(C,.qalkyl)carbamoyl (optionally substituted with hydroxy), N-(C,4alkyl)amino (optionally substituted with hydroxy), N-(Ci4alkyl);amino (optionally substituted with hydroxy), C;4alkylsulphinyl (optionally substituted with hydroxy), C;.salkylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)-piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], pyridylsulphonyl [optionally substituted with halo, amino, N-(C,4alkyl),;amino (optionally substituted with hydroxy), N-(C, 4alkyl);carbamoy] (optionally substituted with hydroxy),

N-(C,4alkyl)carbamoyl (optionally substituted with hydroxy), N-(C,4alkyl)amino (optionally substituted with hydroxy), C,_salkylsulphinyl (optionally substituted with hydroxy),

Ci.salkylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, . 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], N-(Csalkyl)amino (optionally substituted with . hydroxy), morpholino, 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin- 1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], thiomorpholino, phenylsulphanyl (optionally substituted with N-(C,alkyl),carbamoyl) or phenylsulphinyl (optionally substituted with

N-(Cj4alkyl),carbamoyl).

In a further aspect of the invention preferably R? is chloro, fluoro, bromo, iodo, nitro, hydroxy, amino, methylthio, acetylamino, Ci4alkylsulphany! (optionally substituted with hydroxy), C,.4alkylsulphinyl, C,4alkylsulphonyl, N-(C,4alkyl)amino (optionally substituted : with hydroxy, methoxy or dimethylamino), thiomorpholino, phenylsulphany! (optionally substituted with N-(C.salkyl)ocarbamoyl) or phenylsulphinyl (optionally substituted with

N-(Ci4alkyl);carbamoyl).

In another aspect of the invention, more preferably R? is chloro, fluoro, bromo, iodo, nitro, amino, methyIthio, acetylamino, hydroxy, methylsulphanyl, ethylsulphanyl (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with

. k -18- . hydroxy), mesyl, ethylsulphony! (optionally substituted with hydroxy), methylamino, ethylamino (optionally substituted with hydroxy, methoxy or dimethylamino), phenylsulphonyl [optionally substituted with halo, amino, N,N-dimethylcarbamoyl,

N,N-diethylcarbamoyl (optionally substituted with hydroxy), N-methyl-N-ethylcarbamoyl (optionally substituted with hydroxy), N-methylcarbamoyl, N-ethylcarbamoyl (optionally substituted with hydroxy), methylamino, ethylamino (optionally substituted with hydroxy),

N,N-dimethylamino, N,N-diethylamino (optionally substituted with hydroxy),

N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphiny! (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)-piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], pyridylsulphonyl [optionally substituted with halo, amino, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl (optionally substituted with hydroxy), N-methyl-N-ethylcarbamoyl (optionally substituted with hydroxy), N-methylcarbamoyl, N-ethylcarbamoyl (optionally substituted with hydroxy),

N,N-dimethylamino, N,N-diethylamino (optionally substituted with hydroxy),

N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), methylamino, ethylamino (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropy!)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], methylamino, ethylamino (optionally substituted with hydroxy), morpholino, 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethy!)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], thiomorpholino or phenylsulphanyl [optionally substituted with N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl (optionally substituted with hydroxy), N-methyl-N-ethylcarbamoyl (optionally substituted with hydroxy)].

In another aspect of the invention, particularly R? is fluoro, chloro, bromo, iodo, nitro, amino, hydroxy, methylthio, ethylsulphinyl, mesyl, 2-hydroxyethylamino, 2-methoxyethylamino, 2-dimethylaminoethylamino, 2,3-dihydroxypropylamino, 2-hydroxyethylsulphanyl, acetylamino, 4-N,N-dimethylcarbamoylphenylsulphanyl, 4-N,N-dimethylcarbamoylphenylsulphinyl or thiomorpholino.

In another aspect of the invention, preferably R? is chloro, fluoro, bromo, iodo, nitro, amino, methoxy, acetylamino, hydroxy, C;4alkylsulphany! (optionally substituted with hydroxy), C,.salkylsulphinyl, C,_4alkylsulphonyl, N-(C,;4alkyl)amino (optionally substituted

-19- . a with hydroxy, methoxy, dimethylamino or carbamoyl), morpholino, 4-acetylpiperazin-1-yl, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, benzylamino, phenoxy, phenylsulphanyl (optionally substituted with N-(C).4alkyl);carbamoyl) or phenylsulphinyl (optionally substituted with N-(C,4alkyl);carbamoyl).

In another aspect of the invention, more preferably R? is chloro, fluoro, bromo, iodo, nitro, amino, methoxy, acetylamino, hydroxy, methylthio, 2-hydroxyethylthio, methylsulphinyl, mesyl, 2-hydroxyethylamino, 2-methoxyethylamino, carbamoylmethylamino, 2-dimethylaminoethylamino, 2,3-dihydroxypropylamino, morpholino, 4-acetylpiperazin-1-y], thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, benzylamino, phenoxy, 4-(N,N-dimethylcarbamoyl)phenylsulphanyl or 4-(N, N-dimethylcarbamoyl)phenylsulphinyl.

In another aspect of the invention, particularly Ris methylthio, morpholino, 4-acetylpiperazin-1-yl, 1-oxothiomorpholino or 1,1-dioxothiomorpholino.

In a further aspect of the invention more preferably R? is amino, 2-hydroxyethylamino or 2-methoxyethylamino.

In an additional aspect of the invention more preferably R? is fluoro or chloro.

Preferably R® is C;.alkyl optionally substituted with one or more A, phenyl optionally . substituted with one or more D or a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D. .

More preferably R? is C)4alkyl optionally substituted with one or more A, phenyl optionally substituted with one or more D or a carbon-linked pyridyl optionally substituted on a ring carbon by one or more D.

Particularly R? is methyl optionally substituted with one or more A, ethyl optionally substituted with one or more A, phenyl optionally substituted with one or more D or a carbon-linked pyridyl optionally substituted on a ring carbon by one or more D.

Particularly R? is methyl, ethyl optionally substituted with A, phenyl optionally substituted with one or more D or a carbon-linked pyridyl optionally substituted on a ring carbon by one or more D.

Therefore, in another aspect of the invention preferably R® is C;_salkyl optionally substituted with one or more hydroxy, phenyl [optionally substituted with halo, amino,

N-(Ci.salkyl),carbamoy] (optionally substituted with hydroxy), N-(C,.salkyl)carbamoyl (optionally substituted with hydroxy), N-(C,4alkyl)amino (optionally substituted with hydroxy), N-(C;salkyl)»,amino (optionally substituted with hydroxy), C,_4alkylsulphinyl

Lo -20- (optionally substituted with hydroxy), C;4alkylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)- piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl], or carbon-linked pyridyl [optionally substituted with halo, amino, N-(C;4alkyl),amino (optionally substituted with hydroxy), N-(C;.salkyl),carbamoyl (optionally substituted with : hydroxy), N-(C).salkyl)carbamoyl (optionally substituted with hydroxy), N-(Cj4alkyl)amino (optionally substituted with hydroxy), C;4alkylsulphinyl (optionally substituted with hydroxy), C)salkylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropy!)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl].

Particularly R? is methyl, ethyl optionally substituted with hydroxy, phenyl [optionally substituted with halo, amino, N,N-dimethylcarbamoyl, N, N-diethylcarbamoyl (optionally substituted with hydroxy), N-methyl-N-ethylcarbamoy! (optionally substituted with hydroxy),

N-methylcarbamoyl, N-ethylcarbamoy! (optionally substituted with hydroxy), methylamino, ethylamino (optionally substituted with hydroxy), N,N-dimethylamino, N,N-diethylamino (optionally substituted with hydroxy), N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl] or carbon-linked pyridyl [optionally substituted with halo, amino, N, N-dimethylcarbamoyl, N,N-diethylcarbamoyl (optionally substituted with hydroxy), N-methyl-N-ethylcarbamoyl (optionally substituted with hydroxy),

N-methylcarbamoyl, N-ethylcarbamoyl (optionally substituted with hydroxy), methylamino, ethylamino (optionally substituted with hydroxy), N,N-dimethylamino, N,N-diethylamino (optionally substituted with hydroxy), N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl].

More particularly R? is methyl, ethyl optionally substituted with hydroxy, or phenyl (optionally substituted with halo).

Particularly preferred R? is ethyl or 4-fluorophenyl.

Therefore, in another aspect of the invention preferably R? is C;4alkyl (optionally substituted with one or more hydroxy), phenyl [optionally substituted with halo, amino,

N-(C,4alkyl),carbamoyl (optionally substituted with hydroxy), N-(C,.salkyl)carbamoyl (optionally substituted with hydroxy), N-(C)4alkyl)amino (optionally substituted with hydroxy), N-(Ci.salkyl);amino (optionally substituted with hydroxy), C).salkylsulphinyl (optionally substituted with hydroxy), C;salkylsulphony! (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)-piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl, 4-(2-hydroxypropyl)piperazin-1-yl, azetidinylcarbonyl, morpholinocarbonyl or pyrrolidinylcarbonyl (optionally substituted with hydroxy)], or carbon-linked pyridyl [optionally substituted with halo, amino, N-(C,4alkyl),amino (optionally substituted with hydroxy), N-(C,.salkyl).carbamoy! (optionally substituted with hydroxy), N-(C;4alkyl)carbamoy! (optionally substituted with hydroxy), N-(C;4alkyl)amino (optionally substituted with hydroxy), Cialkylsulphinyl (optionally substituted with hydroxy), C4alkylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl].

