WO2008090327A1 - New combination for use in the treatment of cancer - Google Patents

Abstract

There is provided combination products comprising: (a) a compound of formula (I), wherein X, Y, T, W, A1, A2 R1, R5 and R6 have meanings given in the description, and (b) trastuzumab or a tyrosine kinase inhibitor. Combination products according to the invention find particular utility in the treatment of cancer.

Description

NEW COMBINATION FOR USE IN THE TREATMENT OF CANCER

Field of the Invention

This invention relates to a novel pharmaceutical combination.

Background and Prior Art

Elevated plasma free fatty acids (FFAs) stimulate pancreatic β-cells and is one cause of hyperinsulinemia.

Excess adiposity is associated to different degrees with an increased risk of developing cancers, such as colorectal adenomas, breast cancer (postmenopausal), endometrial cancer, kidney cancer, oesophageal adenocarcinoma, ovarian cancer, prostate cancer, pancreatic cancer, gallbladder cancer, liver cancer and cervical cancer (CaIIe and Kaaks (2004), Nature Reviews Cancer, 4, 579-591 ).

Recent studies suggest that hyperinsulinemia is correlated among other things to the incidence of colon and lethal breast and prostate cancer.

In prostate cancer, hyperinsulinemia has been shown to be prospective risk factor for death and data support that the insulin level could be used as a marker of prostate cancer prognosis (Hammarsten and Hδgstedt (2005) European Journal of Cancer, 41 , 2887).

Several mechanisms may link hyperinsulinemia to the incidence and outcome of breast cancer. Firstly, chronic hyperinsulinemia results in increased production of ovarian testosterone and oestrogen and inhibition of hepatic production of sex hormone binding globulin, a sex-hormonal profile that is associated with breast cancer. Secondly, hyperinsulinemia suppresses hepatic production of insulin-like growth factor binding protein-1 (IGFBP-1 ), and thus increases circulating levels of IGF-1 , which has potent mitogenic effect on breast tissue. Thirdly, insulin itself may have a direct mitogenic effect on breast cancer cells. The study by Hardy et al ((2005), J. Biol. Chem. 280, 13285) shows that FFAs directly stimulate the growth of breast cancer cells in a GPR40 dependent manner. Moreover, expression studies performed on tumor tissue isolated from 120 breast cancer patient shows a frequent expression of GPR40 emphasizing the clinical relevance of the findings of Hardy (see, for example, Ma et al, Cancer Cell (2004) 6, 445).

Another expression study on clinical material from colon cancer patients suggests that similar mechanisms could be relevant also in these malignancies (see http://www.ncbi.nlm. nih.gov/projects/geo/gds/gds_browse.cgi?gds=1263).

Cancer cells in general exhibit an aberrant metabolism compared to non- transformed cells. Neoplastic cells synthesise lipids to a much larger extent than their normal counterparts and metabolise glucose differently. It has been suggested that this aberrant metabolism constitutes a therapeutic target. By interfering with one or, preferably, several of the pathways controlling cellular metabolism, cancer cells would be more sensitive than non-transformed cells, thus creating a therapeutic window. Examples of pathways/targets include glycolysis interfering agents, lipid synthesis pathway, AMPK activating agents and agents affecting mitochondrial function.

The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

US 1293741 discloses inter alia thiazolidinones. However, there is no mention of the use of the compounds disclosed therein in the treatment of cancer.

US 4,103,018 and US 4,665,083 disclose inter alia thiazolidinones. However, there is no mention or suggestion of the compounds disclosed in those documents in the treatment of cancer.

WO 2005/051890 discloses inter alia thiazolidinones (which are ultimately substituted with a cyclopropyl group) that may be useful in the treatment of diabetes. However, there is no mention or suggestion in this document of the use of the compounds in the treatment of cancer, nor of thiazolidinones that are substituted in the 5-position with heterocyclyl, heteroaryl or, particularly, aryl group, either directly or via an alkylene linker group.

EP 1 535 915 discloses various furan and thiophene-based compounds. Cancer is mentioned as one of numerous indications.

EP 1 559 422 discloses a huge range of compounds for use in the treatment of inter alia cancer. However, this document does not appear to relate to thiazolidinones.

US patent application US 2006/0089351 discloses various benzothiazole derivatives as neuropeptide Y receptor antagonists, and therefore of use in the treatment of eating disorders. International patent application WO 2006/020680 discloses a vast range of heterocyclic compounds as modulators of nuclear receptors. Further, US 6,353,006 discloses various heterocycles for use in modulating progesterone receptor mediated processes, and therefore of use in the treatment of e.g. diseases relating to deficiencies in bone mineral density, such as osteopenia or osteoporosis.

International patent applications VVO 2005/075471 , WO 2005/116002 and WO 2007/061661 disclose inter alia thiazolidinones and oxazolidinones as 1 1-β- hydroxysteroid dehydrogenase type 1 inhibitors. There is no mention or suggestion of the use of the disclosed compounds for the treatment of cancer, nor a teaching towards such thiazolidinones that are each substituted at the 5- position with a heterocyclyl, heteroaryl or, particularly, aryl group, either directly or via an alkylene linker group.

International patent application WO 2006/040050 discloses certain quinazolinylmethylene thiazolinones as CDK1 inhibitors. Similarly, US patent application US 2006/0004045 discloses quinolinylmethylene thiazolinones. US patent application US 2005/0038098 discloses arylmethylene thiazolidindiones and WO 2005/082363 discloses arylmethylene thiazolones. Compounds that are structurally more unrelated to those described herein are also disclosed in US patent applications US 2005/0250784, US 2002/0165259 and international patent application WO 01/72721.

International patent applications WO 2007/032028 discloses various heterocyclic compounds that may be useful in treating PTP-1B mediated diseases.

Finally, international patent application WO 2007/010273 discloses certain e.g. thiazolidin-4-one compounds that are able to antagonize the stimulatory effect of FFAs on cell proliferation when tested in an assay using a human breast cancer cell line (MDA-MB-231 ). Such compounds are this indicated in the treatment of cancer.

HER2-positive breast cancer is a more aggressive disease with a greater likelihood of recurrence, a poorer prognosis, and a decreased chance of survival compared with HER2 -negative breast cancer.

Herceptin (trastuzumab) is the first humanized monoclonal antibody approved for the treatment of HER2-positive metastatic breast cancer. It is designed to target and block the function of HER2 protein overexpression, and acts by attaching itself to the HER2 protein, thereby blocking the latter's interaction between human epidermal growth factor.

Herceptin also works by attracting the body's own immune cells to help destroy the cancer cells.

The drug, which is typically administered by infusion, is only effective in patients with high levels of the HER2 protein. Presently, only about one in five women with breast cancer (and fewer men) have tumours that are sensitive.

Herceptin may increase effectiveness of certain chemotherapy drugs, particularly paclitaxel (Taxol®) and docetaxel (Taxotere®) and may improve survival.

Herceptin is licensed in many countries as a treatment for patients with early stage breast cancer. In the UK, clinical guidance states that Herceptin should be considered as a possible treatment for women with HER2 positive breast cancer following surgery and adjuvant chemotherapy (and radiotherapy if appropriate).

In certain circumstances, the drug may also be used to treat patients with advanced breast cancer in combination with or without the chemotherapy drugs mentioned above. It may also be given on its own to patients with advanced breast cancer who have already received at least two courses of chemotherapy.

Tyrosine kinases (TKs) are enzymes that catalyse the phosphorylation of tyrosine residues. TKs are involved in cellular signalling pathways and regulate key cell functions such as proliferation, differentiation, anti-apoptotic signalling and neurite outgrowth.

Unregulated activation of TKs, through mechanisms such as point mutations or over-expression, can lead to various forms of cancer as well as benign proliferative conditions.

TK inhibitors (TKIs), such as lapatanib (Tykerb®), have been developed to block the ATP-binding site and prohibit the autophosphorylation of tyrosine residues, thereby preventing activation of the intracellular signalling pathways in tumor cells.

TKIs may stabilize tumor progression, creating in many instances a disease state this is no longer life threatening. Furthermore, side effects are considerably reduced when compared to conventional chemotherapeutic agents, such as those mentioned hereinbefore, and clear synergistic effects have been observed when TKIs are combined with such agents and/or radiotherapy.

There is no disclosure or suggestion in international patent application WO 2007/010273 that the compounds described therein may be used in combination with Herceptin (trastuzumab) or TKIs.

Disclosure of the Invention

According to the invention there is provided a combination product comprising: (a) a compound of formula I,

wherein X represents -[C(R₈)(R₈)],,-; n represents 0, 1 , 2 or 3;

Y represents -C(O)-, -S(O)₂- or =C(R₁₀)-;

T represents -S- or -O-;

W represents -NR₇-, -CR₇R₇-, -NR₇C(O)-, -NR₇S(O)₂-, -NR₇C(O)NR₇-, -NR₇C(O)O- or a bond; one of A₁ or A₂ represents a double bond and the other represents a single bond; when A₁ represents a single bond, A₂ is a double bond and R₆ is absent; when A₂ represents a single bond, A₁ is a double bond and, if present, one R₇

(which is attached α to the requisite ring of the compound of formula I) is absent; R₁ represents -C(O)NR₃R₂, -NR₃R₂, -C(O)OR₂, -NR₄C(O)NR₃R₂, -NR₄C(O)OR₂,

-OC(O)NR₃R₂, -NR₄C(O)R₂, -OC(O)R₂ , -OR₂, -SR₂, H, alky!, cycloalkyl, heterocyclyl, benzyl, aryl or heteroaryl (which latter six groups are optionally substituted by one or more groups selected from B¹, B², B³, B⁴, B⁵ and B⁶, respectively); R₂ and R₅ independently represent, on each occasion when used herein, hydrogen, alkyl, cycloalkyl, heterocyclyl, benzyl, aryl or heteroaryl (which latter six groups are optionally substituted by one or more groups selected from B⁷, B⁸, B⁹,

B¹⁰, B¹¹ and B¹², respectively);

R₃, R₄, R₆ and R₇ independently represent, on each occasion when used herein, hydrogen, alkyl, cycloalkyl, aryl or benzyl (which latter four groups are optionally substituted by one or more groups selected from B¹³, B¹⁴, B¹⁵ and B¹⁶, respectively), or heterocyclyl or heteroaryl (which latter two groups are optionally substituted by one or more groups selected from B¹⁴ and B¹⁵, respectively);

Rs and Rg are independently selected from hydrogen, alkyl and aryl (which latter two groups are optionally substituted by B^16a and B^16b, respectively);

Rio represents hydrogen, alkyl or aryl (which latter two groups are optionally substituted by one or more groups selected from B¹⁷ and B¹⁸, respectively); B¹ to B¹⁸ independently represent cyano, -NO₂, halo, -OR₁₁, -NR₁₂Ri₃, -SR₁₄,

-Si(R₁₅)₃, -C(O)ORi₆, -C(O)NRieaRi6b, -S(O)₂NR_16cRi₆d, aryl or heteroaryl (which aryl and heteroaryl groups are themselves optionally and independently substituted by one or more groups selected from halo and Ri₇); or, alternatively, B⁴, B⁵, B⁶, B¹⁰, B¹¹, B¹², B¹⁵, B¹⁶, B^16b or B¹⁸ independently represent R₁₇;

Rii, Ri2, Ri3, Ri₄. Ri_6> Ri6a, Ri6b, Rise and Ried independently represent H or Ri₇; and

Ri₅ and Ri₇ independently represent, on each occasion when used herein, C_1-6 alkyl optionally substituted by one or more halo atoms, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof, provided that, when n represents O and R₁ represents an optionally substituted alkyl group, then that alkyl group is saturated; and

(b) a compound selected from trastuzumab, or a tyrosine kinase inhibitor, or a pharmaceutically-acceptable salt, solvate or pharmaceutically functional derivative of a tyrosine kinase inhibitor, which combination products are referred to hereinafter as "the combination products according to the invention".

The skilled person will appreciate that the terms "trastuzumab" and "Herceptin" are employed interchangeably in the context of the present invention.

The term "tyrosine kinase inhibitor" (hereinafter "TKI") will be understood by the skilled person to include any compound that blocks (or is capable of blocking), to a measurable degree, the autophosphorylation of tyrosine residues, thereby preventing activation of the intracellular signalling pathways in tumor cells, including those mentioned hereinbefore. Preferred TKIs include inhibitors of the HER-family of TKs, such as HER-1 /Human Epidermal Growth Factor (EGFR; erbB1 ), HER3 (erbB3), HER4 (erbB4) and, more particularly, HER2 (erbB2). Preferred TKIs thus include imatinib, gefitinib, erlotinib, canertinib, sunitinib, zactima, vatalanib, sorafenib, leflunomide and, particularly, lapatinib.

Combination products of the invention that may be mentioned include those that comprise trastuzumab. Pharmaceutically-acceptable salts of compounds of formula I, or TKIs, that may be mentioned include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example by reaction of a free acid or a free base form of e.g. a compound of formula I with one or more equivalents of an appropriate acid or base, optionally in a solvent, or in a medium in which the salt is insoluble, followed by removal of said solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter-ion of e.g. a compound of formula I in the form of a salt with another counter-ion, for example using a suitable ion exchange resin.

Examples of pharmaceutically acceptable addition salts include those derived from mineral acids, such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric and sulphuric acids, and organic acids, such as tartaric, acetic, citric, malic, lactic, fumaric, benzoic, glycolic, gluconic, succinic, alkylsulphonic and arylsulphonic acids. Preferred salts of e.g. imatinib include mesylate salts.

"Pharmaceutically functional derivatives" of compounds of formula I as defined herein, and TKIs, includes ester derivatives and/or derivatives that have, or provide for, the same biological function and/or activity as any relevant compound. Thus, for the purposes of this invention, the term also includes prodrugs of compounds of formula I and TKIs.

The term "prodrug" of a relevant compound includes any compound that, following oral or parenteral administration, is metabolised in vivo to form that compound in an experimentally-detectable amount, and within a predetermined time (e.g. within a dosing interval of between 6 and 24 hours (i.e. once to four times daily)). For the avoidance of doubt, the term "parenteral" administration includes all forms of administration other than oral administration.

