ANALOGUES OF ANTI-FIBROTIC AGENTS Field of the invention The present invention relates to derivatives of the anti-fibrotic drug, Tranilast.

Background of the invention

Fibrosis is a common response to a range of tissue insults that may lead to organ dysfunction. Diseases that are characterised by such pathological fibrosis include hepatic cirrhosis, pulmonary interstitial fibrosis, glomerulonephritis, heart failure (ischaemic and non-ischaemic), diabetic nephropathy, scleroderma, excessive scar tissue post surgery or device insertion, progressive kidney disease, glomerulonephritis, hypertension, heart failure due to ischaemic heart disease, valvular heart disease or hypertensive heart disease and hypertrophic scars. In addition, the elaboration of pathological matrix also has a role in fibroproliferative tumor progression and metastasis. Studies conducted over more than a decade have consistently indicated a major role of TGF-β in organ fibrosis and dysfunction, such that blockade of its expression and action represent an important therapeutic target.

Existing agents for treating fibrosis may have any number of undesirable properties including toxicity, poor solubility or efficacy.

One anti-fibrotic agent is Tranilast (n-[3,4-dimethoxycinnamoyl] anthranilic acid) and is used in Japan for the treatment of fibrotic skin disorders such as keloids and scleroderma. Although the precise mechanisms and mode of action of Tranilast are not completely understood, its ability to inhibit ERK phosphorylation, a major intermediate in the TGF-β signalling pathway, may underlie its antifibrotic effects, with known actions of Tranilast including the inhibition of TGF-β-induced extracellular matrix production in a range of cell types. Tranilast has also been shown to attenuate TGF-β-induced collagen synthesis in cardiac fibroblasts using an experimental model of diabetic cardiac disease.

Tranilast has also been shown to reduce inflammation in allergic diseases, such as allergic rhinitis and bronchial asthma, and to have antiproliferative activity.

However, it has recently been shown that genetic factors in certain patients, specifically the presence of a Gilbert's syndrome UGT1A1 variant, confer susceptibility to Tranilast- induced hyperbilirubinemia. Such hyperbilirubinemia may be associated with inhibition of UGT1 A1 by Tranilast per se and/or the in vivo formation of a Tranilast derivative. Therefore, it would be desirable to provide further alternatives to existing agents with potential anti-fibrotic, anti-inflammatory and anti-proliferative or anti-neoplastic activity for the treatment or prevention of diseases associated with fibrosis.

Summary of the invention

The present invention provides a compound of Formula (I)

or a pharmaceutically acceptable drug or prodrug thereof, wherein;

X ¹ (YZ) is C=O, C(F ₂ ) Or SO ₂ ;

X^ is NR ¹⁰ or (CH ₂ ) _P ;

T is a double bond, a triple bond or when T is a single bond, one pair of R ⁶ and R ⁷ are

fused to form a cyclopropane ring of the formula

A is selected from the group consisting of C ₃ to C ₁₂ cycloalkyl, C ₃ to C ₁₂ cycloalkenyl, C ₁ to C ₁₂ heterocycloalkyl, C ₁ to C ₁₂ heterocycloalkenyl, C ₆ -C ₁₈ aryl and C ₆ to C ₁₈ heteroaryl;

R ¹ , R ⁴ ' and R ⁵ are each independently selected from the group consisting of: H, OH, NO ₂ , CN, NH ₂ , optionally substituted C ₁ -C ₁₂ alkyl, optionally substituted C ₂ -C ₁₂ alkenyl, optionally substituted C ₂ -C ₁₂ alkynyl, optionally substituted C ₁ -C ₁₀ heteroalkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₁ -C ₁₂ alkyloxy, optionally substituted C ₂ -C ₁₂ alkenyloxy, optionally substituted C ₂ -C ₁₂ alkynyloxy, optionally substituted C ₁ -C ₁₀ heteroalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkenyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkenyloxy, optionally substituted C ₆ -C ₁₈ aryloxy, optionally substituted C ₁ -C ₁₆ heteroaryloxy, optionally substituted C ₁ -C ₁₂ alkylamino, SR ¹³ , SO ₃ H, SO ₂ NR ¹³ R ¹⁴ , SO ₂ R ¹³ , SONR ¹³ R ¹⁴ , SOR ¹³ , COR ¹³ , COOH, COOR ¹³ , CONR ¹³ R ¹⁴ , NR ¹³ COR ¹⁴ , NR ¹³ COOR ¹⁴ , NR ¹³ SO ₂ R ¹⁴ , NR ¹³ CONR ¹⁴ R ¹⁵ , NR ¹³ R ¹⁴ , and acyl;

R ² and R ³ , are each independently selected from the group consisting of: H, OH, NO ₂ , CN, NH ₂ , optionally substituted C ₁ -C ₁₂ alkyl, optionally substituted C ₂ -C ₁₂ alkenyl, optionally substituted C ₂ -C ₁₂ alkynyl, optionally substituted C ₁ -C ₁₀ heteroalkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₁ -C ₁₂ alkyloxy, optionally substituted C ₂ -C ₁₂ alkenyloxy, optionally substituted C ₂ -C ₁₂ alkynyloxy, optionally substituted C ₁ -C ₁₀ heteroalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkenyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkenyloxy, optionally substituted C ₆ -C ₁₆ aryloxy, optionally substituted Ci-C ₁₈ heteroaryloxy, optionally substituted C ₁ -C ₁₂ alkylamino, SR ¹³ , SO ₃ H, SO ₂ NR ¹³ R ¹⁴ , SO ₂ R ¹³ , SONR ¹³ R ¹⁴ , SOR ¹³ , COR ¹³ , COOH, COOR ¹³ , CONR ¹³ R ¹⁴ , NR ¹³ COR ¹⁴ , NR ¹³ COOR ¹⁴ , NR ¹³ SO ₂ R ¹⁴ , NR ¹³ CONR ¹⁴ R ¹⁵ , NR ¹³ R ¹⁴ and acyl; or R ² and R ³ may be fused to form a 5 or 6 membered cycloalkyl, heterocycloalkyl, aryl or heteroaryl ring each of which may be optionally substituted;

one pair of R ⁶ and R ⁷ are present when T is a double bond but R ⁶ and R ⁷ are not present when T is a triple bond, each R ⁶ and R ⁷ being independently selected from the group consisting of: H, NO ₂ , CN, optionally substituted C ₁ -C ₁₂ alkyl, optionally substituted C ₂ - C ₁₂ alkenyl, optionally substituted C ₂ -C ₁₂ alkynyl, optionally substituted C ₁ -C ₁₀ heteroalkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₂ -C ₁₂ heterocycloalkyl, optionally substituted C ₂ -C ₁₂ heterocycloalkenyl, optionally substituted C ₆ -C ₁₈ aryl, optionally substituted C ₁ -C ₁₈ heteroaryl, optionally substituted C ₁ -C ₁₂ alkyloxy, optionally substituted C ₂ -C ₁₂ alkenyloxy, optionally substituted C ₂ -C ₁₂ alkynyloxy, optionally substituted C ₁ -C ₁₀ heteroalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkenyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkenyloxy, optionally substituted C ₆ -C ₁₈ aryloxy, optionally substituted C ₁ -C ₁₈ heteroaryloxy, optionally substituted C ₁ -C ₁₂ alkylamino, SR ¹³ , SO ₃ H, SO ₂ NR ¹³ R ¹⁴ , SO ₂ R ¹³ , SONR ¹³ R ¹⁴ , SOR ¹³ , COR ¹⁴ , COOH, COOR ¹³ , CONR ¹³ R ¹⁴ , NR ¹³ COR ¹⁴ , NR ¹³ COOR ¹⁴ , NR ¹³ SO ₂ R ¹⁴ , NR ¹³ CONR ¹⁴ R ¹⁵ , NR ¹³ R ¹⁴ , and acyl;

R ⁸ is selected from the group consisting of H, halogen, OH, NO ₂ , CN, NH ₂ , optionally substituted C ₁ -C ₁₂ alkyl, optionally substituted C ₂ -C ₁₂ alkenyl, optionally substituted C ₂ - C ₁₂ alkynyl, optionally substituted C ₁ -C ₁₀ heteroalkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₁ -C ₁₂ alkyloxy, optionally substituted C ₂ -C ₁₂ alkenyloxy, optionally substituted C ₂ -C ₁₂ alkynyloxy, optionally substituted C ₁ -C ₁₀ heteroalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkenyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkenyloxy, optionally substituted C ₆ -C ₁₈ aryloxy, optionally substituted C ₁ -C ₁₈ heteroaryloxy, optionally substituted C ₁ -C ₁₂ alkylamino, SR ¹³ , SO ₃ H, SO ₂ NR ¹³ R ¹⁴ , SO ₂ R ¹³ , SONR ¹³ R ¹⁴ , SOR ¹³ , COR ¹³ , COOH, COOR ¹³ , CONR ¹³ R ¹⁴ , NR ¹³ COR ¹⁴ , NR ¹³ COOR ¹⁴ , NR ¹³ SO ₂ R ¹⁴ , NR ¹³ CONR ¹⁴ R ¹⁵ and NR ¹³ R ¹⁴ and acyl;

R ⁹ is selected from the group consisting of OH, OR ¹³ , COOR ¹³ , CONR ¹³ R ¹⁴ , NR ¹³ R ¹⁴ , tetrazol-5-yl, SO ₂ R ¹³ , SO ₂ NR ¹³ R ¹⁴ and CONHOR ¹³ ;

R ¹⁰ is selected from the group consisting of H, a N-protecting group, optionally substituted C ₁ -C ₁₂ alkyl, optionally substituted C ₂ -C ₁₂ alkenyl, optionally substituted C ₂ - C ₁₂ alkynyl, optionally substituted C ₁ -C ₁₀ heteroalkyl, optionally substituted C ₃ - C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₁ -C ₁₂ heterocycloalkyl, optionally substituted C ₁ -C ₁₂ heterocycloalkenyl, optionally substituted C _β -C ₁₈ aryl, and optionally substituted CrC^heteroaryl;

R ¹¹ and R ¹² are independently selected from the group consisting of H, halogen, OH, NO ₂ , CN, NH ₂ , optionally substituted C ₁ -C ₁₂ alkyl, optionally substituted C ₂ -C ₁₂ alkenyl, optionally substituted C ₂ -C ₁₂ alkynyl, optionally substituted C ₁ -C ₁₀ heteroalkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₂ -C ₁₂ heterocycloalkyl, optionally substituted C ₂ -C ₁₂ heterocycloalkenyl, optionally substituted C ₆ -C ₁₈ aryl, optionally substituted C ₁ -C ₁₈ heteroaryl, optionally substituted C ₁ -C ₁₂ alkyloxy, optionally substituted C ₂ -C ₁₂ alkenyloxy, optionally substituted C ₂ -C ₁₂ alkynyloxy, optionally substituted C ₁ -C ₁₀ heteroalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkyloxy, optionally substituted C ₃ -C ₁₂ cycloalkenyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkyloxy, optionally substituted C ₁ -C ₁₂ heterocycloalkenyloxy, optionally substituted C ₆ -C ₁₈ aryloxy, optionally substituted C ₁ -C ₁₈ heteroaryloxy, optionally substituted C ₁ -C ₁₂ alkylamino, SR ¹³ , SO ₃ H, SO ₂ NR ¹³ R ¹⁴ , SO ₂ R ¹³ , SONR ¹³ R ¹⁴ , SOR ¹³ , COR ¹³ , COOH, COOR ¹³ , CONR ¹³ R ¹⁴ , NR ¹³ COR ¹⁴ , NR ¹³ COOR ¹⁴ , NR ¹³ SO ₂ R ¹⁴ , NR ¹³ CONR ¹⁴ R ¹⁵ , NR ¹³ R ¹⁴ , and acyl;

each R ¹³ , R ¹⁴ , R ¹⁵ are each independently selected from the group consisting of H, -OH, optionally substituted C ₁ -C ₁₂ alkyl, optionally substituted C ₂ -C ₁₂ alkenyl, optionally substituted C ₂ -C ₁₂ alkynyl, optionally substituted C ₁ -C ₁₀ heteroalkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₁ -C ₁₂ heterocycloalkyl, optionally substituted C ₁ -C ₁₂ heterocycloalkenyl, optionally substituted C ₆ -C ₁₈ aryl, and optionally substituted d-C^heteroaryl;

n is an integer selected from the group consisting of O, 1 , 2, 3, and 4;

m is an integer selected from the group consisting of 1 , 2, 3, and 4;

m + n is an integer selected from the group consisting of 1 , 2, 3, 4, and 5; and

p is an integer selected from the group consisting of O, 1 , 2, 3, 4, and 5;

and when X ¹ (YZ) is C(F ₂ ) or SO ₂ ; or when T is a cyclopropane ring as defined above; or when R ¹ and R ⁵ are H and T is a double bond; or when X ² is (CH ₂ ) _P and p is O or 1 ; or when A is selected from the group consisting of C ₃ to C ₁₂ cycloalkyl, preferably C ₄ to C ₆ cycloalkyl, C ₁ to C ₁₂ heterocycloalkenyl, and C ₆ to C ₁₈ heteroaryl; then R ² and R ³ may also be independently selected from -X ³ -R ¹⁶ or -X ⁴ -R ¹⁷ ;

wherein X ³ and X ⁴ may be the same or different and are selected from the group consisting of a bond C, O, N and S; and

