WO2011061277A1

WO2011061277A1 - Amino aryl acetamides and their use in the treatment of malaria

Info

Publication number: WO2011061277A1
Application number: PCT/EP2010/067806
Authority: WO
Inventors: Julia Castro-Pichel; Esther Pilar Fernandez-Velando; Jose Maria Fiandor-Roman; Maria De La O Gordo-Lopez; Maria Lourdes Rueda-Benede
Original assignee: Glaxo Group Ltd
Current assignee: Glaxo Group Ltd
Priority date: 2009-11-20
Filing date: 2010-11-19
Publication date: 2011-05-26
Anticipated expiration: 2012-05-20

Abstract

Amino phenyl acetamide compounds of Formula (I):and pharmaceutically acceptable salts thereof: wherein R¹, R², R³ and R^a are as defined in the description, use of such compounds in the chemotherapy of certain parasitic protozoal infections such as malaria, pharmaceutical compositions including such compounds and processes for the preparation of such compounds, are provided.

Description

AMINO ARYL ACETAMIDES AND THEIR USE IN THE TREATMENT

OF MALARIA

FIELD OF THE INVENTION

The present invention relates to amino aryl acetamide compounds and their use in chemotherapy. More specifically, this invention is concerned with amino phenyl acetamide compounds and their use in chemotherapy of certain parasitic protozoal infections such as malaria, in particular infection by Plasmodium falciparum, pharmaceutical compositions including such compounds and processes for the preparation of such compounds..

BACKGROUND OF THE INVENTION

Parasitic protozoal infections are responsible for a wide variety of diseases of medical and veterinary importance, including malaria in man and various coccidioses in birds, fish and mammals. Many of the diseases are life-threatening to the host and cause considerable economic loss in animal husbandry, such as diseases caused by infection by species of Eimeria, Theileria, Babesia, Cryptosporidium, Toxoplasma (such as Toxoplasma brucei, African sleeping sickness and Toxoplasma cruzi, Chagas disease) and Plasmodium (such as Plasmodium falciparum), and the Mastigophora such as species of Leishmania (such as Leishmania donovani). Another parasitic organism of increasing concern is Pneumocytis carinii, which can cause an often fatal pneumonia in immunodeficient or immunocompromised hosts, including those infected with HIV.

Malaria is one of the major disease problems of the developing world. The most virulent malaria-causing parasite in humans is the parasite Plasmodium falciparum, which is the cause of hundreds of millions of cases of malaria per annum, and is thought to cause over 1 million deaths each year, Breman, J. G., et al., (2001 ) Am. Trop. Med. Hyg. 64, 1 -1 1. One problem encountered in the treatment of malaria is the build-up of resistance by the parasite to available drugs. Thus, there is a need to develop new antimalarial drugs.

US patent 20090163545 discloses certain amino phenyl acetamide derivatives.

SUMMARY OF THE INVENTION

The present invention is directed to certain amino phenyl acetamide derivatives for use in the chemotherapy of certain parasitic infections such as malaria, and in particular infection by Plasmodium falciparum, processes for their preparation and pharmaceutical compositions comprising such compounds.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a compound of Formula (I):

(I)

Wherein:

R¹ is Ci_-6alkyl, Ci_-6haloalkyl, Ci_-6alkoxy, Ci_-6haloalkoxy or halo;

R² is -NR⁴R⁵;

R⁴ and R⁵ are independently Ci_-6alkyl or, together with the N to which they are attached, form C₅-₇heteroaryl or C_5-7heterocyclyl, either of which i) may optionally contain 1 -3 additional heteroatoms, independently selected from the group consisting of N, O and S and ii) may be optionally substituted with Ci_-3alkyl;

R³ is H;

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷;

R⁶ is H or Ci_-6alkyl;

R⁷ is H or C_1-6alkyl;

or R⁶ and R⁷ together with the carbon atom to which they are attached, form cyclopropyl; Y is a direct bond, -O- or -NR⁸-(CH₂)_P;

R⁸ is H or C_1-4alkyl;

p is 0 or 1 ;

R⁹ is H, Ci_-6alkyl, Ci_-6haloalkyl, Ci_-6alkoxy, Ci_-6haloalkoxy, halo, cyano or optionally substituted C_5-7heteroaryl or optionally substituted C_5-7heterocyclyl, wherein the optional substitutents for C_5-7heteroaryl and C_5-7heterocyclyl are selected from C_1-3alkyl;

R¹⁰ is H, Ci_-6alkyl, Ci_-6haloalkyl, Ci_-6alkoxy, Ci_-6haloalkoxy halo or cyano;

or R⁹ and R¹⁰ together with the carbon atoms to which they are attached form a C_5-6 membered carbocyclyl which may optionally contain 1 -3 heteroatoms, independently selected from the group consisting of N, O and S;

or a pharmaceutically acceptable salt thereof, for use in therapy, for example the treatment of parasitic protozoal infections such as malaria, for example infection by Plasmodium falciparum.

In another aspect, the present invention provides a compound of Formula (ID):

(I)

wherein

R¹ is Ci_-6alkyl, Ci_-6haloalkyl, Ci_-6alkoxy, Ci_-6haloalkoxy or halo;

R² is -NR⁴R⁵;

R⁴ and R⁵ are independently Ci_-6alkyl or, together with the N to which they are attached, form C₅-₇heteroaryl or C_5-7heterocyclyl, either of which may optionally contain 1 -3 additional heteroatoms, independently selected from the group consisting of N, O and S;

R³ is H;

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷;

R⁶ is H or Ci_-6alkyl;

R⁷ is H or Ci_-6alkyl;

R⁸ is H or C_1-4alkyl;

p is 0 or 1 ;

Z is

R⁹ is H, C_1-6alkyl, C_1-6haloalkyl, C_1-6alkoxy, C_1-6haloalkoxy, halo or optionally substituted C_5- ₇heteroaryl or optionally substituted C_5-7heterocyclyl;

R¹⁰ is H, Ci_-6alkyl, Ci_-6haloalkyl, Ci_-6alkoxy, Ci_-6haloalkoxy or halo;

or a pharmaceutically acceptable salt thereof, for use in therapy, for example the treatment of parasitic protozoal infections such as malaria, for example infection by Plasmodium falciparum. Definitions

As used herein, the term "alkyl" refers to straight or branched saturated hydrocarbon chains containing the specified number of carbon atoms. For example, C_1-4alkyl means a straight or branched alkyl containing at least 1 , and at most 4, carbon atoms. Examples of "alkyl" as used herein include, but are not limited to, methyl, ethyl, n-propyl, 1 -methylethyl (isopropyl), n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or hexyl. As used herein, the term "alkoxy" refers to an -O-alkyI group wherein alkyl is as defined herein. Examples of "alkoxy" as used herein include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentoxy or hexoxy.

As used herein, the term "C₅-₆ membered carbocyclyl" refers to a 5- or 6-membered saturated hydrocarbon ring. Examples of "C_5-6 membered carbocyclyl" as used herein include cyclopropyl or cyclohexyl.

As used herein, the term "haloalkyi" refers to an alkyl group as defined herein wherein at least one hydrogen atom is replaced with a halogen atom, for example a fluoro atom. For example, Ci_-4haloalkyl means an alkyl group containing at least 1 , and at most 4, carbon atoms and at least one halogen atom, for example a fluoro atom. Examples of "haloalkyi" as used herein include, but are not limited to, -CF₃, -CHF₂ and -CH₂F.

As used herein, the term "haloalkoxy" refers to an alkoxy group, as defined herein wherein at least one hydrogen atom is replaced with a halogen atom, for example a fluoro atom. For example, Ci_-4haloalkoxy means an alkoxy group containing at least 1 , and at most 4, carbon atoms and at least one halogen atom, for example a fluoro atom. Examples of "haloalkoxy" as used herein include, but are not limited to, -OCF₃, -OCHF₂ and -OCH₂F.

As used herein, the term "halogen" or "halo" refers to a fluorine (fluoro), chlorine (chloro), bromine (bromo) or iodine (iodo) atom. In another embodiment, the term "halogen" or "halo" refers to a fluorine (fluoro), chlorine (chloro) or bromine (bromo) atom.

As used herein, the term "C₅-₇heteroary refers to an aromatic monocyclic group containing 5 to 7 ring-atoms, 1 , 2, 3 or 4 of which are heteroatoms independently selected from nitrogen, oxygen and sulphur and the remaining ring-atoms are carbon, e.g. pyridyl. Examples of "C_5- ₇heteroaryl" as used herein include, but are not limited to, imidazolyl, pyrazolyl, pyrrolyl, pyridyl, pyrazinyl, thiazolyl, thiophenyl, oxadiazolyl, oxazolyl, pyrimidinyl, pyridazinyl, triazolyl and tetrazolyl. All isomers of the above C_5-7heteroaryl groups are within the scope of this invention.

As used herein, the term "C_5-7heterocyclyr' refers to a monocyclic group containing 5 to 7 ring-atoms 1 , 2, 3 or 4 of which are heteroatoms independently selected from nitrogen, oxygen and sulphur and the remaining ring-atoms are carbon, wherein said cyclic group is saturated, partially or fully unsaturated but, which is not aromatic. Where the C_5-7heterocyclyl is pyrrolidinyl, it can optionally be substituted by =0 to form pyrrolidinone. Examples of "C_5- ₇heterocyclyl" as used herein include, but are not limited to, pyrrolidinyl, pyrrolidinone, imidazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, hydantoinyl, valerolactamyl, oxathiolanyl, oxathianyl, diazepanyl, dihydrofuranyl, tetrahydrofuranyl, dioxolanyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, dihydropyranyl, tetrahydropyranyl, dioxanyl and dithianyl. All isomers of the above heterocyclic groups are within the scope of this invention.

For the avoidance of doubt, the term "independently" means that where more than one substituent is selected from a number of possible substituents, those substituents may be the same or different.

As used herein, the term "pharmaceutically acceptable" used in relation to an ingredient (such as an active ingredient, a salt thereof or an excipient) which may be included in a pharmaceutical formulation for administration to a patient, refers to that ingredient being acceptable in the sense of being compatible with any other ingredients present in the pharmaceutical formulation and not being deleterious to the recipient thereof.

Throughout the specification and the claims which follow, unless the context requires otherwise, the word 'comprise', and variations such as 'comprises' and 'comprising', will be understood to imply the inclusion of a stated integer or step or group of integers but not to the exclusion of any other integer or step or group of integers or steps.

Description of the embodiments

All aspects and embodiments of the invention described herein are in respect of compounds of Formula (I) and Formula (ID), unless otherwise specified.

In one aspect of the invention, R¹ is Ci_-6haloalkyl or halo. In a further aspect, R¹ is Ci_ ₄haloalkyl or halo. In another aspect, when R¹ is or contains halo, halo is selected from fluoro, chloro and bromo. In a further aspect, R¹ is -CF₃ or chloro. In a further aspect, R¹ is -CF₃. In a further aspect, R¹ is chloro.

In one aspect of the invention, R² is-NR⁴R⁵ wherein R⁴ and R⁵, together with the N to which they are attached, form C₅-₆heteroaryl or C₅-₆heterocyclyl, which may optionally contain 1-3 additional heteroatoms, independently selected from the group consisting of N and O. In a further aspect, R² is -NR⁴R⁵ wherein R⁴ and R⁵, together with the N to which they are attached, form C₅heteroaryl or C₆heterocyclyl, which may optionally contain 1-3 additional heteroatoms, independently selected from the group consisting of N and O. In a further aspect, R² is -NR⁴R⁵ wherein R⁴ and R⁵, together with the N to which they are attached, forms one of the following:

In another aspect, R is -NR R⁵ wherein R and R⁵, together with the N to which they are attached, forms

In one aspect of the invention, R^a is X-Y-Z.

In one aspect of the invention, R⁶ is H. In a further aspect of the invention, R⁶ is C_1-4alkyl. In a further aspect of the invention, R⁶ is H, methyl or ethyl. In a further aspect of the invention, R⁶ is H or methyl. In a further aspect of the invention, R⁶ is methyl.

In one aspect of the invention, R⁷ is H. In a further aspect of the invention, R⁷ is Ci_-4alkyl. In a further aspect of the invention, R⁷ is H or methyl. In a further aspect of the invention, when R⁶ is H, R⁷ is methyl. In another aspect, when when R⁶ is methyl, R⁷ is H.

In another aspect of the invention, R⁶ and R⁷ together with the carbon atom to which they are attached, form cyclopropyl.

In one aspect of the invention, Y is a direct bond, -0-, -NH- or -N(CH₃)-CH₂. In a further aspect of the invention , Y is a direct bond, -NH- or -N(CH₃)-CH₂. In a further aspect, Y is a direct bond or -0-. In one aspect of the invention, R is H or methyl. In a further aspect of the invention, R is H. In a further aspect of the invention, R⁸ is methyl.

