WO2010133672A1 - Derivatives of quinoline-3-carboxylic acid and their medical use - Google Patents

Abstract

A compound of formula (I) as well as pharmaceutically acceptable salts thereof and pharmaceutical compositions comprising a therapeutically effective amount of the compounds. The compounds are useful in treatment of cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema or psoriasis.

Description

DERIVATIVES OF QUINOLINE-3-CARBOXYLIC ACID AND THEIR MEDICAL USE

FIELD OF THE INVENTION

The present invention relates to quinoline-3-carboxylic acid derivatives and to the use 5 thereof in therapy. Particularly, the present invention relates to their use for the treatment of cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema and psoriasis.

BACKGROUND OF THE INVENTION 0 The following review of the state of art is only provided to aid the understanding of the present invention and neither it nor any of the references cited within it are admitted to be prior art to the present invention.

Angiogenesis, the outgrowth of new capillaries from pre-existing vessels, is essential for embryonic development, organ formation, tissue regeneration, and remodeling [Folkman, J. & 5 Shing, Y. (1992) J. Biol. Chem. 267, 10931-10934]. It also contributes to the development and progression of a variety of pathological conditions, including tumor growth and metastasis, cardiovascular diseases, diabetic retinopathy, rheumatoid arthritis, psoriasis [Folkman, J. Nat.

Med. 1995, 1, 27-30] and age-related macular degeneration [Barakat, M. R.; Kaiser, P. K. Expert

Opin. Investig. Drugs 2009, 18, 637-46; Chappelow, A. V.; Kaiser, P. K. Drugs 2008, 68, 1029- :0 1036].

Angiogenesis and vasculogenesis are complex multistep processes that include proliferation, migration and differentiation of endothelial cells, degradation of the extracellular matrix, tube formation, and sprouting of new capillary branches [Hanahan, D.; Folkman, J. Cell

1996, 86, 353-364; Risau, W. Nature (London) 1997, 386, 671-674]. The complexity of the ,5 angiogenic processes suggests the existence of multiple controls of the system, which can be transiently switched on and off. A switch of the angiogenic phenotype in tissues is thought to depend on a local change of the balance between angiogenic stimulators and inhibitors

[Folkman, J. N Engl. J. Med. 1995, 333, 1757-1763].

Among many described angiogenic factors, vascular endothelial growth factor (VEGF)/ 0 vascular permeability factor is one of the best-characterized positive regulators with its distinct specificity for vascular endothelial cells [Senger, D. R.; Galli, S. J.; Dvorak, A. M.; Perruzzi, C.

A.; Harvey, V. S.; Dvorak, H. F. Science 1983, 219, 983-985; Ferrara, N.; Henzel, W. J.

Biochem. Biophys. Res. Commun. 1989, 161, 851-858; Gospodarowicz, D.; Abraham, J. A.;

Schilling, J. Proc. Natl. Acad. ScL USA 1989, 86, 7311-7315]. The biological actions of VEGF include stimulation of endothelial cell proliferation, migration, differentiation, tube formation, increase of vascular permeability, and maintenance of vascular integrity [Mustonen, T.; Alitalo, K. J Cell Biol. 1995, 129, 895-898; Ferrara, N.; Davis-Smyth, T. Endocr. Rev. 1997, 18, 4-25; Thomas, K. J Biol. Chem. 1996, 271, 603-606; Risau, W. Nature (London) 1997, 386, 671-674; Breier, G.; Risau, W. Trends Cell Biol. 1997, 6, 454-456]. The angiogenic responses induced by VEGF are mediated by tyrosine kinase receptors, which are expressed primarily on vascular cells of the endothelial lineage [Mustonen, T.; Alitalo, K. J Cell Biol. 1995, 129, 895-898; De Vries, C; Escobedo, J. A.; Ueno, H.; Huck, K.; Ferrara, N.; Williams, L. T. Science 1992, 255, 989-99; Terman, B. L; Dougher-Vermazen, M.; Carrion, M. E.; Dimitrov, D.; Armellino, D. C; Gospodorawicz, D.; Bohlen, P. Biochem. Biophys. Res. Commun. 1992, 187, 1579-1586].