R ¹⁶ and R ¹⁷ may be the same or different and are selected from the group consisting of H, NHR ¹³ , NR ¹³ R ¹⁴ , OR ¹³ , halogen, C ₁ to C ₁₀ alkyl, C ₃ to C ₁₀ cyclokalkyl, C ₃ to C ₁₀ cycloalkylmethyl, C ₃ to C ₁₀ alkene, C ₃ to C ₁₀ alkyne, aryl, C ₅ to C ₂₀ alkaryl, fused C ₅ to C ₂₀ aryl or alkaryl and a hydrocarbon chain containing a heterocyclic or fused ring, any of which may be optionally substituted;

providing that when X ¹ (YZ) is C=O, X ² is NH and T is a double bond then A has the general formula:

wherein X ⁵ , X ⁶ , X ⁷ and X ⁸ may be independently C, S, O or N; R ¹⁸ is absent, H or COOR ¹³ and R ⁹ can be H when R ¹⁸ is COOR ¹³ , more preferably COOH; but A cannot be phenyl and R ¹ to R ⁵ cannot be -CF ₃ ;

and further providing that when X ¹ (YZ) is C=O, X ² is NH and T is a cyclopropane ring as defined above then R ⁹ cannot be tetrazol-5-yl; and

and even further providing that when X ¹ (YZ) is SO ₂ , then A has the general formula:

wherein X ⁵ , X ⁶ , X ⁷ and X ⁸ may be independently C, S, O or N and R ⁹ can be H when R ² and R ³ are each independently a Ci-C ₁₂ alkyloxy group containing at least one halogen atom, and more preferably when R ² and R ³ are each -OCHF ₂ .

In some embodiments at least one of R ¹ , R ² , R ³ , R ⁴ , and R ⁵ is selected from the group consisting of C ₁ -C ₁₂ alkyloxy containing at least one halogen atom, C ₁ -C ₁₂ alkenyloxy containing at least one halogen atom, and C ₁ -C ₁₂ alkynyloxy containing at least one halogen atom. In another embodiment at least one of R ² and R ³ is selected from the group consisting of a C ₁ -C ₁₂ alkyloxy group containing at least one halogen atom, a C ₂ -C ₁₂ alkenyloxy containing at least one halogen atom, a C ₂ -C ₁₂ alkynyloxy containing at least one halogen atom and a C ₃ -C ₁₂ cycloalkyloxy containing at least one halogen atom and the other R ² or R ³ is selected from the group consisting of an optionally substituted C ₁ -C ₁₂ alkyloxy group, an optionally substituted C ₂ -C ₁₂ alkenyloxy, an optionally substituted C ₂ - C ₁₂ alkynyloxy and an optionally substituted C ₃ -C ₁₂ cycloalkyloxy.

In some embodiments, the C ₁ -C ₁₂ alkyloxy group is of Formula (A):

Formula (A)

- wherein: R ²⁴ , R ²⁵ , and R ²⁶ are each independently selected from the group consisting of: H, halogen, OH, NO ₂ , CN, NH ₂ , optionally substituted C ₁ -C ₁₂ alkyl, and optionally substituted C ₂ -C ₁₂ alkenyl;

- R ²⁷ , R ²⁸ , R ²⁹ , and R ³⁰ are each independently selected from the group consisting of: H, halogen, OH, NO ₂ , CN ₁ and NH ₂ ; - at least one of R ²⁴ , R ²⁵ , R ²⁶ , R ²⁷ , R ²⁸ , R ²⁹ , and R ³⁰ is or contains a halogen atom;

- q is an integer selected from the group consisting of: O, 1 , 2, 3, 4, 5, 6, 7, 8, 9, and 10; and

- r is an integer selected from the group consisting of: 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

In some embodiments q and r are 0, and at least two of R ²⁴ , R ²⁵ , and R ²⁶ are a halogen.

The halogen may be selected from the group consisting of: fluorine, chlorine, bromine, and iodine. Preferably, the halogen is fluorine. In some embodiments at least one of R ¹ , R ² , R ³ , R ⁴ , and R ⁵ is selected from the group consisting Of-O-CHF _2, -OCF _3, -OCF ₂ CHF ₂ . In some embodiments R ³ is the group -O-CHF ₂ . In some embodiments R ² and R ³ are the group -Q-CHF ₂ . In some aspects, R ¹ and R ⁵ are H and R ² and R ³ are O- R ¹⁶ and O-R ¹⁷ , wherein R ¹⁶ and R ¹⁷ are independently and preferably selected from the group consisting of unsubstituted C ₁ -C ₆ alkyl, preferably methyl or ethyl; C ₁ -C ₆ fluoro substituted alkyl, preferably, F ₃ CO, F ₂ HCO, F ₂ HCF ₂ CO; or are fused to form a 5 or 6 membered ring, preferably R ² and R ³ form a bridging difluoromethylenedioxy group or a bridging tetrafluoroethylenedioxy group.

In some embodiments T is a double bond while in other embodiments T is a triple bond.

In some embodiments R ⁹ is selected from the group consisting of: COOR ¹¹ and CONR ¹¹ R ¹² . In some embodiments R ⁹ is selected from the group consisting of: COOH, CONH ₂ , and CONHCH ₃ .

In some embodiments R ⁹ is NR ¹¹ R ¹² while in some embodiments R ⁹ is NH ₂ .

In some embodiments n is 1.

In some embodiments R ⁸ is halogen.

In one form of the invention, T is a single bond and one pair of R ⁶ and R ⁷ are fused to

form a cyclopropane ring of the formula

In some embodiments R ² and R ³ form a bridging difluoromethylenedioxy group or a bridging tetrafluoroethylenedioxy group.

In some embodiments R ⁶ is CH ₃ . In some embodiments R ⁷ is CH ₃ or CN. In some embodiments R ⁸ is H or Me.

In preferred embodiments m is 1 and R ⁹ is selected from COR ¹³ and CONR ¹³ R ¹⁴ .

In other preferred embodiments R ⁹ is selected from the group consisting of COOH,

CONH ₂ , CONHOH and CONHCH ₃ .

In certain embodiments R ⁹ is the group tetrazol-5-yl. In some embodiments R ⁹ is selected from the group consisting of SO ₂ R , SO ₂ NR ¹ R .

In some more particular embodiments R ⁹ is selected from the group consisting of SO ₂ Me, SO ₂ NH ₂ , SO ₂ NHMe, SO ₂ NMe ₂ .

In some embodiments R ⁹ is NR ¹³ R ¹⁴ and in more particular embodiments R ⁹ is NH ₂ . In some embodiments R ⁸ is a halogen. In some embodiments X ² is NH.

According to a further form of the invention, there is provided a compound of the formula (II)

(H)

Preferred compounds of the invention have the formula (Ma)

(Ha) or a pharmaceutically acceptable salt or prodrug thereof, wherein; A, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ and R ¹² are as defined above in relation to compounds of formula (I).

A further aspect of the present invention is represented by a compound of the Formula (III)

(III) or a pharmaceutically acceptable salt or prodrug thereof, wherein;

A, T, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , and X ² are as defined above in relation to compounds of formula (I).

Yet another aspect of the present invention is represented by a compound of the Formula (IV)

(IV)

or a pharmaceutically acceptable salt or prodrug thereof, wherein;

A, T, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , and X ² are as defined above in relation to compounds of formula (I).

A further aspect of the present invention may be represented by a compound of Formula (V)

(V) or a pharmaceutically acceptable salt or prodrug thereof, wherein;

Het represents a heterocyclic ring and T, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , X ¹ (YZ) and X ² are as defined above in relation to compounds of formula (I).

According to a further aspect of the present invention, there is provided a compound of formula (Vl)

(Vl) or a pharmaceutically acceptable salt or prodrug thereof, wherein; T, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and p are as defined above in relation to compounds of formula (I).

In preferred embodiments, R ⁶ and R ⁷ are H in the compounds of formulas (III), (IV), (V) and (Vl).

In some embodiments T is preferably a double bond in the compounds of formulas (III), (IV), (V) and (Vl).

In the compounds of formula (V), certain preferred embodiments have X ¹ (YZ) as being C=O. In other embodiments X ² is NH or NR ¹³ wherein R ¹³ is preferably C ₁ to C ₆ alkyl, most preferably a methyl group. In preferred embodiments of formula (V), X ¹ (YZ) is C=O and X ² is NH.

In preferred embodiments of the compounds of formulae (II) and (Ha), R ¹¹ and R ¹² are selected from the group consisting of H, CN or halogen. Preferably, R ¹¹ and R ¹² are H. When R ¹¹ and/or R ¹² are halogen, the halogen is preferably fluorine.

When R ⁹ is present in the compounds of formulae (II), (Ma), (III), (IV), (V) and (Vl), R ⁹ is preferably selected from the group consisting of CO ₂ H, CO ₂ R ¹³ , SO ₂ R ¹³ , SO ₂ NH ₂ ,

SONHR ¹³ , SONR ¹³ ₂ , CONH ₂ , CONHR ¹³ , CONHOR ¹³ and 5-tetrazolyl, wherein R ¹³ is preferably C ₁ to C ₆ alkyl, most preferably a methyl group. Alternatively, R ⁹ is selected from the group consisting Of CO ₂ H, CO ₂ R ¹³ , SO ₂ R ¹³ , SO ₂ NH ₂ , SONHR ¹³ , SONR ¹³ ₂ and 5- tetrazolyl.

In preferred embodiments of the formulae (II), (Ha), (III), (IV), (V) and (Vl), R ¹ and R ⁵ are H and R ² and R ³ are 0-R ¹⁶ and O-R ¹⁷ , wherein R ¹⁶ and R ¹⁷ are independently and preferably selected from the group consisting of unsubstituted C ₁ -C ₆ alkyl, preferably methyl or ethyl; C ₁ -C ₆ fluoro substituted alkyl, preferably, F ₃ CO, F ₂ HCO, F ₂ HCF ₂ CO; or are fused to form a 5 or 6 membered ring, preferably a fluoro substituted 1 ,4 dioxane or a fluoro substituted 1 ,3-dioxolane; or a C ₁ to C ₆ alkenyl, preferably -CH ₂ CCH.

In preferred embodiments of the formulae (II), (Ha), (III), (IV), (V) and (Vl), R ¹⁰ is H or a C ₁ to C ₆ alkyl, preferably, methyl.

In preferred embodiments of the formulae (II), (Ha), (III) and (IV), A has the general formula selected from the group consisting of

wherein X ⁵ , X ⁶ , X ⁷ and X ⁸ may be independently C, S, O or N.