Prodrugs of compounds of formula I may be prepared by modifying functional groups present on the compound in such a way that the modifications are cleaved, in vivo when such prodrug is administered to a mammalian subject. The modifications typically are achieved by synthesizing the parent compound with a prodrug substituent. Prodrugs include compounds of formula I wherein a hydroxyl, amino, sulfhydryl, carboxy or carbonyl group in a compound of formula I is bonded to any group that may be cleaved in vivo to regenerate the free hydroxy!, amino, sulfhydryl, carboxy, or carbonyl, group, respectively.

Examples of prodrugs include, but are not limited to, esters and carbamates of hydroxy functional groups, esters groups of carboxyl functional groups, N-acyl derivatives and N-Mannich bases. General information on prodrugs may be found e.g. in Bundegaard, H. "Design of Prodrugs" p. 1-92, Elesevier, New York-Oxford (1985).

Compounds of formula I, as well as pharmaceutically-acceptable salts, solvates and pharmaceutically functional derivatives of such compounds are, for the sake of brevity, hereinafter referred to together as the "compounds of formula I".

Compounds of formula I may contain double bonds and may thus exist as E (entgegen) and Z {zusammen) geometric isomers about each individual double bond. All such isomers and mixtures thereof are included within the scope of the invention.

Compounds of formula I may exist as regioisomers and may also exhibit tautomerism. All tautomeric forms and mixtures thereof are included within the scope of the invention. Specifically, tautomers exist when R⁶ represents H. Such compounds have different point of attachments of R⁶ accompanied by one or more double bond shifts.

Compounds of formula I may also contain one or more asymmetric carbon atoms and may therefore exhibit optica! and/or diastereoisomerism. Diastereoisomers may be separated using conventional techniques, e.g. chromatography or fractional crystallisation. The various stereoisomers may be isolated by separation of a racemic or other mixture of the compounds using conventional, e.g. fractional crystallisation or HPLC, techniques. Alternatively the desired optical isomers may be made by reaction of the appropriate optically active starting materials under conditions which will not cause racemisation or epimerisation (i.e. a 'chiral pool' method), by reaction of the appropriate starting material with a 'chiral auxiliary' which can subsequently be removed at a suitable stage, by derivatisation (i.e. a resolution, including a dynamic resolution), for example with a homochiral acid followed by separation of the diastereomeric derivatives by conventional means such as chromatography, or by reaction with an appropriate chiral reagent or chiral catalyst all under conditions known to the skilled person. All stereoisomers and mixtures thereof are included within the scope of the invention.

Unless otherwise stated, the term "alkyl" refers to an unbranched or branched, cyclic, saturated or unsaturated (so forming, for example, an alkenyl or alkynyl) hydrocarbyl radical, which may be substituted or unsubstituted (with, for example,

B¹, B², B⁷, B⁸, B¹³, B¹⁴, B^16a or B¹⁷). Where the term "alkyl" refers to an acyclic group, it is preferably C_1-I0 alkyl and, more preferably, C₁^ alkyl (such as ethyl, propyl, (e.g. n-propyl or isopropyl), butyl (e.g. branched or unbranched butyl), pentyl or, more preferably, methyl). Where the term "alkyl" is a cyclic group

(which may be where the group "cycloalkyl" is specified), it is preferably C_3--I₂ cycloalkyl and, more preferably, C₅.₁₀ (e.g. C₅.7) cycloalkyl.

When used herein, alkylene refers to C_1-10 (e.g. C_1-6) alkylene and, preferably C_1-3 alkylene, such as pentylene, butylene (branched or unbranched), preferably, propylene (π-propylene or isopropylene), ethylene or, more preferably, methylene (i.e. -CH₂-).

The term "halogen", when used herein, includes fluorine, chlorine, bromine and iodine.

Heterocyclyl groups that may be mentioned include non-aromatic monocyclic heterocyclyl groups in which one or more (e.g. one to four) of the atoms in the ring system is other than carbon (i.e. a heteroatom, which heteroatom is preferably selected from N, O and S), and in which the total number of atoms in the ring system is between three and twelve (e.g. between five and ten). Further, such heterocycloalkyl groups may be saturated or unsaturated containing one or more double and/or triple bonds, forming for example a C_2-q heterocycloalkenyl (where q is the upper limit of the range) or a C₃._q heterocycloalkynyl group. C_2-q heterocycloalkyl groups that may be mentioned include 7- azabicyclo[2.2.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 6-azabicyclo[3.2.1]- octanyl, δ-azabicyclo[3.2.1]octanyl, aziridinyl, azetidinyl, dihydropyranyl, dihydropyridyl, dihydropyrrotyl (including 2,5-dihydropyrrolyl), dioxolanyl (including 1 ,3-dioxolanyl), dioxanyl (including 1 ,3-dioxanyl and 1 ,4-dioxanyl), dithianyl (including 1 ,4-dithianyl), dithiolanyl (including 1 ,3-dithiolanyl), imidazolidinyl, imidazolinyl, morpholinyl, 7-oxabicyclo[2.2.1]heptanyl, 6- oxabicyclo[3.2.1]octanyl, oxetanyl, oxiranyl, piperazinyl, piperidinyl, pyranyl, pyrazolidinyl, pyrrolidinonyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, sulfolanyl, 3- sulfolenyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahyd ropy rid yl (such as 1 ,2,3,4-tetrahydropyridyl and 1,2,3,6-tetrahydropyridyl), thietanyl, thiiranyl, thiolanyl, thiomorpholinyl, trithianyl (including 1 ,3,5-trithianyl), tropanyl and the like. Substituents on heterocycloalkyl groups may, where appropriate, be located on any atom in the ring system including a heteroatom. The point of attachment of heterocycloalkyl groups may be via any atom in the ring system including (where appropriate) a heteroatom (such as a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heterocycloalkyl groups may also be in the N- or S- oxidised form. Preferred heterocycly! groups include cyclic amino groups such as pyrrolidinyl, piperidyl, piperazinyl, morpholinyl or a cyclic ether such as tetrahydrofuranyl, monosaccharide.

The term "aryl" when used herein includes C_6-i₄ (such as C_6-i3 (e.g. C_6-10)) aryl groups. Such groups may be monocyclic, bicyclic or tricyclic and have between 6 and 14 ring carbon atoms, in which at least one ring is aromatic. The point of attachment of aryl groups may be via any atom of the ring system. However, when aryl groups are bicyclic or tricyclic, they are linked to the rest of the molecule via an aromatic ring. C_δ-u aryl groups include phenyl, naphthyl and the like, such as 1 ,2,3,4-tetrahydronaphthyl, indanyl, indenyl and fluorenyl. Most preferred aryl groups include phenyl.

The term "heteroaryl" when used herein refers to an aromatic group containing one or more heteroatom(s) (e.g. one to four heteroatoms) preferably selected from N, O and S (so forming, for example, a mono-, bi-, or tricyclic heteroaromatic group). Heteroaryl groups include those which have between 5 and 14 (e.g. 10) members and may be monocyclic, bicyclic or tricyclic, provided that at least one of the rings is aromatic. However, when heteroaryl groups are bicyclic or tricyclic, they are linked to the rest of the molecule via an aromatic ring. Heterocyclic groups that may be mentioned include benzothiadiazolyl (including 2,1 ,3- benzothiadiazolyl), isothiochromanyl and, more preferably, acridinyl, benzimidazolyl, benzodioxanyl, benzodioxepinyl, benzodioxolyl (including 1 ,3- benzodioxolyl), benzofuranyl, benzofurazanyl, benzothiazolyl, benzoxadiazolyl (including 2,1 ,3-benzoxadiazolyl), benzoxazinyl (including 3,4-dihydro-2H-1 ,4- benzoxazinyl), benzoxazolyl, benzomorpholinyl, benzoselenadiazolyl (including 2,1 ,3-benzoselenadiazolyl), benzothienyl, carbazolyl, chromanyl, cinnolinyl, furanyl, imidazolyl, imidazo[1 ,2-a]pyridyl, indazolyl, indolinyl, indolyl, isobenzofuranyl, isochromanyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiaziolyl, isoxazolyl, naphthyridinyl (including 1 ,6-naphthyridinyl or, preferably, 1,5-naphthyridinyl and 1,8-naphthyridinyl), oxadiazolyl (including 1 ,2,3- oxadiazolyl, 1 ,2,4-oxadiazolyl and 1 ,3,4-oxadiazolyl), oxazolyl, phenazinyl, phenothiazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinolizinyl, quinoxalinyl, tetrahydroisoquinolinyl (including 1 ,2,3,4-tetrahydroisoquinolinyl and 5,6,7,8-tetrahydroisoquinolinyl), tetrahydroquinolinyl (including 1 ,2,3,4- tetrahydroquinolinyl and 5,6,7,8-tetrahydroquinolinyl), tetrazolyl, thiadiazolyl (including 1 ,2,3-thiadiazolyl, 1 ,2,4-thiadiazolyl and 1 ,3,4-thiadiazolyl), thiazolyl, thiochromanyl, thiophenetyl, thienyl, triazolyl (including 1 ,2,3-triazolyl, 1 ,2,4-triazolyl and 1 ,3,4-triazolyl) and the like. Substituents on heteroaryl groups may, where appropriate, be located on any atom in the ring system including a heteroatom. The point of attachment of heteroaryl groups may be via any atom in the ring system including (where appropriate) a heteroatom (such as a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heteroaryl groups may also be in the N- or S- oxidised form. Particularly preferred heteroaryl groups include pyridyl, pyrrolyl, quinolinyl, furanyl, thienyl, oxadiazolyl, thiadiazolyl, thiazolyl, oxazolyl, pyrazolyl, triazolyl, tetrazolyl, isoxazolyl, isothiazolyl, imidazolyl, pyrimidinyl, indolyl, pyrazinyl, indazolyl, pyrimidinyl, thiophenetyl, pyranyl, carbazolyl, acridinyl, quinolinyl, benzoimidazolyl, benzthiazolyl, purinyl, cinnolinyl and pterdinyl.

For the avoidance of doubt, in cases in which the identity of two or more substituents in a compound of formula I may be the same, the actual identities of the respective substituents are not in any way interdependent. For example, in the situation in which R¹ and R² are both aryl groups substituted by one or more C_1-6 alkyl groups, the alkyl groups in question may be the same or different.

For the avoidance of doubt, when a term such as "S* to B¹⁸" is employed herein, this will be understood by the skilled person to mean B¹, B², B³, B⁴, B⁵, B⁶, B⁷, B⁸, B⁹, B¹⁰, B¹¹, B¹², B¹³, B¹⁴, B¹⁵, B¹⁶, B^16a, B^16b, B¹⁷ and B¹⁸ inclusively.

For the avoidance of doubt, when the group 'benzyl' is substituted, then the substituents are preferably on the phenyl ring of the benzyl group, rather than on the methylene (-CH₂-) group.

For the avoidance of doubt, in compounds of formula I, when Y represents =C(R¹⁰)-, this refers to the following compound of formula Ia

Compounds of formula I that may be mentioned include those in which:

Y preferably represents -C(O)-; R₁ represents -C(O)NR₃R₂, -NR₃R₂, -C(O)OR₂, -NR₄C(O)NR₃R₂, -NR₄C(O)OR₂,

-OC(O)NR₃R₂, -NR₄C(O)R₂, -OC(O)R₂ , -OR₂, -SR₂, H, alkyl, haloalkyl cycloalkyl, heterocyclyl, benzyl, aryl or heteroaryl;

R₂ and R₅ independently represent, on each occasion when used herein, hydrogen, alkyl, haloalkyl, cycloalkyl, heterocyclyl, benzyl, aryl or heteroaryl; R₃, R₄, R₆ and R₇ independently represent, on each occasion when used herein, aryl or, more particularly, hydrogen, alkyl, haloalkyl, cycloalkyl or benzyl;

R₈ and Rg are independently selected from hydrogen, alkyl and aryl;

R₁₀ represents hydrogen, alkyl, haloalkyl or aryl.

Further compounds of formula I that may be mentioned include those in which: B¹ to B¹⁸ independently represent halo, -ORn, -NR₁₂R13, -SR₁₄, -Si(Ri₅)_3l -C(O)OR₁₆ or aryl (which aryl group is itself optionally substituted by one or more groups selected from halo or R₁₇, or is preferably unsubstituted); Rn, R₁₂, R₁S₁ Ru and R₁₆ independently represent R₁₇ or, more preferably, H.

B¹ to B¹⁸ may alternatively independently represent functional groups such as hydroxyl, amine, sulfide, silyl, carboxylic acid, halogen, aryl, etc.

Further compounds of formula I that may be mentioned include those in which: Y represents -C(O)-;

T represents -S-; n represents 1 ;

W represents -N-;

A₂ represents a single bond and A-i is a double bond; and/or RQ represents H; Ri and R₅ independently represent aryl or heteroaryl.

Further compounds of formula I that may be mentioned include those in which:

X is alkylene or a bond (i.e. when n represents O);

T represents -S-; Y represents =C(H)- or, preferably -C(O)-;

W represents -NR₇-;

A₁, A₂, Ri, R₂ and R₅ are as hereinbefore defined; and/or

R₃, R₄ and R₆ independently represent hydrogen, alkyl (e.g. optionally substituted by one or more groups selected from B¹³), haloalkyl, cycloalky! (e.g. optionally substituted by one or more groups selected from B¹⁴) or benzyl (e.g. optionally substituted by one or more groups selected from B¹⁶).

More preferred compounds of formula I include those in which:

X represents -CH₂-; Y represents -C(O)-;

R₁ and R₂ independently represent aryl (e.g. phenyl) as hereinbefore defined (i.e.

R₁ represents aryl optionally substituted by one or more B⁵ groups and R₂ represents aryl optionally substituted by one or more B¹¹ groups); when Ri and/or R₂ represent phenyl, it/they is/are substituted para relative to the point of attachment of the Ri or R₂ group to X; B⁵ and B¹¹ independently represent halo; and/or R₅ represents heteroaryl (e.g. pyridyl).