In one aspect of the invention, p is 0. In a further aspect of the invention, p is 1. In one aspect of the invention, Z is

In one aspect of the invention, Z is

In one aspect of the invention, when R⁹ is or contains halo, halo is selected from fluoro, chloro and bromo. In one aspect of the invention for Formula (I), R⁹ is H, Ci_-6alkyl, d. ₆haloalkyl, C_1-6alkoxy, halo, cyano or optionally substituted C_5-7heteroaryl. In another aspect of the invention for Formula (I) R⁹ is H, methyl, CF₃, methoxy, chloro, fluoro, cyano or

In another aspect of the invention, R is CF₃, chloro, fluoro or H. In another aspect, R⁹ is CF₃, chloro or fluoro.

In one aspect of the invention for Formula (ID), R⁹ is H, Ci_-6alkyl, Ci_-6haloalkyl, Ci_-6alkoxy, halo or optionally substituted C_5-7heteroaryl.

In one aspect of the invention, R⁹ is H, Ci_-6alkyl or optionally substituted C₅-₇heteroaryl. In a further aspect of the invention, R⁹ is H, d^alkyl or optionally substituted heteroaryl. In a further aspect of the invention, R⁹ is H, C_1-4alkyl or

H

In a further aspect of the invention, R⁹ is H, methyl or

In one aspect of the invention, when R is or contains halo, halo is selected from fluoro, chloro and bromo. In one aspect of the invention, R¹⁰ is H, C_1-6alkyl, C_1-6alkoxy, halo or cyano. In one aspect of the invention, R¹⁰ is H, C_1-6alkyl, C_1-6alkoxy or halo. In another aspect of the invention, R¹⁰ is H, methyl, methoxy, chloro, fluoro or cyano. In a further aspect of the invention, R¹⁰ is H, C_1-4alkyl or halo. In a further aspect of the invention, R¹⁰ is H, methyl or chloro.

In one aspect of the invention, R⁹ is other than H and R¹⁰ is H. In another aspect of the invention, R¹⁰ is other than H and R⁹ is H.

In one aspect of the invention, R⁹ and R¹⁰ together with the carbon atoms to which they are attached form a C_5-6 membered saturated carbocyclyl which may optionally contain 1 -3 heteroatoms, independently selected from the group consisting of N and O. In a further aspect of the invention, R⁹ and R¹⁰ together with the carbon atoms to which they are attached form a C_5-6 membered saturated carbocyclyl which may optionally contain 1 -2 -O- atoms. In a further aspect of the invention, R⁹ and R¹⁰ together with the carbon atoms to which they are attached form

In one aspect of the invention, Z is

In a further aspect of the invention, Z is

In a further aspect, the present invention provides a compound of Formula (I) as represented by Formula (IA)

(IA)

wherein

R¹ is Ci_-6haloalkyl or halo; R² is-NR⁴R⁵;

R⁴ and R⁵ together with the N to which they are attached, form C_5-6heteroaryl or C_5- ₆heterocyclyl, either of which may optionally contain 1-3 additional heteroatoms, independently selected from the group consisting of N and O;

R³ is H;

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷;

R⁶ is H or Ci_-4alkyl;

R⁷ is H or Ci_-4alkyl;

or R⁶ and R⁷ together with the C to which they are attached, form cyclopropyl;

Y is a direct bond, -0-, -NH- or - N(CH₃)-CH₂;

Z is

R⁹ is H, or optionally substituted heteroaryl;

R¹⁰ is H, C_1-4alkyl or halo;

In a further aspect, the present invention provides a compound of Formula (I) as represented by Formula (IB)

wherein

R¹ is -CF₃ or chloro;

R² is-NR⁴R⁵;

R⁴ and R⁵ together with the N to which they are attached, form

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷; R⁶ is H or methyl;

R⁷ is H or methyl;

or R⁶ and R⁷ together with the C atom to which they are attached, form cyclopropyl;

Y is a direct bond, -0-, -NH- or - N(CH₃)-CH₂;

R¹⁰ is H, methyl or chloro;

or R⁹ and R¹⁰ together with the carbon atoms to which they are attached form a C₅-₆ membered carbocyclyl which may optionally contain 1 -3 heteroatoms, independently selected from the group consisting of N, O and S;

In a further aspect, the present invention provides a compound of Formula (I) as represented by Formula (IC)

(IC)

wherein

R¹ is -CF₃ or chloro;

R² is-NR⁴R⁵;

R⁴ and R⁵ together with the N to which they are attached, form

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷;

R⁶ is H or methyl;

R⁷ is H or methyl;

or R⁶ and R⁷ together with the C atom to which they are attached, form cyclopropyl; Y is a direct bond, -0-, -NH- or - N(CH₃)-CH₂;

Z is

The meaning of any functional group or substituent thereon at any one occurrence in Formula (I), Formula (ID) or any subformula thereof, is independent of its meaning, or any other functional group's or substituent's meaning, at any other occurrence, unless stated otherwise. It is to be understood that the present invention covers all combinations of the groups according to different aspects and embodiments of the invention as described hereinabove. References herein to a compound of Formula (I) will be understood to include compounds of Formula (IA), (IB), (IC) and (ID).

In a further aspect, the present invention provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group (List A) consisting of:

1- phenyl-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]cyclopropanecarboxamide;

/V-[5-chloro-2-(1 H-1 ,2,4-triazol-1 -yl)phenyl]-1-phenylcyclopropanecarboxamide;

2- [(4-chlorophenyl)oxy]-/V-[2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]propanamide;

2-[(3-methylphenyl)oxy]-/V-[2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]propanamide;

/V²-methyl-/V²-(2-naphthalenylmethyl)-/V¹-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl] glycinamide;

2,2,2-trichloro-/V-[2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]acetamide;

N2-(2,3-dihydro-1 H-inden-5-yl)-N1 -[2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]alaninamide; 2-[(4-chlorophenyl)oxy]-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]propanamide; N2-[3-(2-methyl-1 ,3-thiazol-4-yl)phenyl]-N1 -[2-(4-morpholinyl)-5-trifluoromethy alaninamide;

N2-(2,3-dihydro-1 ,4-benzodioxin-6-yl)-N1-[2-(4-morpholinyl)-5-(trifluoromethyl)phen alaninamide and

/V-[2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]-1-phenylcyclopropanecarboxamide

for use in therapy, for example the treatment of parasitic protozoal infections such as malaria, for example infection by Plasmodium falciparum.

In a further aspect, the present invention provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

2-phenyl-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]propanamide

Λ/-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-2-[3-(trifluoromethyl)- phenyl]propanamide

2-(4-chlorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]propanamide

2-phenyl-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]butanamide

2-(4-cyanophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]butanamide

Λ/-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-1 -[3-(trifluoromethyl)- phenyl]cyclopropanecarboxamide

1-(3-chlorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1-yl)-5-

(trifluoromethyl)phenyl]cyclopropanecarboxamide

1-(3-fluorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-

(trifluoromethyl)phenyl]cyclopropanecarboxamide

1-(4-chlorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1-yl)-5- (trifluoromethyl)phenyl]cyclopropanecarboxamide

1-[3-(methyloxy)phenyl]-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-

(trifluoromethyl)phenyl]cyclopropanecarboxamide

1- (3,4-dichlorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1-yl)-5- (trifluoromethyl)phenyl]cyclopropanecarboxamide

2-methyl-2-phenyl-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]propanamide

2- (4-fluorophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-acetamide

1-phenyl-/V-[2-(1 -pyrrolidinyl)-5-(trifluoromethyl)phenyl]cyclopropane-carboxamide

1-(4-methylphenyl)-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]- cylopropanecarboxamide

1-[4-(methyloxy)phenyl]-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)- phenyl]cyclopropanecarboxamide

/V-[2-(1 H-imidazol-1 -yl)-5-(trifluoromethyl)phenyl]-1-phenylcyclopropane-carboxamide

1- phenyl-/V-[2-(1 H-pyrazol-1-yl)-5-(trifluoromethyl)phenyl]cyclopropane-carboxamide

2- [(3-chlorophenyl)oxy]-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-propanamide 2-(phenyloxy)-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)-phenyl]propanamide

/V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]alaninamide; and /V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]glycinamide

In a further aspect, the present invention provides a compound of Formula (I), wherein the compound is selected from the group consisting of:

2-phenyl-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]propanamide

Λ/-[2-(1 H-1 ,2,4-triazoM -yl)-5-(trifluoromethyl)phenyl]-2-[3-(trifluoromethyl)- phenyl]propanamide

2-(4-chlorophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]-propanamide

2-phenyl-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]butanamide

2-(4-cyanophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-butanamide

1-(3-chlorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]- cyclopropanecarboxamide

1-(3-fluorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]- cyclopropanecarboxamide

1-(4-chlorophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]- cyclopropanecarboxamide

1- [3-(methyloxy)phenyl]-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]- cyclopropanecarboxamide

1-(3,4-dichlorophenyl)-N-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]- cyclopropanecarboxamide

2- methyl-2-phenyl-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-propanamide

2-(4-fluorophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-acetamide

/V-[2-(4-methyl-1 -piperazinyl)-5-(trifluoromethyl)phenyl]-1 -phenylcyclo-propanecarboxamide bromohydrate salt

/V-[2-(4-methyl-1 -piperazinyl)-5-(trifluoromethyl)phenyl]-1 -phenylcyclo-propanecarboxamide /V-[2-(1 H-imidazol-1 -yl)-5-(trifluoromethyl)phenyl]-1 -phenyl cyclopropane-carboxamide

2- methyl-2-(phenyloxy)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-propanamide 2-[(3-chlorophenyl)oxy]-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)-phenyl]propanamide

/V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]alaninamide; and /V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]glycinamide;

or a pharmaceutically acceptable salt thereof. Also included in the present invention are pharmaceutically acceptable salt complexes. In certain embodiments of the invention, pharmaceutically acceptable salts of the compounds according to Formula (I) may be preferred over the respective free base because such salts impart greater stability or solubility to the molecule thereby facilitating formulation into a dosage form. Therefore, the present invention also covers the pharmaceutically acceptable salts of the compounds of Formula (I). As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the subject compound and exhibits minimal undesired toxicological effects. For a review on suitable salts see Berge et al, J. Pharm. Sci., 1977, 66, 1-19. The term "pharmaceutically acceptable salt" includes any pharmaceutically acceptable acid addition salts. These pharmaceutically acceptable salts may be prepared in situ during the final isolation and purification of the compound, or by separately reacting the purified compound in its free base form with a suitable acid, respectively. The salt may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent. Therefore, according to a further aspect, the invention provides a pharmaceutically acceptable salt of a compound of Formula (I) thereof.

In certain embodiments, compounds according to Formula (I) may contain a basic functional group and may therefore be capable of forming pharmaceutically acceptable acid addition salts by treatment with a suitable acid. A pharmaceutically acceptable acid addition salt may be formed by reaction of a compound of Formula (I) with a suitable strong inorganic or organic acid (such as hydrobromic, hydrochloric, sulfuric, nitric, phosphoric, perchloric, p- toluenesulfonic, benzenesulfonic, methanesulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, naphthalenesulfonic (e.g. 2-naphthalenesulfonic), optionally in a suitable solvent such as an organic solvent, to give the salt which is usually isolated for example by crystallisation and filtration. Pharmaceutically acceptable acid addition salts include a hydrobromide, hydrochloride, hydroiodide, sulfate, bisulfate, nitrate, phosphate, perchlorate, p- toluenesulfonate, benzenesulfonate, methanesulfonate, ethanesulfonate, 2- hydroxyethanesulfonate or naphthalenesulfonate (e.g. 2-naphthalenesulfonate) salt. In one embodiment, a pharmaceutically acceptable acid addition salt of a compound of Formula (I) is a salt of a strong acid, for example a hydrobromide, hydrochloride, hydroiodide, sulfate, nitrate, perchlorate, phosphate p-toluenesulfonic, benzenesulfonic or methanesulfonic salt.

Suitable pharmaceutically acceptable salts of the compounds of Formula (I) include mono- or di- basic salts with the appropriate acid for example organic carboxylic acids such as acetic, lactic, tartaric, malic, isethionic, lactobionic and succinic acids; organic sulfonic acids such as methanesulfonic, ethanesulfonic, benzenesulfonic and p-toluenesulfonic acids and inorganic acids such as hydrochloric, sulfuric, phosphoric and sulfamic acids and the like.

The invention includes within its scope all possible stoichiometric and non-stoichiometric forms of the salts of the compounds of Formula (I).