In a particularly preferred embodiments, A has the general formula

wherein X ⁴ , X ⁵ , X ⁶ and X ⁷ may be C or N. More preferably, not more than two of X ⁴ , X ⁵ , X ⁶ and X ⁷ may be N. Non-limiting examples of suitable compounds of formula (Ha) include

Particular examples of compounds of formula (III) are as follows

R ⁹ = SO ₂ Me, SO ₂ NH ₂ , 5-tetrazolyl R ⁹ = NH ₂ , CONH ₂ , CONHMe, CONHOH Particular examples of compounds of formula (IV) are as follows

R ⁹ = SO ₂ Me, SO ₂ NH ₂ , 5-tetrazolyl R ⁹ = NH ₂ , CONH ₂ , CONHMe, CONHOH

R ⁹ = SO ₂ Me, SO ₂ NH ₂ , 5-tetrazolyl R ⁹ = NH ₂ , CONH ₂ , CONHMe, CONHOH Specific compounds of formula (V) include

R ⁹ = NH ₂ , CONH ₂ , CONHMe, CONHOH R ⁹ = SO ₂ Me ₁ SO ₂ NH ₂ , SONHMe, SONMe ₂

Some specific examples of compounds of formula Vl are as follows:

R ⁹ = SO ₂ Me, SO ₂ NH ₂ , 5-tetrazolyl R ⁹ = NH ₂ , CONH ₂ , CONHMe, CONHOH wherein p is O or 1.

Further derivatives include

In addition to compounds of formulae (I), (II), (Ma) ₁ (III), (IV), (V) and (Vl), the embodiments disclosed are also directed to pharmaceutically acceptable salts, pharmaceutically acceptable N-oxides, pharmaceutically acceptable prodrugs, and pharmaceutically active metabolites of such compounds, and pharmaceutically acceptable salts of such metabolites.

The compounds of the present invention may have anti-fibrotic, anti-inflammatory, anti- proliferative or anti-neoplastic activity and may, therefore, find use as an alternative and/or adjunct to Tranilast.

Detailed Description

In this specification a number of terms are used which are well known to a skilled addressee. Nevertheless for the purposes of clarity a number of terms will be defined.

As used herein, the term unsubstituted means that there is no substituent or that the only substituents are hydrogen.

The term "optionally substituted" as used throughout the specification denotes that the group may or may not be further substituted or fused (so as to form a polycyclic system), with one or more non-hydrogen substituent groups. In certain embodiments the substituent groups are one or more groups independently selected from the group consisting of halogen, =O, =S, -CN, -NO ₂ , -CF ₃ , -OCF ₃ , -OCHF ₂ , alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, heteroarylalkyl, arylalkyl, cycloalkylalkenyl, heterocycloalkylalkenyl, arylalkenyl, heteroarylalkenyl, cycloalkylheteroalkyl, heterocycloalkylheteroalkyl, arylheteroalkyl, heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxycycloalkyl, alkoxyheterocycloalkyl, alkoxyaryl, alkoxyheteroaryl, alkoxycarbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, phenoxy, benzyloxy, heteroaryloxy, arylalkyloxy, arylalkyl, heteroarylalkyl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, alkylsulfinyl, arylsulfinyl, aminosulfinylaminoalkyl, -COOH, - COR ¹¹ , -C(O)OR ¹¹ , CONHR ¹¹ , NHCOR ¹¹ , NHCOOR ¹¹ , NHCONHR ¹¹ , C(=N0H)R ¹¹ , -SH, -SR ¹¹ , -OR ¹¹ , and acyl, wherein R ¹¹ is H, optionally substituted d-C ₁₂ alkyl, optionally substituted C ₂ -C ₁₂ alkenyl, optionally substituted C ₂ -Ci ₂ alkynyl, optionally substituted C ₁ - C ₁₀ heteroalkyl, optionally substituted C ₃ -C ₁₂ cycloalkyl, optionally substituted C ₃ -C ₁₂ cycloalkenyl, optionally substituted C ₁ -C ₁₂ heterocycloalkyl, optionally substituted C ₁ -C ₁₂ heterocycloalkenyl, optionally substituted C ₆ -C ₁₈ aryl, optionally substituted C ₁ -C ₁₈ heteroaryl, and acyl.

"Alkyl" as a group or part of a group refers to a straight or branched aliphatic hydrocarbon group, such as a C ₁ -C ₁₄ alkyl, a C ₁ -C ₁₀ alkyl or a C ₁ -C ₆ unless otherwise noted. Examples of suitable straight and branched C ₁ -C ₆ alkyl substituents include methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, t-butyl, hexyl, and the like. The group may be a terminal group or a bridging group.

"Alkylamino" includes both mono-alkylamino and dialkylamino, unless specified. "Mono- alkylamino" means a -NH-Alkyl group, in which alkyl is as defined above. "Dialkylamino" means a -N(alkyl) ₂ group, in which each alkyl may be the same or different and are each as defined herein for alkyl. The alkyl group may be a C ₁ -C ₆ alkyl group. The group may be a terminal group or a bridging group.

"Arylamino" includes both mono-arylamino and di-arylamino unless specified.

Mono-arylamino means a group of formula arylNH-, in which aryl is as defined herein. Di-arylamino means a group of formula (aryl) ₂ N- where each aryl may be the same or different and are each as defined herein for aryl. The group may be a terminal group or a bridging group.

"Acyl" means an alkyl-CO- group in which the alkyl group is as described herein. Examples of acyl include acetyl and benzoyl. The alkyl group may be a C ₁ -C ₆ alkyl group. The group may be a terminal group or a bridging group.

"Alkenyl" as a group or part of a group denotes an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched such as a group having 2-14 carbon atoms, 2-12 carbon atoms, or 2-6 carbon atoms, in the normal chain. The group may contain a plurality of double bonds in the normal chain and the orientation about each is independently E or Z. Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl and nonenyl. The group may be a terminal group or a bridging group.

"Alkoxy" refers to an -O-alkyl group in which alkyl is defined herein. The alkoxy may be a C ₁ -C ₆ alkoxy. Examples include, but are not limited to, methoxy and ethoxy. The group may be a terminal group or a bridging group.

"Alkenyloxy" refers to an -O- alkenyl group in which alkenyl is as defined herein. Preferred alkenyloxy groups are C ₂ -C ₆ alkenyloxy groups. The group may be a terminal group or a bridging group.

"Alkynyloxy" refers to an -O-alkynyl group in which alkynyl is as defined herein. Preferred alkynyloxy groups are C ₂ -C ₆ alkynyloxy groups. The group may be a terminal group or a bridging group.

"Alkoxycarbonyl" refers to an -C(O)-O-alkyl group in which alkyl is as defined herein. The alkyl group may be a C ₁ -C ₆ alkyl group. Examples include, but not limited to, methoxycarbonyl and ethoxycarbonyl. The group may be a terminal group or a bridging group.

"Alkylsulfinyl" means a -S(O)-alkyl group in which alkyl is as defined above. The alkyl group is preferably a C ₁ -C ₆ alkyl group. Exemplary alkylsulfinyl groups include, but not limited to, methylsulfinyl and ethylsulfinyl. The group may be a terminal group or a bridging group. "Alkylsulfonyl" refers to a -S(O) ₂ -alkyl group in which alkyl is as defined above. The alkyl group may be a Ci-C ₆ alkyl group. Examples include, but not limited to methylsulfonyl and ethylsulfonyl. The group may be a terminal group or a bridging group.

"Alkynyl" as a group or part of a group means an aliphatic hydrocarbon group containing a carbon-carbon triple bond and which may be straight or branched and may have from 2-14 carbon atoms, 2-12 carbon atoms, or 2-6 carbon atoms in the normal chain. Exemplary structures include, but are not limited to, ethynyl and propynyl. The group may be a terminal group or a bridging group.

"Alkylaminocarbonyl" refers to an alkylamino-carbonyl group in which alkylamino is as defined above. The group may be a terminal group or a bridging group.

"Cycloaikyl" refers to a saturated or partially saturated, monocyclic or fused or spiro polycyclic, carbocycle that may contain from 3 to 9 carbons per ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like, unless otherwise specified. It includes monocyclic systems such as cyclopropyl and cyclohexyl, bicyclic systems such as decalin, and polycyclic systems such as adamantane. The group may be a terminal group or a bridging group.

"Cycloalkenyl" means a non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon double bond and may have from 5-10 carbon atoms per ring. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl or cycloheptenyl. The cycloalkenyl group may be substituted by one or more substituent groups. The group may be a terminal group or a bridging group.

The above discussion of alkyl and cycloaikyl substituents also applies to the alkyl portions of other substituents, such as without limitation, alkoxy, alkyl amines, alkyl ketones, arylalkyl, heteroarylalkyl, alkylsulfonyl and alkyl ester substituents and the like.

"Cycloalkylalkyl" means a cycloalkyl-alkyl- group in which the cycloaikyl and alkyl moieties are as previously described. Exemplary monocycloalkylalkyl groups include cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl and cycloheptylmethyl. The group may be a terminal group or a bridging group. "Halogen" represents fluorine, chlorine, bromine or iodine.

"Heterocycloalkyl" refers to a saturated or partially saturated monocyclic, bicyclic, or polycyclic ring containing at least one heteroatom selected from nitrogen, sulfur, oxygen. The heterocycloalkyl group may have from 1 to 3 heteroatoms in at least one ring. Each ring may be from 3 to 10 membered, such as 4 to 7 membered. Examples of suitable heterocycloalkyl substituents include pyrrolidyl, tetrahydrofuryl, tetrahydrothiofuranyl, piperidyl, piperazyl, tetrahydropyranyl, morphilino, 1 ,3-diazapane, 1 ,4-diazapane, 1 ,4- oxazepane, and 1 ,4-oxathiapane. The group may be a terminal group or a bridging group.

"Heterocycloalkenyl" refers to a heterocycloalkyl as described above but containing at least one double bond. The group may be a terminal group or a bridging group.

"Heterocycloalkylalkyl" refers to a heterocycloalkyl-alkyl group in which the heterocycloalkyl and alkyl moieties are as previously described. Exemplary heterocycloalkylalkyl groups include (2-tetrahydrofuryl)methyl, (2-tetrahydrothiofuranyl) methyl. The group may be a terminal group or a bridging group.

"Heteroalkyl" refers to a straight- or branched-chain alkyl group that may have from 2 to 14 carbons, such as 2 to 10 carbons in the chain, one or more of which has been replaced by a heteroatom selected from S, O, P and N. Exemplary heteroalkyls include alkyl ethers, secondary and tertiary alkyl amines, amides, alkyl sulfides, and the like. The group may be a terminal group or a bridging group. As used herein reference to the normal chain when used in the context of a bridging group refers to the direct chain of atoms linking the two terminal positions of the bridging group.

"Aryl" as a group or part of a group denotes (i) an optionally substituted monocyclic, or fused polycyclic, aromatic carbocycle (ring structure having ring atoms that are all carbon) that may have from 5 to 12 atoms per ring. Examples of aryl groups include phenyl, naphthyl, and the like; (ii) an optionally substituted partially saturated bicyclic aromatic carbocyclic moiety in which a phenyl and a C _5-7 cycloalkyl or C _5-7 cycloalkenyl group are fused together to form a cyclic structure, such as tetrahydronaphthyl, indenyl or indanyl. The group may be a terminal group or a bridging group. "Arylalkenyl" means an aryl-alkenyl- group in which the aryl and alkenyl are as previously described. Exemplary arylalkenyl groups include phenylallyl. The group may be a terminal group or a bridging group.

"Arylalkyl" means an aryl-alkyl- group in which the aryl and alkyl moieties are as previously described. Preferred arylalkyl groups contain a C _1-5 alkyl moiety. Exemplary arylalkyl groups include benzyl, phenethyl and naphthelenemethyl. The group may be a terminal group or a bridging group.

"Heteroaryl" either alone or as part of a group refers to groups containing an aromatic ring (such as a 5 or 6 membered aromatic ring) having one or more heteroatoms as ring atoms in the aromatic ring with the remainder of the ring atoms being carbon atoms. Suitable heteroatoms include nitrogen, oxygen and sulphur. Examples of heteroaryl include thiophene, benzothiophene, benzofuran, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, naphtho[2,3-b]thiophene, furan, isoindolizine, xantholene, phenoxatine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indole, isoindole, 1 H-indazole, purine, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxaline, cinnoline, carbazole, phenanthridine, acridine, phenazine, thiazole, isothiazole, phenothiazine, oxazole, isooxazole, furazane, phenoxazine, 2-, 3- or 4- pyridyl, 2-, 3-, 4-, 5-, or 8- quinolyl, 1-, 3-, 4-, or 5- isoquinolinyl 1-, 2-, or 3- indolyl, and 2-, or 3-thienyl. The group may be a terminal group or a bridging group.