More preferred compounds of formula I include those in which: R₁ represents -C(O)NHR₂; R₂ represents aryl (e.g. phenyl); when R₂ represents phenyl, it is substituted (i.e. with a B¹¹ substituent) at the para position (relative to the point of attachment of the R₂ group to the remainder of the compound of formula I); and/or B¹¹ represents d-C₆ alkyl.

In another preferred embodiment of the present invention:

R₁ is -NHR₂;

R₂ is aryl (e.g. phenyl); when R₂ represents phenyl, it is substituted (i.e. with a B¹¹ substituent) at the para position;

B¹¹ represents C₁-C₆ alkyl;

Y represents =C(H)-;

R₅ represents aryl (e.g. phenyl); and/or when R₅ represents phenyl, it is either unsubstituted or substituted with a halogen

(i.e. B¹¹ represents halo).

In a still another preferred embodiment of the present invention: R₆ represents aryl (e.g. phenyl); when R₅ represents phenyl, it is substituted (i.e. with a B¹¹ substituent) at the para position;

B¹¹ represents R₁7; and/or

Ri₇ represents C₁^ alkyl preferably substituted by one or more halo atoms (so forming a haloalkyl group).

In a still another preferred embodiment of the present invention; Y represents =C(H)-; R₅ represents aryl (e.g. phenyl); when R₅ represents phenyl, it is substituted (i.e. with a B¹¹ substituent) at the para position; B¹¹ represents halo or R₁7; and/or

Rn represents d.₆ alkyl preferably substituted by one or more halo atoms (so forming a haloalkyl group).

In a still another preferred embodiment of the present invention: X represents a single bond (i.e. n represents 0); R₁ is -C(O)NHR₂; R₂ is aryl (e.g. phenyl); when R₂ represents phenyl, it is substituted with B¹¹; B¹¹ represents R₁7; and/or Ru represents C_rC₆ alkyl.

Preferred compounds of formula I include those in which:

T represents -S-; Y represents =C(R₁₀)-, preferably, -S(O)₂- or, more preferably,

-C(O)-;

R₁0 represents H or, more preferably, alkyl (e.g. methyl or trifluoromethyl);

W represents -CR₇R₇-, a bond, or, more preferably,

-NR₇-, -NR₇C(O)-, -NR₇C(O)O-, -NR₇C(O)NR₇- Or -NR₇S(O)₂-; R₅ represents optionally substituted (i.e. by B⁷) alkyl (such as Ci_-3 alkyl, e.g. propylene or, preferably, isopropyl or methyl; so forming, for example, a benzyl group), cycloalkyl (e.g. cyclohexyl) or, more preferably represents optionally substituted (i.e. by B¹¹) aryl (e.g. phenyl) or optionally substituted (i.e. by B¹²) heteroaryl (e.g. 2-pyridyl); n represents 3 or O or, more preferably, 1 or 2;

R₈ and R₉ independently represent C_1-3 (e.g. Ci_-2) alkyl (e.g. methyl) or, more preferably, H;

R₁ represents (e.g. when n represents 1 ) alkyl or, more preferably -NR₃R₂, -OR₂,

-SR₂, -NR₄C(O)R₂, -NR₄C(O)NR₃R₂, -NR₄C(O)OR₂, particularly -C(O)NR₃R₂, -C(O)OR₂, more particularly, optionally substituted (i.e. by B⁶) heteroaryl (e.g. furanyl, such as furan-2-yl or thienyl, such as thien-2-yl) or, especially, optionally substituted (i.e. by B⁵) aryl (e.g. phenyl);

R₄ represents C₁.3 (e.g. C_1-2) alkyl (e.g. methyl) or H;

R₃ represents Ci_-3 (e.g. C_1-2) alkyl (e.g. methyl) or, preferably, H; R₂ represents optionally substituted (i.e. by B⁷) alky! (such as C_1-3 alkyl, e.g. ethyl or, preferably, methyl; so forming, for example, a benzyl group) or, preferably, optionally substituted (i.e. by B¹¹) aryl (e.g. phenyl) or (e.g. when Ri represents

-C(O)OR₂) H; when W represents -NR₇- and R₇ is absent, then R₆ represents alkyl such as

C_1-6 (e.g. C_1-3) alkyl (e.g. methyl) or aryl (e.g. phenyl), both of which may be substituted by one or more of B¹³ or B¹⁵, respectively, or are more preferably unsubstituted, or, more preferably R₆ represents H; when W represents -NR₇- and R₆ is absent, then R₇ represents C_1-3 (e.g. C_1-2) alkyl (e.g. methyl), aryl (e.g. phenyl) or benzyl, all of which may be substituted by one or more B¹³, B¹⁵ and B¹⁶, respectively, or, are more preferably unsubstituted; when W represents -CR₇R₇-, then A₂ represents a double bond; when W represents -CR₇R₇-, then each R₇ independently represents, at each occurrence, C_1-3 (e.g. Ci_-2) alkyl or H; B¹ to B¹⁸ (and, in particular, B⁵, B⁶, B¹¹ and B¹²) independently represent cyano,

NO₂, halo (e.g. chloro, fluoro or bromo), -OR₁₁, -C(O)OR₁₆, -C(O)NR_16aRi₆b or

-S(O)₂NR_16cR_16d; and/or

B⁴ to B⁶, B¹⁰ to B¹², B¹⁵, B¹⁶ and B¹⁸ (and, in particular, B⁵, B¹¹ and B¹²) represents

R₁₇; and/or B¹ to B¹⁸ (and, in particular, B¹ and B⁷) independently represent heteroaryl (e.g. furaπyl, such as furan-2-yl or thienyl, such as thien-2-yl) or, preferably, aryl (e.g. phenyl), both of which may be substituted by one or more groups selected from halo (e.g. fluoro) or Ri₇;

Rn represents Ci_-3 (e.g. Ci-₂) alkyl (e.g. methyl or ethyl) or H; R₁₆ represents H or C₁.₃ (e.g. Ci_-2) alkyl (e.g. ethyl);

Ri6_a, Ri6_b, Rise and R_16d independently represent C_1-2 alkyl or, more preferably, H;

Ri₇ represents Ci_-4 (e.g. C₁.₃) alkyl (e.g. methyl or isopropyl) optionally substituted by one or more halo (e.g. fluoro) atoms (so forming, for example, a trifluoromethyl group).

Other preferred compounds of the invention that may be mentioned include:

W represents -NR₇-, -NR₇C(O)- or -NR₇S(O)₂-;

Ri represents phenyl optionally substituted by B⁵; R₅ represents phenyl optionally substituted by B¹¹; R₆ and R₇ (if present) independently represent hydrogen; B⁵ represents halo or, preferably, Ri₇; B¹¹ represents R₁₇ or, preferably, halo (e.g. chloro);

Ri₇ represents C₁.₃ alkyl (e.g. methyl) optionally substituted by one or more halo (e.g. fluoro) atoms (so forming, for example, a trifluoromethyl group); when R₁ represent an optionally substituted phenyl group, then it is preferably a trifluoromethylphenyl group (e.g. 3-trifluoromethylphenyl); when R₅ represents an optionally substituted phenyl group, then it is preferably a halophenyl group (i.e. one in which the phenyl group is substituted with one or more halo substituents), such as monochlorophenyl (e.g. 2-chlorophenyl, 3- chlorophenyl or, preferably, 4-chlorophenyl) or dichlorophenyl (e.g. 3,4- dichlorophenyl).

It preferred that:

Rio does not represent H; when Y represents

W does not represent -N(R₇)C(O)-; n represents 2, 3 or, more preferably, 1;

R₃, R₄, Re and R₇ independently represent, on each occasion when used herein, hydrogen, alkyl, cycloalkyl, aryl or benzyl (which latter four groups are optionally substituted by one or more groups selected from B¹³, B¹⁴, B¹⁵ and B¹⁶, respectively);

R₁ does not represent H or alkyl as hereinbefore defined;

R₅ does not represent H.

Preferred compounds of formula I include those in which: when X does not represent a single bond (i.e. n does not represent 0), then R₁ does not represent -NR₃R₂, -OR₂, -SR₃, -NR₄C(O)R₂, -NR₄C(O)NR₃R₂ or

-NR₄C(O)OR₂; when X represents -CH₂-, R₁ represents optionally substituted aryl, W represents

-NR₇-, then: (i) R₅ does not represent alkyl or cycloalkyl; or

(W) R₅ does not represent hydrogen; when X represents a single bond (i.e. n represents O) and R₅ represents optionally substituted aryl, then Ri does not represent an optionally substituted alkyl group or hydrogen; when X represents -CH₂- and R₅ represents optionally substituted aryl, then R₁ does not represent -C(O)NR3R2,' when X represents -CH₂- and R₅ represents optionally substituted alkyl or aryl, then R₁ does not represent -C(O)NR₃R₂.

It is further preferred that: R₆ and Rg both represent H; R₆ represents H.

More preferred compounds of formula I include those of the examples described hereinafter and, in particular:

5-(4-fluorobenzyl)-2-(pyridin-2-ylimino)thiazolidin-4-one;

5-(p-methylben2yl)-2-(4-chlorophenylimino)thia2olidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(p-tolylimino)thiazolidin-4-one; 5-(3-(trifluoromethyl)ben2yl)-2-(4-chlorophenylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(4-isopropylphenylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(4-methoxyphenylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(phenylimino)thiazolidin-4-one;

2-(3,4-dichlorophenylimino)-5-(3-(trifluoromethyl)benzyl)thiazolidin-4-one; 2-(2,4-dichlorophenylimino)-5-(3-(trifluoromethyl)benzyl)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(p-tolylimino)-3-methylthiazolidin-4-one;

N-(5-(3-(trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylidene)-4-chlorobenzamide;

5-(3-(trifluoromethyl)benzyl)-2-(4-chlorophenyl)sulfonyliminothiazolidin-4-one; phenyl 5-(3-(trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylidenecarbamate; 5-(4-methoxyphenethyl)-2-(p-tolylimino)thiazolidin-4-one;

5-(4-methoxyphenethyl)-2-(phenylimino)thiazolidin-4-one; and

2-(p-tolylimino)-5-phenethylthiazolidin-4-one.

Particularly preferred compounds of formula I include; 5-(4-fluorobenzyl)-2-(pyridin-2-ylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(4-chlorophenylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(p-tolylimino)thiazolidin-4-one

5-(4-methoxyphenethyl)-2-(p-tolylimino)thiazolidin-4-one;

5-(4-methoxyphenethyl)-2-(phenylimino)thiazolidin-4-one; and 2-(p-tolylimino)-5-phenethylthiazolidin-4-one. Especially preferred compounds of formula I include 5-(3-(trifluoromethyl)benzyl)- 2-(4-chlorophenylimino)thiazolidin-4-one.

Compounds of formula I may be known and/or may be commercially available. Other compounds of formula I (e.g. that are not commercially available) may be prepared in accordance with techniques that are well known to those skilled in the art, for example as described hereinafter.

According to a further aspect of the invention there is provided a process for the preparation of a compound of formula I, which process comprises:

(i) for compounds of formula I in which Y represents -C(O)-, W represents -NR₇, and Ai represents a double bond (and R₇ is therefore absent), reaction of either:

(A) a compound of formula II,

(B) a compound of formula III,

wherein R^a represents Ci_-6 alkyl (e.g. ethyl; so forming an ester group), L¹ represents a suitable leaving group, such as a sulfonate group (e.g. mesylate or, preferably, tosylate) or, for example preferably, halo (e.g. bromo or chloro); or

(C) a compound of formula IV,

wherein, in all cases, X and Ri are as hereinbefore defined,

with, in each case, a compound of formula V,

wherein T^a represents O or, more preferably, S and R5 and R₆ are as hereinbefore defined, under reaction conditions known to those skilled in the art, for example for reaction (A) above conditions such as those described in Blanchet et al, Tetrahedron Letters, 2004, 45, 4449-4452; for reaction (B) above, conditions such as those described in St. Laurent ef al, Tetrahedron Letters, 2004, 45, 1907-1910; K. Arakawa et al., Chem. Pharm. Bull. 1997, 45, 1984- 1993; A. Mustafa, W. Musker, A.F.A.M. Shalaby, A.H. Harhash, R. Daguer, Tetrahedron 1964, 20; 25-31 ; or P. Herold, A. F. Indolese, M. Studer, H. P. Jalett, U. Siegrist, H. U. Blaser, Tetrahedron 2000, 56, 6497-6499, and for reaction (C) above, conditions such as those described in Le Martchalal et al, Tetrahedron 1990, 46, 453-464;

(ii) for compounds of formula I in which Y represents -S(O)₂-, W represents -NR₇-, and A₁ represents a double bond (and R₇ is therefore absent), X represents -[RsRg]_n- in which n represents 0 and R₁ represents H, reaction of a compound of formula Vl,

wherein L² represents a suitable leaving group, such as halo (e.g. chloro), with a compound of formula VII,

R₅-N=C=T³ VII

wherein T^a is as hereinbefore defined but is preferably S and R₅ is as hereinbefore defined under conditions known to those skilled in the art, for example such as those described in Zbirovsky and Seifert, Coll. Czech. Chem. Commun. 1977, 42, 2672-2679 or Von Zaki El-Heweri, Franz Runge, Journal fur praktische Chemie, 4, Band 16, 1962, e.g. in the presence of base (e.g. an aqueous solution of NaOH) in an appropriate solvent (e.g. acetone), for example at elevated temperature (e.g. 50°);

(iii) for compounds of formula I in which Ai represents a double bond (and R₇ is therefore absent), X represents -[ReRg]_n- in which n represents 1 , 2 or 3 and Ri is as hereinbefore defined and, preferably, Y represents -S(O)₂- and/or W represents -NR₇, reaction of a corresponding compound of formula I in which n represents 0 and R₁ represents hydrogen, with a compound of formula VIII,