Therefore, in a further aspect of the invention preferably R}is C;.4alkyl (optionally . substituted with one or more hydroxy), phenyl [optionally substituted with halo,

N-(C,.salkyl),carbamoyl, N-(C;.salkyl)carbamoyl, N-(C;4alkyl)amino (optionally substituted i with hydroxy), Cjsalkylsulphonyl, azetidinylcarbonyl, morpholinocarbonyl or pyrrolidinylcarbony! (optionally substituted with hydroxy)], or carbon-linked pyridyl [optionally substituted with amino].

Particularly R? is methyl, ethyl (optionally substituted with hydroxy), butyl (optionally substituted with hydroxy), phenyl [optionally substituted with halo, amino,

N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl (optionally substituted with hydroxy),

N-methyl-N-ethylcarbamoy] (optionally substituted with hydroxy), N-methylcarbamoyl,

N-ethylcarbamoyl (optionally substituted with hydroxy), methylamino, ethylamino (optionally . substituted with hydroxy), N,N-dimethylamino, N,N-diethylamino (optionally substituted with hydroxy), N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl, 4-(2-hydroxypropyl)piperazin-1-yl, azetidinylcarbonyl, morpholinocarbony! or pyrrolidinylcarbonyl (optionally substituted with hydroxy)] or carbon-linked pyridyl [optionally substituted with halo, amino, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl (optionally substituted with hydroxy), N-methyl-N-ethylcarbamoyl (optionally substituted with hydroxy), N-methylcarbamoyl, N-ethylcarbamoy! (optionally substituted with hydroxy), 5S methylamino, ethylamino (optionally substituted with hydroxy), N, N-dimethylamino,

N,N-diethylamino (optionally substituted with hydroxy), N-methyl-N-ethylamino (optionally substituted with hydroxy), methylsulphinyl, ethylsulphinyl (optionally substituted with hydroxy), mesyl, ethylsulphonyl (optionally substituted with hydroxy), 4-acetylpiperazin-1-yl, 4-methylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-(3-hydroxypropyl)piperazin-1-yl or 4-(2-hydroxypropyl)piperazin-1-yl].

More particularly R? is methyl, ethyl, 2-hydroxyethyl, 2-hydroxybutyl, 4-fluorophenyl, 4-mesylphenyl, 4-(2-hydroxyethylamino)phenyl, 4-(N-methylcarbamoyl)phenyl, 4-(N-ethylcarbamoyl)phenyl, 4-(N, N-dimethylcarbamoyl)phenyl, 4-(N-methyl-N-ethylcarbamoyl)phenyl, 4-(azetidinylcarbonyl)phenyl, 4-(morpholinocarbonyl)phenyl, 4-(3-hydroxypyrrolidinylcarbonyl)phenyl or 6-aminopyrid-2-yl.

In another aspect of the invention particularly Ris methyl, ethyl (optionally substituted with hydroxy), isopropyl, butyl (optionally substituted with hydroxy), phenyl [optionally substituted with halo, N,N-dimethylcarbamoyl, N-methyl-N-ethylcarbamoyl,

N-methylcarbamoyl, N-ethylcarbamoyl, ethylamino (optionally substituted with hydroxy), mesyl, azetidinylcarbonyl, morpholinocarbony! or pyrrolidinylcarbonyl (optionally substituted with hydroxy)] or carbon-linked pyridyl [optionally substituted with amino].

In another aspect of the invention more particularly R? is methyl, ethyl, 2-hydroxyethyl, isopropyl, 2-hydroxybutyl, 4-fluorophenyl, 4-(2-hydroxyethylamino)phenyl, 4-mesylphenyl, 4-(N,N-dimethylcarbamoyl)phenyl, 4-(N-ethylcarbamoyl)phenyl, 4-(N-methyl-N-ethylcarbamoyl)phenyl, 4-(N-methylcarbamoyl)phenyl, 4-(azetidinylcarbonyl)phenyl, 4-(morpholinocarbonyl)phenyl, 4-(3-hydroxypyrrolidinylcarbonyl)phenyl or 2-aminopyrid-6-yl.

In another aspect of the invention more particularly preferred R® is methyl, ethyl or isopropyl.

In a further aspect of the invention more particularly preferred R3 is 4-(N-methylcarbamoyl)phenyl or 4-(N, N-dimethylcarbamoyl)phenyl.

In a further aspect of the invention especially particularly preferred R® is 4-(N,N-dimethylcarbamoyl)phenyl.

In one aspect of the invention, preferably R* is hydrogen.

In another aspect of the invention, preferably R* is fluoro.

At the -C(OH)(Me)(CFs) chiral center, the R-configuration is generally the preferred stereochemistry.

Therefore in another aspect of the invention, there is provided a compound of the formula (I) as depicted above wherein: nis 1or2;

R'is methyl, chloro or fluoro;

R%is chloro, fluoro, bromo, iodo, nitro, amino, methoxy, acetylamino, hydroxy,

Ci4alkylsulphanyl (optionally substituted with hydroxy), C,4alkylsulphinyl,

Ci 4alkylsulphonyl, N-(C,.salkyl)amino (optionally substituted with hydroxy, methoxy, dimethylamino or carbamoyl), morpholino, 4-acetylpiperazin- 1-yl, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, benzylamino, phenoxy, phenylsulphanyl (optionally substituted with N-(C, 4alkyl),carbamoyl) or phenylsulphiny! (optionally substituted with N-(C;_4alkyl),carbamoyl); .

R? is methyl, ethyl (optionally substituted with hydroxy), isopropyl, butyl (optionally substituted with hydroxy), phenyl [optionally substituted with halo, N,N-dimethylcarbamoyl,

N-methyl-N-ethylcarbamoyl, N-methylcarbamoyl, N-ethylcarbamoyl, ethylamino (optionally substituted with hydroxy), mesyl, azetidinylcarbonyl, morpholinocarbonyl or pyrrolidinylcarbonyl (optionally substituted with hydroxy)] or carbon-linked pyridyl {optionally substituted with amino]; and

Ris hydrogen, or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

Therefore in another aspect of the invention, there is provided a compound of the formula (I) as depicted above wherein: . nis 2;

R'is chloro;

R? is methylthio, morpholino, 4-acetylpiperazin-1-yl, 1-oxothiomorpholino or 1,1-dioxothiomorpholino;

R? is methyl, ethyl or isopropyl;

R*is hydrogen; - . or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

A preferred compound of the invention is any one of the Examples or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

More preferred compounds of the invention are Examples 7, 8, 22, 23, 24, 28, 48, 64, 69, 70, 74, 75 or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

In another aspect of the invention, more preferred compounds of the invention are

Examples 32, 35 and 61 or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

In a further aspect of the invention more preferred compounds of the invention are

Examples 17, 18 and 58 or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

Preferred aspects of the invention are those which relate to the compound or a pharmaceutically acceptable salt thereof.

Within the present invention it is to be understood that a compound of the formula (I) or a salt thereof may exhibit the phenomenon of tautomerism and that the formulae drawings } within this specification can represent only one of the possible tautomeric forms. It is to be understood that the invention encompasses any tautomeric form which elevates PDH activity and is not to be limited merely to any one tautomeric form utilized within the formulae drawings. The formulae drawings within this specification can represent only one of the possible tautomeric forms and it is to be understood that the specification encompasses all possible tautomeric forms of the compounds drawn not just those forms which it has been possible to show graphically herein.

It will be appreciated by those skilled in the art that certain compounds of formula (I) contain one or more asymmetrically substituted carbon and/or sulphur atoms, and accordingly may exist in, and be isolated as enantiomerically pure, a mixture of diastereoisomers or as a racemate. Some compounds may exhibit polymorphism. It is to be understood that the present , invention encompasses any racemic, optically-active, enantiomerically pure, mixture of diastereoisomers, polymorphic or stereoisomeric form, or mixtures thereof, which form possesses properties useful in the elevation of PDH activity, it being well known in the art how to prepare optically-active forms (for example, by resolution of the racemic form by recrystallization techniques, by synthesis from optically-active starting materials, by chiral synthesis, by enzymatic resolution, (for example WO 9738124), by biotransformation, or by

- 25 - "w n chromatographic separation using a chiral stationary phase) and how to determine efficacy for the elevation of PDH activity by the standard tests described hereinafter.

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 elevate PDH activity.