"Heteroarylalkyl" means a heteroaryl-alkyl group in which the heteroaryl and alkyl moieties are as previously described. The heteroarylalkyl groups may contain a lower alkyl moiety. Exemplary heteroarylalkyl groups include pyridylmethyl. The group may be a terminal group or a bridging group.

"Lower alkyl" as a group means, unless otherwise specified, an aliphatic hydrocarbon group which may be straight or branched having 1 to 6 carbon atoms in the chain, for example 1 to 4 carbons such as methyl, ethyl, propyl (n-propyl or isopropyl) or butyl (n-butyl, isobutyl or tertiary-butyl). The group may be a terminal group or a bridging group.

As would be understood by the skilled person, throughout the synthesis of the compounds of Formula (I) it may be necessary to employ a protecting group on the amino group and/or on the carboxyl group in order to reversibly preserve a reactive amino or carboxyl functionality while reacting other functional groups on the compound. In such a case, the free amino group and/or the free carboxyl groups of the compounds of Formula (I) can be liberated either by deprotection of the amino group followed by deprotection of the acid moieties or vice versa.

Examples of suitable amino protecting groups that may be used include formyl, trityl, phthalimido, trichloroacetyl, chloroacetyl, bromoacetyl, iodoacetyl, and urethane-type blocking groups such as benzyloxycarbonyl ( ¹ CBz ¹ ), 4-phenylbenzyloxycarbonyl, 2- methylbenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl, 4- chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, 2,4- dichlorobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl, 3-bromobenzyloxycarbonyl, 4- nitrobenzyloxycarbonyl, 4cyanobenzyloxycarbonyl, t-butoxycarbonyl ('tBoc'), 2-(4-xenyl)- isopropoxycarbonyl, 1 ,1-diphenyleth-i-yloxycarbonyl, 1 ,1-diphenylprop-i-yloxycarbonyl, 2-phenylprop-2-yloxycarbonyl, 2-(p-toluyl)-prop-2-yloxycarbonyl, cyclopentanyloxy- carbonyl, i-methylcyclopentanyloxycarbonyl, cyclohexanyloxycarbonyl, 1- methylcyclohexanyloxycarbonyl, 2-methylcyclohexanyloxycarbonyl, 2-(4-toluylsulfono)- ethoxycarbonyl, 2-(methylsulfono)ethoxycarbonyl, 2-(triphenylphosphino)- ethoxycarbonyl, fluorenylmethoxycarbonyl ("FMOC"), 2-(trimethylsilyl)ethoxycarbonyl, allyloxycarbonyl, 1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl, 5- benzisoxalylmethoxycarbonyl, 4-acetoxybenzyloxycarbonyl, 2,2,2- trichloroethoxycarbonyl, 2-ethynyl-2-propoxycarbonyl, cyclopropylmethoxycarbonyl, 4- (decycloxy)benzyloxycarbonyl, isobornyloxycarbonyl, 1-piperidyloxycarbonlyl and the like; benzoylmethylsulfono group, 2-nitrophenylsulfenyl, diphenylphosphine oxide, and the like. The actual amino protecting group employed is not critical so long as the dehvatised amino group is stable to the condition of subsequent reaction(s) and can be selectively removed as required without substantially disrupting the remainder of the molecule including any other amino protecting group(s). Preferred amino-protecting groups are t-butoxycarbonyl (Boc), and benzyloxycarbonyl (Cbz). Further examples of these groups are found in: Greene, T. W. and Wuts, P. G. M., Protective Groups in Organic Synthesis, Second edition; Wiley-lnterscience: 1991; Chapter 7; McOmie, J. F. W. (ed.), Protective Groups in Organic Chemistry, Plenum Press, 1973; and Kocienski, P. J., Protecting Groups, Second Edition, Theime Medical Pub., 2000.

Examples of carboxyl protecting groups that may be used include methyl, ethyl, n-propyl, i-propyl, p-nitrobenzyl, p-methylbenzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4- dimethoxybenzyl, 2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl, 3,4- methylenedioxybenzyl, benzhydryl, 4,4'-dinnethoxybenzhyclryl, 2,2'4,4'- tetramethoxybenzhydryl, t-butyl, t-amyl, trityl, 4-methoxytrityl, 4 _I 4'-dimethoxytrityl, 4,4,'4"- trimethoxytrityl, 2-phenylprop-2-yl, trimethylsilyl, t-butyldimethylsilyl, phenacyl, 2,2,2- trichloroethyl, β-(di(n-butyl)methylsilyl)ethyl, p-toluenesulfonoethyl, A- nitrobenzylsulfonoethyl, allyl, cinnamyl, 1-(trimethylsilylmethyl)prop-1-en-3-yl, and the like. Preferred carboxyl protecting groups are methyl and t-butyl. Further examples of these groups are found in: Greene, T. W. and Wuts, P. G. M., Protective Groups in Organic Synthesis, Second edition; Wiley-lnterscience: 1991 ; McOmie, J. F. W. (ed.), Protective Groups in Organic Chemistry, Plenum Press, 1973; and Kocienski, P. J., Protecting Groups, Second Edition, Theime Medical Pub., 2000.

It is understood that included in the family of compounds of Formula (I) are isomeric forms including diastereoisomers, enantiomers, tautomers, and geometrical isomers in "E" or "Z" configurational isomer or a mixture of E and Z isomers. It is also understood that some isomeric forms such as diastereomers, enantiomers, and geometrical isomers can be separated by physical and/or chemical methods and by those skilled in the art.

Some of the compounds of the disclosed embodiments may exist as single stereoisomers, racemates, and/or mixtures of enantiomers and/or diastereomers. All such single stereoisomers, racemates and mixtures thereof, are intended to be within the scope of the subject matter described and claimed.

In the cyclopropane derivatives of the present invention, it is understood that the general formula and representative compounds refer to both cis and trans isomers as either single isomers or any mixture thereof and all isomers and mixtures thereof are intended to fall within the scope of the subject matter described and claimed. Where R ⁶ and R ⁷ are different, it will be understood that either the R or S configurations or any mixture thereof is included within the scope of the subject matter described and claimed.

Additionally, formulae (I), (II), (Ma), (III), (IV), (V) and (Vl) are intended to cover, where applicable, solvated as well as unsolvated forms of the compounds. Thus, each formula includes compounds having the indicated structure, including the hydrated as well as the non-hydrated forms.

In addition to compounds of the formulae (I), (II), (Ma), (III), (IV), (V) and (Vl), the compounds of the various embodiments include pharmaceutically acceptable salts, prodrugs, N-oxides and active metabolites of such compounds, and pharmaceutically acceptable salts of such metabolites.

The term "pharmaceutically acceptable salts" refers to salts that retain the desired biological activity of the above-identified compounds, and include pharmaceutically acceptable acid addition salts and base addition salts. Suitable pharmaceutically acceptable acid addition salts of compounds of Formula (I) may be prepared from an inorganic acid or from an organic acid. Examples of such inorganic acids are hydrochloric, sulfuric, and phosphoric acid. Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, heterocyclic carboxylic and sulfonic classes of organic acids, examples of which are formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, fumaric, maleic, alkyl sulfonic, arylsulfonic. Suitable pharmaceutically acceptable base addition salts of compounds of Formula (I) include metallic salts made from lithium, sodium, potassium, magnesium, calcium, aluminium, and zinc, and organic salts made from organic bases such as choline, diethanolamine, morpholine. Other examples of organic salts are: ammonium salts, quaternary salts such as tetramethylammonium salt; amino acid addition salts such as salts with glycine and arginine. Additional information on pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 19th Edition, Mack Publishing Co., Easton, PA 1995. In the case of agents that are solids, it is understood by those skilled in the art that the inventive compounds, agents and salts may exist in different crystalline or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulae.

"Prodrug" means a compound which is convertible in vivo by metabolic means (e.g. by hydrolysis, reduction or oxidation) to a compound of formula (I). For example an ester prodrug of a compound of formula (I) containing a hydroxyl group may be convertible by hydrolysis in vivo to the parent molecule. Suitable esters of compounds of formula (I) containing a hydroxyl group, are for example acetates, citrates, lactates, tartrates, malonates, oxalates, salicylates, propionates, succinates, fumarates, maleates, methylene-bis-β-hydroxynaphthoates, gestisates, isethionates, di-p-toluoyltartrates, methanesulphonates, ethanesulphonates, benzenesulphonates, p-toluenesulphonates, cyclohexylsulphamates and quinates. As another example an ester prodrug of a compound of formula (I) containing a carboxy group may be convertible by hydrolysis in vivo to the parent molecule. (Examples of ester prodrugs are those described by F.J. Leinweber, Drug Metab. Res., 18:379, 1987). The term "pharmaceutically acceptable" refers generally to a substance or composition that is compatible chemically and/or toxicologically with the other ingredients including a formulation, and/or the subject being treated.

The term "compounds of the present invention" (unless specifically identified otherwise) refers generally to compounds, prodrugs thereof, pharmaceutically acceptable salts of the compounds and/or prodrugs, and hydrates or solvates of the compounds, salts, and/or prodrugs, as well as all stereoisomers (including diastereoisomers and enantiomers), tautomers and isotopically labelled compounds. The compounds of the present invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms.

The term "derivative thereof when used in reference to compounds of the present invention refers generally to prodrugs, pharmaceutically acceptable salts of the compounds and/or prodrugs, and hydrates or solvates of the compounds, salts, and/or prodrugs.

The compounds of the present invention may be prepared using the reaction routes and synthesis schemes as described below, employing the techniques available in the art using starting materials that are commercially available or can be synthesised using known procedures or adaptations thereof. Whilst the preparation of particular compounds is outlined below, the skilled person will also recognize that the chemical reactions described may be readily adapted to prepare a number of other agents of the various embodiments. For example, the synthesis of non-exemplified compounds may be successfully performed by modifications apparent to those skilled in the art, e.g. by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by making routine modifications of reaction conditions. A list of suitable protecting groups in organic synthesis can be found in T. W. Greene's Protective Groups in Organic Synthesis, 3 ^rd Edition, John Wiley & Sons, 1991. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the various embodiments.

Reagents useful for synthesizing compounds may be obtained or prepared according to techniques known in the art. To produce compounds of the present invention in which T is a single bond the cinnamoyl benzamide (1) can be reduced by hydrogenation with a suitable catalyst, such as palladium on carbon, RhCI(PPh ₃ ) ₃ , or by any other methods known in the art (see J. March, Advanced Organic Chemistry, John Wiley & Sons, New York 1985, pp. 694).

The compounds of the invention and intermediates in their synthesis can be isolated from a reaction mixture using standard work-up and purification procedures. Suitable procedures include solvent extraction, chromatography (thin or thick layer chromatography, HPLC, flash chromatography, MPLC ₁ etc.), recrystallisation etc.

The present invention includes salts of the compounds of Formula (I). The salts may serve as intermediates in the purification of compounds or in the preparation of other, for example pharmaceutically acceptable, acid addition salts, or they may be useful for identification, characterisation or purification. The salts can exist in conjunction with the acidic or basic portion of the molecule and can exist as acid addition, primary, secondary, tertiary, or quaternary ammonium, alkali metal, or alkaline earth metal salts. Generally, acid addition salts are prepared by the reaction of an acid with a compound of Formula (I). The alkali metal and alkaline earth metal salts are generally prepared by the reaction of the hydroxide form of the desired metal salt with a compound of Formula (I).

Acid addition salts are preferably the pharmaceutically acceptable, non-toxic addition salts with suitable acids, such as those with inorganic acids, for example hydrochloric, hydrobromic, nitric, sulphuric or phosphoric acids, or with organic acids, such as organic carboxylic acids, for example, glycollic, maleic, hydroxymaleic, fumaric, malic, tartaric, citric, salicyclic, o-acetoxybenzoic, or organic sulphonic, 2-hydroxyethane sulphonic, toluene-p-sulphonic, or naphthalene-2-sulphonic acid.