R_1a-X^a-L³ VIII

wherein X^a represents -[RsRg]_n- in which n represents 1 , 2 or 3 and Ri₃ represents R₁ as hereinbefore defined, or n represent 0 and R-i_a represents R₁ as hereinbefore defined provided that it does not represent hydrogen, aryl or heteroaryl, and L³ represents a suitable leaving group (e.g. a halo, such as chloro, iodo or, preferably, bromo, or a sulfonate group), under reaction conditions known to those skilled in the art, for example, in the presence of a suitable base (e.g. an organometallic base (e.g. an organolithium), an alkali metal base (e.g. sodium hydride) or an amide salt (e.g. (Me₃Si)₂NNa) and the like) and a suitable solvent (e.g. tetrohydrofuran, dimethylformamide, dimethlysulfoxide or the like) at room temperature or below (such as at sub-zero temperatures (e.g. -78⁰C). For example, for the synthesis of compounds of formula I in which Y represents -S(O)₂- and/or W represents -NR₇, reaction conditions include those described in the journal article mentioned in respect of process step (ii) above;

o? (iv) for compounds of formula I in which n represents 0 and R-i represents alkenyl optionally substituted as hereinbefore defined (i.e. by B¹) in which one double bond of the alkenyl group is directly attached to the requisite ring or R₁ represents alkyl substituted with a -OH group α to the point of attachment of the said alkyl group to the requisite and which alkyl group is optionally further substituted as hereinbefore defined (i.e. by B¹) and, in both cases, W represents -NR₇C(O)-, -NR₇S(O)₂-, -NR₇C(O)NR₇-, -NR₇C(O)O- or -NR₇-, -CR₇R₇- or a bond, reaction of a corresponding compound of formula I in which n represents O and R₁ represents H with a compound of formula IX,

R_1b=0 IX

wherein R^ represents alkyl optionally substituted by B¹ as hereinbefore defined, under standard reactions conditions known to those skilled in the art. For example for the preparation of compounds in which Ri represents alkenyl as defined above, under standard dehydration conditions, e.g. in the presence of a suitable base (such as NaOAc or an appropriate base described hereinafter in respect of process step (vii)) in the presence of a suitable solvent (e.g. glacial acetic acid), e.g. as described in A. Mustafa, W. Musker, A.F.A.M. Shalaby, A.H. Harhash, R. Daguer, Tetrahedron 1964, 20, 25-31. For the preparation of compounds in which Ri represents alkyl substituted by -OH as defined above, reaction in the presence of a suitable base (e.g. lithium diisopropylamide or another suitable base described in process step (vii) below) in the presence of an appropriate solvent (e.g. anhydrous THF) at room temperature or below (e.g. about O⁰C) under an inert atmosphere. The skilled person will appreciate that for preparation of compounds in which R₁ represents optionally substituted alkenyl as described above, this may involve an intermediate which is the above- mentioned compound of formula I in which R₁ represents alkyl substituted by -OH as defined above (which intermediate may be isolable), which intermediate may need to be transformed to the alkenyl group separately, for example by converting the -OH group to a better leaving group, for example by reaction with trifluoroacetic anhydride or the like optionally in the presence of a suitable base (e.g. triethylamine) and a catalyst (e.g. DMAP) in an appropriate solvent (e.g. dichloromethane) at below room temperature (such as at about O⁰C) e.g. employing conditions described in Zbirovsky and Seifert, Coll. Czech. Chem. Commun. 1977, 42, 2672-2679;

(v) for compounds of formula I in which n represents 0 and R₁ represents saturated alkyl optionally substituted (i.e. by B¹) as hereinbefore defined, Y represents -S(O)₂ or, preferably, -C(O)- or =C(R₁o)- as hereinbefore defined, reduction of a corresponding compound of formula I in which Ri represents optionally substituted unsaturated alkyl, under standard reaction conditions, for example in the presence of a suitable (e.g. chemoselective) reducing agent such as LiBH₄ or NaBH₄ optionally in the presence of a suitable solvent such as a THF or pyridine (or a mixture thereof, e.g. as described in R.G. Giles, NJ. Lewis, J. K. Quick, MJ. Sasse, M.W.J. Urquhart, L. Youssef, Tetrahedron 2000; 56, 4531 - 4537). The skilled person will appreciate that the choice of the reducing agent is important in order to achieve the desired reduction selectively (i.e. whilst not reducing other functional groups, such as carbonyl groups, in the relevant compound of formula I). Alternative methods include reduction by hydrogenation under standard conditions, for example in the presence of hydrogen gas or nascent hydrogen, an appropriate solvent (e.g. an alcoholic solvent) and catalyst (e.g. Pd/C);

(vi) for compounds of formula I in which Re is alkyl, cycloalkyl or benzyl, all of which are optionally substituted as hereinbefore defined, reaction of a corresponding compound of formula I in which Re represents H, with a compound of formula X,

R₆₃L⁴ X

wherein R_6a represents alkyl, cycloalkyl or benzyl (e.g. which are optionally substituted by one or more groups selected from B¹³, B¹⁴ or B¹⁶, respectively) and L⁴ represents a suitable leaving group such as halo (e.g. iodo or bromo) or a sulfonate group, under standard reaction conditions, for example at around room temperature, in the presence of a suitable base (e.g. sodium hydride, sodium bicarbonate, potassium carbonate, pyrrolidinopyridine, pyridine, triethytamine, tributylamine, trimethylamine, dimethylaminopyridine, diisopropylamine, 1 ,8- diazabicyclo[5.4.0]undec-7-ene, sodium hydroxide, or mixtures thereof), an appropriate solvent (e.g. pyridine, dichloromethane, chloroform, tetrahydrofuran, dimethylformamide, triethylamine, dimethylsulfoxide, water or mixtures thereof) and, in the case of biphasic reaction conditions, optionally in the presence of a phase transfer catalyst;

(vii) for compounds of formula I that are substituted with at least one of B¹ to B¹⁸ that represents a -C(O)NR_16aRieb group, reaction of a corresponding compound of formula I in which that/those (as appropriate) B¹ to B¹⁸ substituents represent -C(O)OR₁₆, with a compound of formula Xl,

or a protected derivative (e.g. a salt) thereof, wherein R_16a and Ri₆b are as hereinbefore defined, for example under standard coupling reaction conditions. For example, in the case where R^ represents H, in the presence of a suitable coupling reagent (e.g. 1 ,1 '-carbonyldiimidazole, Λ/,Λ/'-dicyclohexylcarbodiimide, 1- (3-dimethylamino-propyl)-3-ethylcarbodiimide (or hydrochloride thereof), N, N'- disuccinimidyl carbonate, benzotriazol-1 -yloxytris(dimethylamino)phosphonium hexafluorophosphate, 2-(1 /-/-benzotriazol-1 -yl)-1 ,1 , 3, 3-tetramethyluronium hexa- fluorophosphate, benzotriazol-1 -yloxytris-pyrrolidinophosphonium hexafluorophosphate, bromo-tris-pyrrolidinophosponium hexafluorophosphate, 2-(1 H- benzotriazol-1-yl)-1 , 1 ,3, 3-tetramethyluronium tetra-fluorocarbonate) or 1- cyclohexylcarbodiimide-3-propyloxymethyl polystyrene, a suitable base (e.g. sodium hydride, sodium bicarbonate, potassium carbonate, pyrrolidinopyridine, pyridine, triethylamine, tributylamine, trimethylamine, dimethylaminopyridine, diisopropylamine, 1 ,8-diazabicyclo[5.4.0]undec-7-ene, sodium hydroxide, N- ethyldiisopropylamine, Λ/-(methylpolystyrene)-4-(methylamino)pyridine, potassium bis(trimethylsilyl)amide, sodium bis(trimethylsilyl)amide, potassium ferf-butoxide, lithium diisopropylamide, lithium 2,2,6,6-tetramethylpiperidine or mixtures thereof) and an appropriate solvent (e.g. tetrahydrofuran, pyridine, toluene, dichloromethane, chloroform, acetonitrile or dimethylformamide). Alternatively, for example in the case where Ri₆ is other than H (i.e. -C(O)ORi6 represents an ester group), the reaction may be performed in the presence of an appropriate reagent (e.g. trimethylaluminium) in the presence of a suitable solvent (e.g. benzene), for example at elevated temperature (e.g. about 60⁰C), e.g. as described in Hwang, K.-J.; O'Neil, J.-P.; Katzenellenbogen, J. A. J. Org. Chem. 1992, 57, 1262;

(viii) for compounds of formula I in which W represents -NR₇C(O)-, -NR₇S(O)₂-, -NR₇C(O)NR₇- or -NR₇C(O)O-, reaction of a corresponding compound of formula I in which W represents -NR₇ and R₅ represents H, with a compound of formula XII,

L⁵W^XR₅ XII

wherein W^x represents -C(O)-, -S(O)₂, -C(O)NR₇- or -C(O)O-, L⁵ represents a suitable leaving group such as halo (e.g. chloro) and R₅ is as hereinbefore defined, under reaction conditions known to those skilled in the art, for example in the presence of a suitable base (e.g. NaH, NaOH, triethylamine, pyridine, another suitable base mentioned at process step (vii) above or mixtures thereof) and solvent (e.g. pyridine (which may serve as the base and solvent) DMF or dichloromethane (e.g. further in the presence of water and, optionally, a phase transfer catalyst)) for example at room temperature e.g. as described in Hurst, D. T.; Stacey, A. D., Nethercleft, M., Rahim, A., Harnden, M. R. Aust. J. Chem. 1998, 41 , 1221 ; or

(ix) for compounds of formula in which W represents -NR₇C(O)NH-, reaction of a corresponding compound of formula I in which W represents -NR₇ and Rs represents H, with a compound of formula XIII,

R₅-N=C=O XIII

wherein R₅ is as hereinbefore defined, under standard conditions, for example, in the presence of a suitable solvent (e.g. a polar aprotic solvent such as toluene) and at elevated temperature (e.g. reflux), for example as described in the journal article mentioned in respect of process (viii) above.

Compounds of formula Il may be prepared by reaction of a compound of formula XIV, R₁-X-C(O)H XIV

wherein Ri and X are as hereinbefore defined, with trichloroacetic acid under standard conditions known to those skilled in the art, for example such as those described in the journal article mentioned in respect of process step (i) (part (A)) above.

Compounds of formula III may be commercially available, prepared under standard conditions or, for those compounds in which X represents -CH2-, Ri represents aryl or heteroaryl optionally substituted as hereinbefore defined and L¹ represents a halo group, reaction of a compound of formula XV,

R₁₀NH₂ XV

wherein R_{1 c} represents aryl or heteroaryl (e.g. optionally substituted by B⁵ and B⁶) to form the corresponding diazonium salt (for example by reaction with sodium nitrite at low temperatures such as at about 0⁰C) followed by reaction with a compound of formula XVI,

R^a-OC(O)CH=CH₂ XVI

wherein R^a is as defined above, in the presence of a suitable solvent (e.g. acetone) and a hydrohalic acid which is preferably concentrated (e.g. in the case where L¹ represents chloro, concentrated hydrochloric acid) optionally in the presence of an agent that aids the Michael addition of the halide onto the acrylate/enone such as cuprous oxide.

Compounds of formula III in which L¹ represents a sulfonate group (e.g. a toslyate or mesylate) may be prepared by reaction of a compound corresponding to a compound of formula III but in which L¹ represents -OH with an appropriate sulfonyl chloride (e.g. tosyl chloride or mesyl chloride) under standard conditions known to those skilled in the art, such as those described in respect of preparation of compounds of formula I above (process step (vi) above). Compounds of formula Vl may be prepared by reaction of a compound of formula XVII,

wherein L⁶ represents a suitable leaving group such as halo (e.g. chloro) and L² is as hereinbefore defined, with ammonia (e.g. in gaseous or other form) for example under standard conditions known to those skilled in the art, such as those described in respect of preparation of compounds of formula I above (process step (vi) above) or, preferably, in the presence of diethyl ether at low temperature (e.g. about O⁰C) in which case the skilled person will appreciate that the ammonia additionally serves as a base.

Compounds of formulae IV, V, VII, VIII, IX, X, Xl, XII, XIII₁ XIV, XV, XVI and XVII (and also certain compounds of formulae I₁ II, III and Vl) are either commercially available, are known in the literature, or may be obtained either by analogy with the processes described herein (or processes described in references contained herein), or by conventional synthetic procedures, in accordance with standard techniques, from available starting materials using appropriate reagents and reaction conditions.

Substituents, such as R-i, R₅, Re, X, W and Y in final compounds of formula I or relevant intermediates may be modified one or more times, after or during the processes described above by way of methods that are well known to those skilled in the art. Examples of such methods include substitutions, reductions, oxidations, alkylations, acylations, hydrolyses, esterifications, and etherifications. The precursor groups can be changed to a different such group, or to the groups defined in formula I, at any time during the reaction sequence.

Compounds of formula I may be isolated from their reaction mixtures using conventional techniques. It will be appreciated by those skilled in the art that, in the processes described above and hereinafter, the functional groups of intermediate compounds may need to be protected by protecting groups.

The protection and deprotection of functional groups may take place before or after a reaction in the above-mentioned schemes.

Protecting groups may be removed in accordance with techniques that are well known to those skilled in the art and as described hereinafter. For example, protected compounds/intermediates described herein may be converted chemically to unprotected compounds using standard deprotection techniques.

The type of chemistry involved will dictate the need, and type, of protecting groups as well as the sequence for accomplishing the synthesis.

The use of protecting groups is fully described in "Protective Groups in Organic Chemistry", edited by J W F McOmie, Plenum Press (1973), and "Protective Groups in Organic Synthesis", 3^rd edition, T.W. Greene & P.G.M. Wutz, Wiley- lnterscience (1999).

As used herein, the term "functional groups" means, in the case of unprotected functional groups, hydroxy-, thiolo-, aminofunction, carboxylic acid and, in the case of protected functional groups, lower alkoxy, N-, O-, S- acetyl, carboxylic acid ester.

Combination products according to the invention provide for the administration of compounds of formula I in conjunction with trastuzumab or TKI (or salt or other derivative thereof), and may thus be presented either as separate formulations, wherein at least one of those formulations comprises compound of formula I₁ and at least one comprises trastuzumab or TKI, or may be presented (i.e. formulated) as a combined preparation (i.e. presented as a single formulation including a compound of formula I and trastuzumab or TKI).