A compound of the formula (I), or salt thereof, and other compounds of the invention (as hereinafter defined) may be prepared by any process known to be applicable to the preparation of chemically-related compounds. Such processes include, for example, those illustrated in European Patent Applications, Publication Nos. 0524781, 0617010, 0625516, and in GB 2278054, WO 9323358 and WO 9738124.

Another aspect of the present invention provides a process for preparing a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof, which process (in which variable groups are as defined for formula (I) unless otherwise stated) comprises of: (a) deprotecting a protected compound of formula (II): .

R#4 3 .

R S(O) o

ALE

R N

) H CF,

R OPg am where Pg is an alcohol protecting group; (b) oxidising a compound of formula (III):

RES

0 ’ Me

R N

, H CF,

R OH

(c) coupling compounds of formula (IV):

R*

R- ae!

R' i

RE 0) with an acid of formula (V): 0

AL

X oH wv) wherein X is OH; (d) coupling an aniline of formula (IV) with an activated acid derivative of formula (V); and thereafter if necessary: ; i) converting a compound of the formula (I) into another compound of the formula (I); ii) removing any protecting groups; or : iii) forming a pharmaceutically acceptable salt or in vivo hydrolysable ester.

Suitable values for Pg are a benzyl group, a silyl group (for example a trialkylsilyl group or an alkyldiphenylsilyl group) or an acetyl protecting group.

Where formula (V) is an activated acid derivative, suitable values for X include halo (for example chloro or bromo), anhydrides, aryloxys (for example 4-nitrophenoxy or pentafluorophenoxy) or imidazol-1-yl.

Specific conditions of the above reactions are as follows:

Process a)

Examples of suitable reagents for deprotecting an alcohol of formula (II) are: : 1) when Pg is benzyl: (i) hydrogen in the presence of palladium/carbon catalyst, i.e. hydrogenolysis; or (ii) hydrogen bromide or hydrogen iodide; 2) when Pg is a silyl protecting group: (i) tetrabutylammonium fluoride; or

(ii) aqueous hydrofluoric acid; 3) when Pg is acetyl: i) mild aqueous base for example lithium hydroxide; or ii) ammonia or an amine such as dimethylamine.

The reaction can be conducted in a suitable solvent such as EtOH, MeOH, acetonitrile, or DMSO and may conveniently be performed at a temperature in the range of -40 to 100°C.

Compounds of formula (IT) may be prepared according to the following scheme:

EO Me Standard

M i M

HO WM¢ CF, E-OH, H,SO, CF, Protecting EO Me CF,

EtOAc Group

O OH 0 OH Conditions (3 OPg (Ila) AcCL (tb) (Ie)

Toluene Aq LiOH (For Pg = THF

Acetyl) i) (COCl),, DMF, HO Me CE wm kM ii) (IV), Jd ope 2,6-di-r-butylpyridine, a1 d)

DCM )

Scheme 1

E is a carboxy protecting group. Suitable values for E include C;.salkyl, such as ) methyl and ethyl.

Compounds of formula (IIa) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art. The synthesis of compounds of formula (IV) is described below.

Process b) .

Suitable oxidising agents include potassium permanganate, OXONE, sodium periodate, tert-butyl hydroperoxide (as solution in toluene), peracids (such as for example 3-chloroperoxybenzoic acid), hydrogen peroxide, TPAP (tetrapropylammonium perruthenate) or oxygen. The reaction may be conducted in a suitable solvent such as ether,

DCM, MeOH, EtOH, water, acetic acid, or mixtures of two or more of these solvents. The reaction may conveniently be performed at a temperature in the range of -40 to 100°C.

Compounds of formula (III) may be prepared according to the following schemes:

R* R* (V) (X=Cl), -_ 0 2,6-di-t-butylpyridine, Ae

R N ] , NH pcm RY N Fy

R R OH

(IIa) (IIIb)

Je

Either i) R3SH, CuCl (or Cu,0), R*

NMP, A I 0 al) —-->-----— e ii) R3SH, Pd(0), gH Oy

NaOMe, DMF, A (IIIc)

Scheme 2

The skilled reader will appreciate that the order of steps 1 and 2 in Scheme 2 may be reversed.

R* R*

X RSM, EtOH R’S - Fr NO, R NO,

R' R . (ITId) (Ille)

Fe, HCI,

EtOH.

R? (V) (X=CD), R’S am -— 2,6-di-t-butylpyridine,

DCM R™ Tq, NH,

R

(ITI)

Scheme 3 wherein M is an alkali metal. Suitable values for M include lithium, sodium or potassium. }

X is a leaving group, suitable values for X include halo, mesyl and tosyl.

Re Ie

NaSCN S ———————————————

R NH, Br,, MeOH, NaBr R NH,

R' R (IIIa) (Ig) (V) X=C1), . 2,6-di-t-butylpyridine,

DCM

N

(IS

R3X, Cu,0, Na,S

BE —————— EE. ai) S 0 e rR H om : (ITTh)

Scheme 4

X is a leaving group, suitable values for X include halo, mesyl and tosyl.

Compounds of formula (ITla) and (ITId) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art.

Process ¢) ’

The reaction can be conducted in the presence of a suitable coupling reagent. Standard peptide coupling reagents known in the art can be employed as suitable coupling reagents, for : example conditions such as those described above for the coupling of (IId) and (IV), or for example dicyclohexyl-carbodiimide, optionally in the presence of a catalyst such as dimethylaminopyridine or 4-pyrrolidinopyridine, optionally in the presence of a base for example triethylamine, pyridine, or 2,6-di-alkyl-pyridines (such as 2,6-lutidine or 2,6-di-tert-butylpyridine) or 2,6-diphenylpyridine. Suitable solvents include DMA, DCM, benzene, THF, and DMF. The coupling reaction may conveniently be performed at a temperature in the range of -40 to 40°C.

Compounds of formula (IV) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art, for example they may be prepared by oxidising compounds of formula (IIIf) (with the aniline protected with a suitable protecting group) under standard oxidation conditions, for example with hydrogen peroxide or meta-chloroperoxybenzoic acid (followed by de-protection), or they may be prepared according to the following scheme:

R? Either

Re I i) R3SH, CuCl (or Cu,0),

ICI ol NMP, A VI)

J RY NL or

R' R ii) R3SH, Pd(0), (IVa) (IVb) NaOMe, DMF, A

Scheme §

Compounds of formula (IVa) and (V) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art.

If the resolved acid of formula (V) is required it may be prepared by any of the known methods for preparation of optically-active forms (for example, by recrystallization of the chiral salt {for example WO 9738 124}, by enzymatic resolution or by chromatographic separation using a chiral stationary phase). For example if an (R)-(+) resolved acid is required it may be prepared by the method of Scheme 2 in World Patent Application Publication No.

WO 9738124 for preparation of the (S)-(-) acid, i.e. using the classical resolution method described in European Patent Application Publication No. EP 0524781, also for preparation of the (S)-(-) acid, except that (1S,2R)-norephedrine may be used in place of ' (S)-(-)-1-phenylethylamine. The chiral acid may also be prepared by using the enzymatic resolution method as described in Tetrahedron Asymmetry, 1999, 10, 679.

Process d)

This coupling may be achieved optionally in the presence of a base for example triethylamine, pyridine, 2,6-di-alkyl-pyridines (such as 2,6-lutidine or 2,6-di-tert-butylpyridine) or 2,6-diphenylpyridine. Suitable solvents include DMA, DCM, benzene, THF, and DMF. The coupling reaction may conveniently be performed at a temperature in the range of -40 to 40°C.

If not commercially available, the necessary starting materials for the procedures such as that described above may be made by procedures which are selected from standard organic chemical techniques, techniques which are analogous to the synthesis of known, structurally similar compounds, or techniques which are analogous to the above described procedure or the procedures described in the examples.

For example, it will be appreciated that certain of the optional aromatic substituents in the compounds of the present invention may be introduced by standard aromatic substitution reactions or generated by conventional functional group modifications or interconversions either prior to or immediately following the processes mentioned above, and as such are

-31- v . included in the process aspect of the invention. Such reactions and modifications include, for example, introduction of a substituent by means of an aromatic substitution reaction, reduction of substituents, alkylation of substituents and oxidation of substituents. The reagents and reaction conditions for such procedures are well known in the chemical art. Particular examples of aromatic substitution reactions include the introduction of a nitro group using concentrated nitric acid, the introduction of an acyl group using, for example, an acylhalide and Lewis acid (such as aluminium trichloride) under Friedel Crafts conditions; the introduction of an alkyl group using an alkyl halide and Lewis acid (such as aluminium trichloride) under Friedel Crafts conditions; and the introduction of a halogeno group.

Particular examples of modifications include the reduction of a nitro group to an amino group by, for example, catalytic hydrogenation with a nickel catalyst or treatment with iron in the presence of hydrochloric acid with heating; oxidation of alkylthio to alkylsulphinyl or alkylsulphonyl using, for example, hydrogen peroxide in acetic acid with heating or 3-chloroperbenzoic acid. Particular examples of functional group interconversions are for example conversion of an aniline into a halophenyl by, for example, diazotization in the presence of cupurous halides.