As used herein, the term "compounds of the invention" means the compounds according to Formula (I) and the pharmaceutically acceptable salts, solvates and pro-drugs thereof, especially pharmaceutically acceptable salts and solvates thereof. The term "a compound of the invention" means any one of the compounds of the invention as defined above. For the avoidance of doubt the phrase "a compound of the invention" as used herein encompasses a compound of Formula (I), a pharmaceutically acceptable solvate of a compound of Formula (I), a pharmaceutically acceptable salt of a compound of Formula (I), and a pharmaceutically acceptable solvate of a pharmaceutically acceptable salt of a compound of Formula (I).

The compounds of the invention may exist as solids or liquids, both of which are included in the invention. In the solid state, the compounds of the invention may exist as either amorphous material or in crystalline form, or as a mixture thereof. It will be appreciated that pharmaceutically acceptable solvates of compounds of the invention may be formed wherein solvent molecules are incorporated into the crystalline lattice during crystallisation. Solvates may involve non-aqueous solvents such as ethanol, isopropanol, dimethylsulfoxide (DMSO), acetic acid, ethanolamine, and ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent that is incorporated into the crystalline lattice are typically referred to as "hydrates." Solvates of the compound of Formula (I) are within the scope of the invention. Therefore, in one aspect of the present invention, there is provided solvates of the compounds of Formula (I), for example hydrates.

The salts of a compound of Formula (I) may be prepared by contacting appropriate stoichiometric amounts of the free base with the appropriate acid in a suitable solvent. The free base of a compound of Formula (I) may for example be in solution with the appropriate acid added as a solid or both the free base of a compound of Formula (I) and the appropriate acid may independently be in solution. Suitable solvents for solubilising a compound of Formula (I) free base include for example alcohols such as isopropanol; ketones such as acetone; acetonitrile or toluene. If the base is to be added as a solution in a solvent, the solvent used may include acetone, methanol or water. The salts of a compound of Formula (I) may be isolated in solid form by conventional means from a solution thereof obtained as above. For example, a non-crystalline salt may be prepared by precipitation from solution, spray drying or freeze drying of solutions, evaporating a solution to a glass, or vacuum drying of oils, or solidification of melts obtained from reaction of the free base and the acid.

The salts of a compound of Formula (I) may be prepared by directly crystallising from a solvent in which the salt has limited solubility, or by triturating or otherwise crystallising a noncrystalline salt. For example, organic solvents such as acetone, acetonitrile, butanone, 1- butanol, ethanol, 1-propanol or tetrahydrofuran or mixtures of such solvents may be used. An improved yield of the salts may be obtained by the evaporation of some or all of the solvent or by crystallisation at elevated temperature followed by controlled cooling, for example in stages. Careful control of the precipitation temperature and seeding may be used to improve the reproducibility of the production process and the particle size distribution and form of the product.

Salts and solvates of compounds of Formula (I) which are suitable for use in medicine are those wherein the counterion or associated solvent is pharmaceutically acceptable. However, salts and solvates having non-pharmaceutically acceptable counterions or associated solvents are within the scope of the present invention, for example, for use as intermediates in the preparation of other compounds of Formula (I) or salts, solvates or amides thereof and their pharmaceutically acceptable salts and solvates.

It will be appreciated by those skilled in the art that certain protected derivatives of compounds of Formula (I), which may be made prior to a final deprotection stage, may not possess pharmacological activity as such, but may, in certain instances, be administered orally or parenterally and thereafter metabolised in the body to form compounds defined in the first aspect which are pharmacologically active. Such derivatives may therefore be described as "prodrugs". All protected derivatives and prodrugs of compounds defined in the first aspect are included within the scope of the invention. Examples of suitable prodrugs for the compounds of the present invention are described in Drugs of Today, Volume 19, Number 9, 1983, pp 499 - 538 and in Topics in Chemistry, Chapter 31 , pp 306 - 316 and in "Design of Prodrugs" by H. Bundgaard, Elsevier, 1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as "pro-moieties", for example as described by H. Bundgaard in "Design of Prodrugs" (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within the compounds of Formula (I). Suitable prodrugs for compounds of Formula (I) or salts or solvates thereof include : amides, carbamates, azo-compounds, phosphamides, glycosides. Therefore, in one aspect of the present invention, there is provided prodrugs of the compounds of Formula (I), for example amides. The present invention also relates to pharmaceutically acceptable amides of the compounds of Formula (I), for example amides from carboxylic acids -COOR, in which R is selected from straight or branched chain alkyl, for example n-propyl, n-butyl, alkoxyalkyl (e.g. methoxymethyl), aralkyl (e.g. benzyl), aryloxyalkyl (e.g. phenoxymethyl), aryl (e.g. phenyl optionally substituted by halogen, -Ci_-4alkyl or -Ci_-4alkoxy or amino); or for example -CH₂OC(0)R' or -CH₂OC0₂R' in which R' is alkyl (e.g. R' is i-butyl). Unless otherwise specified, any alkyl moiety present in such amides suitably contains 1 to 18 carbon atoms, particularly 1 to 4 carbon atoms. Any aryl moiety present in such amides suitably comprises a phenyl group.

As described above, the compounds of the present invention may be in the form of their free base or a pharmaceutically acceptable salt, solvate, or prodrug e.g. an amide of a compound of Formula (I), which upon administration to the recipient is capable of providing (directly or indirectly) a compound of Formula (I), or an active metabolite or residue thereof. Such pharmaceutically acceptable salts, solvates, and prodrugs are recognizable to those skilled in the art, without undue experimentation. Nevertheless, reference is made to the teaching of Burger's Medicinal Chemistry and Drug Discovery, 5th Edition, Vol 1 : Principles and Practice, which is incorporated herein by reference to the extent of teaching such derivatives. In one aspect of the invention, the compounds of the invention are pharmaceutically acceptable salts, solvates or amides. In a further aspect, the compounds of the present invention are pharmaceutically acceptable salts or solvates. In another aspect, the compounds of the present invention are pharmaceutically acceptable salts.

Furthermore, some of the crystalline forms of the compounds of Formula (I) or salts and solvates thereof may exist in one or more polymorphic form, which are included in the present invention. It will further be appreciated that certain compounds of the present invention may exist in different tautomeric forms. All possible tautomers are contemplated to be within the scope of the present invention.

The compounds of the invention can be useful in the treatment of certain parasitic infections such as parasitic protozoal infections by the malarial parasite Plasmodium falciparum, species of Eimeria, Pneumocytis carinii, Trypanosoma cruzi, Trypanosoma brucei or Leishmania donovani. In particular, the compounds of the invention can be useful for treatment of infection by Plasmodium falciparum. Accordingly, the invention is directed to methods of treating such conditions.

In one aspect of the invention there are provided certain compounds of Formula (I), which can be useful for the treatment of a parasitic protozoal infection, such as malaria.

In another aspect of the invention, there is provided a compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of a parasitic protozoal infection such as malaria, for example infection by Plasmodium falciparum. In a further aspect of the invention, there is provided a compound of List A or a pharmaceutically acceptable salt thereof, for use in the treatment of parasitic protozoal infections such as malaria, for example a condition caused by infection by Plasmodium falciparum.

In another aspect of the invention, there is provided the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a parasitic protozoal infection such as malaria, for example infection by Plasmodium falciparum. In a further aspect of the invention, there is provided the use of a compound of List A or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of parasitic protozoal infections such as malaria, for example a condition caused by infection by Plasmodium falciparum.

In a further aspect of the invention, there is provided a method for the treatment of a human or animal subject suffering from a parasitic protozoal infection such as malaria, for example infection by Plasmodium falciparum, which method comprises administering to said human or animal subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In a further aspect of the invention there is provided a method for the treatment of a human or animal subject suffering from a parasitic protozoal infection such as malaria, for example infection by Plasmodium falciparum, comprising administering to said human or animal subject an effective amount of a compound of List A or a pharmaceutically acceptable salt thereof.

The methods of treatment of the invention comprise administering a safe and effective amount of a compound according to Formula (I), and a pharmaceutically acceptable salt thereof to a patient in need thereof.

As used herein, "treatment" means: (1 ) the amelioration or prevention of the condition being treated or one or more of the biological manifestations of the condition being treated, (2) the interference with (a) one or more points in the biological cascade that leads to or is responsible for the condition being treated or (b) one or more of the biological manifestations of the condition being treated, or (3) the alleviation of one or more of the symptoms or effects associated with the condition being treated. The skilled artisan will appreciate that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or severity of a condition or biological manifestation thereof, or to delay the onset of such condition or biological manifestation thereof.

As used herein, "safe and effective amount" means an amount of the compound sufficient to significantly induce a positive modification in the condition to be treated but low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment. A safe and effective amount of a compound of the invention will vary with the particular compound chosen (e.g. depending on the potency, efficacy, and half-life of the compound); the route of administration chosen; the nature of the infection and/or condition being treated; the severity of the infection and/or condition being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient to be treated; the duration of the treatment; the nature of concurrent therapy; the desired therapeutic effect; and like factors, but can nevertheless be routinely determined by the skilled artisan.

As used herein, "patient" refers to a human or other animal.

The compounds of the invention may be administered by any suitable route of administration, including systemic administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages.

The compounds of the invention may be administered once only, or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. The dosage will also vary according to the nature of the intended treatment, wherein "treatment" is as hereinbelow defined, for example a greater dose of compound may be given for amelioration as compared with prevention of a condition being treated. Suitable dosing regimens for a compound of the invention depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half-life, which can be determined by the skilled artisan. In addition, suitable dosing regimens for a compound of the invention, including the duration such regimens are administered, depend on the route of administration of the compound, on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of any concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change. It will also be appreciated that if the compounds of the present invention are administered in combination with one or more additional active therapeutic agents as discussed further hereinbelow, the dosing regimen of the compounds of the invention may also vary according to the nature and amount of the one or more additional active therapeutic agents as necessary.

Typical daily dosages may vary depending upon the particular route of administration chosen. Typical daily dosages for oral administration range from about 0.01 to about 25 mg/kg, in one embodiment from about 0.1 to about 14 mg/kg. Typical daily dosages for parenteral administration range from about 0.001 to about 10 mg/kg; in one embodiment from about 0.01 to about 6 mg/kg. In one embodiment, the daily dose range of the compounds is from 100-1000 mg per day.

The compounds of the invention may also be used in combination with other active therapeutic agents. The invention thus provides, in a further aspect, a combination comprising a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof together with a further active therapeutic agent. When a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof is used in combination with a second active therapeutic agent which is active against the same disease state the dose of each compound may differ from that when the compound is used alone. Appropriate doses will be readily appreciated by those skilled in the art. It will be appreciated that the amount of a compound of the invention required for use in treatment will vary with the nature of the condition being treated and the age and the condition of the patient and will be ultimately at the discretion of the attendant physician or veterinarian.

The compounds of the present invention may be used alone or in combination with one or more additional active therapeutic agents, such as other antiparasitic drugs, for example antimalarial drugs.

Such other active therapeutic agents include antimalarial drugs such as (e.g. chloroquine, mefloquine, primaquine, pyrimethamine, quinine, artemisinin, halofantrine, doxycycline, amodiaquine, atovaquone ,tafenoquinedapsone, proguanil, sulfadoxine, cycloguanil) and fansidar.

The combinations referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above together with a pharmaceutically acceptable carrier and/or excipient comprise a further aspect of the invention. The individual components of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations by any convenient route.

When administration is sequential, either the compound of the present invention or the one or more additional active therapeutic agent(s) may be administered first. When administration is simultaneous, the combination may be administered either in the same or different pharmaceutical composition. When combined in the same formulation it will be appreciated that the compound of the present invention and the one or more additional active therapeutic agent(s) must be stable and compatible with each other and the other components of the formulation. When formulated separately the compound of the present invention and the one or more additional active therapeutic agent(s) may be provided in any convenient formulation, conveniently in such manner as are known for such compounds in the art. Compositions

The compounds of the invention will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient. In one aspect, the invention is directed to pharmaceutical compositions comprising a compound of the invention. In another aspect the invention is directed to a pharmaceutical composition comprising (a) a compound of the invention and (b) one or more pharmaceutically acceptable carriers and/or excipients. In another aspect, the invention provides a pharmaceutical composition comprising a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In a further aspect, the invention provides a pharmaceutical composition comprising (a) a compound of Formula (I) or a pharmaceutically acceptable salt thereof and (b) one or more pharmaceutically acceptable carriers and/or excipients.

The carrier and/or excipient must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be extracted and then given to the patient such as with powders or syrups. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains a safe and effective amount of a compound of the invention. When prepared in unit dosage form, the pharmaceutical compositions of the invention typically contain from about 0.1 to 1000mg, in another aspect 0.1 mg to about 500 mg of a compound of the invention.