Thus, there is further provided: (1 ) a pharmaceutical formulation including a compound of formula I, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof; trastuzumab or a TKI, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof; and a pharmaceutically-acceptable adjuvant, diluent or carrier (which formulation is hereinafter referred to as a "combined preparation"); and

(2) a kit of parts comprising components:

(a) a pharmaceutical formulation including a compound of formula I, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof, in admixture with a pharmaceutically- acceptable adjuvant, diluent or carrier; and

(b) a pharmaceutical formulation including trastuzumab, or a TKI, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof, in admixture with a pharmaceutically- acceptable adjuvant, diluent or carrier, which components (a) and (b) are each provided in a form that is suitable for administration in conjunction with the other.

According to a further aspect of the invention, there is provided a method of making a kit of parts as defined above, which method comprises bringing component (a), as defined above, into association with a component (b), as defined above, thus rendering the two components suitable for administration in conjunction with each other.

By bringing the two components "into association with" each other, we include that components (a) and (b) of the kit of parts may be:

(i) provided as separate formulations (i.e. independently of one another), which are subsequently brought together for use in conjunction with each other in combination therapy; or

(ii) packaged and presented together as separate components of a "combination pack" for use in conjunction with each other in combination therapy.

Thus, there is further provided a kit of parts comprising: (I) one of components (a) and (b) as defined herein; together with (II) instructions to use that component in conjunction with the other of the two components.

The kits of parts described herein may comprise more than one formulation including an appropriate quantity/dose of compound of formula I, and/or more than one formulation including an appropriate quantity/dose of trastuzumab or

TKI, in order to provide for repeat dosing. If more than one formulation

(comprising either active compound) is present, such formulations may be the same, or may be different in terms of the dose of either compound, chemical composition(s) and/or physical form(s).

The combination products according to the invention find utility in the treatment of cancer.

The term "cancer" will be understood by those skilled in the art to include one or more diseases in the class of disorders that is characterized by uncontrolled division of cells and the ability of these cells to invade other tissues, either by direct growth into adjacent tissue through invasion, proliferation or by implantation into distant sites by metastasis.

In a preferred embodiment, combination products according to the invention are capable of inhibiting the proliferation of cancer cells. By "proliferation" we include an increase in the number and/or size of cancer cells.

Alternatively, or preferably in addition, combination products according to the invention are capable of inhibiting metastasis of cancer cells.

By "metastasis" we mean the movement or migration (e.g. invasiveness) of cancer cells from a primary tumour site in the body of a subject to one or more other areas within the subject's body (where the cells can then form secondary tumours). Thus, in one embodiment the invention provides combinations and methods for inhibiting, in whole or in part, the formation of secondary tumours in a subject with cancer. It will be appreciated by skilled persons that the effect of a combination product according to the invention as described herein on "metastasis" is distinct from any effect such a combination product may or may not have on cancer cell proliferation.

Advantageously, combination products according to the invention may be capable of inhibiting the proliferation and/or metastasis of cancer cells selectively.

By "selectively" we mean that the combination product inhibits the proliferation and/or metastasis of cancer cells to a greater extent than it modulates the function (e.g. proliferation) of non-cancer cells. Preferably, the combination product inhibits the proliferation and/or metastasis of cancer cells only.

The combination products according to the invention are suitable for use in the treatment of any cancer type, including all solid tumors. For example, the cancer cells may be selected from the group consisting of cancer cells of the breast, bile duct, brain, colon, stomach, reproductive organs, thyroid, hematopoetic system, lung and airways, skin, gallbladder, liver, nasopharynx, nerve cells, kidney, prostate, lymph glands and gastrointestinal tract. Preferably, the cancer is selected from the group of colon cancer (including colorectal adenomas), breast cancer (e.g. postmenopausal breast cancer), endometrial cancer, cancers of the hematopoetic system (e.g. leukemia, lymphoma, etc), thyroid cancer, kidney cancer, oesophageal adenocarcinoma, ovarian cancer, prostate cancer, pancreatic cancer, gallbladder cancer, liver cancer and cervical cancer. More preferably, the cancer is selected from the group of colon, prostate and, particularly, breast cancer.

Preferably, the cancer cells are breast cancer cells. Combination products according to the invention are particularly useful in the treatment of HER2- positive cancers.

Combination products according to the invention may also find utility for example in adjuvant therapy (i.e. reducing the risk of the cancer coming back after surgery), in neo-adjuvant therapy (before surgery, to shrink a large breast cancer so that a lumpectomy is possible), in the control of breast cancers that have come back after initial treatment, or in the control of breast cancers that cannot be removed when first diagnosed. According to a further aspect of the invention there is provided a method of treatment of cancer, which method comprises the administration of a combination product according to the invention to a patient in need of such treatment.

For the avoidance of doubt, in the context of the present invention, the terms "treatment", "therapy" and "therapy method" include the therapeutic, or palliative, treatment of patients in need of, as well as the prophylactic treatment and/or diagnosis of patients which are susceptible to, cancer.

With respect to the kits of parts as described herein, by "administration in conjunction with", we include that respective formulations comprising compound of formula I and trastuzumab or TKI (or salt/solvate/derivative thereof) are administered, sequentially, separately and/or simultaneously, over the course of treatment of the relevant condition.

Thus, in respect of the combination product according to the invention, the term "administration in conjunction with" includes that the two components of the combination product (compound of formula I and trastuzumab or TKI) are administered (optionally repeatedly), either together, or sufficiently closely in time, to enable a beneficial effect for the patient, that is greater, over the course of the treatment of the relevant condition, than if either a formulation comprising compound of formula I₁ or a formulation comprising trastuzumab or TKI, are administered (optionally repeatedly) alone, in the absence of the other component, over the same course of treatment. Determination of whether a combination provides a greater beneficial effect in respect of, and over the course of treatment of, a particular condition will depend upon the condition to be treated or prevented, but may be achieved routinely by the skilled person.

Further, in the context of a kit of parts according to the invention, the term "in conjunction with" includes that one or other of the two formulations may be administered (optionally repeatedly) prior to, after, and/or at the same time as, administration with the other component. When used in this context, the terms "administered simultaneously" and "administered at the same time as" include that individual doses of compound of formula I and trastuzumab or TKI are administered within 48 hours (e.g. 24 hours) of each other.

"Patients" include mammalian (including human) patients.

The term "effective amount" refers to an amount of a compound, which confers a therapeutic effect on the treated patient (e.g. sufficient to treat or prevent the disease). The effect may be objective (i.e. measurable by some test or marker) or subjective (i.e. the subject gives an indication of or feels an effect).

In accordance with the invention, compounds of formula I may be administered alone, but are preferably administered orally, intravenously, intramuscularly, cutaneously, subcutaneously, transmucosally (e.g. sublinguaily or buccally), rectally, transdermal^, nasally, pulmonarily (e.g. tracheally or bronchially), topically, by any other parenteral route, in the form of a pharmaceutical preparation comprising the compound in a pharmaceutically acceptable dosage form. Preferred modes of delivery include oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, or intraperitoneal delivery.

Compounds of formula I will generally be administered as a pharmaceutical formulation in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier, which may be selected with due regard to the intended route of administration and standard pharmaceutical practice. Such pharmaceutically acceptable carriers may be chemically inert to the active compounds and may have no detrimental side effects or toxicity under the conditions of use. Suitable pharmaceutical formulations may be found in, for example, Remington The Science and Practice of Pharmacy, 19th ed., Mack Printing Company, Easton, Pennsylvania (1995). For parenteral administration, a parenterally acceptable aqueous solution may be employed, which is pyrogen free and has requisite pH, isotonicity, and stability. Suitable solutions will be well known to the skilled person, with numerous methods being described in the literature. A brief review of methods of drug delivery may also be found in e.g. Langer, Science 249, 1527 (1990). Suitable formulations for use in administering trastuzumab and TKIs are described in the literature (see for example Martindale - The Complete Drug Reference (34^th Edition) (e.g. at pages 589 to 590, 557 to 558 and 562), and the documents referred to therein, the relevant disclosures in all of which documents are hereby incorporated by reference). Otherwise, the preparation of suitable formulations, and in particular combined preparations including both compound of formula I and trastuzumab or TKI may be achieved non-inventively by the skilled person using routine techniques and/or in accordance with standard and/or accepted pharmaceutical practice. For example suitable formulations of trastuzumab for infusion include solutions and/or suspensions of the active ingredient in an appropriate isotonic solution (e.g. NaCI 0.9%).

The amount of active ingredients in the formulation(s) will depend on the severity of the condition, and on the patient, to be treated, as well as the compound(s) which is/are employed, but may be determined non-inventively by the skilled person.

Depending on the disorder, and the patient, to be treated, as well as the route of administration, active ingredients may be administered at varying therapeutically effective doses to a patient in need thereof.

However, the dose administered to a mammal, particularly a human, in the context of the present invention should be sufficient to effect a therapeutic response in the mammal over a reasonable timeframe. One skilled in the art will recognize that the selection of the exact dose and composition and the most appropriate delivery regimen will also be influenced by inter alia the pharmacological properties of the formulation, the nature and severity of the condition being treated, and the physical condition and mental acuity of the recipient, as well as the potency of the specific compound, the age, condition, body weight, sex and response of the patient to be treated, and the stage/severity of the disease.

Administration of active ingredients may be continuous or intermittent (e.g. by bolus injection). The dosage may also be determined by the timing and frequency of administration. In the case of oral or parenteral administration of compounds of formula I, the dosage can vary from about 0.01 mg to about 1000 mg per day of the relevant compound of formula I (or, if employed, a corresponding amount of a pharmaceutically acceptable salt or prodrug thereof).

Suitable doses of trastuzumab are known to those skilled in the art (e.g. infusion doses of about 4 mg/kg body weight followed by about 2 mg/kg doses as weekly intervals).

Suitable doses of TKIs may be found in the literature (see, for example, Steeghs et a/, Ann. Surg. Oncol., Epub ahead of print, 14 November 2006; see www.ncbi.nlm. nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=Abstra ctPlus&list_uids=17103252&itool=iconabstr&query_hl=0&itool=pubmed_DocSu) and are in the range 10 mg to 2,000 mg, such as 30 to 1 ,500, e.g. 50 to 1250 mg daily oral doses.

In any event, the medical practitioner, or other skilled person, will be able to determine routinely the actual dosage, which will be most suitable for an individual patient. The above-mentioned dosages are exemplary of the average case; there can, of course, be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention.

Combination products according to the invention may be further combined in combination therapy with other chemotherapeutic agents known in the art to be of use in combination with e.g. trastuzumab, such as paclitaxel and docetaxel.

Combination products according to the invention may also comprise, in place of a compound selected from trastuzumab, or a tyrosine kinase inhibitor, or a pharmaceuticalfy-acceptable salt, solvate or pharmaceutically functional derivative of a tyrosine kinase inhibitor, a compound selected from the following: (i) a glitazone (such as rosiglitazone); (ii) metformin;

(iii) a statin, such as fluvastatin, simvastatin, rosuvastatin, pravastatin, atorvastatin and, particularly, lovastatin; (iv) an antibody (other than trastuzumab) that is useful in the treatment of cancer, such as bevacizumab, cetuximab or panitumumab; and/or

(v) an inhibitor of activity of the mammalian target of rapamycin (mTOR), such as rapamycin or a pharmaceutically functional derivative (as hereinbefore defined) thereof.

Compounds as listed in above-mentioned classes (iv) and (v) are well known to be of potential use in the treatment of cancer, as described herein. It has recently been suggested in the literature (see, for example, MoI. Cancer Ther., 5, 430 (2006), Cancer Res., 66, 10269 (2006) and Int. J. Cancer, 118, 773 (2006)) that the above mentioned compound classes (i) to (iii) may be used in the treatment of cancer, as described herein. There is therefore provided, as a further aspect of the invention, a combination product comprising (a) a compound of formula I as hereinbefore defined, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof; and (b) a compound selected from a glitazone, metformin, a statin, an antibody (other than trastuzumab) that is useful in the treatment of cancer, or an inhibitor of activity of the mammalian target of rapamycin, or a pharmaceutically-acceptable salt, solvate or pharmaceutically functional derivative (as hereinbefore defined) of a glitazone, metformin, a statin or an inhibitor of activity of the mammalian target of rapamycin.

It will be appreciated that all other aspects of the invention, and preferred features of the invention, as defined herein may be incorporated into the latter aspect of the invention.

The combination products/methods described herein may have the advantage that, in the treatment of cancer, they may be more convenient for the physician and/or patient than, be more efficacious than, be less toxic than, have a broader range of activity than, be more potent than, produce fewer side effects than, or that they may have other useful pharmacological properties over, similar methods (treatments) known in the prior art for use in the treatment of cancer or otherwise.

The invention is illustrated by the following examples in which error bars denote SEM and the following abbreviations are employed: LA - linolenic acid

DMSO - dimethyl sulfoxide.

Figures 1a to 1e are representative examples of cell cycle analysis using Flow Cytometer. Cells were incubated with or without linolenic acid and the compound of Example 95 below (Compound X) for 24 hours. Histograms represent accumulated events and their distribution in the cell cycle by intensity of Pl staining (FL3). (a) untreated, (b) LA 100 μM, (c) LA 100 μM + Compound X 10 μM, (d) Compound X 10 μM, (e) DMSO 0.2%.

Figure 2A is a histogram summarizing 4 experiments where one compound is identified and verified as an FFA antagonist. Cells were incubated with or without linolenic acid and the Compound X for 24 hours at indicated concentrations. Cells in S-phase from untreated sample were set to 100% in each experiment.

Figures 2B and 2C are histograms where compounds are identified and verified as FFA antagonists. Cells were incubated with or without linolenic acid and the compound of Examples 4 and 6 below (Compound 2 and Compound Y, respectively) for 24 hours at indicated concentrations. Cells in S-phase from untreated sample were set to 100% in each experiment (n=2).

Figures 3A to 3F show hematoxylin stained sections from tumors dissected from vehicle or test compound treated mice.

Examples

The following examples were prepared in accordance with correspondng procedures described in international patent application WO 2007/010273.