It is noted that many of the starting materials for synthetic methods as described above } are commercially available and/or widely reported in the scientific literature, or could be made from commercially available compounds using adaptations of processes reported in the scientific literature.

It will also be appreciated that in some of the reactions mentioned herein it may be necessary/desirable to protect any sensitive groups in the compounds. The instances where protection is necessary or desirable and suitable methods for protection are known to those skilled in the art. Thus, if reactants include groups such as amino, carboxy or hydroxy it may be desirable to protect the group in some of the reactions mentioned herein.

A suitable protecting group for an amino or alkylamino group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or z-butoxycarbonyl group, an arylmethoxycarbonyl group, for example benzyloxycarbonyl, or an aroyl group, for example benzoyl. The deprotection conditions for the above protecting groups necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or alkoxycarbonyl group or an aroyl group may be removed for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide. Alternatively an acyl group such as a t-butoxycarbonyl group may be removed, for example, by treatment with a suitable acid such as, for example hydrochloric, sulphuric or phosphoric acid or TFA and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group may be removed, for example, by hydrogenation in the presence of a catalyst such as palladium-on-carbon, or by treatment with a Lewis acid for example boron tris(trifluoroacetate). A suitable alternative protecting group for a primary amino group is, for example, a phthaloyl group which may be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or with hydrazine.

A suitable protecting group for a hydroxy group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an aroyl group, for example benzoyl, or an arylmethyl group, for example benzyl. The deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or an aroyl group may be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide.

Alternatively an arylmethyl group such as a benzyl group may be removed, for example, by hydrogenation in the presence of a catalyst such as palladium-on-carbon.

A suitable protecting group for a carboxy group is, for example, an esterifying group, } for example a methyl or an ethyl group which may be removed, for example, by hydrolysis with a base such as sodium hydroxide, or for example a t-butyl group which may be removed, for example, by treatment with an acid, for example an organic acid such as TFA, or for example a benzyl group which may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.

The protecting groups may be removed at any convenient stage in the synthesis using conventional techniques well known in the chemical art.

In cases where compounds of formula (I) are sufficiently basic or acidic to form stable acid or basic salts, administration of the compound as a salt may be appropriate, and pharmaceutically acceptable salts may be made by conventional methods such as those described following. Examples of suitable pharmaceutically acceptable salts are organic acid . addition salts formed with acids which form a physiologically acceptable anion, for example, tosylate, methanesulphonate, acetate, tartrate, citrate, succinate, benzoate, ascorbate, o-ketoglutarate, and o-glycerophosphate. Suitable inorganic salts may also be formed such as sulphate, nitrate, and hydrochloride.

Pharmaceutically acceptable salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound of formula (I) (or its ester) with a suitable acid affording a physiologically acceptable anion. It is also possible with most compounds of the invention to make a corresponding alkali metal (e.g. sodium, potassium, or lithium) or alkaline earth metal (e.g. calcium) salt by treating a compound of formula (I) (and in some cases the ester) with one equivalent of an alkali metal or alkaline earth metal hydroxide or alkoxide (e.g. the ethoxide or methoxide) in aqueous medium followed by conventional purification techniques.

The compounds of the formula (I) may be administered in the form of a prodrug which is broken down in the human or animal body to give a compound of the formula (I). Examples of prodrugs include in vivo hydrolysable esters of a compound of the formula (I).

An in vivo hydrolysable ester of a compound of the formula (I) containing carboxy or hydroxy group is, for example, a pharmaceutically acceptable ester which is hydrolysed in the human or animal body to produce the parent acid or alcohol.

Suitable in vivo hydrolysable esters for a compound of the formula (I) containing a carboxy group include C,.salkoxymethyl esters for example methoxymethyl,

C,.salkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidyl esters,

Cs.scycloalkoxycarbonyloxyCi alkyl esters for example 1-cyclohexylcarbonyloxyethyl; : 1,3-dioxolen-2-onylmethyl esters for example 5-methyl-1,3-dioxolen-2-onylmethyl; and

Ci.salkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxyethyl and may be : formed at any carboxy group in the compounds of this invention.

Suitable in vivo hydrolysable esters of a compound of the formula (I) containing a hydroxy group includes inorganic esters such as phosphate esters and a-acyloxyalkyl ethers.

Examples of a-acyloxyalkyl ethers include acetoxymethoxy and 2,2-dimethylpropionyloxymethoxy. Other in vivo hydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl and

N-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates), dialkylaminoacety! and carboxyacetyl. Examples of substituents for benzoyl include morpholino and piperazino linked from a ring nitrogen atom via a methylene group to the 3- or 4- position of the benzoyl ring.

In vivo cleavable prodrugs of compounds of formula (I) also include in vivo hydrolysable amides of compounds of the formula (I) containing a carboxy group, for

. 0 . -34- example, a N-C, alkyl or N-di-C, alkyl amide such as N-methyl, N-ethyl, N-propyl,

N-dimethyl, N-ethyl-N-methyl or N-diethyl amide.

The identification of compounds which elevate PDH activity is the subject of the present invention. These properties may be assessed, for example, using one or more of the procedures set out below: (a) In vitro elevation of PDH activity

This assay determines the ability of a test compound to elevate PDH activity. cDNA encoding PDH kinase may be obtained by Polymerase Chain Reaction (PCR) and subsequent cloning. This may be expressed in a suitable expression system to obtain polypeptide with

PDH kinase activity. For example rat PDHkinasell (rPDHKII) obtained by expression of recombinant protein in Escherichia coli (E. Coli), was found to display PDH kinase activity.

In the case of the tPDHKII (Genbank accession number U10357) a 1.3kb fragment encoding the protein was isolated by PCR from rat liver cDNA and cloned into a vector (for example pQE32 - Quiagen Ltd.). The recombinant construct was transformed into E. coli (for example M15pRep4 - Quiagen Ltd.). Recombinant clones were identified, plasmid DNA was isolated and subjected to DNA sequence analysis. One clone which had the expected nucleic } acid sequence was selected for the expression work. Details of the methods for the assembly of recombinant DNA molecules and the expression of recombinant proteins in bacterial systems can be found in standard texts for example Sambrook et al, 1989, Molecular Cloning - A Laboratory Manual, 2" edition, Cold Spring Harbour Laboratory Press. Other known

PDH kinases for use in assays, may be cloned and expressed in a similar manner.

For expression of rPDHKII activity, E. coli strain M15pRep4 cells were transformed with the pQE32 vector containing rPDHKII cDNA. This vector incorporates a 6-His tag onto the protein at its N-terminus. E. coli were grown to an optical density of 0.6 (600 nM) and protein expression was induced by the addition of 10 pM isopropylthio-B-galactosidase. Cells were grown for 18 hours at 18°C and harvested by centrifugation. The resuspended cell paste was lysed by homogenisation and insoluble material removed by centrifugation at 24000xg for 1 hour. The 6-His tagged protein was removed from the supernatant using a nickel chelating nitrilotriacetic acid resin (Ni-NTA: Quiagen Ltd.) matrix (Quiagen) which was washed with 20 mM tris(hydroxymethyl)aminomethane-hydrogen chloride, 20 mM imidazole, 0.5 M sodium chloride pH 8.0, prior to elution of bound protein using a buffer containing 20 mM tris(hydroxymethyl)aminomethane-hydrogen chloride, 200 mM imidazole, 0.15 M sodium chloride pH 8.0. Eluted fractions containing 6-His protein were pooled and stored in aliquots at -80°C in 10% glycerol.

Each new batch of stock enzyme was titrated in the assay to determine a concentration giving approximately 90% inhibition of PDH in the conditions of the assay. For a typical batch, stock enzyme was diluted to 7.5pug/ml.

For assay of the activity of novel compounds, compounds were diluted with 10%

DMSO and 10pl transferred to individual wells of 96-well assay plates. Control wells contained 20pl 10% DMSO instead of compound. 40u! Buffer containing 50mM potassium phosphate buffer pH 7.0, 10mM ethylene glycol-bis(B-aminoethyl ether)-N,N-tetracetic acid (EGTA), 1mM benzamidine, mM phenylmethylsulphonyl fluoride (PMSF), 0.3mM tosyl-L-lysine chloromethyl ketone (TLCK), 2mM dithiothreitol (DTT), recombinant rPDHKII and compounds were incubated in the presence of PDH kinase at room temperature for 45 minutes. In order to determine the maximum rate of the PDH reaction a second series of control wells were included containing 10% DMSO instead of compound and omitting rPDHKII PDH kinase activity was then initiated by the addition of 5 uM ATP, 2 mM magnesium chloride and 0.04 U/ml PDH (porcine heart PDH Sigma P7032) in a total volume of 50 ul and plates incubated at ambient temperature for a further 45 minutes. The residual : activity of the PDH was then determined by the addition of substrates (2.5mM coenzyme A, 2.5mM thiamine pyrophosphate (cocarboxylase), 2.5mM sodium pyruvate, 6mM NAD in a : total volume of 80ul and the plates incubated for 90 minutes at ambient temperature. The : production of reduced NAD (NADH) was established by measured optical density at 340nm using a plate reading spectrophotometer. The EDs for a test compound was determined in the usual way using results from 12 concentrations of the compound. (b) In vitro elevation of PDH activity in isolated primary cells

This assay determines the ability of compounds to stimulate pyruvate oxidation in primary rat hepatocytes.