Inhibition of cell adhesion to the endothelial cell membrane (ECM), the fundamental step for activation, survival, targeting and migration of activated endothelial cells, might be one of the most promising target mechanisms for anti-angiogenesis. Not only VEGF is involved in these mechanisms but many of these interactions are also mediated by integrins, a family of multifunction cell adhesion receptors [Stupack, D. G. Oncology (Williston Park) 2007, 21 (9 Suppl 3), 6-12; Avraamides, C. J.; Garmy-Susini, B.; Varner, J. A. Nat. Rev. Cancer 2008, 8, 604-17.]. Members of the integrin family are non-covalent alpha/beta heterodimers that mediate cell-cell, cell-extracellular matrix and cell-pathogen interactions. They are also are believed to modulate the effect of receptors for vascular endothelial growth factor (VEGFRs) [Napione, L.; Cascone, L; Mitola, S.; Serini, G.; Bussolino, F. Autoimmun. Rev. 2007, 7, 18-22].

Until now, 19 different integrin alpha subunits and 8 different beta subunits are known that combine to form at least 24 different alpha/beta heterodimers with different ligand specificity [Silva, R.; D'Amico, G.; Hodivala-Dilke, K. M.; Reynolds, L. E. Arterioscler Thromb Vase Biol, 2008, 28, 1701-1713]. Of the presently approximately 24 known integrins, 16 have been reported to have involvement in some aspects of vascular biology. Of these αlβl, α2βl, α3βl, α5βl, α6βl, α6β4, αvβ3, and αvβ5 are known to be present in endothelial cells [Rupp, P. A.; Little, C. D. Circ. Res., 2001, 566-572; Stupack, D. G.; Cheresh, D. A. Sci. STKE, 2002, PE7], while vascular smooth muscle cells have been reported to have αlβl, α2βl, α3βl, α4βl, α5βl, α6βl, α7βl, α8βl,α9βl, αvβl, αvβ3, αvβ5, and α6β4 [Moiseeva, E. P. Cardivasc. Res., 2001, 372- 386].

The ligands for the extracellular domain of many integrins are the proteins of the extracellular matrix and the intracellular domain of the integrins are either directly or indirectly connected to intracellular components such as kinases and the cytoskeleton. Integrins serve as bidirectional signalling receptors, whereby protein activities and gene expression are changed by integrins in response to ligand binding to the extracellular domain thereof, which is also referred to as outside-in-signalling. On the other hand, the affinity of the integrins is modulated in response to intracellular changes such as binding of proteins to the extracellular domain of the integrin, which is referred to as inside-out signalling [Humphries, M. J. Biochem. Soc. Trans.

5 2000, 28, 311-339; Hynes, R. O. Cell, 2002, 110, 673-687].

Several studies on the integrin pattern on activated endothelial cells, mice gene knockouts and inhibition studies in angiogenic animal models with antibodies, peptides and small molecules have provided information about integrins and ECM proteins involved in critical steps of angiogenesis [Brooks, P. C; Clark, R. A.; Cheresh, D. A. Science, 1994, 264, 569-571;

0 Brooks, P. C. Eur. J. Cancer, 1996, 32A, 2423-2429; Mousa, S. A. Curr Opin Chem Biol, 2002, 6, 534-541; Hynes, R. O. Nature Medicine 2002, 8, 918-21; Kim, S.; Bell, K.; Mousa, S. A.; Varner, J. A.; Am. J. Pathol. 2000, 156, 1345-1362].