Example 1 5-Benzyl-2-(phenylimino)thiazolidin-4-one

Example 2 δ-^-MethylbenzvO^-^-chlorophenylirninoHhiazolidin^-one

3S Example 3 5-(4-Chlorobenzvπ-2-(4-chlorophenyliminoHhiazolidiπ-4-one

Example 4

5-(3-(Trifluoromethyl)benzvπ-2-(p-tolylimino)thiazolidin-4-one

Example 5

5-(3-(Trifluoromethyl)benzyl)-2-(4-isopropylphenylimino)thiazolidin-4-one

Example 6 5-(3-(Trifluoromethyl)benzyl)-2-(4-chlorophenylimino)thiazolidin-4-one

Example 7 5-(3-(Trifluoromethyl)benzyl)-2-(4-methoxyphenylimino)thiazolidin-4-one

Example 8 5-(3-(Trifluoromethyl)benzyl)-2-(phenylimino)thiazolidin-4-one

Example 9

5-(4-Fluorobenzyl)-2-(phenylimino)thiazolidin-4-one

Example 10

5-(4-Fluorobenzyl)-2-(p-tolylimino)thiazolidin-4-one

Example 11 2-('4-Chlorophenylimino)-5-(4-fluorobenzyl)thiazolidiπ-4-one

Example 12 5-(4-Fluorobenzyπ-2-(4-methoxyphenylimino)thiazolidin-4-one

Example 13 5-(4-FluorobenzylV2-(4-isopropylphenylimino')thiazolidin-4-one Example 14 5-(4-(Trifluoromethyl)benzyπ-2-(p-tolylimino)thiazolidin-4-one

Example 15

5-(4-Methoxybeπzyl)-2-(p-tolylimino)thiazolidin-4-one

Example 16

5-Benzyl-2-(phenylimino)thiazolidin-4-one

Example 17 5-(3-(Trifluoromethyl)benzyl)-2-(4-fluorophenylimino)thiazolidin-4-one

Example 18 5-(3-(Trifluoromethyl)benzyl)-2-(4-bromophenylimino)thiazolidin-4-one

Example 19 2-(3,4-DichlorophenyliminoV5-(3-(trifluoromethyl)benzyl)thiazolidin-4-one

Example 20

2-(2,4-Dichlorophenylimino)-5-(3-(trifluoromethyl)benzyl)thiazolidin-4-one

Example 21

4-(5-(3-(Trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylideneamino)benzonitrile

Example 22

Ethyl 4-(5-(3-(trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylideneamino)benzoate

Example 23 4-(5-(3-(Trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylideneamino)beπzoic acid

Example 24 4-(5-f3-(Trifluoromethyl)benzvπ-4-oxothiazolidin-2-ylideπeamino)benzamide Example 25 5-(3-(Trifluoromethyl)benzvπ-2-(m-tolylimino)thiazolidin-4-one

Example 26 2-(4-Chlorophenylimino)-5-(4-fluoro-3-(trifluoromethyl)benzyl)thiazoliclin-4-one

Example 27 5-(3-(Trifluoromethyl)benzyl)-2-(p-tolylimino)-3-methylthiazolidin-4-one

Example 28

5-(3-(Trifluoromethyl)benzyl)-2-(Λ/-methyl-Λ/-phenylamino)thiazol-4(5H)-one

Example 29

5-(3-(Trifluoromethyl)benzyl)-2-(Λ/-methyl-Λ/-p-tolylamino)thiazol-4(5H)-one

Example 30 5-(4-Fluorobenzyl)-2-(Λ/-methyl-Λ/-(pyridin-2-vπamino)thiazol-4(5H)-one

Example 31 2-(2-(Λ/-Methyl-Λ/-p-tolylaminoV4,5-dihydro-4-oxothiazol-5-yl)-Λ/-p-tolylacetamide

Example 32 5-(3-(Trifluoromethyl)benzyl)-2-(Λ/-benzyl-Λ/-p-tolylamino)thiazol-4(5H)-one

Example 33

5-(4-Fluorobenzyl)-2-(Λ/-benzyl-A/-(pyridin-2-yl)amino)thiazol-4(5H)-one

Example 34

2-(2-(Λ/-Benzyl-Λ/-p-tolylamino)-4,5-dihvdro-4-oxothiazol-5-vπ-Λ/-p-tolylacetamide

Example 35 5-(3-(Trifluoromethyl)benzvπ-2-(Λ/-phenyl-Λ/-p-tolylamino)thiazol-4(5H)-one

Example 36 5-(4-Fluorobenzvn-2-(A/-phenyl-A/-(pyridin-2-yl)amino')thiazol-4(5H)-one Example 37 2-(2-(A/-pheny[-A/-p-tolylannino^')-4,5-dihvdro-4-oxothiazol-5-yl)-A/-p-tolylacetamicle

Example 38

5-(3-(Trifluoromethyl)benzylidene)-2-(phenylimino)thiazolidin-4-one

Example 39

5-(3-(Trifluoromethvπbenzylidene)-2-(p-tolv]imino)thiazolidiπ-4-one

Example 40 5-(4-Fluorobenzylidene)-2-(pheπylimino)thiazolidin-4-one

Example 41 5-(4-Fluorobenzylidene)-2-(p-tolylimino)thiazolidin-4-one

Example 42 5-Benzylidene-2-(phenylimino)thiazolidin-4-one

Example 43

2-(p-Tolylimino)-5-benzylidenethiazolidin-4-one

Example 44

5-(3-(Trifluoromethyl)beπzylidene)-2-(4-chlorophenylimino)thiazolidin-4-one

Example 45 2-(4-Chlorophenylimino)-5-benzylidenethiazolidin-4-one

Example 46 2-(4-Chlorophenylimino)-5-(4-fluoro-3-(trifluoromethyl)benzylidene)thiazolidin-4- one

Example 47 2-(4-Methylbenzyl)-5-(3-trifluoromethyl-benzyl)-thiazol-4-one Example 48 5-(4-Fluorobeπzyl)-2-pyridin-2-ylmethylthiazol-4-one

Example 49 2-f2-(4-Methylbenzyl)-4-oxo-4,5-dihydrothiazol-5-vn-Λ/-p-tolyl-acetamide

Example 50 2-(1-p-Tolylethyl)-5-(3-trifluoromethylbenzyl)-thiazol-4-one

Example 51

5-(4-Fluorobenzvπ-2-(1-pyridin-2-yl-ethyl)thiazol-4-one

Example 52

2-r4-Oxo-2-(1-p-tolylethyl)-4,5-dihvdro-thiazol-5-yl1-Λ/-p-tolylacetannide

Example 53 2-Phenyl-5-(3-trifluoromethylbenzyl)thiazol-4-one

Example 54 5-(4-FluorobenzyD-2-pyridin-2-yl-thiazol-4-one

Example 55

2-(4-Oxo-2-phen yl-4, 5-dihyd roth iazol-5-yl )-Λ/-p-tolylaceta mide

Example 56

2-p-Tolylimino-5-ri -(3-trifluoromethylphenyl)ethyl1-thiazolidin-4-one

Example 57

5-f1-(4-Fluorophenyl)ethyl1-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 58 5-f1-Methyl-1-(3-trifluoromethylphenvπethyl]-2-p-tolyliminothiazolidin-4-one

Example 59 5-f1-(4-Fluorophenyl)-1-methylethyl1-2-(pyridin-2-ylimino)thiazolidin-4-one Example 60 5-(4-MethoxyphenethyO-2-(p-tolylimino)thiazolidin-4-one

Example 61

5-(4-Methoχyphenethyl)-2-(phenylimino)thiazolidin-4-one

Example 62

2-(p-Tolylimino)-5-phenethylthiazolidin-4-one

Example 63 2-p-Tolylimino-5-[2-(3-trifluoromethyl-phenyl)-ethyl1-thiazolidin-4-one

Example 64 5-[2-(4-Fluorophenyl)-ethyl1-2-(pyridin-2-ylimino)-thiazolidin-4-one

Example 65 2-(p-Tolylimino)-5-(3-pheπylpropyl)thiazolidin-4-one

Example 66

2-p-Tolylimino-5-[3-(3-trifluoromethylphenyl)propyηthiazolidin-4-one

Example 67

5-[3-(4-Fluorophenyl)propyn-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 68 5-(3-Phenylallylidene)-2-(phenylimino)thiazolidin-4-one

Example 69 2-p-Tolylimino-5-[(3-trifluoromethylphenylamino)methyl1thiazolidin-4-one

Example 70 5-|^"(4-Fluorophenylamino)methvn-2-(pyridin-2-yliminoHhiazolidin-4-one Example 71 5-{fMethyl-(3-trifluoromethylphenyl)anriinolrnethyl}-2-p-tolylirnino-thiazolidin-4-one

Example 72 5-{[(4-Fluorophenyl)methylamino1methyl)-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 73 2-p-Tolylimino-5-(3-trifluoromethyl-phenoxymethyl)-thiazolidin-4-one

Example 74

5-(4-F[uorophenoxymethvπ-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 75

2-p-Tolylimino-5-(3-trifluoromethylphenylsulfanylmethyl)thiazolidin-4-one

Example 76 5-(4-Fluorophenylsulfanylmethyl)-2-(pyridiπ-2-ylimino)thiazolidin-4-one

Example 77 2-p-Tolylimino-5-[(3-thfluoromethylbeπzylamino)methyl1thiazolidin-4-one

Example 78 5-[(4-Fluorobenzylamino)methyl^'l-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 79

5-{rMethyl-(3-trifluoromethylbenzyl)amino]methyl}-2-p-tolylimino-thiazolidin-4-one

Example 80

5-{[(4-Fluorobenzyl)methylaminolmethyl|-2-(pyridin-2-yliminoHhiazolidin-4-one

Example 81 Λ/-(4-Oxo-2-p-tolylimino-thiazolidin-5-ylmethyl)-3-trifluoromethyl-benzamide

Example 82 4-Fluoro-Λ/-f4-oxo-2-(pyridin-2-ylimino)thiazolidin-5-ylmethvnbenzamide 3

Example 83

Λ/-Methyl-/\/-(4-oxo-2-p-tolylimino-thiazolidiπ-5-ylmethyl)-3-trifluoromethyl- benzamide

Example 84

4-Fluoro-Λ/-methyl-Λ/-[4-oxo-2-(pyridin-2-ylimino^')thiazolidin-5-ylmethvπ- benzamide

Example 85

N-(4-Oxo-2-p-tolylimino-thiazolidin-5-ylmethyl)-2-(3-trifluoromethyl-phenyl)- acetamide

Example 86 2-(4-Fluorophenvπ-Λ/-r4-oxo-2-(pyridin-2-ylimino)thiazolidin-5-ylmethyll-acetamide

Example 87 1-(4-Oxo-2-p-tolyliminothiazolidin-5-ylmethyl)-3-(3-trifluoromethylphenyl)urea

Example 88

1-(4-Fluorophenyl)-3-[4-oxo-2-(pyridin-2-ylimino)thiazolidin-5-ylmethyl1urea

Example 89

(4-0x0-2 -p-tolyliminothiazolidin-δ-ylmethyD-carbamic acid 3-trifluoromethyl- phenyl ester

Example 90 r4-Oxo-2-(pyridin-2-ylimino)thiazolidin-5-ylmethyl1carbamic acid 4-fluorophenyl ester

Example 91

(3-Trifluoromethylphenyl)carbamic acid 4-oxo-2-p-tolyliminothiazolidin-5-ylmethyl ester Example 92

(4-FluorophenvOcarbamic acid 4-oxo-2-(pyridin-2-ylimino)thiazolidin-5-ylmethyl ester

Example 93

5-(4-Chlorobenzyl)-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 94

5-(4-MethoxybenzylV2-(pyridin-2-ylimino)thiazolidin-4-one

Example 95 5-(4-Fluorobenzyl)-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 96 5-(2-MethylbenzvO-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 97 5-(4-Methylbeπzyl)-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 98

5-(2,3-Dichlorobenzyl)-2-(pyridin-2-ylimino)thiazolidin-4-one

Example 99

5-(4-BromobenzylV2-(pyridin-2-ylimino)thiazolidin-4-one

Example 100 5-(3-(Trifluoromethyl)benzyl)-2-(pyridiπ-2-ylimino)thiazolidin-4-one

Example 101 5-(4-Fluorobenzyl)-2-(benzylamino)thiazol-4(5H)-one

Example 102 5-(3-(Trifluoromethyl)benzyl)-2-(benzylimino)thiazolidin-4-one Example 103 2-((Pyridin-2-vπmethylamino)-5-(4-fluorobenzyl)thiazol-4(5H)-one

Example 104 /V-(5-(3-(Trifluoromethyl)benzylH-oxothiazolidin-2-ylidene)benzamide

Example 105 Λ/-(5-(3-(Trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylidene)-4-chlorobenzamide

Example 106

Λ/-(5-(3-(Trifluoromethyl)beπzyl)-4-oxothiazolidin-2-ylidene)-4-methylbenzamide

Example 107

Λ/-(5-(4-Fluorobenzyl)-4,5-dihvdro-4-oxothiazol-2-yl)picolinamide

Example 108

Phenyl 5-(3-(trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylidenecarbamate

Example 109 Pyridin-2-yl 5-(4-fluorobenzyl)-4,5-dihydro-4-oxothiazol-2-ylcarbamate

Example 110 1-(5-(3-(Trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylidene)-3-phenylurea

Example 111

1-(5-(3-(Trifluoromethyl)benzyl)-4-oxothiazolidiπ-2-ylidene)-3-p-tolylurea

Example 112

1-(5-(3-(Trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylidene)-3-(4-chlorophenyl)- urea

Example 113 1-(5-(4-Fluorobenzyl)-4,5-dihvdro-4-oxothiazol-2-yl)-3-(pyridin-2-yl)urea Example 114 5-(3-(Trifluoromethyl)beπzyl)-2-tosyliminothiazolidin-4-one

Example 115 5-(3-(Trifluoroιnethyl)beπzyl)-2-phenylsulfonyliminothiazolidin-4-one

Example 116 5-(3-(Trifluoromethyl)beπzvπ-2-(4-chlorophenvπsulfonyliminothiazolidin-4-one