The pharmaceutical compositions of the invention typically contain one compound of the invention. However, in certain embodiments, the pharmaceutical compositions of the invention contain more than one compound of the invention. For example, in certain embodiments the pharmaceutical compositions of the invention contain two compounds of the invention. In addition, the pharmaceutical compositions of the invention may optionally further comprise one or more additional active therapeutic compounds. The pharmaceutical compositions of the invention typically contain more than one pharmaceutically acceptable excipient. However, in certain embodiments, the pharmaceutical compositions of the invention contain one pharmaceutically acceptable excipient.

The compound of the invention and the pharmaceutically acceptable excipient or excipients will typically be formulated into a dosage form adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, caplets, pills, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; and (5) inhalation such as aerosols and solutions.

Suitable pharmaceutically acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically acceptable excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the carriage or transport of the compound or compounds of the invention from one organ, or portion of the body, to another organ, or portion of the body, once administered to the patient. Certain pharmaceutically acceptable excipients may be chosen for their ability to enhance patient compliance.

Suitable pharmaceutically acceptable excipients include the following types of excipients: binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavouring agents, flavour masking agents, coloring agents, anticaking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. The skilled artisan will appreciate that certain pharmaceutically acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.

Skilled artisans possess the knowledge and skill in the art to enable them to select suitable pharmaceutically acceptable excipients in appropriate amounts for use in the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically acceptable excipients and may be useful in selecting suitable pharmaceutically acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press). The pharmaceutical compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

In one aspect, the invention is directed to a solid or liquid oral dosage form such as a liquid, tablet, lozenge or a capsule, comprising a safe and effective amount of a compound of the invention and a carrier. The carrier may be in the form of a diluent or filler. Suitable diluents and fillers in general include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. A liquid dosage form will generally consist of a suspension or solution of the compound or pharmaceutically acceptable derivative in a liquid carrier for example, ethanol, olive oil, glycerine, glucose (syrup) or water (e.g. with an added flavouring, suspending, or colouring agent). Where the composition is in the form of a tablet or lozenge, any pharmaceutical carrier routinely used for preparing solid formulations may be used. Examples of such carriers include magnesium stearate, terra alba, talc, gelatin, acacia, stearic acid, starch, lactose and sucrose. Where the composition is in the form of a capsule, any routine encapsulation is suitable, for example using the aforementioned carriers or a semi solid e.g. mono di-glycerides of capric acid, Gelucire™ and Labrasol™, or a hard capsule shell e.g. gelatin. Where the composition is in the form of a soft shell capsule e.g. gelatin, any pharmaceutical carrier routinely used for preparing dispersions or suspensions may be considered, for example aqueous gums or oils, and may be incorporated in a soft capsule shell.

An oral solid dosage form may further comprise an excipient in the form of a binder. Suitable binders include starch (e.g. corn starch, potato starch, and pre-gelatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g. microcrystalline cellulose). The oral solid dosage form may further comprise an excipient in the form of a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise an excipient in the form of a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and talc.

There is further provided by the present invention a process of preparing a pharmaceutical composition, which process comprises mixing at least one compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof, together with a pharmaceutically acceptable carrier and/or excipient. In a further aspect of the invention, there is provided a process of preparing a pharmaceutical composition, which process comprises mixing at least one compound of Formula (I) or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier and/or excipient.

Preparations for oral administration may be suitably formulated to give controlled/extended release of the active compound.

PROCESSES

A general procedure for acid chloride formation and amide formation is described in the literature (J. Chem. Res. 2008 (22), 530-533) using appropriate commercially available acids and anilines as starting materials.

Compounds of Formula (I) may be prepared by reacting a compound of Formula (II) CI Ra

Ύ o

(II)

wherein R^a is as herein described

with a compound of Formula (III)

wherein R¹ and R² are as herein described, in the presence of a suitable solvent and under suitable conditions, for example ACN under elevated temperature, optionally in the presence of a base such as NaHC0₃, optionally in the presence of microwaves.

Compounds of Formula (II) may be commercially available or prepared by reacting a compound of Formula (IV)

HO^ _^Ra

T O

(IV)

wherein R^a is as herein described, with a chlorinating agent e.g. COCI₂ (phosgene) or oxalyl chloride and a suitable solvent and under suitable conditions, for example DMF and/or DCM, at room temperature under an inert atmosphere.

Compounds of Formula (IV) may be commercially available or prepared by hydrolysis of a compound of Formula (V)

R_{i i}O— Ra

Y O

(V)

wherein R¹¹ is methyl or ethyl and R^a is X-Y-Z,

by reacting with an inorganic base such as sodium hydroxide or lithium hydroxide in the presence of an alcohol solvent such as methanol or ethanol, for example at room temperature under an inert atmosphere.

Certain compounds of Formula (IV) may be prepared from certain other compounds of Formula (IV). For example, compounds of Formula (IV) wherein R^a is -X-Y-Z, in which X is -CH(Ci_-6alkyl)-, Y is a direct bond and Z is as defined for Formula (I), may be prepared from compounds of Formula (IV) wherein R^a is -X-Y-Z, in which X is -CH₂-, Y is a direct bond and Z is as defined for Formula (I), in the presence of an alkylating agent such as C_1-6alkyl iodide, in the presence of a strong base such as LDA, in a suitable solvent such as THF, at reduced temperature, such as -78°C, as illustrated below.

_HO alkylating agent

O strong base

(IV) (IV) Compounds of Formula (V) are commercially available when

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷;

R⁶ is H or Ci_-6alkyl;

R⁷ is H or C_1-6alkyl;

R⁸ is H or C^alkyl;

p is 1 ;

and Z is

Compounds of Formula (V) wherein

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷;

R⁶ is H;

R⁷ is H or Ci_-6alkyl;

or R⁶ and R⁷ together with the carbon atom to which they are attached, form cyclopropyl; Y is -NR⁸-(CH₂)_P;

R⁸ is H or C_1-4alkyl;

p is 0

and Z is

may be prepared as described in WO 2008/01 1551 A1 by reacting a compound of Formula (VI) which is commercially available

(VI)

wherein R⁶, R⁷ and R¹¹ are as herein described,

with

which are commercially available, in the presence of a suitable base.

Compounds of Formula (III) and Formula (VI) are commercially available.

Compounds of Formula (I) may alternatively be prepared by reaction of a compound of Formula (IV) as described above, with a compound of Formula (III) as described above, in the presence of a suitable activating agent such as i) a mixture of 1-hydroxy-1 H-benzotriazol hydrate (HOBt) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDCI), or ii) N,N,N',N'-tetramethyl-0-(7-azabenzotriazol-1 -yl)uronium hexafluorophosphate (HATU), optionally in the presence of a suitable base such as NaHC0₃ or DIPEA, DI PEA or Et₃N, in a suitable solvent such as DMF or DCM, under suitable conditions such as elevated temperature and/or microwaves.

Compounds of Formula (I) may alternatively be prepared by reaction of a compound of Formula (VII), wherein R¹ and R^a are rmula (I),

(VII)

with a compound R²H, wherein R² is as described for Formula (I), in the presence of a suitable base such as Cs₂C0₃, also in the presence of a suitable activating agent such as copper (I) iodide, optionally in the presence of a suitable catalyst such as 8-hydroxyquinoline, in a suitable solvent such as DMF, under suitable conditions such as elevated temperature and/or microwaves.

Compounds of Formula (VII) may be prepared by reaction of a compound of Formula (II) as described above, with a compound of Formula (VIII) which is commercially available

(VI II)

in the presence of a suitable solvent and under suitable conditions for example ACN under elevated temperature, optionally in the presence of a base such as NaHC0₃, optionally in the presence of microwaves.

Compounds of Formula (I), wherein Y is -NR⁸-(CH₂)_P may alternatively be prepared by reaction of a compound of Formula (IX), wherein R¹, R², R³ and X are as described for Formula (I)

(IX)

compound of Formula (X), wherein R⁸, p and Z are as described for Formula (I)

H-N-(CH₂)— Z

R⁸

(X)

under suitable conditions, such as i) in a suitable solvent such as MeOH, in the presence of a suitable base such as Et₃N, under suitable conditions such as elevated temperature in the presence of microwaves, or ii) in a suitable solvent such as DCM in the presence of a suitable base such as DIPEA. Compounds of Formula (IX) may be prepared by reaction of a compound of Formula (III) as described above, with a commercially available compound of Formula (XI), wherein X is as described for Formula (I)

(XI)

i) when D is OH, in the presence of a suitable coupling agent such as DCC, in a suitable solvent such as DCM; ii) when D is Br or CI, in the presence of a suitable base such as Et₃N, in a suitable solvent such as DCM. Compounds of Formula (XI), wherein D is CI may be preparedfrom compounds (XI) wherein D is OH by treatment with oxalyl chloride or phosgene under standard conditions.

Compounds of Formula (X) are either commercially available or may be prepared according to standard conditions, for example using a reductive amination reaction when p is 1 : O reductive

H-N-H ₊ amination _{¾ H}_N-_(CH₂)-_Z

R _R8

(X)

It will be readily apparent to those skilled in the art that other compounds of Formula (I) may be prepared using methods analogous to those outlined above, or by reference to the experimental procedures detailed in the Examples provided herein. Further details for the preparation of compounds of Formula (I) are found in the Examples.

With appropriate manipulation and protection of any chemical functionality, the synthesis of compounds of Formula (I) is accomplished by methods analogous to those above. In any particular case, particular protecting groups may be required. Suitable protecting groups can be found, but are not restricted to, those found in T W Greene and P G M Wuts 'Protective Groups in Organic Synthesis', 3^rd Ed (1999), J Wiley and Sons.

EXPERIMENTAL

Abbreviations

In describing the invention, chemical elements are identified in accordance with the Periodic Table of the Elements. Abbreviations and symbols utilised herein are in accordance with the common usage of such abbreviations and symbols by those skilled in the chemical arts. The following abbreviations are used herein:

ACN acetonitrile

anh anhydrous

APAD 3-Acetylpyridine adenine dinucleotide

aq. aqueous

AWCI Antimalarial whole cell screening

COCI₂ phosgene

CDCI₃ deuterated chloroform

DCC dicyclohexylcarbodiimide

DCM dichloromethane

DIPEA diisoproylethylamine

DME dimethoxyethane

DMF dimethylformamide

DMSO dimethylsulfoxide

DMSO-d₆ deuterated dimethylsulfoxide

EDCI /V-(3-dimethylaminopropyl)-/V-ethylcarbodiimide hydrochloride

EtOAc ethyl acetate

Et₂0 diethyl ether

ES MS Electrospray mass spectrometry

Ex Example

h hours HATU N , N , N', N'-Tetramethyl-0-(7-azabenzotriazol-1 -yl)u

hexafluorophosphate

HCI hydrochloric acid

HOBt 1 -hydroxy-1 H-benzotriazol hydrate

HPLC high performance liquid chromatography

HTS high throughput screen

Int. Intermediate

LDA Lithium diisopropylamide

Mel Methyl iodide

MeOH methanol

min(s) minutes

MW microwave

¹H NMR proton Nuclear Magnetic Resonance spectroscopy

Pmax maximum pressure

sat. saturated

TFA trifluoroacetic acid

THF tetrahydrofuran

Compound Preparation

Examples

The following Examples illustrate the invention. These Examples are not intended to limit the scope of the invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the invention. While particular embodiments of the invention are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention.

Where materials were commercially available, this is indicated in parentheses after the compound name, in capitals. For example, in the preparation of Intermediate 1 , 3- (trifluoromethyl)phenylacetic acid was purchased from ALDRICH, so it is stated "3- (trifluoromethyl)phenylacetic acid (ALDRICH)".

Intermediates Intermediate 1 : 2-[3-(trifluoromethyl)phenyl]propanoic acid

A solution of 3-(trifluoromethyl)phenylacetic acid (ALDRICH, 300 mg, 1 .470 mmol) in THF (2 mL) was added dropwise to a solution of lithium diisopropylamide, 2M solution in heptane/THF/ethylbenzene (ALDRICH, 2.94 mL, 5.88 mmol) in THF (5 mL) at -78 °C. After 1 h at -78 °C, methyl iodide (ALDRICH, 0.735 mL, 1 1.76 mmol) was added and the solution stirred for 1 hour. The mixture was hydrolyzed with 2N HCI until acidic pH and extracted with DCM, dried over MgS0₄, filtered and dried under vacuum to give a crude. It was purified by flash column chromatography using hexane/EtOAc 85/15 to 0/100 to give: 2-[3- (trifluoromethyl)phenyl]propanoic acid (304 mg, 1.39 mmol, 95% yield). ¹H NMR (400 MHz, CDCI₃) δ ppm: 7.44-7.58 (m, 4H), 3.82 (q, 1 H), 1.56 (d, 3H).