The present invention also includes esters of the compounds of Formula (I), such esters being for example aliphatic esters such as alkyl esters. The esters of the compounds of Formula (I) may be pharmaceutically acceptable metabolically labile esters. These are ester derivatives of compounds of Formula (I) that are hydrolysed in vivo to afford the compound of Formula (I) and a pharmaceutically acceptable alcohol. Examples of metabolically labile esters include esters formed with alkanols in which the alkanol moiety may be optionally substituted by an alkoxy group, for example methanol, ethanol, propanol and methoxyethanol. The compounds of the various embodiments may be prepared using the reaction routes and synthesis schemes as described below, employing the techniques available in the art using starting materials that are readily available. The person skilled in the art will recognise that the chemical reactions described may be readily adapted to prepare a number of other compounds. For example, the synthesis of non-exemplified compounds may be successfully performed by modifications apparent to those skilled in the art, e.g. by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by making routine modifications of reaction conditions. A list of suitable protecting groups in organic synthesis can be found in T. W. Greene's Protective Groups in Organic Synthesis, 3 ^rd Edition, John Wiley & Sons, 1991. Reagents useful for synthesizing compounds may be obtained or prepared according to techniques known in the art.

The utility of compounds of Formula (I) can be tested using any of the following methods: (i) In a renal cell line by measuring proline incorporation after transforming growth factor-β stimulation;

(ii) Matrix synthesis may be stimulated by platelet derived growth factor (PDGF). Accordingly, mesangial cells incubated with PDGF can be used to demonstrate proline incorporation, which is an indicator of matrix synthesis and thereby a model for fibrosis; or

(iii) Matrix synthesis may be stimulated by both angiotensin Il or transforming growth factor beta (TGF-β). Accordingly, neonatal cardiac fibroblasts incubated with angiotensin Il or TGF-β can be used to demonstrate proline incorporation, which is an indicator of matrix synthesis and thereby a model for fibrosis.

Compounds of formula (II) may suitably be prepared according to scheme I;

Scheme 1

In a preferred form of the preparation according to Scheme 1 , the cyclopropanation step is performed with with excess CH ₂ N ₂ and a catalytic quantity of Pd(OAc) ₂ in a 1 :1 mixture of CH ₂ CI ₂ and diethyl ether. It is also preferred that an esterification step in which the cinnamic acid derivative is reacted with sulphuric acid in methanol to provide the corresponding methyl ester precedes the cyclopropanation step. Preferably, a hydrolysis step in which the cyclopropanated derivative is hydroiysed with aqueous NaOH/MeOH to provide the cyclopropanated carboxylic acid derivative follows the cyclopropanation step.

Thus according to one form of the invention, there is provided a method of preparing a compound of formula II, the method including;

(a) cyclopropanation of a compound of the formula

to obtain a compound of the formula and

(b) condensing the compound obtained in step (a) with a compound of formula

An exemplary preparation is shown below:

An alternative method for preparing a compound of formula Ma is shown below in Scheme 2.

Scheme 2

According to a further aspect of the invention, there is provided a method of preparing a compound of Formula Ma including the steps of: (a) reacting a compound of the formula

to obtain an acid chloride of the formula

and (b) condensing the acid chloride prepared in step (a) with a compound of the formula

H ₅ N An example of preparing a suitable compound of formula Ma is shown below:

A further method of preparing a compound of formula Ma is shown in scheme 3 below:

Scheme 3

Compounds of formula III may be prepared according to scheme 4 below;

Scheme 4

Thus, according to a further aspect of the invention, there is provided a method for preparing a compound of formula III including the steps of; (a) reacting a terminal alkene of the formula

to provide a sulfonyl chloride compound of the formula

and

(b) condensing the sulfonyl chloride prepared in step (a) with a compound of formula

An example of preparing a compound of formula III is as follows:

An alternative method for preparing a compound of formula III is shown below in Scheme 5.

Scheme 5

Thus, according to a further aspect of the invention, there is provided a method of preparing a compound of Formula III including the steps of: (a) reacting a sulfonate compound of the formula

to provide a sulfonyl chloride compound of the formula

(b) condensing the sulfonyl chloride compound prepared in step (a) with a compound of the formula

If desired, the method of preparing the compound of Formula III as described immediately above may also include an initial step of reacting an aldehyde compound of the formula ό to provide the sulfonate compound of the formula

Compounds of Formula IV in which X ² is NH may be prepared according to scheme 6 below.

Scheme 6

Thus according to a further aspect of the invention, there is provided a method for preparing a compound of formula IV in which X ² is NH that includes the steps of; (a) reacting a terminal alkene of the formula

to provide a compound of formula and (b) condensing the compound prepared in step (a) with a compound of the formula

A specific example for the preparation of a compound of formula IV is as follows:

Compounds of formula IV in which X ² is CH ₂ may be prepared by forming the °c,β- unsaturated ketone and then converting the ketone to the difluorovinyl species using morpholino-SF3 (See Tetrahedron Letters (2004), 45(7), 1527; Synthesis (1963), 787) according to scheme 7 below:

Scheme 7

Thus, according to a further aspect of the invention, there is provided a method for the preparation of compounds of formula IV in which X ² is CH ₂ that includes the steps of; (a) reacting a compound of the formula

to provide a ketone of the formula

(b) converting the ketone prepared in step (a) to a compound of the formula

A specific example of such a method is shown below:

Compounds of formula V in which X ² is NH may be made by reaction with Meldrum's acid followed by condensation of the malonic-type acid with a suitable aldehyde as shown in the following Scheme 8:

Meldrum's acid toluene

Scheme 8

A specific example of such a preparation is shown below:

piperidine toluene

Compounds of formula V may also be prepared by Knoevenagel condensation of a suitable aldehyde with malonic acid then coupling of the cinnamic acid with an aryl amine according to scheme 9 below.

malonic acid piperidine

Scheme 9

A specific example of such a synthesis is shown below: malonic acid piperidine

Another specific example of such a synthesis in which the 3,4-dimethoxycinnamic acid is converted to the corresponding acid chloride prior to reaction with a heterocyclic amine is shown below:

malonic acid piperidine oxalyl chloride CH ₂ CI ₂

Compounds of the formula Vl may be prepared by a method as shown in Schemes and 10 and 11 below:

Scheme 10

Scheme 11

Thus according to a further broad form of the invention there is provide a method of preparing a compound of formula (Vl) including the step of: reacting a compound of the formula

with a compound of the formula

to obtain a compound of the formula

A further form of the invention provides a method of preparing a compound of formula

(Vl) including the step of reacting a compound of formula

with I a compound of formula

to provide a compound of formula

Specific examples of such a synthesis are shown below:

Examples of materials and methods for use with the compounds of the present invention will now be provided. In providing these examples, it is to be understood that the specific nature of the following description is not to limit the generality of the above description. Examples

Experimental

Electrospray ionization (ESI) high resolution mass spectra (HRMS) were obtained on a Finnigan hybrid LTQ-FT mass spectrometer (Thermo Electron Corp.). Proton nuclear magnetic resonance ( ¹ H NMR) and proton decoupled carbon nuclear magnetic resonance ( ¹³ C NMR) spectra were obtained on Unity 400, Innova 400 or Innova 500 instruments (Melbourne, Australia) operating at 400 or 500 MHz for ¹ H and at 100 or 125 MHz for ¹³ C. All signals were referenced to solvent peaks (CDCI ₃ : 7.26 ppm for ¹ H and 77.0 ppm for ¹³ C; DMSO-cfe: 2.49 ppm for ¹ H and 39.5 ppm for ¹³ C). Infrared (IR) spectra were obtained using a PerkinElmer Spectrum One FT-IR spectrometer with zinc selenide/diamond Universal ATR Sampling Accessory. Melting points were obtained using a Reichert-Jung hot stage apparatus and are corrected. Analytical thin layer chromatography (TLC) was conducted on 2 mm thick silica gel GF ₂₅₄ . Compounds were visualised with solutions of 20% w/w phosphomolybdic acid in ethanol, 20% w/w potassium permanganate in water or under UV (365 nm). Flash chromatography was performed with Merck Silica Gel 60. Petrol refers to the fraction boiling at 40-60 ⁰ C. All other reagents were used as received.

Example 1 - Synthesis of compounds of Formula (I)

(E)-1-[[3-(3, 4-dimethoxyphenyl)-1-oxo-2-propenyl]amino]cyclohexanecarboxy lic acid (FT101)

A suspension of (E)-3-(3,4-dimethoxyphenyl)-2-propenoic acid (0.25 g, 1.2 mmol) in CH ₂ CI ₂ (5 mL) was treated with oxalyl chloride (0.41 mL, 4.8 mmol). The solution was stirred at rt for 1 h and the solvent was removed under reduced pressure to give the acid chloride as a yellow solid. 1-Aminocyclohexanecarboxylix acid (0.21 g, 1.4 mmol) was added to a solution of the acid chloride (1.2 mmol) in pyridine (2.0 mL) and the suspension was stirred at rt for 16 h. The solution was diluted with water and the precipitate was collected by filtration. The crude product was recrystallised from EtOAc providing (£)-1-[[3-(3,4-dimethoxyphenyl)-1-oxo-2-propenyl]amino]cycl ohexanecarboxylic acid (40 mg, 10%) as a colourless crystalline solid; mp 171-173 ⁰ C; δ _H (500 MHz, DMSO-cfe) 1.23 (m, 2H, CH ₂ ), 1.45-1.53 (m, 4H, CH ₂ ), 1.67 (m, 2H, CH ₂ ), 2.00 (m, 2H, CH ₂ ), 3.77 (S, 3H ₁ OCH ₃ ), 3.79 (s, 3H, OCH ₃ ), 6.67 (d, J = 15.5 Hz, 1 H, CH=CHCO), 6.98 (d, J ₅ , ₆ = 8.0 Hz, 1 H, H5), 7.09 (d, J ₅ , ₆ = 8.0 Hz, 1H, H6), 7.13 (s, 1H, H2), 7.28 (d, J = 15.5 Hz, 1 H ₁ CH=CHCO), 7.92 (s, 1 H, NH), 12.05 (s, 1 H, CO ₂ H); δ _c (125 MHz, DMSO- d ₆ ) 21.0, 25.0, 31.8, 55.3, 55.5, 57.8, 109.7, 111.7, 120.0, 121.4, 127.7, 138.6, 148.9, 150.1 , 164.8, 175.5; HRMS (ESI) calculated for C ₁₈ H ₂₃ NO ₅ [M+Na] ⁺ 351.1468, found 351.1467; v _max 1137, 1239, 1260, 1516, 1599, 1653, 1731, 2932, 3236, 3334 cm ^"1 .

(E)-[6-[[3-(3,4-Dimethoxyphenyl)-1-oxo-2-propenyl]amino]] -2-pyhdinecarboxylic acid (FT103)

A suspension of (E)-3-(3,4-dimethoxyphenyl)-2-propenoic acid (0.25 g, 1.2 mmol) in CH ₂ CI ₂ (5 mL) was treated with oxalyl chloride (0.41 ml_, 4.8 mmol). The solution was stirred at rt for 1 h and the solvent was removed under reduced pressure to give the acid chloride as a yellow solid. 3-Aminopicolinic acid (0.25 g, 1.4 mmol) was added to a solution of the acid chloride (1.2 mmol) in pyridine (2.0 mL) and the suspension was stirred at rt for 4 d. The solution was diluted with water and the precipitate was collected by filtration providing (£)-2-(3,4-dimethoxystyryl)-4H-pyrido[2,3-d][1 ,3]oxazin-4-one (80 mg, 20%) as a brown solid; mp 206-207 ⁰ C; δ _H (500 MHz, DMSO-Cl ₆ ) 3.80 (s, 3H, OCH ₃ ), 3.83 (S, 3H, OCH ₃ ), 6.84 (d, J = 15.5 Hz, 1 H, CH=CHCO), 6.99 (d, J _s - _tV = 8.0 Hz, 1H, H5 ¹ ), 7.25 (d, J ₅₇ ,. = 8.0 Hz, 1H, H6 ¹ ), 7.38 (s, 1H, H2"), 7.58 (d, J = 15.5 Hz, 1 H, CH=CHCO), 7.71 (dd, J ₃ , ₄ = 8.0, J _4,5 = 5.0 Hz, 1H, H4), 7.87 (d, J ₄ , ₅ = 5.0 Hz, 1 H, H5), 8.47 (d, J ₃ , ₄ = 8.0 Hz, 1H, H3), 11.49 (s, 1H, NH); δ _c (125 MHz, DMSO-(Z ₆ ) 55.5, 55.6, 110.4, 111.6, 119.2, 122.8, 127.1 , 127.9, 129.8, 134.0, 137.9, 141.5, 142.3, 149.0, 150.8, 164.8, 166.9; HRMS (ESI) calculated for C ₁₇ H ₁₆ N ₂ O ₅ [M+H] ⁺ 329.1132, found 329.1131 ; v _max 808, 1141 , 1259, 1508, 1667, 2938 cm ^"1 .