Example 117

5-(4-Fluorobenzyl)-2-(2-pyridylsulfonylamino)thiazol-4(5H)-one

Example 118

5-(3-(Trifluoromethyl)benzyl)-2-(isopropylamino)thiazol-4(5H)-one

Example 119 5-(3-(Thfluoromethyl)benzyl)-2-(cvclohexylamino)thiazol-4(5H)-one

Example 120 5-(3-(Trifluoromethyl)benzyl)-2-(methylamino)thiazol-4(5H)-one

Example 121 2-(p-Tolylimino)-5-methylthiazolidin-4-one

Example 122

2-(p-Tolylirnino)thiazolidin-4-one

Example 123

5-(3-(Trifluoromethyl)benzyl)-2-aminothiazol-4(5H)-one

Example 124

2-(2-(4-Carboxyphenylimino)-4-oxothiazolidin-5-vπ-Λ/-(3-methoxyphenyl)- acetamide Example 125 2-(2-(4-Hvdroxyphenylimino)-4-oxothiazolidin-5-yl)-Λ/-(4-bromophenyl)-acetamide

Example 126 2-(2-(4-Ethoxyphenylimino)-4-oxothiazolidin-5-yl)-Λ/-(4-bromophenyl)acetamide

Example 127 2-(2-(3-Hvdroxyphenylimino)-4-oxothiazolidin-5-yl)-A/-(4-bromophenyl)-acetamide

Example 128

2-(2-(4-Hvdroxypheπylimino)-4-oxothiazolidiπ-5-yl)-A/-phenylacetamide

Example 129

2-(2-(4-Hvdroxypheπylimino)-4-oxothiazolidin-5-yπ-Λ/-(4-fluorophenvπ-acetamide

Example 130 2-(2-(p-Tolylimino)-4-oxothiazolidin-5-yl)-^-p-tolylacetamide

Example 131 2-(2-(4-MethoxyphenyliminoV4-oxothiazolidin-5-yπ-Λ/-(4-methoχyphenyl)- acetamide

Example 132

2-(2-(4-Ethoxyphenylimino)-4-oxothiazolidin-5-yl)-Λ/-phenylacetamide

Example 133

Ethyl 4-(2-f2-(4-ethoxyphenylimino)-4-oxothiazolidin-5-yl)acetamido)benzoate

Example 134 2-(2-(3-(Trifluoromethyl)phenylimiπo)-4-oxothiazolidin-5-yl)acetic acid

Example 135 N-(2,4-DimethylphenylV2-(4-oxo-2-(phenylimino)thiazolidin-5-yl)acetamide Example 136 Λ/-(2,4-Dimethoxyphenyl)-2-(4-oxo-2-(phenylimino)thiazolidin-5-yl)acetamide

Example 137 2-(4-Oxo-2-(4-sulfonylamidophenylimino)thiazolidin-5-yl)-Λ/-p-tolylacetamide

Example 138 A/-(4-Fluorophenyl)-2-(4-oxo-2-(phenylimino)thiazolidin-5-yl)acetamide

Example 139

2-(2-(m-TolyliminoV4-oxothiazolidin-5-yl)-Λ/-(2-chloropheπyl)acetamide

Example 140

2-(2-(2,5-Dimethylphenylimino)-4-oxothiazolidin-5-yl)-Λ/-(2,4-dichlorophenyl)- acetamide

Example 141 2-(4-Oxo-3-phenyl-2-(phenylimino)thiazolidin-5-yl)-A/-p-tolylacetamide

Example 142

2-(2-(Cvclohexylimino)-4-oxothiazolidin-5-yl)-Λ/-phenylacetamide

Example 143

2-(2-(Methylimino)-4-oxothiazolidin-5-yl)-A/-(2,4-dimethylphenyl)acetamide

Example 144 Λ/-Ethyl-2-(2-(methylimino)-4-oxothiazolidin-5-yl)acetamide

Example 145 2-(2-(Allylimino)-4-oxothiazolidin-5-yl)-Λ/-(2-nitrophenyl)acetamide

Example 146

1 , 1 -Dioxo-1 λ ⁶-[1 ,4.21dithiazolidin-3-ylidene1-p-tolyl-amine Example 147 π .i-Dioxo-δ-O-αrifluoromethvDphenylVhvdroxyimethylViλ^ri Λ.ΣIdithiazo-idiπ-S- ylidenei-p-tolyl-amine

Example 148 ri ,1-Dioxo-5-(3-(trifluoromethyl)benzylidene)-1A⁶-[1 ,4,2ldithiazolidin-3-ylid-enel-p- tolyl-amine

Example 149 ri ,1-Dioxo-5-(3-trifluoromethylbenzvn-1λ⁶-π ,4,2ldithiazolidin-3-ylidenel-p- tolylamine

Example 150 π ,1-Dioxo-5-(4-(fluoro)phenyl)fhvdroxy)methvh-1λ⁶-π ,4,2ldithiazolidin-3-ylidene1- p-tolyl-amine

Example 151

[1 ,1-Dioxo-5-(4-(fluoro)benzylidene)-1λ⁶-[1 ,4,21dithiazolidiπ-3-ylidenel-p-tolyl- amine

Example 152 π ,1-Dioxo-5-(3-(trifluoromethyl)phenyl)(hvdroxy)methyl)-1λ⁶-π ,4,21dithiazol-idiπ-

3-ylidene1-4-chlorophenyl-amine

Example 153 r5-(4-Fluoro-benzyl)-1 ,1-dioxo-1A⁶-ri ,4,21dithiazolidin-3-ylidene1-pyhdin-2-yl- amine

Example 154 2-(1 ,1-Dioxo-3-)D-tolylimino-1λ⁶-π ,4.21dithiazolidin-5-yl^')-A/-p-tolyl-acetamide

Example 155 5-(3-(Trifluoromethyl)benzyl)-4-methyl-/V-p-tolylthiazol-2-amine Example 156 Λ/-(5-(4-Fluorobenzyl)-4-methylthiazol-2-yQpyridin-2 -amine

Example 157 5-(3-(Trifluoromethyl)benzyl)-4-(trifluoromethvπ-N-p-tolylthiazol-2-amine

Example 158 A/-(5-(4-Fluorobenzyl)-4-(trifluoromethy])thiazol-2-yl)pyridin-2 -amine

Example 159

2-(4-Chlorophenylimino)-5-((5-methylfuran-2-yl)methylene)thiazolidin-4-one

Example 160

2-(4-Chlorophenylimino)-5-((5-methylfuran-2-yl)methyl)thiazolidin-4-one

Example 161 2-(4-Chlorophenylimino)-5-((5-methylthiophen-2-yl)methylene)thiazolidin-4-one

Example 162 2-(4-Chlorophenylimino)-5-((5-methylthiophen-2-yl)methyl)thiazolidin-4-one

Example 163 5-(3-(Trifluoromethyl)benzyl)-2-(p-tolylimino)oxazolidin-4-one

Example 164

[5-(3-Trifluoromethylbenzyl)-1 ,1-dioxo-1λ⁶-l^'1 ,4,21dithiazolidin-3-ylidenel-(4- chloro)phenyl-2-amine

Example 165 r5-(3-Trifluoromethylbenzyl)-1 ,1 -dioxo-1λ⁶-π ,4,21dithiazolidin-3-ylidene1-2- benzamide

Example 166 5-(3-(Trifluoromethyπbenzyl)-4-methyl-N-(4-chlorophenvhthiazol-2-amine Biological Tests

Test A

Cell Proliferation Assay

Reagents

Dulbecco's modified Eagle's medium (D-MEM) +1000mg/L Glucose +GlutaMAX™1 + Pyruvate (Gibco #21885-025) VA/ Foetal Bovine Serum (Gibco 10500-064) PEST (100 U/ml penicillin, 100ug/ml streptomycin, Gibco 15140-122) CyStain Pl absolute T Kit (Partec # 05-5023) Linolenic acid 99%, L2376 from Sigma Aldrich Dimethyl sulfoxide (DMSO)

Equipment

Cytomics™ FC500 Flow Cytometer with CXP software (Beckman Coulter)

MDA-MB-231 cells

MDA-MB-231 cells were cultured in the propagation media D-MEM +1000mg/L Glucose +GlutaMAX™1 ÷Pyruvate supplemented with 10% VA/ Foetal Bovine Serum and PEST (100 U/mL penicillin, 100 μg/mL streptomycin). Cells were seeded in 6 well plates to a density of 300 000 cells/well in propagation media. After 24 hours, media was replaced with serum free D-MEM media.

Linolenic acid was diluted in DMSO to a concentration of 100 mM and added to the culture media to a final concentration of 100 μM.

Compounds were as dissolved in DMSO to a concentrations of 10 mM (Compounds of Examples 95 and 6 (Compound X and Compound Y, respectively)) and 40 mM (Compound of Example 4 (Compound Z)) and added to the culture media to a final concentration of 10 μM (X and Y) and 40 μM (Z) respectively.

After 24 hours in serum free media DMEM, linolenic acid (to a final concentration of 10 μM) and compounds to be screened for activity were added to a final concentration of 10 μM (Compounds X and Y) and 40 μM (Compound Z) respectively. Final DMSO concentration was kept at 0.2% in all wells. After 24 hours of stimulation, cells were harvested and propidium iodine stained using a CyStain Pl absolute T Kit according to manufacturer's recommendations. Cells were subsequently analyzed using a Cytomics™ FC500 Flow Cytometer with CXP software (Beckman Coulter) for cell cycle distribution. Cells were incubated with or without linolenic acid (LA) and the Compounds X, Y and Z for 24 hours at indicated concentrations. Cells in S-phase from untreated sample were set to 100% in each experiment.

Results

The described method was shown to exhibit the sensitivity required to detect an antagonist to free fatty acid stimulation. The measurement of DNA synthesis for quantification of cell proliferation minimizes errors inherent in several other assays.

It was observed that FFA stimulation of MDA-MB-231 cells leads to an increased proliferation as demonstrated in Figure 1a and 1b, where the proportion of cells in S-phase of the cell cycle is increased in b versus a as measured by propidium iodine incorporation. This stimulatory effect of FFA could be attenuated by Compound X in a 10:1 molar ratio (Figure 1 c). These results indicate that Compound X is able to antagonize free fatty acid stimulated cell proliferation.

The experiment described was repeated 4 times and the results are summarized in Figure 2A. Compounds Z and Y were also able to antagonize free fatty acid stimulated cell proliferation, as shown Figures 2B and 2C, respectively.

Thus, the relevant compounds attenuate the FFA induced cell proliferation in a human breast cancer cell line. The ability of Compounds X, Y and Z to inhibit such proliferation may be expressed as percentage antagonist activity as follows: Compound X - 70% at a concentration of 10 μM Compound Y - 100% at a concentration of 10 μM Compound Z - 50% at a concentration of 10 μM. Similar experiments were conducted in respect of compounds of the examples above, which were also found to exhibit percentage antagonist activities at least 20% at a concentration of 10 μM.

Test B

In vivo Mouse Model

5 week old Athymic BALB/cA nude mice were delivered from Taconic (Denmark) and kept under barrier conditions for 1 week acclimatisation. At 6 weeks, 17 mice were injected subcutaneously on the flank with 1.8 x 10⁶ MDA-MB-231 human breast cancer cells (LGC Promochem-ATCC) in a 50/50 v/v solution of phosphate buffered saline (PBS) (Gibco 10010-015, Invitrogen) Matrigel HC (BD Biosciences).

After 11 days, palpable tumors were observed in 16 mice. 2 mice were sacrificed and the tumors dissected and examined. 2 groups of 7 mice each were treated once daily by intraperitoneal injections of 1 mg/kg bodyweight of the compund of Example 6 (Compound Y) in PBS/1 %v/v dimethylsufoxide or vehicle control respectively for 9 days. The mice were sacrificed by cervical dislocation and tumors were dissected.

Histology

The tumor tissue were fixated overnight in PBS (containing 4% w/v paraformaldehyde (Scharlau PA0095, Sharlau Chemie SA, Spain) at +4⁰C. The tumor tissue were then cryopreserved by 24 hour incubation in PBS containing 30% w/v sucrose (BDH #102745C (www.vwr.com)) at +4⁰C and embedded in Tissue-Tek embedding media (Sakura Finetek Europa BV, Netherlands). 10 μm cryosections were generated an stained with Mayers Hematoxylin (Dako) for 5 min and destained for 3 x 10 minutes in tap water. Slides were mounted using Dako faramount aqueous mounting medium and examined using a Nikon Eclipse TS 100 microscope documented using a Nikon coolpix 4500. Results

The tumors from mice treated with test compound and vehicle were analyzed for morphology by microscopic examination of hematoxylin stained cryosections. The results are shown in Figures 3A to 3F.

Figure 3A shows a hematoxylin stained section from a tumor dissected from a vehicle treated mouse at 1Ox magnification. It is to be noted that there is a relative abundance of cells in the interior of the section as well as the relative thickness of the uninterrupted zone of cell in the periphery of the section.

Figure 3B shows a hematoxylin stained section from a tumor dissected from a vehicle treated mouse at 2Ox magnification. It is to be noted that the cells in the interior of the section display morphology consistent with adenocarcinoma.

Figure 3C shows a hematoxylin stained section from a tumor dissected from a vehicle treated mouse at 4Ox magnification. It is to be noted that no cell displaying morphology indicative of macrophage/monocyte could be found.

Figure 3D shows a hematoxylin stained section from a tumor dissected from a mouse treated with the Compound Y at 10x magnification. The low cell density in the interior of the section and the thin layer of cells displaying morphology is to be noted, which is consistent with poorly differentiated adenocarcinoma.

Figure 3E shows a hematoxylin stained section from a tumor dissected from mouse treated with the Compound Y at 2Ox magnification. The lack of cells displaying fibroblast morphology in the interior of the section is to be noted.

Figure 3F shows a hematoxylin stained section from a tumor dissected from a mouse treated with the compound of Compound Y at 4Ox magnification. The accumulation of cells displaying morphology indicative of macrophage/monocyte in the interior of the section (black arrows) is to be noted.

Thus, the main finding was that the cell-density in the interior of the tumors was markedly reduced in tumors dissected from test compound treated mice as compared to tumors from vehicle treated mice. Moreover, the majority of the cells found in the interior of the sections from the treated group displayed a morphology inconsistent with adenocarcinoma while cells displaying macrophage/monocyte morphology was a frequent finding. In contrast, only one of seven tumors from the vehicle treated group showed indication of macrophage/monocyte infiltration.