Hepatocytes were isolated by the two-step collagenase digestion procedure described by Seglen (Methods Cell Biol. (1976) 13, 29-33) and plated out in 6-well culture plates (Falcon Primaria) at 600000 viable cells per well in Dulbecco’s Modified Eagles Medium (DMEM, Gibco BRL) containing 10% foetal calf serum (FCS), 10% penicillin/streptomycin (Gibco BRL) and 10% non-essential amino acids (NEAA, Gibco BRL). After 4 hours incubation at 37°C in 5% CO, the medium was replaced with Minimum Essential Medium

. o -36- (MEM, Gibco BRL) containing NEAA and penicillin/streptomycin as above in addition to 10nM dexamethasone and 10nM insulin.

The following day cells were washed with phosphate buffered saline (PBS) and medium replaced with 1m! HEPES-buffered Krebs solution (25mM HEPES, 0.15M sodium chloride, 25 mM sodium hydrogen carbonate, SmM potassium chloride, 2mM calcium chloride, 1mM magnesium sulphate, 1 mM potassium dihydrogen phosphate) containing the compound to be tested at the required concentration in 0.1% DMSO. Control wells contained 0.1% DMSO only and a maximum response was determined using a 10 uM treatment of a known active compound. After a preincubation period of 40 minutes at 37°C in 5% CO,, cells were pulsed with sodium pyruvate to a final concentration of 0.5mM (containing 1-C sodium pyruvate (Amersham product CFA85) 0.18Ci/mmole) for 12 minutes. The medium was then removed and transferred to a tube which was immediately sealed with a bung containing a suspended centre well. Absorbent within the centre well was saturated with 50% phenylethylamine, and CO; in the medium released by the addition of 0.2pl 60% (w/v) perchloric acid (PCA). Released “CO, trapped in the absorbent was determined by liquid scintillation counting. The EDs for a test compound was determined in the usual way using . results from 7 concentrations of the compound. (c) In vivo elevation of PDH activity ) The capacity of compounds to increase the activity of PDH in relevant tissues of rats may be measured using the test described hereinafter. Typically an increase in the proportion of PDH in its active, nonphosphorylated form may be detected in muscle, heart, liver and adipose tissue after a single administration of an active compound. This may be expected to lead to a decrease in blood glucose after repeated administration of the compound. For example a single administration of DCA, a compound known to activate PDH by inhibition of

PDH kinase (Whitehouse, Cooper and Randle (1974) Biochem. J. 141, 761-774) 150 mg/kg, intraperitoneally, increased the proportion of PDH in its active form (Vary et al. (1988) Circ.

Shock 24, 3-18) and after repeated administration resulted in a significant decrease in plasma glucose (Evans and Stacpoole (1982) Biochem. Pharmacol.31, 1295-1300).

Groups of rats (weight range 140-180g) are treated with a single dose or multiple doses of the compound of interest by oral gavage in an appropriate vehicle. A control group of rats is treated with vehicle only. At a fixed time after the final administration of compound, animals are terminally anaesthetised, tissues are removed and frozen in liquid nitrogen. For determination of PDH activity, muscle samples are disrupted under liquid nitrogen prior to homogenisation by one thirty-second burst in a Polytron homogenizer in 4 volumes of a buffer containing 40 mM potassium phosphate pH 7.0, 5 mM EDTA, 2mM DTT, 1% Triton X-100, 10mM sodium pyruvate, 10uM phenylmethylsulphonyl chloride (PMSF) and 2pg/ml each of leupeptin, pepstain A and aprotinin. Extracts are centrifuged before assay. A portion of the extract is treated with PDH phosphatase prepared from pig hearts by the method of Siess and

Wieland (Eur. J. Biochem (1972) 26, 96): 20 pl extract, 40 pl phosphatase (1:20 dilution), in a final volume of 125 pl containing 25 mM magnesium chloride, 1 mM calcium chloride. The activity of the untreated sample is compared with the activity of the dephosphorylated extract thus prepared. PDH activity is assayed by the method of Stansbie et al., (Biochem. J. (1976) 154, 225). 50 wl Extract is incubated with 0.75 mM NAD, 0.2 mM CoA, 1.5 mM thiamine pyrophosphate (TPP) and 1.5mM sodium pyruvate in the presence of 20 pg/ml p-(p-amino-phenylazo) benzene sulphonic acid (AABS) and 50 mU/ml arylamine transferase (AAT) in a buffer containing 100 mM tris(hydroxymethyl)aminomethane, 0.5 mM EDTA, 50mM sodium fluoride, 5SmM 2-mercaptoethanol and 1mM magnesium chloride pH 7.8. AAT is prepared from pigeon livers by the method of Tabor et al. (J. Biol. Chem. (1953) 204, 127).

The rate of acetyl CoA formation is determined by the rate of reduction of AABS which is . indicated by a decrease in optical density at 460 nm.

Liver samples are prepared by an essentially similar method, except that sodium : pyruvate is excluded from the extraction buffer and added to the phosphatase incubation to a final concentration of SmM.

Treatment of an animal with an active compound results in an increase in the activity of PDH complex in tissues. This is indicated by an increase in the amount of active PDH (determined by the activity of untreated extract as a percentage of the total PDH activity in the same extract after treatment with phosphatase).

According to a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the formula (I) as defined hereinbefore or a : pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof, in association with a pharmaceutically acceptable excipient or carrier.

The composition may be in a form suitable for oral administration, for example as a tablet or capsule, for parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion) for example as a sterile solution, suspension or emulsion, for topical administration for example as an ointment or cream or for rectal administration for example as a suppository. In general the above compositions may be prepared in a conventional manner using conventional excipients.

The compositions of the present invention are advantageously presented in unit dosage form. The compound will normally be administered to a warm-blooded animal at a unit dose within the range 5-5000 mg per square metre body area of the animal, i.e. approximately 0.1-100 mg/kg. A unit dose in the range, for example, 1-100 mg/kg, preferably 1-50 mg/kg is envisaged and this normally provides a therapeutically-effective dose. A unit dose form such as a tablet or capsule will usually contain, for example 1-250 mg of active ingredient.

According to a further aspect of the present invention there is provided a compound of the formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof as defined hereinbefore for use in a method of treatment of the human or animal body by therapy.

We have found that compounds of the present invention elevate PDH activity and are therefore of interest for their blood glucose-lowering effects.

A further feature of the present invention is a compound of formula (I) and pharmaceutically acceptable salts or in vivo hydrolysable esters thereof for use as a medicament.

Conveniently this is a compound of formula (I), or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof, for use as a medicament for producing an elevation of

PDH activity in a warm-blooded animal such as a human being.

Particularly this is a compound of formula (I), or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof, for use as a medicament for treating diabetes mellitus in a warm-blooded animal such as a human being.

In another aspect of the invention, particularly this is a compound of formula (I), or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof, for use as a medicament for treating diabetes mellitus, peripheral vascular disease and myocardial ischaemia 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 the formula (I), or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof in the manufacture of a medicament for use in the production of an elevation of

PDH activity in a warm-blooded animal such as a human being.

-39. . iN

Thus according to a further aspect of the invention there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof in the manufacture of a medicament for use in the treatment of diabetes mellitus 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 the formula (I), or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof in the manufacture of a medicament for use in the treatment of diabetes mellitus, peripheral vascular disease and myocardial ischaemia in a warm-blooded animal such as a human being.

According to a further feature of the invention there is provided a method for producing an elevation of PDH 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) or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof as defined hereinbefore.

According to a further feature of the invention there is provided a method of treating diabetes mellitus 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) or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof as defined hereinbefore. .

According to a further feature of the invention there is provided a method of treating diabetes mellitus, peripheral vascular disease and myocardial ischaemia 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) or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof 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.-

The elevation of PDH activity described herein may be applied as a sole therapy or may involve, in addition to the subject of the present invention, one or more other substances and/or treatments. Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate administration of the individual components of the treatment. For example in the treatment of diabetes mellitus chemotherapy may include the following main categories of treatment: i) insulin; ii) insulin secretagogue agents designed to stimulate insulin secretion (for example glibenclamide, tolbutamide, other sulphonylureas); iii) oral hypoglycaemic agents such as metformin, thiazolidinediones; iv) agents designed to reduce the absorption of glucose from the intestine (for example acarbose); v) agents designed to treat complications of prolonged hyperglycaemia; vi) other agents used to treat lactic acidaemia; vii) inhibitors of fatty acid oxidation; viii) lipid lowering agents; ix) agents used to treat coronary heart disease and peripheral vascular disease such as aspirin, pentoxifylline, cilostazol; and/or x) thiamine.