Intermediates 2-3 were prepared by a method analogous to that described for Intermediate 1 but replacing the phenylacetic acid 3-(trifluoromethyl)phenylacetic acid with that indicated in Table A.

Intermediate 4:

2-bromo-5-(trifluoromethyl)aniline (ALDRICH, 0.036 mL, 0.250 mmol) was dissolved in ACN

(3 mL). Sodium bicarbonate (21 mg, 0.250 mmol) was added followed by 1- phenylcyclopropanecarbonyl chloride (CHEMCOLLECT, 49.7 mg, 0.275 mmol). Reaction was heated at 60°C under MW irradiation during 30 min. Solids were filtered and the resulting solution was concentrated to dryness and purified by silica chromatography using hexane/EtOAc as eluents. Title compound was obtained 63.3 mg (65.9 % yield).¹ H NMR (400 MHz, DMSO) δ ppm: 8.45 (m, 1 H), 8.14 (s, 1 H), 7.85-7.79 (m, 1 H), 7.60-7.53 (m, 2H), 7.52-7.44 (m, 2H), 7.43-7.37 (m, 2H), 1 .67 (q, 2H), 1.15 (q, 2H). [ES+ MS] m/z 384 (M+H). Intermediate 5: 2-methyl-2-(phenyloxy)propanoyl chloride

2-methyl-2-(phenyloxy)propanoic acid (CHEMBRIDGE, 82 mg, 0.432 mmol) was dissolved in DCM under N₂ atmosphere and was cooled down in an ice bath. Two drops of DMF were added followed by oxalyl chloride (ALDRICH, 0.041 ml_, 0.432 mmol). Reaction was stirred during 5 hours at room temperature. Solvent was evaporated to dryness, and the crude material was used as it is in the next step of the synthesis without further purification. ¹ H NMR (400 MHz, DMSO-d₆) δ ppm: 7.27 (t, 2H), 6.96 (t, 1 H), 6.82 (d, 2H), 1.50 (s, 6H). Intermediate 6: 2-bromo-/V-[2-(1 H-1 ,2, (trifluoromethyl)phenyl]-propanamide

2-bromopropanoic acid (ALDRICH, 134 mg, 0.877 mmol) was added to a solution of DCC (ALDRICH, 253 mg, 1.227 mmol) in 15 mL of anhydrous DCM cooled to -15°C. Reaction mixture was stirred for 20 minutes at this temperature and then [2-(1 H-1 ,2,4-triazol-1 -yl)-5- (trifluoromethyl)phenyl]amine (purchased from ENAMINE, 200 mg, 0.877 mmol) was added dropwise. Reaction mixture was stirred at room temperature for 4 days, was then diluted with Et₂0 and stirred for further 10 minutes. The mixture was then filtered through Celite, washed through with Et₂0 and the filtrate was concentrated under vacuum. Residue was purified by silica chromatography using DCM/EtOAc as eluents to afford the title compound (75 mg, 23% yield) as a pale yellow solid. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 10.52 (br s, 1 H), 8.86 (s, 1 H), 8.55 (s, 1 H), 8.30 (s, 1 H), 7.57-7.50 (m, 2H), 4.51 (q, 1 H), 1.94 (d, 3H). [ES+ MS] m/z 363 (MH⁺).

Intermediate 7: 2-bromo-/V-[2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]propanamide

Title compound was prepared by a method analogous to that described for Intermediate 6 but replacing [2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]amine with [2-(4-morpholinyl)-5- (trifluoromethyl)phenyl]amine (ALFAAESAR, 200 mg, 0.812 mmol). 2-bromo-/V-[2-(4- morpholinyl)-5-(trifluoromethyl)phenyl]propanamide was obtained (189 mg, 61 % yield) as a white solid. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 9.51 (br s, 1 H), 8.68 (d, 1 H), 7.39 (dd, 1 H), 7.28 (d, 1 H), 4.64 (q, 1 H), 3.96-3.92 (m, 4H), 2.98-2.88 (m, 4H), 2.01 (d, 3H). [ES+ MS] m/z 381 (MH⁺). Intermediate 8: 2-bromo-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]acetamide

Bromoacetyl bromide (ALDRICH, 265 mg, 1 14 μΙ, 1.315 mmol) was added dropwise to a solution of [2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 300 mg, 1.315 mmol) in 5ml_ of anhydrous DCM at 0°C. Et₃N (182 μΙ) was added and the reaction was stirred at room temperature overnight. The reaction was diluted with DCM and washed with water and brine. Organic layer was dried with Na₂S0₄ (anh.), filtered and concentrated under reduced pressure. Purification by silica chromatography using DCM/EtOAc as eluents afforded the title compound (368 mg, 80% yield) as a pale yellow solid. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 10.60 (br s, 1 H), 8.83 (s, 1 H), 8.56 (s, 1 H), 8.29 (s, 1 H), 7.57-7.51 (m, 2H), 4.00 (s, 2H). [ES+ MS] m/z 349 (MH⁺).

Examples The following compounds disclosed in Table 1 are commercially available unless stated otherwise or may be prepared using methods described in the literature.

Table 1 provides Example compounds of the invention (List A):

Table 1

Method of preparation of Example 7: /V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(4-morpholinyl) 5-(trifluoromethyl)-phenyl]alaninamid

Title compound was prepared by a method analogous to that described for Example 46, replacing 2-Bromo-N-[2-(1 /-/-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]propanamidewith

Intermediate 7 (65 mg, 0.171 mmol). Crude was purified twice by silica chromatography using hexane/EtOAc and DCM/EtOAc as eluents. Another purification by preparative HPLC was needed (XBridge 19x150 mm, gradient 60% to 100% NH4HCO3 10mM:ACN). Λ/²-(2,3- dihydro-1 H-inden-5-yl)-/V¹-[2-(4-morpholinyl)-5-(trifluoromethyl)-phenyl]alaninamide was obtained ( mg, %). ¹ H NMR (400 MHz, DMSO-d₆) δ ppm: 9.74 (br s, 1 H), 8.65 (d, 1 H), 7.41 (dd, 1 H), 7.34 (d, 1 H), 6.96 (d, 1 H), 6.50 (s, 1 H), 6.42 (dd, 1 H), 6.15 (d, 1 H), 3.80-3.71 (m, 1 H), 3.47-3.36 (m, 4H), 2.75-2.56 (m, 8H), 1 .97-1 .85 (m, 2H), 1 .45 (d, 3H). [ES+ MS] m/z 434 (MH⁺).

Method of preparation of Example 8: 2-[(4-chlorophenyl)oxy]-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5- (trifluoromethyl)-phenyl]propanamid

A solution of 2-[(4-chlorophenyl)oxy]propanoic acid (ENAMI NE, 100 mg, 0.498 mmol), HATU (ABCHEM, 190 mg, 0.498 mmol) and triethylamine (0.139 ml_, 0.997 mmol) in 5 ml. of anhydrous DCM, was stirred at room temperature for 20 minutes. [2-(1 H-1 ,2,4-triazol-1 -yl)-5- (trifluoromethyl)phenyl]amine (ENAMI NE, 1 14 mg, 0.498 mmol) was added. Reaction was stirred at room temperature until completion. Reaction mixture was diluted with DCM and washed with water and brine. Organic layer was dried with Na₂S0₄ (anh.), filtered and concentrated to dryness. The residue was purified by silica chromatography using DCM/EtOAc as eluents giving the title compound (160mg, 78%) as a pale yellow solid. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 10.71 (br s, 1 H) , 8.97 (s, 1 H), 8.45 (s, 1 H), 8.10 (s, 1 H), 7.52-7.46 (m, 2H), 7.24 (d, 2H), 6.84 (d, 2H), 4.74 (q, 1 H), 1 .64 (d, 3H). [ES+ MS] m/z 41 1 (MH⁺).

Example 11

1 - Phenyl-1 -cyclopropanecarboxylic acid ALDRICH, 37 mg, 0.221 mmol) was dissolved in DCM (2 ml.) under nitrogen. 1 -Hydroxy-1 H-benzotriazol (ALDRICH, 34 mg, 0.222 mmol), N-

(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (ALDRICH, 43 mg, 0.224 mmol),

2- (4-morpholinyl)-5-(trifluoromethyl)aniline (ALFAAESAR, 55 mg, 0.223 mmol), and finally DI PEA (0.096 mL, 0.670 mmol) were added. The reaction was run for 24 hours at room temperature. Reaction mixture was diluted with DCM and washed with water, brine, dried over anhydrous Mg₂S0₄, filtered, concentrated to dryness and purified by chromatography using a 2g Sil l cartridge and a linear gradient of AcOEt/Hexane in 25 min. After combining the appropriated fractions and concentrated to dryness the title compound was obtained (N- [2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]-1 -phenylcyclopropane carboxamide) (12 mg, 13% yield). ¹ H N MR (300 MHz, CDCI₃) ppm: 8.85 (br s, 1 H), 8.67 (br s, 1 H), 7.47-7.54 (m, 4H), 7.28 (m, 1 H), 7.15 (m, 1 H), 3.31 (m, 4H, 2.58 (m, 4H), 1 .74-1 .77 (m, 2H), 1 .16-1 .20 (m, 2H). [ES+ MS] m/z 391 (MH⁺) Table 2

Example 23: 2-phenyl-/V-[2-(1 H-1 ,2 oromethyl)phenyl]propanamide

[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 48 mg, 0.206 mmol) was dissolved in ACN. Sodium bicarbonate (30 mg, 0.357 mmol) was added followed by 2- phenylpropionyl chloride (ALDRICH, 43 mg, 0.242 mmol). Reaction was heated 40 min at 1 10°C under MW irradiation. Reaction crude was filtered and solvent was evaporated to dryness to obtain a crude that was purified by flash column chromatography using DCM/MeOH as eluents from 99:1 to 1 :99. 2-phenyl-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5- (trifluoromethyl)phenyl]propanamide (68.3 mg, 0.190 mmol, 92 % yield) was obtained. ¹ H NMR (400 MHz, DMSO-d₆) δ ppm: 9.96 (s, 1 H), 8.87 (s, 1 H), 8.30 (s, 1 H), 8.19 (s, 1 H), 7.81 (d, 1 H), 7.68 (d, 1 H), 7.35-7.26 (m, 5H), 3.83 (q, 1 H), 1.39 (d, 3H). [ES+ MS] m/z 361 (MH+).

Example 24: Λ/-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-2-[3-(trifluoromethyl)- phenyl]propanamide

To a solution of Intermediate 1 (105 mg, 0.481 mmol) in 2 mL of DCM, was added oxalyl chloride, 98% (ALDRICH, 0.083 mL, 0.963 mmol). Mixture of reaction was stirred at room temperature overnight. Solvent was removed and co-evaporated with toluene. The residue was dissolved in dry ACN(3 mL) and [2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 84 mg, 0.370 mmol) was added. The mixture was stirred at 90 °C for 2 hour. Reaction was allowed to reach room temperature and the mixture was concentrated. The crude material was dissolved in DCM and washed with NH₄CI (aq. sat.), NaHC0₃ and brine. The crude was purified first by chromatography on silica gel with hexane:EtOAc 100:0 to 40:60 as eluents and second by preparative HPLC (column: Sunfire 19x150 mm, gradient 40%-100% ACN/water) to obtain 47 mg of a white solid as the desired product (29.7% yield). 1 H NMR (400 MHz, DMSO-d₆) δ ppm: 10.03 (br s, 1 H), 8.89 (s, 1 H), 8.23 (m, 1 H), 8.12 (s, 1 H), 7.79 (d, 1 H), 7.70 (dd, 1 H), 7.55-7.65 (m, 4H), 3.97 (q, 1 H), 1.41 (d, 3H). [ES+ MS] m/z 429 (MH+).

Examples 25-27 were prepared by methods analogous to that described for Example 24 but replacing Intermediate 1 with the carboxylic acid indicated in Table 3. Reaction time: from 1 hour to 2 hours. Other modifications are also indicated.