In summary, these findings show a correlation between treatment with test compound and reduction of cancer cells in the xenograft tumors.

Test C

Proliferation Assay by BrdU Incorporation.

5,000 SK-BR-3 human breast cancer cells/well are plated in 96-well plates containing complete culture media (DMEM 10% Fetal Calf Serum, Gibco).

After 24 hours, the media are changed to serum free media. 24 hours later, trastuzumab (Herceptin) and Compound Y are added to a final concentration of 10 μg/mL and 10 μM, respectively.

After 18 hours of incubation, bromodeoxyuridine (5-bromo-2-deoxyuridine; BrdU) is added according to the manufacturer's recommendation (Cell proliferation ELISA, BrdU, Roche).

6 hours later, media are removed and cells are fixed and BrdU incorporation is measured according to the manufacturer's recommendation.

The trastuzumab treated cells and the Compound Y treated cells show a significant reduction in proliferation. The combination of 10 μg/mL trastuzumab and 10 μM Compound Y displays a synergistic effect on cell proliferation.

5S

Claims

Claims

1. A combination product comprising: (a) a compound of formula I,

wherein

X represents -[C(R₈)(R₉)I_n-; n represents 0, 1 , 2 or 3; Y represents -C(O)-, -S(O)₂- or =C(R₁₀)-;

T represents -S- or -O-;

W represents -NR₇-, -CR₇R₇-, -NR₇C(O)-, -NR₇S(O)₂-, -NR₇C(O)NR₇-,

-NR₇C(O)O- or a bond; one of A₁ or A₂ represents a double bond and the other represents a single bond; when A₁ represents a single bond, A₂ is a double bond and R₆ is absent; when A₂ represents a single bond, A₁ is a double bond and, if present, one R₇

(which is attached α to the requisite ring of the compound of formula I) is absent;

R₁ represents -C(O)NR₃R₂, -NR₃R₂, -C(O)OR₂, -NR₄C(O)NR₃R₂, -NR₄C(O)OR₂,

-OC(O)NR₃R₂, -NR₄C(O)R₂, -OC(O)R₂ , -OR₂, -SR₂, H, alkyl, cycloalkyl, heterocyclyl, benzyl, aryl or heteroaryl (which latter six groups are optionally substituted by one or more groups selected from B¹, B², B³, B⁴, B⁵ and B⁶, respectively);

R₂ and R₅ independently represent, on each occasion when used herein, hydrogen, alkyl, cycloalkyl, heterocyclyl, benzyl, aryl or heteroaryl (which latter six groups are optionally substituted by one or more groups selected from B⁷, B⁸, B⁹,

B¹⁰, B¹¹ and B¹², respectively);

R₃, R₄, R₆ and R₇ independently represent, on each occasion when used herein, hydrogen, alkyl, cycloalkyl, aryl or benzyl (which latter four groups are optionally substituted by one or more groups selected from B¹³, B¹⁴, B¹⁵ and B¹⁶, respectively), or heterocyclyl or heteroaryl (which latter two groups are optionally substituted by one or more groups selected from B¹⁴ and B¹⁵, respectively); R₈ and R₉ are independently selected from hydrogen, alkyl and aryl (which latter two groups are optionally substituted by B^16a and B^16b, respectively); Rio represents hydrogen, alkyl or aryl (which latter two groups are optionally substituted by one or more groups selected from B¹⁷ and B¹⁸, respectively); B¹ to B¹⁸ independently represent cyano, -NO₂, halo, -ORn, -NR₁₂Ri3. -SRu, -Si(Ri₅)3, -C(O)OR₁₆, -C(O)NR_16aRi6b, -S(O)₂NRi6cRi6d, aryl or heteroaryl (which aryl and heteroaryl groups are themselves optionally and independently substituted by one or more groups selected from halo and Ri₇); or, alternatively, B⁴, B⁵, B⁶, B¹⁰, B¹¹, B¹², B¹⁵, B¹⁶, B^16b or B¹⁸ independently represent R₁₇; Ri₁, R₁₂, Ri3, Ri4, Ri6, Riea, Ri6b, Rise and R₁₆C independently represent H or R₁₇; and

R₁₅ and Ri₇ independently represent, on each occasion when used herein, C₁^ alkyl optionally substituted by one or more halo atoms, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof, provided that, when n represents 0 and Ri represents an optionally substituted alkyl group, then that alkyl group is saturated; and

(b) a compound selected from trastuzumab, or a tyrosine kinase inhibitor, or a pharmaceutically-acceptable salt, solvate or pharmaceutically functional derivative of a tyrosine kinase inhibitor.

2. A combination product as claimed in Claim 1 wherein, in the compound of formula I, T represents -S-,

3. A combination product as claimed in Claim 1 or Claim 2 wherein, in the compound of formula I, Y represents -C(O)-.

4. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, Ri₀ represents H or alkyl.

5. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, W represents -NR₇-, -NR₇C(O)-, -NR₇C(O)O-, -NR₇C(O)NR₇- or -NR₇S(O)₂-.

6. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ R₅ represents optionally substituted C₁.3 alkyl, cycloalkyl or optionally substituted phenyl or optionally substituted heteroaryl.

7. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ n represents 1 , 2 or 3.

8. A combination product as claimed in Claim 7, wherein n represents 1 or 2.

9. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ R₈ and R₉ independently represent C₁.3 alkyl or H.

10. A combination product as claimed in Claim 9, wherein Rs and Rg both represent H.

1 1. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ R₁ represents alkyl, -NR₃R₂, -OR₂, -SR₂, -NR₄C(O)R₂, -NR₄C(O)NR₃R₂, -NR₄C(O)OR₂, -C(O)NR₃R₂, -C(O)OR₂, optionally substituted heteroaryl or optionally substituted phenyl.

12. A combination product as claimed in Claim 1 1 , wherein R₁ represents aryl optionally substituted by B⁵.

13. A combination product as claimed in Claim 11 wherein R₁ represents optionally substituted furanyl, thienyl or phenyl.

14. A combination product as claimed in Claim 12 wherein R₁ represents phenyl substituted by B⁵.

15. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ R₄ or R₃ independently represent Ci_-3 alkyl or H.

16. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, R₂ represents optionally substituted d.₃ alkyl, optionally substituted phenyl or H;

17. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, when W represents -NR₇- and R₇ is absent, then R₆ represents H, Ci_-6 alkyl or phenyl, which latter two groups may be substitutued by one or more of B¹³ and B¹⁵, respectively.

18. A combination product as claimed in any one of Claims 1 to 16 wherein, in the compound of formula I, when W represents -NR₇- and R₆ is absent, then R₇ represents Ci_-3 alkyl, phenyl or benzyl, all of which may be substituted by one or more B¹³, B¹⁵ and B¹⁶, respectively.

19. A combination product as claimed in any one of Claims 1 to 16 wherein, in the compound of formula I, when W represents -CR₇R₇-, then A₂ represents a double bond.

20. A combination product as claimed in any one of Claims 1 to 16 wherein, in the compound of formula I, when W represents -CR₇R₇-, then each R₇ independently represents Ci_-3 alkyl or H.

21. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, R₆ represents H.

22. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ B¹ to B¹⁸ independently represent cyano, NO₂, halo, -ORn, -C(O)ORi₆, -C(O)NRi_6aR_16b or -S(O)₂NRi_6cRi6d; and/or B⁴ to B⁶, B¹⁰ to B¹², B¹⁵, B¹⁶ and B¹⁸ independently represent Ri₇; and/or B¹ to B¹⁸ independently represent heteroaryl or phenyl, both of which may be substituted by one or more groups selected from halo or Ri₇.

23. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, B⁵ represents R₁₇.

24. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, R₁₇ represents C1.4 alkyl optionally substituted by one or more halo atoms.

25. A combination product as claimed in Claim 24 wherein Ri₇ represents C_1-3 alkyl substituted by one or more halo atoms.

26. A combination product as claimed in Claim 25 wherein Ri₇ represents Ci_-3 alkyl substituted by one or more fluoro atoms.

27. A combination product as claimed in Claim 26 wherein Ri₇ represents methyl substituted by one or more fluoro atoms.

28. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ Rn represents C_1-3 alkyl or H.

29. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I₁ R₁₆ represents H or C_1-3 alkyl.

30. A combination product as claimed in any one of the preceding claims wherein, in the compound of formula I, R_16a, Ri6t» Ri6c and Riβd independently represent C_1-2 alkyl or H.

31. A combination product as claimed in any one of the preceding claims where the compound of formula I is selected from:

5-(4~fluorobenzyl)-2-(pyridin-2-ylimino)thiazolidin-4-one;

5-(p-methylbenzy!)-2-(4-chlorophenylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(p-tolylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(4-chlorophenylimino)thiazolidin-4-one; 5-(3-(trifluoromethyl)benzyl)-2-(4-isopropylphenylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(4-methoxyphenylimino)thiazolidin-4-one;

5-(3-(trifluoromethyl)benzyl)-2-(phenylimino)thiazolidin-4-one;

2-(3,4-dichlorophenylimino)-5-(3-(trifluoromethyl)benzyl)thiazolidin-4-one;

2-(2,4-dichlorophenylimino)-5-(3-(trifluoromethyl)benzyl)thiazolidin-4-one; 5-(3-(trifluoromethyl)benzyl)-2-(p-tolylimino)-3-methylthiazolidin-4-one; N-(5-(3-(tnfluoromethyl)benzyl)-4-oxothiazoIidin-2-ylidene)-4-chlorobenzamide; 5-(3-(trifluoromethyl)benzyl)-2-(4-chlorophenyl)sulfonyliminothiazolidiπ-4-one; phenyl 5-(3-(trifluoromethyl)benzyl)-4-oxothiazolidin-2-ylidenecarbamate; 5-(4-methoxyphenethyl)-2-(p-tolylimino)thiazolidin-4-one; 5-(4-methoxyphenethyl)-2-(phenylimino)thiazolidin-4-one; and 2-(p-tolylimino)-5-phenethylthiazolidin-4-one.

32. A combination product as claimed in Claim 31 wherein the compound is 5-(3- (trifluoromethyl)benzyl)-2-(4-chlorophenylimino)thiazolidin-4-one.

33. A combination product as claimed in any one of the preceding claims which comprises a pharmaceutical formulation including a compound of formula I, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof; trastuzumab or a tyrosine kinase inhibitor, or a pharmaceutically-acceptable salt, solvate or pharmaceutically functional derivative of a tyrosine kinase inhibitor; and a pharmaceutically-acceptable adjuvant, diluent or carrier.

34. A combination product as claimed in any one of Claims 1 to 32 which comprises a kit of parts comprising components:

(A) a pharmaceutical formulation including a compound of formula I, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof, in admixture with a pharmaceutically- acceptable adjuvant, diluent or carrier; and (B) a pharmaceutical formulation including trastuzumab or a tyrosine kinase inhibitor, or a pharmaceutically-acceptable salt, solvate, or pharmaceutically functional derivative of a tyrosine kinase inhibitor, in admixture with a pharmaceutically-acceptable adjuvant, diluent or carrier, which components (A) and (B) are each provided in a form that is suitable for administration in conjunction with the other.

35. A combination product as defined in any one of the preceding claims comprising a compound of formula I, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof, and trastuzumab.

36. A combination product comprising (a) a compound of formula I as defined in any one of the preceding claims, or a pharmaceutically-acceptable salt or solvate, or a pharmaceutically functional derivative thereof; and (b) a compound selected from a glitazone, metformin, a statin, an antibody (other than trastuzumab) that is useful in the treatment of cancer, or an inhibitor of activity of the mammalian target of rapamycin, or a pharmaceutically-acceptable salt, solvate or pharmaceutically functional derivative of a glitazone, metformin, a statin or an inhibitor of activity of the mammalian target of rapamycin.

37. A combination product as claimed in Claim 36 wherein the glitazone is rosiglitazone.

38. A combination product as claimed in Claim 36 wherein the statin is selected from fluvastatin, simvastatin, rosuvastatin, pravastatin, atorvastatin and lovastatin.

39. A combination product as claimed in Claim 38 wherein the statin is lovastatin.

40. A combination product as claimed in Claim 36 wherein the antibody is bevacizumab, cetuximab or panitumumab.

41. A combination product as claimed in Claim 36 wherein the inhibitor of activity of the mammalian target of rapamycin is rapamycin or a pharmaceutically- acceptable salt, solvate or pharmaceutically functional derivative thereof.

42. A combination product as claimed in Claim 41 wherein the inhibitor of activity of the mammalian target of rapamycin is rapamycin.

43. A kit of parts comprising:

(I) one of components (A) and (B) as defined in Claim 34 or any one of Claims 35 to 42 (as dependent on Claim 34); together with

(II) instructions to use that component in conjunction with the other of the two components.

44. A method of making a kit of parts as defined in Claim 34 or any one of Claims 35 to 42 (as dependent on Claim 34), which method comprises bringing a component (A) into association with a component (B), thus rendering the two components suitable for administration in conjunction with each other.

45. A kit of parts as claimed in Claim 34, any one of Claims 35 to 42 (as dependent on Claim 34) or Claim 43, wherein components (A) and (B) are suitable for sequential, separate and/or simultaneous use in the treatment of cancer.

46. The use of a combination product as claimed in any one of Claims 1 to 42, or a kit of parts as defined in Claim 43 or Claim 45, for the manufacture of a medicament for the treatment of cancer.

47. A method of treatment of cancer, which method comprises the administration of a combination product as claimed in any one of Claims 1 to 42, or a kit of parts as defined in Claim 43 or Claim 45, to a patient in need of such treatment.

48. A kit of parts as claimed in Claim 45, a use as claimed in Claim 46, or a method as claimed in Claim 47, wherein the cancer is of the colon, the breast or the prostate.

49. A kit of parts, use or method as claimed in Claim 48 wherein the cancer is of the breast.

50. A kit of parts, use or method as claimed in Claim 48 or Claim 49 wherein the cancer is an HER2-positive cancer.