(E)-[6-[[3-(3,4-Dimethoxyphenyl)-1-oxo-2-propenyl]amino]] -3-pyhdinecarboxylic acid (FT104)

A suspension of (£)-3-(3,4-dimethoxyphenyl)-2-propenoic acid (1.0 g, 4.8 mmol) in CH ₂ CI ₂ (10 mL) was treated with oxalyl chloride (1.6 ml_, 19 mmol). The solution was stirred at rt for 1 h and the solvent was removed under reduced pressure to give the acid chloride as a yellow solid. 4-Aminonicotinic acid (0.73 g, 5.3 mmol) was added to a solution of the acid chloride (4.8 mmol) in pyridine (5.0 mL) and the suspension was heated to reflux and stirred for 5 d. The solution was diluted with water and the precipitate was collected by filtration providing (£)-[6-[[3-(3,4-Dimethoxyphenyl)-1-oxo-2- propenyl]amino]]-3-pyridinecarboxylic acid (0.25 g, 16%) as a colourless solid; mp 258- 260 ⁰ C; δ _H (500 MHz, DMSO-d ₆ ) 3.80 (s, 3H, OCH ₃ ), 3.83 (s, 3H, OCH ₃ ), 6.82 (d, J = 15.5 Hz, 1H ₁ CH=CHCO), 7.00 (d, J ₅ . _iβ . = 8.0 Hz, 1 H, H5'), 7.23 (dd, J ₆ . _>β . = 8.0, J ₂ , ₆ . = 2.0 Hz, 1 H ₁ H6'), 7.33 (s, J ₂ , ₆ . = 2.0 Hz, 1H, H2'), 7.62 (d, J = 15.5 Hz, 1H, CH=CHCO), 8.60 (d, J _5,6 = 8.0 Hz, 1 H, H5), 8.63 (d, J _5j6 = 8.0 Hz, 1H, H6), 9.03 (s, 1H, H2); HRMS (ESI) calculated for C ₁₇ H ₁₄ N ₂ O ₄ [M+H] ⁺ 329.1132, found 329.1129; v _max 1140, 1262, 1488, 1623, 2938, 3543 cm ^"1 .

2-(3, 4-dimethoxyphenyl)cyclopropanecarboxylic acid

Concentrated sulfuric acid (1.0 mL) was added to a solution of (£)-3,4- dimethoxycinnamic acid (2.0 g, 9.6 mmol) in MeOH (50 mL). The solution was heated to reflux for 4 h, cooled to rt and then quenched with saturated aqueous NaHCO ₃ . The aqueous phase was extracted with EtOAc and the combined organic fractions were washed with water, brine, dried and concentrated providing methyl ester (2.2 g, 95%). A cooled solution of the methyl ester (0.11 g, 0.49 mmol) and Pd(OAc) ₂ (6 mg, 0.027 mmol) in CH ₂ CI ₂ (3 mL) and ether (3 mL) was treated with excess CH ₂ N ₂ at 0 ⁰ C The suspension was filtered and the filtrate was concentrated. The crude residue was dissolved in MeOH (5 mL) and treated with 1 M NaOH (5 mL). The mixture was stirred at rt for 16 h and then concentrated under reduced pressure to remove the MeOH. The aqueous phase was washed with 50% EtOAc/petrol, acidified with 1 M HCI and then extracted with EtOAc. The combined organic fractions were washed with water, brine, dried and concentrated providing 2-(3,4-dimethoxyphenyl)cyclopropanecarboxylic acid (96 mg, 87%) as a colourless crystalline solid; δ _H (500 MHz, DMSO-cfe) 1.36 (ddd, J = 5.0, J = 4.0, J = 2.0 Hz, 1H, CH), 1.62 (ddd, J = 2.0, J = 4.0, J = 2.0 Hz, 1H, CH) ₁ 1.84 (ddd, J = 2.0, J = 4.0, J = 2.0 Hz, 1 H, CH), 2.57 (ddd, J = 2.0, J = 4.0, J = 3.0 Hz, 1 H, CH), 3.85 (s, 3H, OCH ₃ ), 3.87 (s, 3H, OCH ₃ ), 6.65 (d, J _5β = 8.0, 1H, H6), 6.66 (s, 1 H, H2), 6.78 (d, J _5i6 = 8.0 Hz, 1H, H5); δ _c (125 MHz, DMSO-Cf ₆ ) 7.1 , 23.6, 26.9, 55.8, 55.9, 110.1 , 111.3, 118.2, 131.9, 147.9, 148.9, 179.8.

2-(2-(3, 4-dimethoxyphenyl)cyclopropanecarboxamido)benzoic acid (FT111)

A suspension of 2-(3,4-dimethoxyphenyl)cyclopropanecarboxylic acid (96 mg, 0.41 mmol) in CH ₂ CI ₂ (1 mL) was treated with oxalyl chloride (0.14 ml_, 1.6 mmol). The solution was stirred at rt for 1 h and the solvent was removed under reduced pressure to give the acid chloride. Anthranilic acid (0.11 g, 0.82 mmol) was added to a solution of the acid chloride (0.41 mmol) in pyridine (2.0 mL) and the suspension was stirred at rt for 16 h. The solution was acidified with 1 M HCI and the solvent was decanted and discarded. The crude residue was dissolved in EtOAc and the organic phase was extracted into 1 M NaHCO ₃ and discarded. The aqueous phase was then acidified with 1 M HCI and extracted with EtOAc. The combined organic fractions were washed with water, brine, dried and concentrated providing 2-(2-(3,4- dimethoxyphenyl)cyclopropanecarboxamido)benzoic acid (65 mg, 46%) as a pale brown solid; mp 159-163 ⁰ C; δ _H (500 MHz, DMSO-cfe) 1.39 (m, 1H, CH), 1.48 (m, 1 H, CH), 1.94 (m, 1H, CH), 2.40 (m, 1 H, CH), 3.72 (s, 3H, OCH ₃ ), 3.76 (s, 3H, OCH ₃ ), 6.72 (d, J ₅₆ = 8.0, 1H, H6 ¹ ), 6.81 (s, 1H, H2"), 6.87 (d, J ₅ , ₆ = 8.0 Hz, 1H, H5 ¹ ), 7.13 (t, J ₃ , ₄ = J _4|5 = 7.9 Hz, 1 H, H4), 7.56 (t, J _4,5 = J _5,6 = 7.9 Hz, 1H, H5), 7.97 (d, J ₃₄ = 7.9 Hz, 1H, H3), 8.46 (d, J _S A = 7.9 Hz, 1H, H6), 11.58 (s, 1H, NH); δ _c (125 MHz, DMSO-Cf ₆ ) 15.9, 25.2, 27.5, 55.5, 55.6, 110.0, 110.3, 112.0, 117.8, 120.0, 122.5, 131.0, 132.8, 133.6, 140.6, 147.4, 148.8, 169.4, 170.1 ; HRMS (ESI) calculated for C ₁₉ H ₁₉ NO ₅ [M-H] ^" 340.1191 , found 340.1186; v _max 1023, 1140, 1255, 1516, 1671, 2836, 2921, 3326 cm ^"1 .

2-(3, 4-bis(difluoromethoxy)phenyl)cyclopropanecarboxylic acid

Concentrated sulfuric acid (0.25 mL) was added to a solution of (£)-3,4- dimethoxycinnamic acid (0.50 g, 1.8 mmol) in MeOH (15 mL). The solution was heated to reflux for 16 h, cooled to rt and then quenched with saturated aqueous NaHCO ₃ . The aqueous phase was extracted with EtOAc and the combined organic fractions were washed with water, brine, dried and concentrated providing methyl ester (0.51 g, 97%). A solution of the methyl ester (0.13 g, 0.44 mmol) and Pd(OAc) ₂ (5 mg, 0.022 mmol) in CH ₂ CI ₂ (3 ml.) and ether (3 mL) was treated with excess CH ₂ N ₂ . The suspension was filtered and the filtrate was concentrated. The crude residue was dissolved in MeOH (5 mL) and treated with 1 M NaOH (5 mL). The mixture was stirred at rt for 16 h and then concentrated under reduced pressure to remove the MeOH. The aqueous phase was washed with 50% EtOAc/petrol, acidified with 1 M HCI and then extracted with EtOAc. The combined organic fractions were washed with water, brine, dried and concentrated providing 2-(3,4-bis(difluoromethoxy)phenyl)cyclopropanecarboxylic acid (0.12 g, 92%) as a colourless oil; δ _H (500 MHz, DMSO-d ₆ ) 1.38 (ddd, J = 5.0, J = 4.0, J = 2.0 Hz, 1 H, CH), 1.69 (ddd, J = 2.0, J = 4.0, J = 2.0 Hz, 1 H, CH), 1.89 (ddd, J = 2.0, J = 4.0, J = 2.0 Hz, 1H, CH), 2.59 (ddd, J = 2.0, J = 4.0, J = 3.0 Hz, 1H, CH), 6.49 (t, J = 73 Hz, 1H, OCHF ₂ ), 6.51 (t, J = 73 Hz, 1 H, OCHF ₂ ), 6.97 (d, J _5β = 8.0, 1 H, H6), 7.01 (s, 1 H, H2), 7.18 (d, J _5β = 8.0 Hz, 1 H, H5); δ _c (125 MHz, DMSO-d ₆ ) 17.4, 24.0, 26.1 , 115.7 (t, J = 261 Hz), 115.7 (t, J = 261 Hz), 120.6, 122.6, 124.4, 138.7, 140.9, 142.4, 179.1.