From studies referred to herein above it appeared that the vitronectin receptors αvβ3, αvβ5 and the fibronectin receptor α5βl play a critical role in angiogenesis. Integrin α5βl expression

5 is significantly upregulated in blood vessels in human tumors and after stimulation with growth factors and, once expressed, α5βl regulates the survival and migration of endothelial cells in vitro and in vivo. Integrin α5βl is poorly expressed on quiescent endothelium but its expression is significantly upregulated on endothelium during tumor angiogenesis in both mice and humans, which makes α5βl a viable target for anti-angiogenic therapy [Kim, S.; Bell, K.; Mousa, S. A.;

!0 Varner, J. A.; Am. J. Pathol. 2000, 156, 1345-1362; Bhaskar, V.; Zhang, D.; Fox, M.; Seto, P.; Wong, M. H.; Wales, P. E.; Powers, D.; Chao, D. T; Dubridge, R. B.; Ramakrishnan, V. J. Transl. Med. 2007, 27, 61]. Expression of this integrin is also upregulated during corneal angiogenesis [Muether, P. S.; Dell, S.; Kociok, N.; Zahn, G.; Stragies, R.; Vossmeyer, D.; Joussen, A. M.; Exp. Eye. Res. 2007, 85, 356-365].

'5 Combination of anti-angiogenetic therapy and other therapeutic approaches, such as chemotherapy, radiotherapy and gene therapy has also been applied and suggested for cancer treatment. Mounting evidence suggests that there is potentially synergistic effect of combined therapeutic approaches over single modality alone [Huveneers, S.; Truong, H.; Danen, H. J. Int. J. Radial Biol. 2007, 83, 743-751; Huber, P. E.; Bischof, M.; Jenne, J.; Heiland, S.; Peschke, P.;

0 Saffrich, R.; Grδne, H. J.; Debus, J.; Lipson, K. E.; Abdollahi, A. Cancer Res. 2005, 65, 3643- 3655]. SUMMARY OF THE INVENTION

The present inventors now have found that novel quinoline derivatives with certain side- chain patterns, compared to similar analogs in the field have improved in vitro or in vivo properties when used in therapy.

Consequently, according to one aspect, the present invention relates to a compound of formula (I)

(I) wherein:

LO n is 0 (zero) or 1;

R and R are independently selected from hydrogen; branched or unbranched Cj-C₈ alkyl, C₂-C₈ alkenyl, or C₂-C₈ alkynyl; monocyclic or bicyclic, saturated or unsaturated C₃-C₈ carbocyclyl; and monocyclic or bicyclic, saturated or unsaturated C₁-C₇ heterocyclyl wherein the heteroatoms are independently selected from N, O and S;

[5 said alkyl, alkenyl, alkynyl, carbocyclyl or heterocyclyl optionally being substituted with 1,

2, or 3 groups R^a;

R³ is selected from monocyclic or bicyclic C₆-C₁₀ aryl; and monocyclic or bicyclic C₁-C₉ heteroaryl, wherein the heteroatoms independently are selected from N, O and S; said aryl or heteroaryl optionally being substituted with 1, 2, 3, 4 or 5 groups R ; >0 Y is selected from -C(O)-; -S(O)-; and -S(O)₂-;

X is selected from -NR^C-; -0-; and -S-; each R is independently selected from halogen; hydroxy; carbonyl; methoxy; halomethoxy; dihalomethoxy; and trihalomethoxy; each R is independently selected from halogen, branched or unbranched C₁-C₄ alkyl, C₂- .5 C₄ alkenyl or C₂-C₄ alkynyl; branched or unbranched C₁-C₄ alkyloxy, C₂- C₄ alkenyloxy or C₂- C₄ alkynyloxy; branched or unbranched C₁-C₄ alkylthio, C₂-C₄ alkenylthio or C₂-C₄ alkynylthio; wherein any alkyl, alkenyl, alkynyl, alkyloxy, alkenyloxy alkynyloxy, alkylthio, alkenylthio or alkynylthio group optionally is substituted with 1, 2 or 3 halogens; R is selected from hydrogen and branched or unbranched Cj-C₄ alkyl, C₂-C₄ alkenyl or C₂-C₄ alkynyl; or a pharmaceutically acceptable salt thereof.