As stated above the compounds defined in the present invention are of interest for their ability to elevate the activity of PDH. Such compounds of the invention may therefore be useful in a range of disease states including diabetes mellitus, peripheral vascular disease, (including intermittent claudication), cardiac failure and certain cardiac myopathies, myocardial ischaemia, cerebral ischaemia and reperfusion, muscle weakness, hyperlipidaemias, Alzheimer’s disease and/or atherosclerosis. Alternatively such compounds of the invention may be useful in a range of disease states including peripheral vascular disease, (including intermittent claudication), cardiac failure and certain cardiac myopathies, myocardial ischaemia, cerebral ischaemia and reperfusion, muscle weakness, hyperlipidaemias, Alzheimer’s disease and/or atherosclerosis in particular peripheral vascular disease and myocardial ischaemia.

In addition to their use in therapeutic medicine, the compounds of formula (I) and their pharmaceutically acceptable salts 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 elevators of PDH activity in laboratory animals such as cats, dogs, rabbits, monkeys, rats and mice, as part of the search for new therapeutic agents. - The invention will now be illustrated by the following non-limiting 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 sulphate; evaporation of solvent was carried out using a rotary evaporator under reduced pressure (600-4000 Pascals; 4.5-30 mm Hg) with a bath temperature of up to 60°C; (iii) chromatography means flash chromatography on silica gel; thin layer chromatography (TLC) was carried out on silica gel plates; where a silica Mega Bond Elut column is referred to, this means a column containing 1Q g or 20 g or 50 g of silica of 40 micron particle size, the silica being contained in a 60 ml disposable syringe and supported by a porous disc, obtained from Varian, Harbor City, California, USA under the name “Mega Bond Elut ST”; “Mega

Bond Elut” is a trademark; where a Biotage cartridge is referred to this means a cartridge containing KP-SIL™ silica, 604, particle size 32-63mM, supplied by Biotage, a division of

Dyax Corp., 1500 Avon Street Extended, Charlottesville, VA 22902, USA; (iv) where a Chem Elut column is referred to, this means a “Hydromatrix™ extraction cartridge for adsorption of aqueous material, i.e. a polypropylene tube containing a special grade of flux-calcined, high purity, inert diatomaceous earth, pre-buffered to pH 4.5 or 9.0, incorporating a phase-separation filtering material, used according to the manufacturers instructions, obtained from Varian, Harbor City, California, USA under the name of “Extube,

Chem Elut”; “Extube” is a registered trademark of International Sorbent Technology Limited; (v) where an ISOLUTE column is referred to, this means an “ion exchange” extraction cartridge for adsorption of basic or acid material, i.e. a polypropylene tube containing a special grade of ion exchange sorbent, high purity, surface to pH ~7, incorporating a phase-separation filtering material, used according to the manufacturers instructions, obtained from Varian, Harbor City, California, USA under the name of “Extube, Chem Elut,

ISOLUTE”; “Extube” is a registered trademark of International Sorbent Technology Limited; (vi) in general, the course of reactions was followed by TLC and reaction times are given for illustration only;

(vii) final products had satisfactory proton nuclear magnetic resonance (NMR) spectra and/or mass spectral data; (viii) 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; (ix) where given, NMR data is in the form of delta values for major diagnostic protons, given in parts per million (ppm) relative to tetramethylsilane (TMS) as an internal standard, : determined at 300 MHz using perdeuterio dimethyl sulphoxide (DMSO-8) as solvent unless otherwise indicated, other solvents (where indicated in the text) include deuterated chloroform - CDCl; coupling constants (J) are given in Hz; Ar designates an aromatic proton when such an assignment is made; (x) chemical symbols have their usual meanings; SI units and symbols are used; (xi) reduced pressures are given as absolute pressures in Pascals (Pa); elevated pressures are given as gauge pressures in bars; (xii) solvent ratios are given in volume : volume (v/v) terms; (xiii) mass spectra (MS) were run with an electron energy of 70 electron volts in the chemical : ionisation (CI) mode using a direct exposure probe; where indicated ionisation was effected by electron impact (EI), fast atom bombardment (FAB) or electrospray (ESP); values for m/z are given; generally, only ions which indicate the parent mass are reported and unless otherwise stated the value quoted is (M-H)’; (xiv) Oxone is a Trademark of E.I. du Pont de Nemours & Co., Inc., and refers to potassium peroxymonosulphate; : (xv) The following abbreviations are used: ether diethyl ether;

DMF N,N-dimethylformamide;

DMA N,N-dimethylacetamide;

TFA trifluoroacetic acid;

NMP N-methylpyrrolidin-2-one

SM starting material;

DMSO dimethylsulphoxide;

EtOAc ethyl acetate;

MeOH methanol;

EtOH ethanol;

DCM dichloromethane; and

THF tetrahydrofuran; and (xvi) where (R) or (S) stereochemistry is quoted at the beginning of a name, unless further clarified, it is to be understood that the indicated stereochemistry refers to the -NH-C(0)-C*(Me)(CF3)(OH) centre as depicted in formula (I).

Example 1 (R)-N-(2,3-Dichloro-4-ethylsulphinylphenyl)-2-hydroxy-2-methyl-3.3 3- trifluoropropanamide t-Butyl hydrogen peroxide (2.4 ml of a 5.5M solution in decane) was added to a solution of (R)-N-[2,3-dichloro-4-ethylsulphanylphenyl]-2-hydroxy-2-methy}-3,3,3- trifluoropropanamide (Method 1) (0.23 g) and d-10-camphorsulphonic acid (0.018 g) in chloroform (10 ml) and the mixture was stirred for 18 hours. Volatile material was removed by evaporation and the residue was purified by chromatography on a silica gel Mega Bond

Elut column eluting with 0-50% EtOAc / isohexane to give the title compound (0.22 g) as a white solid. NMR (CDCl; + 1 drop DMSO): 1.21-1.28 (m, 3H), 1.71 (s, 3H), 2.77-2.89 (m, 1H), 3.04-3.16 (m, 1H), 7.14 (s, 1H), 7.78 (d, 1H), 8.66 (d, 1H), 9.73 (s, 1H); m/z: 376.

Examples 2-8

Following the procedure of Example 1 and using the appropriate starting materials the X following compounds were made. -

I CE Co (R)-N-[4-Methylsulphinyl-3- 1.62 (s, 3H), 2.86 (s, 3H), 346 Meth 12 fluoro-2-chlorophenyl]-2- 7.71-7.76 (m, 1H), 8.04-8.10 hydroxy-2-methyl-3,3,3- (m, 1H), 9.94 (brs, 1H) trifluoropropanamide (R)-N-[4-Ethylsulphinyl-3- 1.08 (t, 3H), 1.61 (s, 3H), 360 Meth 27 fluoro-2-chlorophenyl]-2- 2.83-2.94 (m, 1H), 3.08-3.22 hydroxy-2-methyl-3,3,3- (m, 1H), 7.64 (d, 1H), 7.97 trifluoropropanamide (brs, 1H), 8.05-8.09 (m, 1H),

9.94 (brs, 1H)

(R)-N-[4-Ethylsulphinyl-3-iodo- | 1.09 (s, 3H), 1.61 (s, 3H), 468 Meth 28

2-chlorophenyl]-2-hydroxy-2- 2.71-2.85 (m, 1H), 3.07-3.19 methyl-3,3,3- (m, 1H), 7.62 (d, 1H), 8.27 trifluoropropanamide (d, 1H)

(R)-N-[4-Methylsulphinyl-2,3- | 1.6 (s, 3H), 2.8 (s, 3H), 7.8 | 362 Meth 33 . dichlorophenyl]-2-hydroxy-2- (d, 1H), 7.9 (s, 1H), 8.2 (d,

methyl-3,3,3- 1H), 9.9 (s, 1H)

(R)-N-[2-Chloro-3-(1- 1.61 (s, 3H), 2.83-3.02 (m, | 461 Ex 72 oxothiomorpholino)-4-(methyl- | 6H), 3.38 (s, 3H), 4.19-4.28 sulphonyl)phenyl]-2-hydroxy-2- | (m, 2H), 7.94 (d, 1H), 8.07 methyl-3,3,3- «| (brs, 1H), 8.23 (d, 1H), 9.95 trifluoropropanamide

7% | (R)-N-[2-Chloro-3-(1- 1.13 (t, 3H), 1.62 (s, 3H), 475 Ex 65 oxothiomorpholino)-4-(ethyl- 2.85-2.89 (m, 6H), 3.43-3.50 sulphonyl)phenyl]-2-hydroxy-2- | (gq, 2H), 4.17-4.27 (m, 2H), methyl-3,3,3- 7.93 (d, 1H), 8.11 (brs, 1H), trifluoropropanamide 8.27 (d, 1H), 9.96 (brs, 1H)

(R)-N-[2-Chloro-3-(1- 1.17-1.25 (m, 6H), 1.62 (s, | 489 Ex 73 : oxothiomorpholino)-4- 3H), 2.87 (brm, 6H), (isopropylsulphonyl)phenyl]-2- | 3.71-3.80 (m, 1H), 4.19-4.23 hydroxy-2-methyl-3,3,3- (m, 2H), 7.91 (d, 1H), 8.11 trifluoropropanamide (brs, 1H), 8.26-8.31 (m, 1H), 9.95 (brs, 1H)

Product obtained by addition of DCM to residue after evaporation followed by filtration. 2 Product was a mixture of two diastereoisomers, Example 4 is the less polar diastereoisomer. ’ Residue was purified on a 8g silica Biotage cartridge eluting 3% MeOH / DCM. “Residue was purified on a 8g silica Biotage cartridge eluting 10% MeOH / EtOAc.