2-(2-(3, 4-bis(difluoromethoxy)phenyl)cyclopropanecarboxamido)benzoic acid (FT112)

A suspension of 2-(3,4-dimethoxyphenyl)cyclopropanecarboxylic acid (0.12 g, 0.41 mmol) in CH ₂ CI ₂ (5 mL) was treated with oxalyl chloride (0.14 mL, 1.6 mmol). The solution was stirred at rt for 2 h and the solvent was removed under reduced pressure to give the acid chloride. Anthranilic acid (0.11 g, 82 mmol) was added to a solution of the acid chloride (0.41 mmol) in pyridine (2.0 mL) and the suspension was stirred at rt for 16 h. The solution was acidified with 1 M HCI and the resulting precipitate was collected by filtrated. The crude solid was triturated with 75% CH ₂ CI ₂ /petrol providing 2-(2-(3,4- bis(difluoromethoxy)phenyl)cyclopropanecarboxamido)benzoic acid (90 mg, 53%) as a colourless solid; mp 177-178 ⁰ C; δ _H (500 MHz, DMSO-Cf ₆ ) 1.43 (m, 1H, CH), 1.53 (m, 1 H, CH), 2.09 (m, 1 H, CH), 2.49 (m, 1H, CH), 6.98-7.35 (m, 6H ₁ H4, HZ, H5', H6', OCHF ₂ ), 7.57 (t, J ₄ ,5 = J ₅ ,6 = 7.9 Hz, 1 H, H5), 7.96 (d, J ₃ , ₄ = 7.9 Hz, 1 H, H3), 8.42 (d, J ₅ , ₆ = 7.9 Hz, 1 H, H6), 11.31 (s, 1H, NH); δ _c (125 MHz, DMSO-d ₆ ) 16.4, 24.5, 27.7, 116.4 (t, J = 258 Hz), 116.5 (t, J = 258 Hz), 116.9, 118.6, 120.3, 121.3, 122.8, 123.9, 131.0, 133.9, 139.5, 139.9, 140.5, 142.0, 169.4, 169.7; HRMS (ESI) calculated for C ₁₉ H ₁₅ F ₄ NO ₅ [M-H]- 412.0814, found 412.0808; v _max 1053, 1143, 1378, 1511 , 1537, 1661 , 3010 cm ^"1 . (E)-2-[3-(3, 4-Dimethoxyphenyl)-1-oxo-2-propenyl]benzoic acid (FT114)

A solution of 3,4-dimethoxybenzldehyde (0.37 g, 2.2 mmol) and 2-acetylbenzoic acid (0.33 g, 2.0 mmol) was dissolved in MeOH (10 mL) and treated with 50% aqueous KOH (1 mL). The mixture was heated to reflux for 16 h and then concentrated under reduced pressure to remove the MeOH. Water was added and the aqueous phase was acidified with 1 M HCI. The crude product was collected by filtration and recrystallised from EtOH providing (E)-2-[3-(3,4-dimethoxyphenyl)-1-oxo-2-propenyl]benzoic acid (0.45 g, 72%) as a yellow crystalline solid; mp 221-223 ⁰ C; δ _H (500 MHz, DMSO-d ₆ ) 3.79 (s, 3H, OCH ₃ ), 3.82 (s, 3H, OCH ₃ ), 6.96 (d, J ₅ , ₆ . = 8.0 Hz, 1 H, H5 ¹ ), 7.09 (d, J = 17.0 Hz, 1H, CH=CHCO), 7.13 (d, J = 17.0 Hz, 1 H, CH=CHCO), 7.20 (d, = 8.0 Hz, 1H, H6 ¹ ), 7.32 (s, 1 H, H2 ¹ ), 7.45 (d, J _3,4 = 8.0 Hz, 1H, H3), 7.60 (t, J ₄₅ = J _5β = 8.0 Hz, 1H, H5), 7.67 (t, •AM = As = 8.0 Hz, 1 H, H4), 7.90 (d, J _5β = 8.0 Hz, 1H, H6), 13.10 (br s, 1H, CO ₂ H); δ _c (125 MHz ₁ DMSO-cfc) 55.5, 55.6, 110.5, 111.5, 123.2, 125.1 , 127.0, 127.4, 129.6, 130.3, 131.8, 141.8, 144.6, 149.0, 151.1, 167.5, 195.5; HRMS (ESI) calculated for C ₁₈ H ₁₆ O ₅ [M+H] ⁺ 313.1071, found 313.1071 ; v _max 1138, 1251 , 1584, 1707, 2844, 2913 cm ^"1 .

(E)-2-[3-Methoxy-4-(difluoromethoxy)phenyl)-1-oxo-2-prope nyl]benzoic acid (FT115)

A solution of 3,4-bis(difluoromethoxy)benzaldehyde (0.26 g, 1.1 mmol) and 2- acetylbenzoic acid (0.16 g, 1.0 mmol) was dissolved in MeOH (10 mL) and treated with 50% aqueous KOH (1 mL). The mixture was heated to reflux for 16 h and then concentrated under reduced pressure to remove the MeOH. Water was added and the aqueous phase was acidified with 1 M HCI. The crude product was collected by filtration and recrystallised from CH ₂ CI ₂ /petrol providing (£)-2-[3-methoxy-4- (difluoromethoxy)phenyl)-1-oxo-2-propenyl]benzoic acid (40 mg, 11%) as a colourless solid; mp 161-168 . ⁰ C; δ _H (500 MHz, DMSO-Cf ₆ ) 3.90 (s, 3H, OCH ₃ ), 7.09-7.18 (m, 4H, OCHF ₂ , CH=CHCO, CH=CHCO, H5 ¹ ), 7.46 (d, J ₃₄ = 8.0 Hz, 1 H, H3), 7.56-7.62 (m, 3H, H5, H2\ H6'), 7.68 (t, J _3,4 = J _4,5 = 8.0 Hz, 1H, H4), 7.89 (d, J _5,6 = 8.0 Hz, 1H, H6), 13.10 (br s, 1H, CO ₂ H); δ _c (125 MHz, DMSO-d ₆ ) 56.1 , 113.3, 116.6 (t, J = 256 Hz), 120.1 , 126.1 , 127.2, 127.5, 127.8, 129.6, 129.8, 130.3, 130.7, 131.9, 141.6, 143.0, 152.4, 167.5, 195.3; HRMS (ESI) calculated for C ₁₈ H ₁₄ F ₂ O ₅ [M+H] ⁺ 349.0882, found 349.0882; v _max 1048, 1114, 1250, 1270, 1590, 1711, 2619, 2942 cm ^'1 .

(E)-2-[3,4-Bis(difluoromethoxy)phenyl)-1-oxo-2-propenyl]b enzoic acid (FT116)

A solution of 3,4-bis(difluoromethoxy)benzaldehyde (0.52 g, 2.2 mmol) and 2- acetylbenzoic acid (0.33 g, 2.0 mmol) was dissolved in MeOH (10 ml_) and treated with 50% aqueous KOH (1 ml_). The mixture was stirred at rt for 16 h and then concentrated under reduced pressure to remove the MeOH. Water was added and the aqueous phase was acidified with 25% AcOH. The crude product was collected by filtration and recrystallised from toluene/petrol providing (£)-2-[3,4-bis(difluoromethoxy)phenyl)-1-oxo- 2-propenyl]benzoic acid (0.31 g, 40%) as a colourless crystalline solid; mp 118-119 ⁰ C; 7.18 (d, J = 15.6 Hz, 1H, CH=CHCO), 7.23 (d, J = 15.6 Hz, 1H, CH=CHCO), 7.24 (t, J = 73 Hz, 1H, OCHF ₂ ), 7.26 (t, J = 73 Hz, 1 H, OCHF ₂ ), 7.35 (d, J ₅ ',* = 8.0 Hz, 1H, H5'), 7.45 (d, J _3,4 = 8.0 Hz, 1 H, H3), 7.61-7.71 (m, 3H, H4, H5, H6 ¹ ), 7.75 (s, 1H, H2"), 7.91 (d, J _5|6 = 8.0 Hz, 1H, H6), 13.10 (br s, 1 H, CO ₂ H); δ _c (125 MHz, DMSO-d ₆ ) 55.5, 55.6, 116.2 (t, J = 268 Hz), 116.4 (t, J = 268 Hz) ₁ 120.2, 120.6, 126.8, 127.5, 128.4, 129.7, 129.9, 130.2, 132.0, 132.6, 141.4, 141.6, 141.8, 143.0, 167.4, 195.5; HRMS (ESI) calculated for C ₁₈ H ₁₂ F ₄ O ₅ [M+H] ⁺ 385.0694, found 385.0694; v _max 1072, 1035, 1277, 1667, 1688, 2833, 3011 cm ^"1 .

(E)-2-[3-Methoxy-4-(propargyloxy)phenyl)-1-oxo-2-propenyl ]benzoic acid (FT117)

A solution of 3-methoxy-4-propargyloxybenzaldehyde (0.21 g, 1.1 mmol) and 2- acetylbenzoic acid (0.16 g, 1.0 mmol) was dissolved in MeOH (10 ml.) and treated with 50% aqueous KOH (1 mL). The mixture was stirred at rt for 16 h and then concentrated under reduced pressure to remove the MeOH. Water was added and the aqueous phase was acidified with 25% AcOH. The crude product was collected by filtration and recrystallised from toluene providing (E)-2-[3-Methoxy-4-(propargyloxy)phenyl)-1-oxo-2- propenyl]benzoic acid (0.17 g, 52%) as a colourless crystalline solid; mp 146-147 ⁰ C; δ _H (500 MHz, DMSO-Cf ₆ ) 3.67 (t, J = 2.4 Hz, 1 H, C≡CH), 3.90 (s, 3H, OCH ₃ ), 4.93 (d, J = 2.4 Hz, 2H, OCH ₂ ), 7.12 (d, J _{5 ,6} . = 8.0 Hz, 1 H, H5'),7.23 (s, 2H, CH=CHCO, CH=CHCO), 7.31 (dd, J _{5 , 6} . = 8.0 Hz, J _{2 , 6} . = 2.0 Hz), 7.45 (d, J ₂₆ . = 2.0 Hz, 1H, H2 ¹ ), 7.56 (d, J ₃ , ₄ = 8.0 Hz, 1H, H3), 7.70 (t, J ₄ , ₅ = J ₅ , ₆ = 8.0 Hz, 1H, H5), 7.78 (t, J ₃ , ₄ = J ₄ , ₅ = 8.0 Hz, 1 H, H4), 8.00 (d, J ₅₆ = 8.0 Hz ₁ 1H, H6), 13.12 (br s, 1 H, CO ₂ H); δ _c (125 MHz, DMSO-cfe) 55.7, 55.9, 78.5, 78.9, 111.0, 113.5, 122.6, 125.6, 127.4, 128.0, 129.6, 129.7, 130.3, 131.9, 141.7, 144.3, 148.7, 149.3, 167.5, 195.5; HRMS (ESI) calculated for C ₂₀ H ₁₆ O ₅ [M+H] ⁺ 337.1070, found 337.1070; v _max 1141, 1253, 1507, 1585, 1701, 2960, 3290 cm ^"1 .

(E)-2-[4-(3, 4-Dimethoxyphenyl)-2-oxobut-3-enyl]benzoic acid (FT118)

A solution of 3,4-dimethoxybenzldehyde (0.18 g, 1.1 mmol) and 2-(2-oxopropyl)benzoic acid (0.18 g, 2.0 mmol) was dissolved in MeOH (10 mL) and treated with 50% aqueous KOH (1 mL). The mixture was stirred at rt for 16 h and then concentrated under reduced, diluted with water and the solution was was acidified with 25% AcOH. The crude product was collected by filtration and recrystallised from toluene providing (E)-2-[4-(3,4- dimethoxyphenyl)-2-oxobut-3-enyl]benzoic (54 mg, 17%) as a yellow crystalline solid; mp 152-154 ⁰ C; δ _H (500 MHz, DMSO-cfe) 3.80 (s, 3H, OCH ₃ ), 3.81 (s, 3H, OCH ₃ ), 4.41 (s, 2H, CH ₂ ), 6.88 (d, J = 16.0 Hz, 1H, CH=CHCO), 7.00 (d, J ₅ , ₆ . = 8.0 Hz, 1 H, H5 ¹ ), 7.25- 7.30 (m, 2H, H3, H6 ¹ ), 7.35 (s, 1 H ₁ H2'), 7.37 (t, J ₄ , ₅ = J ₅ , ₆ = 8.0 Hz, 1 H ₁ H5), 7.51 (t, J ₃ , ₄ = J ₄₅ = 8.0 Hz ₁ 1 H ₁ H4), 7.51 (d, J = 16.0 Hz, 1H ₁ CH=CHCO), 7.90 (d, J _5|6 = 8.0 Hz, 1 H, H6), 12.78 (br s, 1 H, CO ₂ H); δ _c (125 MHz, DMSO-Cf ₆ ) 46.1, 55.5, 55.6, 110.4, 111.6, 123.0, 124.4, 126.7, 127.2, 130.3, 130.7, 131.7, 132.5, 137.0, 142.0, 149.0, 150.9, 168.2, 196.5; HRMS (ESI) calculated for C ₁₉ H ₁₈ O ₅ [M+H] ⁺ 327.1227, found 327.1227; v _max 1140, 1260, 1510, 1687, 2835, 2933 cm ^"1 .

(E)-Ethyl 2-(3, 4-bis(difluoromethoxy)phenyl)ethenesulfonate

A solution of 1.6 M nBuLi in hexane (1.6 mL, 2.5 mmol) was added to a cooled solution of ethyl diethylphosphorylmethanesulfonate (0.68 g, 2.6 mmol) in THF (25 mL) at -78 ⁰ C.