Another aspect of the invention relates to a compound of formula (I) as defined herein above, or a pharmaceutically acceptable salt thereof, for use in therapy.

Another aspect of the invention relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) as defined herein above, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable excipient. hi one embodiment of this aspect said pharmaceutical composition comprises at least one further, pharmaceutically active compound. Said further pharmaceutically active compound may have anti-tumor activity.

Another aspect of the invention provides compounds of formula (I) or pharmaceutically acceptable salts thereof, for use in the treatment of diseases or disorders such as cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema and psoriasis.

Another aspect of the invention provides the use of the compounds of formula (I) or pharmaceutically acceptable salts thereof, in the manufacture of a medicament for the treatment of disorders such as cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema and psoriasis. Another aspect of the invention provides a method of treating a mammal suffering from cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema or psoriasis, comprising administering to said mammal in need thereof, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment of this aspect, said mammal is a human.

Another aspect of the invention provides a method of treating a mammal suffering from a disease or disorder related to VEGFR tyrosine kinase or integrin activity, comprising administering to said mammal in need thereof, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment of this aspect, said mammal is a human.

Another aspect of the invention provides a method of treating a mammal suffering from cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema or psoriasis, comprising administering to said patient in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a second therapeutic agent that inhibits VEGF, VEGFR tyrosine kinase or integrin. In another embodiment of this aspect, said second therapeutic agent is a therapeutic antibody. In one embodiment of this aspect, said second

5 therapeutic agent is selected from an alkylating agent; a folic acid antagonist; an antimetabolite of nucleic acid metabolism; a pyrimidine analog; 5-fluorouracil; and a purine nucleoside. In another embodiment of this aspect, said mammal is a human. In another embodiment of this aspect, said second therapeutic agent is administered in combination or sequentially with the first therapeutic agent.

10 Another aspect of the invention provides a method of treating a patient suffering from cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema or psoriasis, comprising administering to said patient in need thereof a therapeutically effective amount of a compound of formula (I) in combination with radiological treatment, including irradiation and/or administration of a

5 radioactive substance.

Another aspect of the invention provides a method of treating a patient suffering from cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema or psoriasis, comprising administering to said patient in need thereof a therapeutically effective amount of a compound of formula (I) or a

!0 pharmaceutically acceptable salt thereof in combination with at least two of the treatment mentioned above. Such a method can involve the combination a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with any antiangiogenic agent, radiological treatment and/or chemotherapy.

Further aspects and embodiments of the invention are as defined in the claims.

'.5

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to substituted quinoline derivatives, which can be utilized to treat diseases and conditions such as cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema,

0 psoriasis, and the like in mammals.

The preparation of the compounds of the invention lies well within the capability of the person skilled in the art. As an example, a quinoline-3-carboxylic acid ester of the invention may be formed in a six step procedure wherein, first, a suitable halo aniline derivative is reacted with a suitable mono- or diethylester, the formed intermediate cyclized to give a 4-halo-quinoline-3- carboxylic acid ester, which is then coupled with a suitable amine, H(R^c)N-(CH₂)n-R³ to form a substituted secondary or tertiary 4-amino quinoline-3 -carboxylic acid ester. The halogen can then be carbonylated, to yield the corresponding amide, -C(O)-NR¹R². It this context, it should be obvious for the one skilled in the art that a substituted sulphonamide, -S(O)₂-NR R , can be prepared via reaction of the halogen with sulfite ion, followed by further manipulation to yield the corresponding sulphonamide or corresponding sulfoxide. The quinoline-3 -carboxylic acid ester can then be hydrolysed to give the corresponding inventive compounds. The entire synthesis is illustrated by Reaction Scheme 1. With regard to the below reaction sequence, it should be well within the capability of the person skilled in the art to select suitable reaction components as well as reaction conditions.