Example 28: A -[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-1-[3-(trifluoromethyl^ phenyl]cyclopropanecarboxamid

1-[3-(trifluoromethyl)phenyl]cyclopropanecarboxylic acid (CHEMIZON, 100 mg, 0.434 mmol) was dissolved in anhydrous DCM (2 mL) under N₂ atmosphere. Solution was cooled down in an ice bath and two drops of anhydrous DMF. were added followed by oxalyl chloride (0.239 mL, 0.478 mmol). Reaction was stirred at room temperature for 3 hours and was evaporated to dryness to afford 1 -[3-(trifluoromethyl)phenyl]-cyclopropanecarbonyl chloride (108 mg, 0.434 mmol). [2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 53.6 mg, 0.235 mmol) was dissolved in ACN (2 mL) and sodium bicarbonate (21.9 mg, 0.261 mmol) was added followed by 1 -[3-(trifluoromethyl)phenyl]cyclopropanecarbonyl chloride (108 mg, 0.434 mmol). Reaction was heated at 75°C during 2 hours and at room temperature overnight. Solid was filtered off and the resulting solution was concentrated, dissolved in DCM and washed with NaHC0₃ (aq. sat.), NaCI (aq. sat.), dried over anhydrous Mg₂S0₄, filtered and concentrated to dryness. Crude obtained was purified twice by silica chromatography using a linear gradient of hexane/EtOAc. Title compound (Λ/-[2-(1 Η-1 ,2,4- triazol-1 -yl)-5-(trifluoromethyl)phenyl]-1-[3-(trifluoromethyl)phenyl]cyclopropanecarboxamide (55.8 mg, 0.127 mmol, 29.2 % yield)) was obtained. ¹ H NMR (400 MHz, DMSO) δ ppm: 9.64 (s, 1 H), 9.08 (s, 1 H), 8.75 (m, 1 H), 7.82-7.87 (m, 2H), 7.73 (m, 2H), 7.62-7.69 (m, 3H), 1.55 (q, 2H), 1.23 (q, 2H). [ES+ MS] m/z 441 (MH+).

Examples 29-34 were prepared by methods analogous to that described for Example 28, replacing 1-[3-(trifluoromethyl)phenyl]cyclopropanecarboxylic acid with the carboxylic acid indicated in Table 4. Reaction time: from 30 minutes to 2h 30min. In most cases only one silica chromatography was necessary. Other modifications are also indicated.

a) Preparative HPLC purification needed.

b) Solid was dissolved in a mixture MeOH/DCM. A precipitate appeared that was filtered-off obtaining the title product.

c) Amide formation under MW irradiation 1 h at 100°C

Example 2-(4-fluorophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]- acetamide

[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 30 mg, 0.131 mmol) was dissolved in acetonitrile (2 ml). Sodium bicarbonate (12 mg, 0.146 mmol) was added followed by (4-fluorophenyl)acetyl chloride (ALDRICH, 0.020 mL, 0.146 mmol). Reaction was heated 45 min at 55°C under MW irradiation. Solution was concentrated to dryness and crude was purified by silica chromatography using hexane/EtOAc as eluents. 2-(4- fluorophenyl)-/V-[2-(1 /-/-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]-acetamide (18 mg, 0.049 mmol, 33.8% yield) was obtained. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 9.62 (s, 1 H), 8.93 (s, 1 H), 8.41 (s, 1 H), 7.90 (s, 1 H), 7.45 (t, 1 H), 7.43 (t, 1 H),7.21-7.25 (m, 1 H), 7.09 (t, 2H), 7.07 (t, 1 H), 3.73 (s, 2H). [ES+ MS] m/z 365 (MH+).

Example 36: 1-phenyl-/V-[2-(1 -pyrrolidinyl)-5-(trifluoromethyl)phenyl]cyclopropane- carboxamide

Title compound was prepared by a method analogous to that described for Example 35 replacing (4-fluorophenyl)acetyl chloride with 1 -phenylcyclopropanecarbonyl chloride (CHEMCOLLECT, 52 mg, 0.288 mmol) and the corresponding aniline with (1-pyrrolidinyl)-5- (trifluoromethyl)aniline (APOLLO, 68 mg, 0.295 mmol) to yield 1 -phenyl-A -[2-(1 -pyrrolidinyl)- 5-(trifluoromethyl)phenyl]cyclopropanecarboxamide (21 .1 mg, yield 19.8 %). ¹ H NMR (400 MHz, DMSO-d₆) δ ppm: 8.24 (s, 1 H), 8.09 (s, 1 H), 7.54-7.48 (m, 2H), 7.48-7.35 (m, 3H), 7.34-7.27 (m, 1 H), 7.09-7.04 (m, 1 H), 2.82 (m, 4H), 1.64 (m, 4H), 1.50 (q, 2H),1.13 (q, 2H). [ES+ MS] m/z 375 (M+H).

Example 37: 1-(4-methylphenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]- cylopropanecarboxamide

1-(4-methylphenyl)cyclopropanecarboxylic acid (ACROS, 34.3 mg, 0.195 mmol) was dissolved in DMF (2 mL) under N₂ atmosphere. 1-Hydroxy-1 H-benzotriazol hydrate (ALDRICH, 29.8 mg, 0.195 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDCI) (ALDRICH, 37.3 mg, 0.195 mmol) and sodium bicarbonate (16.4 mg, 0.195 mmol) were added. This mixture was heated at 80°C under MW irradiation during 10 min. [2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 40 mg, 0.175 mmol) was added to the mixture and it was heated in the MW at 120°C during 30 min. Solvent was evaporated to dryness and the crude obtained was dissolved in DCM and purified firstly by silica chromatography using DCM/MeOH as eluents and secondly by preparative HPLC (column: XBridge 19x150mm, 25% to 100% ACN/water) to obtain the desired product (6 mg 8% yield). ¹ H NMR (400 MHz, CDCI₃) δ ppm: 9.41 (br s, 1 H), 8.98 (s, 1 H), 8.29 (s, 1 H), 7.50 (s, 1 H), 7.40 (dd, 1 H), 7.34 (d, 1 H),7.25 (d, 2H), 7.21 (d, 2H), 2.47 (s, 3H), 1.68 (q, 2H), 1.15 (q, 2H). [ES+ MS] m/z 387 (MH+).

Example 38: 1-[4-(methyloxy)phenyl]-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)- phenyl]cyclopropanecarboxamide

1-[4-(methyloxy)phenyl]cyclopropanecarboxylic acid (ACROS, 33.7 mg, 0.175 mmol) was dissolved in DMF (2 mL) under N₂ atmosphere. HATU (ABCHEM, 123 mg, 0.200 mmol), DIPEA and [2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 40 mg, 0.175 mmol) were added. Reaction was heated at 100°C in MW and at room temperature 24 hours. Reaction media was diluted with DCM and washed with water, brine, dried over anhydrous Mg₂S0₄, filtered and concentrated to dryness. Crude was purified by silica chromatography using DCM/MeOH as eluents. Title compound was obtained (6 mg, 8% yield). ¹ H NMR (400 MHz, CDCI₃) δ ppm: 9.44 (br s, 1 H), 8.99 (s, 1 H), 8.30 (s, 1 H), 7.59 (s, 1 H), 7.33-7.41 (m, 2H), 7.29 (dd, 2H), 6.93 (dd, 2H), 3.91 (s, 3H) 1.68 (q, 2H), 1 .15 (q, 2H). [ES+ MS] m/z 403 (MH+).

Example 39: /V-[2-(4-methyl-1-piperazinyl)-5-(trifluoromethyl)phenyl]-1 -phenylcyclo- propanecarboxamide bromohydrate salt

N-Methylpiperazine (ALDRICH, 0.032 mL, 0.292 mmol) was added to a mixture of N-[2- bromo-5-(trifluoromethyl)phenyl]-1 -phenylcyclopropanecarboxamide (Intermediate 4, 80 mg, 0.208 mmol), caesium carbonate (ALDRICH, 136 mg, 0.416 mmol) and copper (I) iodide (ALDRICH, 7.93 mg, 0.042 mmol) in DMF (3 mL). The mixture was heated at 80 °C under nitrogen overnight. More caesium carbonate (ALDRICH, 136 mg, 0.416 mmol), copper(l) iodide (ALDRICH, 7.93 mg, 0.042 mmol) and N-methylpiperazine (ALDRICH, 0.032 mL, 0.292 mmol) were added. The reaction mixture under N₂ was heated at 80°C during the weekend. Solids were filtered off and the solvent was removed under vacuum. Crude was purified by preparative HPLC (column: XBRIDGE 30x150 mm. gradient: 40% to 100% water- ACN) to obtain 3 mg (yield 3%) of /V-[2-(4-methyl-1 -piperazinyl)-5-(trifluoromethyl)-phenyl]-1 - phenylcyclopropanecarboxamide bromohydrate salt. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 8.88 (s, 1 H), 8.55 (s, 1 H), 7.66-7.58 (m, 2H), 7.58-7.50 (m, 2H), 7.49-7.43 (m, 1 H), 7.35-7.29 (m, 2H), 3.47 (m, 2H), 3.27 (m, 2H), 2.79-2.57 (m, 5H), 2.26-2.04 (m, 2H), 1.80 (q, 2H), 1.21 (q, 2H). [ES+ MS] m/z 404 (M+H).

Example 40: /V-[2-(1 H-imidazol-1 -yl)-5-(trifluoromethyl)phenyl]-1-phenylcyclopropane- carboxamide

Title compound was prepared by a method analogous to that described for Example 39, replacing N-methylpiperazine with imidazole (MERCK, 28.2 mg, 0.414 mmol). Also 8- hydroxyquinoline (ALDRICH, 4.01 mg, 0.028 mmol) was used as catalyst and reaction was heated under MW irradiation at 120°C during 3 hours. Reaction was diluted with EtOAc and washed twice with NH₄CI (aq., sat.) and once with NaCI, dried over Mg₂S0₄, filtered and concentrated to dryness. Crude was purified by silica chromatography using hexane/EtOAc as eluents. to obtain 23 mg (yield 44.9%) of /V-[2-(1 H-imidazol-1-yl)-5- (trifluoromethyl)phenyl]-1 -phenylcyclopropane-carboxamide was obtained. ¹ H NMR (400 MHz, DMSO-d₆) δ ppm: 8.23 (m, 1 H), 8.13 (br s, 1 H), 7.73 (m, 1 H), 7.68-7.63 (m, 1 H), 7.60- 7.55 (m, 1 H), 7.32 (m, 5H), 7.18 (m, 1 H), 6.98 (m, 1 H), 1.39 (q, 2H), 1 .08 (q, 2H). [ES+ MS] m/z 372 (M+H).

Example 41 : 1-phenyl-/V-[2-(1 /-/-pyrazol-1-yl)-5-(trifluoromethyl)phenyl]cyclopropane- carboxamide

Title compound was prepared by a method analogous to that described for Example 40, replacing N-methylpiperazine with pyrazole (ALDRICH, 36 mg, 0.518 mmol). Also 8- hydroxyquinoline (ALDRICH, 3.02 mg, 0.021 mmol) was used as catalyst and reaction was heated under MW irradiation at 130°C during 3 h. 14 mg (yield 36.2%) of 1 -phenyl-/V-[2-(1 H- pyrazol-1-yl)-5-(trifluoromethyl)phenyl]cyclopropane-carboxamide was obtained. ¹ H NMR (400 MHz, CDCI₃) 5 ppm: 10.27 (br s, 1 H), 8.99 (s, 1 H), 7.67 (m, 1 H), 7.42 (m, 5H), 7.38-7.26 (m, 2H), 7.10 (m, 1 H), 6.33 (m, 1 H), 1.72 (q, 2H), 1.18 (q, 2H). [ES+ MS] m/z 372 (M+H). Example 42: 2-methyl-2-(phenyloxy)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5- (trifluoromethyl)phenyl]propanamide

To a solution of [2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]amine (ENAMINE, 70 mg, 0.307 mmol) in 2 mL of ACN was added NaHC0₃ (22.1 mg, 0.263 mmol) followed by 2- phenoxypropionyl chloride (Intermediate 5, 75 mg, 0.378mmol). Reaction was heated 45 minutes at 60°C under MW irradiation. Reaction media was filtered and the resulting solution was evaporated to dryness and the residue was purified by silica chromatography using DCM/MeOH as eluents giving the title compound (28 mg, 23%) as a brown oil. ¹H NMR (400 MHz, DMSO-d₆) δ ppm: 10.88 (s, 1 H), 9.1 1 (s, 1 H), 8.61 (s, 1 H), 8.22 (s, 1 H), 7.96 (d, 1 H), 7.75 (d, 1 H), 7.28 (t, 2H), 7.05 (t, 1 H), 6.92 (d, 2H), 1.49 (s, 6H). [ES+ MS] m/z 391 (MH+).

Examples 43-45 were prepared by methods analogous to that described for Example 8 or 42 replacing the carboxylic acid 2-[(4-chlorophenyl)oxy]propanoic acid in Example 8, or the acid chloride 2-phenoxypropionyl chloride (Intermediate 5) in Example 42 with that indicated in Table 5. Modifications in the reaction conditions are also indicated.