Example 9 (R)-N-(2,3-Dichloro-4-ethylsulphonylphenyD)-2-hydroxy-2-methy}-3.3.3- trifluoropropanamide

Hydrogen peroxide (15 ml of a 30 wt. % solution in water) was added to a solution of (R)-N-[4-ethylsulphanyl-2,3-dichlorophenyl]-2-hydroxy-2-methyl-3,3,3- trifluoropropanamide . (Method 1) (1.88 g) in glacial acetic acid (26 ml) and the mixture was heated at 95°C for 1.5 hours then cooled. EtOAc (200 ml) was added and the mixture was washed with saturated : aqueous sodium hydrogen carbonate solution (4 x 200 ml) and brine (250 ml) then was dried.

Volatile material was removed by evaporation and the residue was purified by chromatography eluting with 0-50% EtOAc / isohexane to give the title compound (1.71 g) as a white solid. NMR: 1.1 (t, 3H), 1.61 (s, 3H), 3.5 (q, 2H), 8.02 (d, 1H), 8.31 (d, 1H); m/z: 392.

Examples 10-26

Following the procedure of Example 9 and using the appropriate starting materials the : following compounds were prepared:

I CT Cc CoC (R)-N-(3-Acetamido-2-chloro- | 1.61 (s, 3H), 1.9 (s, 3H), 747 | 481 Meth 4-{4-fluorophenylsulphonyl} (t, 2H), 7.86 (m, 2H), 8.09 (brs, 11 phenyl)-2-hydroxy-2-methyl-3, 1H), 8.22 (d, 1H), 8.40 (d, 1H), 3,3-trifluoropropanamide 9.81 (brs, 1H), 9.9-(brs, 1H)

11 | (R)-N-(2-Chloro-3-fluoro-4- 1.6 (s, 3H), 7.53 (t, 2H), 442 Meth {4-fluorophenylsulphonyl} 7.99-8.2 (m, SH), 10.0 (brs, 2 phenyl)-2-hydroxy-2-methyl- | 1H) 3,3,3-trifluoropropanamide

12 | (R)-N-[4-(2-Hydroxyethyl- 1.62 (s, 3H), 3.66-3.74 (m, 408 Ex 44

! | sulphonyl)-2,3-dichloro- 4H), 4.82 (t, 2H), 8.03 (d, 1H), phenyl}-2-hydroxy-2-methyl- 8.29 (d, 1H), 9.99 (brs, 1H) 3,3,3-trifluoropropanamide (R)-N-[4-(2-Hydroxy-n-butyl- | 0.81 (t, 3H), 1.31-1.52 (m, 436 Meth sulphonyl)-2,3-dichloro- 2H), 1.62 (s, 3H), 3.50-3.69 34

. phenyl]-2-hydroxy-2-methyl- - | (m, 2H), 3.76-3.84 (m, 1H), 3,3,3-trifluoropropanamide 0.81 (t, 1H), 8.02 (d, 1H), 8.08

) (s, 1H), 8.25-8.29 (m, 1H),

10.01 (s, 1H)

(R)-N-(4-Mesyl-3-fluoro-2- 1.62 (s, 3H), 3.35 (s, 3H), 362 | Ex2 chlorophenyl)-2-hydroxy-2- 7.84-7.90 (m, 1H), 8.06 (s, methyl-3,3,3- 1H), 8.13 (d, 1H), 10.01 (brs, trifluoropropanamide 1H) (R)-N-(4-Ethylsulphonyl-3- 1.16 (t, 3H), 1.62 (s, 3H), 376 Meth fluoro-2-chlorophenyl)-2- 3.38-3.46 (q, 2H), 7.85 (t, 1H), 27 hydroxy-2-methyl-3,3,3- 8.16 (d, 1H), 10.08 (brs, 1H) triflsoropropanamide

(R)-N-(4-Ethylsulphonyl-3- 1.12 (t, 3H), 1.61 (s, 3H), 484 iodo-2-chlorophenyl)-2- 3.51-3.58 (q, 2H), 8.07 (d, 1H), hydroxy-2-methyl-3,3,3- 8.36 (d, 1H) trifluoropropanamide

17 | (R)-N-{2,3-Dichloro-4-[4- 1.60 (s, 3H), 2.85 (s, 3H), 2.95 | 511 Meth (N,N-dimethylcarbamoyl) (s, 3H), 7.60 (d, 2H); 7.95 (d, 36 phenylsulphonyllphenyl}-2- | 2H), 8.10 (s, 1H), 8.40 (dd, hydroxy-2-methyl-3,3,3- 2H), 10.0 (s, 1H) trifluoropropanamide

18 | (R)-N-{2-Chloro-3-fluoro-4- 1.60 (s, 3H), 2.95 (s, 3H), 3.00 | 495 Meth [4-(N,N-dimethylcarbamoyl) (s, 3H), 7.65 (d, 2H); 8.05 (d, 5 phenylsulphonyl]phenyl}-2- 2H), 8.05-8.15 (m, 2H), 8.20 hydroxy-2-methyl-3,3,3- (d, 1H), 9.95 (s, 1H) trifluoropropanamide

19 | (R)-N-[2-Methyl-3-fluoro-4- 1.61 (s, 3H), 2.05 (s, 3H), 7.50 | 422 Meth

~ (4-fluorophenylsulphonyl) (t, 2H), 7.62 (d, 1H), 7.66 (brs, 43 phenyl]-2-hydroxy-2-methyl- 1H), 7.90 (t, 1H), 8.02 (m, 2H), 3,3,3-trifluoropropanamide 9.94 (brs, 1H) (R)-N-(2-Methyl-3-chloro-4- 1.60 (s, 3H), 2.20 (s, 3H), 7.46 | 438 Meth [4-fluorophenyl}sulphonyl ~~ | (t, 2H), 7.60 (brs, 1H), 7.70 (d, 44 phenyl)-2-hydroxy-2-methyl- | 1H), 7.99 (m, 2H), 8.20 (d, 3,3,3-trifluoropropanamide 1H), 10.03 (brs, 1H) :

21 | (R)-N-(2-Methyl-3-fluoro-4- 1.60 (s, 3H), 2.07 (s, 3H), 2.85 | 475 Meth [4-N,N-dimethylcarbamoyl- (s, 3H), 3.00 (s, 3H), 7.66 (m, 52 phenyl]sulphonylphenyl)-2- 4H), 7.98 (m, 3H), 9.90 (brs, hydroxy-2-methyl-3,3,3- 1H) trifluoropropanamide

Claims

Claims.

1. A compound of formula (I): R* AL R N RT on ) wherein: nis 1or2; R! is chloro, fluoro, bromo, methyl or methoxy; R? is selected from one of the following three groups: i) halo, nitro, hydroxy, amino or cyano; ii) -X'-R® wherein X' is a direct bond, -O-, -S-, -SO-, -SO;-, -NR®-, -CO-, -CONR®-, -NR®CO-, -NR®SO;- or NR’ CONR’-; wherein R® and R” are independently hydrogen or