Reaction Scheme 1

Preparation of a compound of formula (I) wherein X is -NR^C-

(I, X is -NR⁰-) Another synthetic method useful for preparing the inventive compounds is illustrated in Reaction Scheme 2. In this case the synthesis is started from a suitable 6-aniline derivative, -Y-

NR . l Rr.2 , and the amine group, -(R )N(CH₂)_nR , is introduced in a later step. The entire synthesis is illustrated by Reaction Scheme 2.

Reaction Scheme 2

Preparation of a compound of formula (I) wherein X is -NR -

(I; X is -NR^C-)

Numerous methods exist in the literature for the synthesis of ethers and sulfides from aryl halides, which should be contemplated when X is O (oxygen) or S. A summary of this work can be found in, for example, Jerry March in Advanced Organic Chemistry, 4* Ed, John Wiley & Sons Inc, New York, 1992, p654-656. An example of a modern synthetic procedure that directly leads to compounds as biaryl ethers can be found in: Evans, D. A.; et al., Tetrahedron Lett. 1998, 39, 2937-2940. 1 9 "-Ϊ C

In summary, there are several ways in which order to introduce the groups R , R , R , R , Y and X, all well known for the one skilled in the art, in order to arrive at the compounds of the invention, and the synthetic routes mentioned herein are not limiting for the invention.

The term "alkyl" as employed herein, alone or as part of another group, refers to an acyclic

5 straight or branched chain radical, unless otherwise specified containing 1, 2, 3, 4, 5, 6, 7 or 8 carbons in the normal chain, which includes methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl and n-octyl. Examples of branched chain radicals, not excluding any of the possible isomers not mentioned, are iso-propyl, sec-butyl, iso-pentyl, 3-methylpentyl, 2,3-dimethylhexyl, 3-ethylhexyl, and the like. Unless otherwise specified, the term alkyl also includes a straight or

10 branched alkyl group that contains or is interrupted by a carbocyclyl, exemplified by cyclopropane, as exemplified below:

In case the alkyl is interrupted or terminated by a carbocyclyl, the alkyl portions can be attached at any variable point of attachment to the carbocyclyl, including the same ring carbon, as exemplified below:

5 When the alkyl chain is interrupted or terminated by a carbocyclyl, the total number of carbon atoms of the alkyl chain and the carbocyclyl is at most 8. hi other words, in the above given example, the sum of z and w is at most 5.

When substituted alkyl is present, this refers to a straight or branched alkyl group as defined above, substituted with 1, 2 or 3 groups of R^a. The alkyl group preferably contains 1, 2, 3

'0 or 4 carbons in the normal chain that also can be substituted with 1, 2 or 3 groups of R^a, which groups may be the same or different at any available point, as defined with respect to each variable. When such a substituted alkyl group is present, the preferred substitution is halogen such as in -CH₂Cl, -CF₃, -CH₂I, -CHF₂, -CH₂Br, -CH₂F, -CHFCH₂F, -CHFCH₂Cl, - CHFCHClCH₃, -CHClCHBrCH₂CF₃, -CHClCBrICH₂CF₃, -CH₂CH₂CH₂CH₂I, and the like.

!5 The term "alkenyl" as used herein, alone or as part of another group, refers to a straight or branched chain radical, unless otherwise specified containing 2, 3, 4, 5, 6, 7 or 8 carbons, which contains at least one carbon to carbon double bond. Preferably only one carbon to carbon double bond is present, such as in the normal chain vinyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 3-octenyl, and the like.

0 The alkenyl group preferably contains 2, 3 or 4 carbons in the normal chain. As described above with respect to the "alkyl", the straight or branched portion of the alkenyl group may be optionally substituted when a substituted alkenyl group is provided. Furthermore, unless otherwise specified, the chain may be interrupted or terminated by a carbocyclyl group, in which case the total number of carbon atoms of the chain and the carbocyclyl is at most 8.