(a) The reaction mixture was heated at 80°C

Example 46: /V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]alaninamide

To a 5 ml. microwave-reaction vial was added 2-bromo-N-[2-(1 H-1 ,2,4-triazol-1-yl)-5- (trifluoromethyl)phenyl]propanamide (Intermediate 6, 73 mg, 0.201 mmol) in MeOH (2 ml_). To the resulting solution 2,3-dihydro-1 H-inden-5-amine (ALDRICH, 40.2 mg, 0.302mmol) and Et₃N (0.028mL) were added. The vial was sealed and heated at 1 10°C in a microwave reactor for 15 minutes. The resulting suspension was evaporated under reduced pressure and the residue was purified twice by silica using DCM/EtOAC and Hexane/EtOAc as eluents. Title compound was obtained (76 mg, 91 % yield) as a pale brown solid. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 10.81 (br s, 1 H), 8.87 (d, 1 H), 8.26 (s, 1 H), 7.75 (s, 1 H), 7.47 (dd, 1 H), 7.41 (d, 1 H), 6.99 (d, 1 H), 6.38 (br s, 1 H), 6.36 (dd, 1 H), 3.84 (q, 1 H), 2.85-2.66 (m, 4H), 2.07-1.97 (m, 2H), 1.58 (d, 3H). [ES+ MS] m/z 416 (MH⁺).

Example 47: /V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]glycinamide

2,3-dihydro-1 H-inden-5-amine (ALDRICH, 38.2 mg, 0.286 mmol) and DIPEA (0.146 ml, 0.859 mmol) were added under stirring to a solution of Intermediate 8 (100 mg, 0.286 mmol) in 5 ml. of dry DCM. Reaction was stirred under reflux overnight. Solvents were removed, and the residue was purified by silica chromatography using hexane/EtOAc as eluents to yield the title compound (71 .9 mg, 62% yield) as a pale yellow solid. ¹ H NMR (400 MHz, CDCI₃) δ ppm: 10.81 (br s, 1 H), 8.89 (s, 1 H), 8.32 (s, 1 H), 7.68 (s, 1 H), 7.50-7.41 (m, 2H), 7.02 (d, 1 H), 6.45-6.37 (m, 2H), 3.91 (s, 2H), 2.85-2.74 (m, 4H), 2.08-1 .99 (m, 2H). [ES+ MS] m/z 402 (MH⁺).

The following compounds in Table 6 (Examples 48 to 50) were commercially available and also can be prepared using the procedures described above. Table 6.

Biological Assays

The compounds of this invention may be tested in one of several biological assays to determine the concentration of compound which is required to have a given pharmacological effect. Three assays are described below. It should be noted that the results obtained using any one of the following three assays is directly comparable with those obtained using either of the other two assays. Assays 1 and 2

The following two assays (XC₅₀ and IC₅₀) determine potency of hits from antimalarial whole cell screening against P. falciparum using parasite lactate dehydrogenase (LDH) as surrogate of parasitic growth. Results for the assays are provided in Table 1 above. Literature References

• Desjardins, R.E., Canfield, C. J., Haynes, J.D. and Chulay J.D. (1979) Quantitative assessment of antimalarial activity in vitro by a semiautomated microdilution technique. Antimicrob. Agents Chemother. 16:710-718.

• Gomez, M.S., Piper, R.C., Hunsaker, L.A., Royer, R.E., Deck, L.M., Makler, M.T. and Vander Jagt D.L. (1997) Substrate and cofactor specificity and selective inhibition of lactate dehydrogenase from the malarial parasite P. falciparum. Mol. Biochem. Parasitology 90:235-246.

Delhaes, L, Lazaro, J.E., Gay, F., Thellier, M., Danis, M. (1999) The microculture tetrazolium assay (MTA) another colorimetric method of testing Plasmodium falciparum chemosensitivity. Annals of Tropical Medicine and Parasitology 93:1 :31-

40.

• Basco, L.K., Marquet; F., Makler; M.M., Le Bras, J. (1995) Plasmodium falciparum and Plasmodium vivax: Lactate Dehydrogenase Activity and its Application for in vitro drug susceptibility assay. Experimental Parasitology 80: 260-271.

· Goodyer, I.D., Taraschi, T.F. (1997) Plasmodium falciparum: A simple, rapid method for detecting parasite clones in microtiter plates. Experimental parasitology 86:158- 160.

General information

Target:

Cell Target Name and Origin: Protozoa

Cell derivation: Cell Stabilized / Transformed cell line

Cell Line Name: 3D7 P. falciparum strain. Solvent Tolerance of the assay:

0.5% (v/v) DMSO allowed, concentrations above 0.5% abolish parasite growth

Assay Data:

Absorbance using Spectramax Plus. NBT reduction at 650 nm.

Instruments:

SpectraMax Plus supplied by Molecular Devices.

Multidrop Combi supplied by Thermo Electron Company.

PS Microplate 384 Well Cell Culture plates mClear,flat bottom, rounded square well, physical surface treatment, with lid, plate black, bottom clear, sterile, supplied by Greiner bio-one, Catalog # 781091

Biological and Molecular Reagents

• Red Blood Cells (Storage 4 °C) - sourced from Spanish Red Cross Blood Bank · Plasmodium falciparum strains, - source MR4

• Hypoxanthine - Supplier FLUKA, Catalog Number 56700, Molecular Weight 136.1 1

• RPMI-1640 medium - Supplier SIGMA, Catalog Number R5886, 25 mM HEPES and NaHCO³ without L-glutamine

ALBUMAX II - Supplier GIBCO, Catalog Number 1 1021-037

· L-GLUTAMINE - Supplier MERCK, Catalog Number 1.00229

• Sodium Lactate - Supplier Sigma, Catalog Number L7022, Molecular Weight 1 12.1 • NitroBlue Tetrazolium - Supplier Sigma, Catalog Number N-6876, Molecular Weight 817.7

• 3-Acetylpyridine adenine dinucleotide - Supplier Sigma, Catalog Number A- 5251 , Molecular Weight 662.4

· Diaphorase from Clostridium Kluyveri - Supplier Sigma, Catalog Number D-5540

• Tween 20 - Supplier Sigma, Catalog Number P-1379

• Triton X-100 - Supplier Sigma, Catalog Number T8787

• Lauryl Sulfate - Supplier Sigma, Catalog Number L4390, Molecular Weight 288.4 Solutions

• Albumax 20x - Albumax II 100 g/l, D-sucrose 40 g/l, L-glutamine 6g/l

• Hypoxanthine 15 mM (100x) - hypoxanthine 2 g/l, add NaOH to solve

• Dilution buffer - 100mM Tris-HCI pH 8.0

• Reaction buffer - 100mM Tris-HCI pH 8.0, 0.5% Tween 20

· Sodium L-Lactate - Prepare a solution of 1 M in Tris-HCI pH 8.0, Stored in appropriate aliquots at -20 °C

• NBT (NitroBlue Tetrazolium) - Prepare a solution of 5 mM in mili-Q water, Stored in appropriate aliquots at -20 °C

• APAD (3-Acetylpyridine adenine dinucleotide) - Prepare a solution of 10 mM in mili-Q water, Stored in appropriate aliquots at -20 °C

• Diaphorase - Prepare a solution of 5 mg/ml in Tris-HCI pH 8.0, 0.5 % Tween 20, freshly prepared before initiation of the assay.

• Lauryl sulphate (SDS) - 10% SDS Assay 1 : XCgn (LDH) assay

Antimalarial whole cell screening (AWCI) - Absorbance - TCS - XC₅₀ determination P. falciparum 3D7 - to determinate XC50 for antimalarial whole cell screening hits.

Dilutions are done using an inter-plate approach (5 concentrations assayed, 5-fold dilution).

Outcome - Concentration Response Curves (CRC) Experiment

Type of Activity or Affinity Measured: IC₅₀

Starting Concentration of Test Compounds (M): 2 μΜ

Dilution factor 1 :5

Number of Points per Curve: 5

Method for Analysis of CRC / Data Reduction: excel worksheet XC50 Data analysis

Assay Capacity

Plate Format: 384

Parameters Measured per Well: Lactate dehydrogenase activity

Reaction mix solution (1.43x final assay concentration) 143 mM Sodium L-Lactate - 143 μΜ APAD, 178.75 μΜ NBT, 286 μς/πιΙ Diaphorase (2.83 U/ml), 0.7% Tween 20, 100 mM Tris-HCI pH 8.0

Final concentrations:

100 mM Sodium L-Lactate - 100 μΜ APAD, 125 μΜ NBT, 200 μςΛτιΙ (1 U/ml) Diaphorase, 0.5% Tween 20, 100mM Tris-HCI pH 8.0, must be freshly prepared before initiation of the assay and protected from the light.

Protocol

Preparation of the compounds to be tested.

Compounds are prepared from stock solutions at 1 mM in DMSO. Master plates of serial dilutions (5-fold dilution, 5 different concentrations) are prepared using an inter-plate approach.

Preparation of assay plates

Assay plates (384 well plates) are prepared by stamping 0.05 μΐ of compound from master plates in each well. Final volume assay is 25 μ\- and final compound concentrations are 2 μΜ, 0.4 μΜ, 0.08 μΜ, 0.016 μΜ and 0.0032 μΜ. In the 6th column 0.05 uL of DMSO are dispensed (positive controls). In the 18th column 0.05 mL of a mix of a 50 μΜ chloroquine, 50 μΜ artemisinin stock solutions are dispensed (negative controls).

Preparation of parasite inoculum.

P. falciparum 3D7 strain is cultivated using standard procedures as described in Trager et al. (1976). Science 193:673-675. An inoculum of parasitized red blood cells (pRBC) with a parasitaemia of 0.25% and 2% haematocrit in RPMI-1640, albumax and 150 μΜ hypoxanthine is prepared and used for the assay.

Whole cell assay

25 μΙ of parasite inoculum is dispensed in assay plates with compounds using a multidrop combi dispenser. Plates are shaken for 10 sec. to ensure mixing of the compound with the parasites. Plates are incubated at 37 °C for 72 h in an atmosphere of 5% C0₂, 5% 0₂, 95% N₂.

Evaluation of LDH activity as surrogate of parasite growth

After 72 hours of incubation, plates are frozen at -70 °C overnight. Next morning, plates are thawed at room temperature for at least 4 hours. To evaluate LDH activity, 70 μ\- of reaction mix solution (143 mM Sodium L-Lactate - 143 μΜ APAD, 178.75 μΜ NBT, 286 μg/ml Diaphorase (2.83 U/ml), 0.7% Tween 20, 100 mM Tris-HCI pH 8.0) freshly made is dispensed using a multidrop. Plates are shaken to ensure mixing and absorbance at 650nm is monitorized after 10 minutes of incubation at room temperature. Data are normalized to percentage of growth inhibition using positive and negative controls. XC₅₀ is calculated as compound concentration giving 50% inhibition of positive control growth using excel XC₅₀ data analysis worksheet. QA

HTS QA (target is value at which assay meets quality requirements): Z'-Factor Target 0.7- 0.8.

Assay 2: IC_sn (LDH) Assay

Plasmodium falciparum whole cell time extended assay - Absorbance - % Inhibition - IC₅₀ - TCS - The assay determines the percentage of inhibition of in vitro P. falciparum growth. Level of P. falciparum LDH is proportional to P. falciparum growth.

Concentration Response Curves (CRC) Experiment Outcome - ICm

Starting Concentration of Test Compounds (M): 20 μΜ

Dilution factor 1 :3

Number of Points per Curve 1 1

Method for Analysis of CRC / Data Reduction: excel worksheet and Grafit 5.0

Assay Capacity

Plate Format is 384

Parameters Measured per Well: Lactate dehydrogenase activity

Reaction mix solution (1.43x final assay concentration)

• 143 mM Sodium L-Lactate - 143 μΜ APAD, 178.75 μΜ NBT, 286 μς/πιΙ Diaphorase (2.83 U/ml), 0.7% Tween 20

100 mM Tris-HCI pH 8.0, 2.86% Triton X-100

Final concentrations:

100 mM Sodium L-Lactate - 100 μΜ APAD, 125 μΜ NBT, 200 μςΛτιΙ (1 U/ml) Diaphorase, 0.5% Tween 20, 100mM Tris-HCI pH 8.0, 2% Triton X-100. Must be freshly prepared before initiation of the assay and protected from the light.

Protocol

Preparation of the compounds to be tested.

Compounds are prepared from stock solutions at 10 mM in DMSO. Master plates of serial dilutions (3-fold dilution, 1 1 different concentrations) are prepared using an intra-plate approach.

Preparation of assay plates

Assay plates (384 well plates) are prepared by stamping 0.05 μΐ of compound from master plates in each well. Final volume assay is 25 μ\- and higher compound concentration tested is 20 μΜ. In the 6th column 0.05 μ\- of DMSO are dispensed (positive controls). In the 18th column 0.05 mL of a mix of a 50 μΜ chloroquine, 50 μΜ artemisinin stock solutions are dispensed (negative controls).

Preparation of parasite inoculum.