Ci.4alkyl optionally substituted with one or more A; and R’ is selected from C,galkyl optionally substituted with one or more A, Cs.;cycloalkyl optionally substituted with one or more A, Cs;cycloalkylC,galkyl optionally substituted with one or more A, Cs.salkenyl optionally substituted with one or more A, C; alkynyl optionally substituted with one or more A, phenyl optionally substituted with one or more D, phenylC, salky! optionally substituted with one or more D, heteroaryl ring optionally substituted on a ring carbon by one or more D or (heteroaryl ring)C,salkyl optionally substituted on a ring carbon with one or more D; wherein said heteroaryl ring is a carbon linked 6-membered ring containing 1-2 nitrogen atoms or a carbon linked 5-membered ring containing 1-3 heteroatoms selected independently from O, N and S; and wherein if said 5-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G; iii) a nitrogen-linked 4-8 membered heterocyclic group optionally substituted on a ring carbon by one or more D and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G; R3 is C)_salkyl optionally substituted with one or more A, Cs.scycloalkyl optionally substituted with one or more A, phenyl optionally substituted with one or more D, a carbon-linked 6-membered heteroaryl ring containing 1-2 nitrogen atoms optionally substituted on a ring carbon by one or more D, or a carbon linked 5-membered heteroaryl ring containing 1-3 heteroatoms selected independently from O, N and S optionally substituted on a ring carbon by one or more D and wherein if said 5-membered heteroaryl ring contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G; - A is selected from hydroxy, amino, halo, carboxy, N-(C;4alkyl)amino, N,N-di-(C 1alkyl)amino, carbamoyl and C, salkoxy; - ~ Dis selected from: i) -X2-R® wherein X" is a direct bond, -O-, -S-, -SO-, -SO;-, -CO-, -NR*SO,-, -NR’CO-, -NRCONR®-, -NR'- or -CONR?-; wherein R? and R® are independently hydrogen or C;alkyl optionally substituted with one or more hydroxy or Cj4alkoxy; and R° is selected from oe hydrogen or C;.alkyl optionally substituted with one or more hydroxy or C,4alkoxy; - ii) a 4-8 membered Het which is optionally substituted on a ring carbon with one or more groups selected from hydroxy, halo, C;.salkoxy, Ci4alkyl or cyano and wherein if said 4-8 membered Het contains an -NH- moiety that nitrogen may be optionally substituted with.a group selected from G; iif) -X*-C, ¢alkyl-X"-R® wherein X® and R° are as defined hereinbefore and X” is -S-, -SO- or -SO,-; iv) cyano or halo; and . v) -X°-Rf wherein X® is -C(O)- or -SO- and Rf is a nitrogen-linked 4-8 membered heterocyclic group optionally substituted on a ring carbon by one or more groups selected from hydroxy, halo, C;4alkoxy, C;.salkyl or cyano and wherein if said heterocyclic group contains an -NH- moiety that nitrogen may be optionally substituted with a group selected from G; G is selected from C;.salkyl optionally substituted with one or more A, Cy.¢alkanoyl optionally substituted with one or more A, C;.¢alkylsulphony! optionally substituted with one or more A, Cj alkoxycarbonyl optionally substituted with one or more A, carbamoyl, . N-(C6alkyl)carbamoy! optionally substituted with one or more A, N-(C;.salkyl)2carbamoyl optionally substituted with one or more A and benzoyl optionally substituted with one or more A;and R'is hydrogen or fluoro; or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

2. A compound of formula (I) according to claim 1 wherein n is 2 or a pharmaceutically acceptable salt or an in vive hydrolysable ester thereof. :

3. A compound of formula (I) according to either of claims 1 or 2 wherein R' is methyl, chloro or fluoro or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

4, A compound of formula (I) according to any one of claims 1 to 3 wherein R? is chloro, fluoro, bromo, iodo, nitro, amino, methoxy, acetylamino, hydroxy, C,.salkylsulphanyl (optionally substituted with hydroxy), Ci4alkylsulphinyl, C;_salkylsulphonyl, N-(C;4alkyl)amino (optionally substituted with hydroxy, methoxy, dimethylamino or carbamoyl), morpholino, 4-acetylpiperazin-1-yl, thiomorpholino, 1-oxothiomorpholino, 1,1-dioxothiomorpholino, benzylamino, phenoxy, phenylsulphanyl (optionally substituted with N-(C,.salkyl),carbamoyl) or phenylsulphiny! (optionally substituted with N-+(Cj.alkyl),carbamoyl) or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

5. A compound of formula (I) according to any one of claims 1 to 4 wherein R? is methyl, ethyl (optionally substituted with hydroxy), isopropyl, butyl (optionally substituted with hydroxy), phenyl [optionally substituted with halo, N,N-dimethylcarbamoy], N-methyl-N-ethylcarbamoy!l, N-methylcarbamoyl, N-ethylcarbamoyl, ethylamino (optionally substituted with hydroxy), mesyl, azetidinylcarbony!l, morpholinocarbonyl or pyrrolidinylcarbonyl (optionally substituted with hydroxy)] or carbon-linked pyridyl [optionally substituted with amino] or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

6. A compound of formula (I) according to any one of claims 1 to 5 wherein R* is hydrogen or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

7. A compound of formula (I) selected from: (R)-N-[2-Chloro-3-(1-oxothiomorpholino)-4-(ethylsulphonyl)phenyl]-2-hydroxy-2-methyl- 3,3,3-trifluoropropanamide;

(R)-N-[2-Chloro-3-(1-oxothiomorpholino)-4-(isopropylsulphonyl)phenyl]-2-hydroxy-2- methyl-3,3,3-trifluoropropanamide; (R)-N-[2-Chloro-3-(1,1-dioxothiomorpholino)-4-(methylsulphonyl)phenyl]-2-hydroxy-2- methyi-3,3,3-trifluoropropanamide; . (R)-N-[2-Chloro-3-(1,1-dioxothiomorpholino)-4-(ethylsulphonyl)phenyl}-2-hydroxy-2- methyl-3,3,3-trifluoropropanamide; (R)-N-[2-Chloro-3-(1,1-dioxothiomorpholino)-4-(isopropylsulphonyl)phenyl}-2-hydroxy-2- methyl-3,3,3-trifluoropropanamide; (R)-N-(2-Chloro-4-ethylsulphonyl-3-methylsulphanylphenyl)-2-hydroxy-2-methyl-3,3,3- trifluoropropanamide; (R)-N-(4-Mesyl-3-methylsulphanyl-2-chlorophenyl)-2-hydroxy-2-methyl-3,3,3- trifluoropropanamide; (R)-N-[2-Chloro-3-(4-acetylpiperazin- 1-yl)-4-(ethylsulphonyl)phenyl]-2-hydroxy-2-methyl- 3,3,3-trifluoropropanamide; (R)-N-{2-Chloro-3-[1-(4-acetyl)piperazinyl]-4-(methylsulphonyl)phenyl}-2-hydroxy-2- methyl-3,3,3-triflucropropanamide; (R)-N-{2-Chloro-3-morpholino-4-(methylsulphonyl)phenyl]-2-hydroxy-2-methyl-3,3,3- } trifluoropropanamide; (R)-N-{2-Chloro-3-(4-acetylpiperazin- 1-yl)-4-(isopropylsulphonyl)phenyl]-2-hydroxy-2- methyl-3,3,3-trifluoropropanamide; and (R)-N-[2-Chloro-3-morpholino-4-(isopropylsulphonyl)phenyl]-2-hydroxy-2-methyl-3,3,3- trifluoropropanamide; or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

8. A process for preparing a compound of formula (I) according to claim 1 or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof, which process (in which variable groups are as defined for formula (I) unless otherwise stated) comprises of: : (a) deprotecting a protected compound of formula (II): :

{

R4 3 R S(O), 0 AL N 1 H CF, R OPg an where Pg is an alcohol protecting group; (b) oxidising a compound of formula (III): R4 RES 0) Me R N 1 H CF, R OH Im) (c) coupling compounds of formula (IV): R?4 R*—S5(0)s R NH, . R! av) with an acid of formula (V): 0) ANE X CF, OH m wherein X is OH; (d) coupling an aniline of formula (IV) with an activated acid derivative of formula (V); and thereafter if necessary: i) converting a compound of the formula (I) into another compound of the formula (I); ii) removing any protecting groups; or iii) forming a pharmaceutically acceptable salt or in vivo hydrolysable ester.

PCT/GB00/03314

9. A pharmaceutical composition which comprises a compound of formula (I) according to any one of claims 1 - 7, or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof in association with a pharmaceutically acceptable diluent or carrier.

10. A substance or composition for use in a method of treatment of the human or animal body by therapy, said substance or composition comprising a compound of the formula (I) according to any one of claims 1 - 7, or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, and said method comprising administering said substance or composition.

11. The use of a compound of the formula (I) according to any one of claims 1 - 7, or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, in the manufacture of a medicament for use in the treatment of diabetes mellitus, peripheral vascular disease and myocardial ischaemia in a warm-blooded animal such as a human being.

12. A compound of the formula (I) according to any one of claims 1 - 7, or a ~ pharmaceutically acceptable salt or in vivo hydrolysable ester thereof for use as a medicament.

13. The use of a compound of formula (I) according to any one of claims 1 - 7, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in the treatment of diabetes mellitus, peripheral vascular disease, (including intermittent claudication), cardiac failure and certain cardiac myopathies, myocardial ischaemia, cerebral ischaemia and reperfusion, muscle weakness, hyperlipidaemias, Alzheimer’s disease and/or atherosclerosis.

14. A compound according to claim 1, or claim 7, substantially as herein described and illustrated. AMENDED SHEET

: PCT/GB00/03314

15. A process according to claim 8, substantially as herein described and illustrated.

16. A composition according to claim 9, substantially as herein described and illustrated.

17. A substance or composition for use in a method of treatment according to claim 10, substantially as herein described and illustrated.

18. Use according to claim 11 or claim 13, substantially as herein described and illustrated.

19. A new compound, a new process for preparing a compound, a new composition, a substance or composition for a new use in a method of treatment, or a new use of a compound according to any one of claims 1 - 7, substantially as herein described. AMENDED SHEET