The solution was stirred at -78 ⁰ C for 10 mins, warmed to at 0 ⁰ C ₁ stirred for 10 mins and then cooled back to -78 ⁰ C. A solution of 3,4-bis(difluoromethoxy)benzaldehyde (0.50 g, 2.1 mmol) in THF (5.0 mL) was added to the cooled mixture. The reaction was stirred at - 78 ⁰ C for 1 h, warmed to 0 ⁰ C and stirred for 16 h. The reaction was quenched with water and the aqueous phase was extracted with EtOAc. The combined organic fractions were washed with water, brine, dried and concentrated. The crude residue was purified by column chromatography, eluting with 20% EtOAc/petrol, to afford (E)-ethyl 2-(3,4- bis(difluoromethoxy)phenyl)ethenesulfonate (0.69 g, 94%) as a colourless oil; δ _H (500 MHz, CDCI ₃ ) 1.41 (t, J = 7.0 Hz, 3H, CH ₃ ), 4.41 (q, J = 7.0 Hz, 2H, CH ₂ ), 6.56 (t, J = 73 Hz, 1H, OCHF ₂ ), 6.58 (t, J = 73 Hz, 1 H, OCHF ₂ ), 6.73 (d, J = 16.0 Hz, 1H, CH=CHCO), 7.32 (d, J _{5 , 6} . = 8.0 Hz, 1H, H5'), 7.39 (d, J ₅ ; _& = 8.0 Hz, 1H, H6 ¹ ), 7.42 (s, 1H, H2'), 7.54 (d, J = 16.0 Hz, 1H, CH=CHCO); δ _c (125 MHz, CDCI ₃ ) 14.9, 67.1 , 115.4 (t, J = 255 Hz), 115.5 (t, J = 255 Hz), 122.2, 122.4, 123.1 , 126.9, 130.6, 142.0, 142.4, 144.5.

(E)-2-[3, 4-Bis(difluoromethoxy)phenyl]-N-phenylethenesulfonamide (FT119)

A solution of (E)-ethyl 2-(3,4-bis(difluoromethoxy)phenyl)ethenesulfonate (100 mg, 0.29 mmol) and tetrabutylammonium iodide (160 mg, 0.44 mmol) in acetone (5.0 mL) was heated to reflux for 16 h. The solution was concentrated under reduced pressure and dissolved in CH ₂ CI ₂ (5.0 mL) and triphenylphosphine (300 mg, 1.2 mmol) was added. The solution was cooled to 0 ⁰ C and sulfuryl chloride (47 μL, 0.58 mmol) was added to the reaction mixture. The solution was allowed to warm to rt and stirred for 16 h. The solvent was removed under reduced pressure and the crude chloride was purified by column chromatography, eluting with 5-20% EtOAc/petrol, providing the sulfonyl chloride (90 mg, 90%) as a colourless oil. A solution of the sulfonyl chloride (90 mg, 0.26 mmol) in CH ₂ CI ₂ (3.0 mL) was added to a cooled solution of aniline (52 μL, 0.58 mmol) in pyridine (2.0 mL) at 0 ⁰ C. The reaction mixture was warmed to rt, stirred for 4 h and acidified with 1 M HCI. The aqueous phase was extracted with CH ₂ CI ₂ and the combined organic fractions were washed with water, brine, dried and concentrated to provide (£)-2-[3,4- bis(difluoromethoxy)phenyl]-N-phenylethenesulfonamide (55 mg, 54%) as a dark purple coloured oil; (δ _H (500 MHz, CDCI ₃ ) 6.55 (t, J = 73 Hz, 1 H, OCHF ₂ ), 6.57 (t, J = 73 Hz, 1 H, OCHF ₂ ), 6.82 (d, J = 16.0 Hz, 1 H, CH=CHCO), 6.93 (s, 1H, NH), 7.20 (t, J _3|4 = J _A,S = 8.0 Hz, 1H, H4), 7.28-7.36(m, 7H, H2, H3, H5, H6, H2'H5\ H6'), 7.43 (d, J = 16.0 Hz, 1 H, CH=CHCO); δ _c (125 MHz, CDCI ₃ ) 115.4 (t, J = 255 Hz), 115.5 (t, J = 255 Hz), 121.1 , 122.0, 122.3, 125.5, 126.0, 126.7, 129.5, 130.9, 136.1 , 140.5, 142.3, 144.1 ; HRMS (ESI) calculated for C ₁₆ H ₁₃ F ₄ NO ₄ S [M+H] ⁺ 392.0574 found 392.0575; v _max 1043, 1136, 1274, 1497, 1599, 3057, 3261 crrT ¹ .

(E)-Ethyl 2-(3, 4-dimethoxyphenyl)ethenesulfonate

A solution of 1.6 M nBuLi in hexane (1.4 ml_, 2.2 mmol) was added to a cooled solution of ethyl diethylphosphorylmethanesulfonate (0.58 g, 2.2 mmol) in THF (25 mL) at -78 ⁰ C. The solution was stirred at -78 ⁰ C for 10 mins, warmed to at 0 ⁰ C, stirred for 10 mins and then cooled back to -78 ⁰ C. A solution of 3,4-dimethoxybenzaldehyde (0.30 g, 1.8 mmol) in THF (5.0 mL) was added to the cooled mixture. The reaction was stirred at -78 ⁰ C for 1 h, warmed to 0 ⁰ C and stirred for 16 h. The reaction was quenched with water and the aqueous phase was extracted with EtOAc. The combined organic fractions were washed with water, brine, dried and concentrated. The crude residue was purified by column chromatography, eluting with 20% EtOAc/petrol, to afford (£)-ethyl 2-(3,4- dimethoxyphenyl)ethenesulfonate (0.47 g, 96%) as a colourless crystalline solid; δ _H (500 MHz, CDCI ₃ ) 1.40 (t, J = 7.0 Hz, 3H, CH ₃ ), 3.92 (s, 3H, OCH ₃ ), 3.93 (a, 3H, OCH ₃ ), 4.22 (q, J = 7.0 Hz, 2H, CH ₂ ), 6.59 (d, J = 16.0 Hz, 1 H, CH=CHCO), 6.90 (d, J ₅ - _? = 8.0 Hz, 1H, H5'), 7.00 (s, 1H, H2'), 7.11 (d, J ₅ - _β - = 8.0 Hz, 1 H, H6'), 7.53 (d, J = 16.0 Hz, 1 H, CH=CHCO); δ _c (125 MHz, CDCI ₃ ) 14.9, 55.9, 56.0, 66.6, 109.9, 111.1, 118.5, 123.3, 124.8, 144.6, 149.4, 152.0.

(E)-2-(2-(3, 4-Dimethoxyphenyl)vinylsulfonamido)benzoic acid (FT120)

A solution of (£)-ethyl 2-(3,4-dimethoxyphenyl)ethenesulfonate (200 mg, 0.73 mmol) and tetrabutylammonium iodide (320 mg, 1.1 mmol) in acetone (10 mL) was heated to reflux for 16 h. The solution was concentrated under reduced pressure and the sodium salt was dissolved in CH ₂ CI ₂ (5.0 mL) and added to a cooled solution of sulfuryl chloride (210 μL, 2.6 mmol) and triphenylphosphine (710 mg, 3.1 mmol) in CH ₂ CI ₂ (5.0 mL) at 0 ⁰ C. The solution was allowed to warm to rt and stirred for 16 h. The solvent was removed under reduced pressure and the crude chloride was purified by column chromatography, eluting with 20% EtOAc/petrol, providing the sulfonyl chloride (150 mg, 78%) as a yellow oil. A solution of the sulfonyl chloride (150 mg, 0.57 mmol) in CH ₂ CI ₂ (2.0 mL) was added to a cooled solution of anthranilic acid (160 mg, 1.14 mmol) in pyridine (2.0 mL) at 0 ⁰ C. The reaction mixture was warmed to rt, stirred for 16 h and diluted with water. The aqueous phase was ashed with 50% EtOAc/petrol and the organic phase was discarded. The aqueous phase was acidified with 1 M HCI and extracted with CH ₂ CI ₂ . The combined organic fractions were washed with water, brine, dried and concentrated. The crude product was recrystallised from CH ₂ CI ₂ to provide (£)-2-(2-(3,4- dimethoxyphenyl)vinylsulfonamido)benzoic acid (30 mg, 14%) as a colourless crystalline solid; mp 194-196 ⁰ C; δ _H (400 MHz, CDCI ₃ ) 3.88 (s, 3H, OCH ₃ ), 3.90 (s, 3H, OCH ₃ ), 6.70 (d, J = 16.0 Hz, 1 H, CH=CHCO), 6.84 (d, J ₅ ^ = 8.0 Hz, 1 H, H5'), 6.94 (s, 1H, H2'), 7.05 (d, J ₅ -V = 8.0 Hz, 1 H, H6 ¹ ), 7.11 (t, J ₃ , ₄ = J ₄ , ₅ = 8.0 Hz, 1 H, H4), 7.57 (t, J ₄ ,s = J _Sfi = 8.0 Hz, 1H, H5), 7.58 (d, J = 16.0 Hz, 1H, CH=CHCO), 7.72 (d, J ₃₄ = 8.0 Hz, 1 H, H3), 8.10 (d, J _5,6 = 8.0 Hz, 1H, H6), 10.36 (s, 1H, NH); δ _c (100 MHz, CDCI ₃ ) 55.9, 56.0, 109.9, 111.1, 114.0, 117.9, 121.9, 122.6, 123.3, 124.9, 132.3, 135.7, 141.2, 143.5, 149.3, 151.9, 171.5; HRMS (ESI) calculated for C ₁₇ H ₁₇ NO ₆ S [M-H] ^" 362.0704, found 362.0699; v _max 1136, 1267, 1513, 1682, 2975, 3277 cm ^"1 .

Proline Incorporation

A well-characterized cloned rat mesangial cell line [30] (gift of D Nikolic-Patterson) is cultured in DMEM with FBS, 100U/ml_ penicillin, and 100ug/mL streptomycin in a humidified 5% CO ₂ atmosphere at 37°C. Cells are plated into 24-well culture dishes in DMEM/10%FBS at low density and allowed to adhere overnight. Cells are used between passages 20 and 40. The subconfluent cells are starved overnight in DMEM/0.1%FBS containing 15OuM L-ascorbic acid, prior to 4 hours of pre-treatment with or without tranilast or the FT compounds, followed by the addition of 5ng/mL ThTGF-P ₁ I (R&D Systems) and 1uCi/mL of L-(2,3,4,5- ³ H)-proline. Control wells have the compounds but no TGF-P ₁ added. Cells are incubated for a further 44 hours during which time their appearance is visually monitored. The cells are then washed three times in ice-cold PBS, twice in ice cold 10% TCA and solubilized in 75OuL 1M NaOH for 45 minutes at 37 ⁰ C or overnight at 4 ⁰ C. A 50OuL aliquot is neutralized with 50OuL 1M HCI and 1OmL scintillation fluid (Instagel Plus - Perkin-Elmer) added. Counts are performed on a beta counter.

To normalize the praline incorporation counts to take into account the proliferative effects of TGF-βi, a BioRad protein assay is performed on a 100-15OuL aliquot of the remaining solubilized cells. The sample is neutralized with an equal amount of 1 M HCI prior to the assay. The BSA standards used to construct the standard curve have the same amount of 1 M NaOH and 1 M HCI added as is present in the samples for assay.

Proline incorporation is expressed as cpm/ug protein. In order to compare inter-assay results, the incorporation is expressed as percentage reduction of TGF stimulated proline incorporation, where TGF alone gives 0% reduction and the zero control gives 100% reduction.

Mesangial cells

Compounds in bold have minimal effect on cell appearance and viability N. B suppressed MTT result indicates reduced cell viability

Ppt - compound precipitated during the assay.

The details of specific embodiments described in this invention are not to be construed as limitations. Various equivalents and modifications may be made without departing from the essence and scope of this invention, and it is understood that such equivalent embodiments are part of this invention.