P. falciparum 3D7 strain is cultivated using standard procedures as described in Trager et al. (1976). Science 193:673-675. An inoculum of parasitized red blood cells (pRBC) with a parasitaemia of 0.25% and 2% haematocrit in RPMI-1640, albumax and 150 μΜ hypoxanthine is prepared and used for the assay. Whole cell assay

Evaluation of LDH activity as surrogate of parasite growth

After 72 hours of incubation, plates are frozen at -70 °C overnight. Next morning, plates are thawed at room temperature for at least 4 hours. To evaluate LDH activity, 70 μ\- of reaction mix solution (143 mM Sodium L-Lactate - 143 μΜ APAD, 178.75 μΜ NBT, 286 μg/ml Diaphorase (2.83 U/ml), 0.7% Tween 20, 100 mM Tris-HCI pH 8.0) freshly made is dispensed using a multidrop. Plates are shaken to ensure mixing and absorbance at 650nm is monitorized after 10 minutes of incubation at room temperature. Data are normalized to percentage of growth inhibition using positive and negative controls. IC₅₀ is calculated as compound concentration giving 50% inhibition of positive control growth using excel and Grafit 5.0 programs.

OA

HTS QA (target is value at which assay meets quality requirements): Z'-Factor Target 0.7-0.8 Assay 3: IC_sn (hypoxanthine) Assay

The following assay (IC₅₀) determines potency of hits from antimalarial whole cell screening against P. falciparum using incorporation of radiolabelled hypoxanthine as surrogate of parasitic growth. Results for the assays are provided in Table 1 above. Literature References

Equipment and Materials

Instrument Cell Harvester TOMTEC

Supplier Perkin Elmer Instrument Microbeta counter Wallac

Supplier Perkin Elmer

Article 96 Well Cell Culture plates,flat bottom with Lid, Tissue culture Treated, Polystyrene, sterile

Supplier Costar

Catalog # 3599

Article 96 Well Cell Culture plates,V-bottom with Lid, Tissue Culture Treated, Polystyrene, sterile

Supplier Costar

Catalog # 3894

Article Printed Filtermat A

Glass fibre filter

Supplier Wallac

Catalog * 1450-421

Article Melti Lex A

Melt-on Scintillator Sheets

Supplier PerkinElmer

Catalog * 1450-441 Biological and Molecular Reagents

Reagent Red Blood Cells ABStorage 4°C

Source Spanish Red Cross Blood Bank

Reagent Plasmodium falciparum 3D7A strain

Source MR4

Chemical Reagents

Component Hypoxanthine

Supplier FLUKA

Catalog Number 56700

Molecular Weight 136.1 1

Component 3H-Hypoxanthine

Supplier Amersham Biosciences

Catalog Number TRK74

Safety relevant information (known hazards, MSDS)

radiolabeled

Component RPMI-1640 medium

Supplier SIGMA

Catalog Number R5886

Additional Info 25mM HEPES and NaHC03 without L-glutamine

Component ALBUMAX II

Supplier GIBCO

Catalog Number 1 1021-037

Component L-GLUTAMINE Supplier MERCK

Catalog Number 1.00229

Solutions

Solution Albumax 20x

Components Albumax II 10Og/l

D-sucrose 40g/l

L-glutamine 6g/l

Solution Hypoxanthine 15 mM (100x)

Components hypoxanthine 2 g/l

Guidelines Add NaOH to solve

Protocol Steps

Experimental protocol

1. Make serial dilutions of each compound in a v-bottom 96 well plate with DMSO from 10 mM stock

(DMSO) to 0.039 mM (from 1 to 9 wells) (dilution factor 2). Add DMSO to wells 10 to 12. Distribute 20μΙ of RMPI 1640 medium to a new v-botton 96 well plate. In this new plate, dispense 20 ul of compound from the

above serial dilution plate (compound dilution factor 2).

To generate the assay plate, in a sterile flat bottom 96 well plate distribute 50 ul of RMPI 1640 and dispense 1 ul of compound diluted in RPMI-1640 from previous plate.

2. Prepare a culture of parasitized red blood cells (RBC) of a parasitaemia of 0.5% and a 4% haematocrit in RPMI-1640, 2X albumax, 30 uM hypoxanthine.

Distribute 50 ul of this culture to the assay plate from well 1 1 to 1.

Prepare a culture of 4% haematocrit RBC in RPMI-1640, 2X albumax, 30 uM hypoxanthine, and distribute 50 ul to well 12.

This culture represents the negative control of the assay.

3. Incubate the plate for 24h at 37°C, 5% C02, 5% 02, 95% N2.

4. After 24h, add to each well, from colum 1 1 to 1 , 0.2 uCi (8 ul) of a stock solution of 3H-hypoxanthine 0.025 uCi/ul in RMPI -1640.

Incubate the plate for another 24h at 37°C, 5% C02, 5% 02, 95% N2 .

5. After 24h, freeze the plates overnight at -70°C.

6. Thaw the plates, shake and harvest the cells on a glass fiber filter in a TOMTEC Cell Harvester 96.

7. Dry the filters, melt heating with melt-on scintillator sheets and measure the incorporation of 3H-hypoxanthine in a microbeta counter.

Data Management

Plate Setup / Control Values

Concentration Response Curve - Control Curves (up to 4 CRCs)

Name of test agent Chloroquine diphosphate salt Supplier / Cat. # Sigma C-6628

Starting Cone. (M) 50 ng/ml

Dilution Factor Applied dilution factor:2

No. Points in each CRC 9

Locations on test plate variable

Expected activity value IC50= 15 ng/ml

Name of test agent Atovaquone

Compound Number GR 151218X

Starting Cone. (M) 60 ng/ml

Dilution Factor Applied dilution factor: 3

No. Points in each CRC 9 dilutions

Locations on test plate variable

Expected activity value Ic50=0.2 ng/ml Results of the biological assays

All Examples of the invention, other than Examples 12, 15, 16, 17, 18, 20, 21 and 22 (Table 2, list B) were tested in at least one of the three biological assays described hereinabove.

Examples 1-1 1 , which are the compounds shown in Table 1 (compounds of List A) were all found to give an XC₅₀ of less than 1.ΟμΜ in Assay 1 .

Examples 14 and 19, which are the tested compounds shown in Table 2 (List B) were found to give an XC₅₀ of greater than 1.ΟμΜ in Assay 1 . Examples 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 40, 41 , 43, 45, 46 and 47 were found to give an IC₅₀ of less than 5.0μΜ in Assay 3. Of these, Examples 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 37, 38, 40, 41 , 43, 45 and 46 were found to give an IC₅₀ of less than 2.0μΜ in Assay 3. In particular, Examples 23, 24, 25, 28, 29, 30, 31 , 32, 33, 37, 38, 40, 41 , 43, 45 and 46 were found to give an IC₅₀ of less than 1.ΟμΜ in Assay 3. Example 28 was found to give an IC₅₀ of 0.001 μΜ in Assay 3.

Examples 39, 42, 44, 48, 49 and 50 gave an IC₅₀ of greater than 5.0μΜ in Assay 3. These Examples may have utility in the preparation of other compounds of Formula (I) described herein.

All publications, including but not limited to patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference as though fully set forth. The application of which this description and claims forms part may be used as a basis for priority in respect of any subsequent application. The claims of such subsequent application may be directed to any feature or combination of features described herein. They may take the form of product, composition, process, or use claims and may include, by way of example and without limitation, the following claims.

Claims

1. A compound of Formula (I)

(I)

Wherein:

R¹ is C_1-6alkyl, C_1-6haloalkyl, C_1-6alkoxy, C_1-6haloalkoxy or halo;

R² is -NR⁴R⁵;

R⁴ and R⁵ are independently Ci_-6alkyl or, together with the N to which they are attached, form

C_5-7heteroaryl or C_5-7heterocyclyl, either of which i) may optionally contain 1 -3 additional heteroatoms, independently selected from the group consisting of N, O and S and ii) may be optionally substituted with d_-3alkyl;

R³ is H;

R^a is -X-Y-Z or -CCI₃;

X is -CR⁶R⁷;

R⁶ is H or C_1-6alkyl;

R⁷ is H or Ci_-6alkyl;

R⁸ is H or C_halky!;

p is 0 or 1 ;

R⁹ is H, C_1-6alkyl, C_1-6haloalkyl, C_1-6alkoxy, C_1-6haloalkoxy, halo, cyano or optionally substituted C₅-₇heteroaryl or optionally substituted C_5-7heterocyclyl, wherein the optional substitutents for C_5-7heteroaryl and C_5-7heterocyclyl are selected from Ci_-3alkyl;

R¹⁰ is H, C_1-6alkyl, C_1-6haloalkyl, C_1-6alkoxy, C_1-6haloalkoxy halo or cyano;

or a pharmaceutically acceptable salt thereof, for use in therapy, for example the treatment of parasitic protozoal infections such as malaria, for example infection by Plasmodium falciparum. or a pharmaceutically acceptable salt thereof, for use in therapy.

2. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy according to claim 1 , wherein R¹ is C_1-6haloalkyl or halo.

3. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy according to claim 1 or claim 2, wherein R² is -NR⁴R⁵ wherein R⁴ and R⁵, together with the N to which they are attached, form C₅heteroaryl or C₆heterocyclyl, which may optionally contain 1-3 additional heteroatoms, independently selected from the group consisting of N and O.

4. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy according to any one of claims 1 to 3, wherein R^a is X-Y-Z and R⁶ and R⁷ together with the carbon atom to which they are attached, form cyclopropyl.

5. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy according to any one of claims 1 to 4, wherein R^a is X-Y-Z and is a direct bond, -0-, -NH- or - N(CH₃)-CH₂.

6. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy according to any one of claim is

7. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy according to claim 6, wherein R⁹ is H, Ci_-6alkyl, Ci_-6haloalkyl, Ci_-6alkoxy, halo, cyano or optionally substituted C_5-7heteroaryl.

8. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy according to claim 6, wherein R¹⁰ is H, C_1-6alkyl, C_1-6alkoxy, halo or cyano.

9. A compound of Formula (I) or a pharmaceutically acceptable salt thereof as defined in any one of claims 1 to 8, wherein the compound is selected from the group consisting of: /V-[2-(4-morpholinyl)-5-(trifluoromethyl)phenyl]-1-phenylcyclopropanecarbox-amide

2-phenyl-/V-[2-(1 H-1 ,2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]propanamide

2-phenyl-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]butanamide

2-(4-cyanophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-butanamide Λ/-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-1 -[3-(trifluoromethyl)- phenyl]cyclopropanecarboxamide

l-iS-chloropheny -N-p-il H-l ^^-triazol-l-y -S-itrifluoromethy phenyl]- cyclopropanecarboxamide

1-(4-chlorophenyl)-/V-[2-(1 H-1 _!2,4-triazol-1-yl)-5-(trifluoromethyl)phenyl]- cyclopropanecarboxamide

HS^-dichloropheny -N-p-O H-l ^^-triazol-l^^

cyclopropanecarboxamide

2- methyl-2-phenyl-N-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-propanamide 2-(4-fluorophenyl)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-acetamide

1 -phenyl-/V-[2-(1 -pyrrolidinyl)-5-(trif luoromethyl)phenyl]cyclopropane-carboxamide

2-methyl-2-(phenyloxy)-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)phenyl]-propanamide

2- [(3-chlorophenyl)oxy]-/V-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoromethyl)-phenyl]propanamide /V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]alaninamide; and

/V²-(2,3-dihydro-1 H-inden-5-yl)-/V¹-[2-(1 H-1 ,2,4-triazol-1 -yl)-5-(trifluoro- methyl)phenyl]glycinamide;

or a pharmaceutically acceptable salt thereof.

10. A compound of Formula (I) or a pharmaceutically acceptable salt thereof as defined in any of claims 1 to 9, for use in the treatment of a parasitic protozoal infection.

1 1 . Use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof as defined in any of claims 1 to 9, in the manufacture of a medicament for the treatment of a parasitic protozoal infection.

12. A method for the treatment of a human or animal subject suffering from a parasitic protozoal infection, comprising administering to said human or animal subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, as defined in any of claims 1 to 9.

13. A compound of Formula (I) or a pharmaceutically acceptable salt thereof for use according to claim 10, use of a compound Formula (I) or a pharmaceutically acceptable salt thereof according to claim 1 1 or a method according to claim 12 wherein the parasitic protozol infection is malaria.

14. A pharmaceutical composition comprising (a) a compound of Formula (I) or a pharmaceutically acceptable salt thereof as defined in any one of claims 1 to 9 and (b) one or more pharmaceutically acceptable carriers and/or excipients.

15. A process for the preparation of a compound of Formula (I) or a pharmaceutically acceptable salt thereof as defined in claim 1 , comprising reacting a compound of Formula (II) with a compound of Formula (III), wherein R^a, R¹ and R² are as defined in claim 1 for Formula (I), in the presence of a suit ions.