The term "alkynyl" as used herein by itself or as part of another group refers to a straight or branched chain radical, unless otherwise specified containing 2, 3, 4, 5, 6, 7 or 8 carbons, which contains at least one carbon to carbon triple bond. Preferably, only one carbon to carbon triple bond is present, such as in the normal chain 2-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3- pentynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, and the like. The alkynyl group preferably contains 1, 2, 3 or 4 carbons in the normal chain. As described above with respect to the "alkyl", the straight or branched portion of the alkynyl group may be optionally substituted when a substituted alkynyl group is provided. Furthermore, unless otherwise specified, the chain may be interrupted or terminated by a carbocyclyl group, in which case the total number of carbon atoms of the chain and the carbocyclyl is at most 8.

In one embodiment, in a compound according to formula (I), any alkyl, alkenyl, or alkynyl group having a number of p (p being an integer of 4 to 8) carbon atoms, optionally and independently from any other alkyl, alkenyl or alkynyl group present in the compound, includes a carbocyclic portion of a number of q (q being an integer of 3 to 7 and q being less than p) carbon atoms, which carbocyclic portion may be located so as to interrupt or terminate the straight or branched chain of the alkyl, alkenyl, or alkynyl group, whereby the number of carbon atoms in the straight or branched chain of the alkyl, alkenyl or alkynyl group equals p-q. In another embodiment, in a compound according to formula (I), any alkyl, alkenyl, or alkynyl group having p carbon atoms has all p carbon atoms in the straight or branched chain portion, i.e. does not include any terminating or interrupting carbocyclic portion.

The term "carbocyclyl" as employed herein alone or as part of another group includes saturated cyclic hydrocarbyl groups or unsaturated (at least 1 double bond) cyclic hydrocarbyl groups, containing at least one ring of in total of 3, 4, 5, 6, 7 or 8 ring carbons, which includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, and the like. The cyclic hydrocarbyl may be monocyclic or bicyclic (i.e. containing two rings of 3 to 8 ring carbons each). For example, the carbocyclyl may be monocyclic and 3- to 6-membered. As described above with respect to the "alkyl", the carbocyclyl group may be optionally substituted by 1, 2 or 3 halogens, which may be the same or different.

As used herein, and unless otherwise specified, the term "heterocyclyl" mean a non- aromatic cyclic group that optionally might be unsaturated, containing one or more heteroatom(s) preferably selected from N, O and S, such as a 4 to 10-membered ring system containing at least one heteroatom, e.g. 1-4 heteroatoms. For example, a suitable heterocyclyl may be a non-aromatic cyclic group that optionally might be unsaturated, containing one or more heteroatom(s) preferably selected from N, O and S, such as a 5 to 6-membered monocyclic ring system containing at least one heteroatom, e.g. 1-4 heteroatoms, 1-3 heteroatoms, or 1 or 2 heteroatoms. A heterocyclyl e.g. may be, but is not limited to, aziridinyl, azetidinyl, dihydropyranyl, dihydropyridyl, dihydropyrrolyl, dioxolanyl, dioxanyl, dithianyl, dithiolanyl, imidazolidinyl, imidazolinyl, morpholinyl, oxetanyl, oxiranyl, pyrrolidinyl, pyrrolidinonyl, piperidyl, piperazinyl, piperidinyl, pyrazolidinyl, quinuclidinyl, sulfalonyl, 3-sulfolenyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridyl, thietanyl, thiiranyl, thiolanyl, thiomorpholinyl, trithianyl, tropanyl, lH-indazolyl and monosaccharide.

By the term "unsaturated", when referring to a bicyclic system, is meant a ring system comprising at least one double or triple bond in at least one ring. Thus, it is contemplated that both rings may be unsaturated or only one ring may be unsaturated, and the other one being saturated. Furthermore, the term "unsaturated bicyclic" also is intended to refer to a non- aromatic bicyclic system comprising a ring that is either unsaturated or saturated fused to a ring that by itself would be aromatic, such as in indane or 4,5-dihydro-l -indole.

The term "halogen" refers to fluorine, chlorine, bromine and iodine, where the preferred halogen radicals are fluorine and chlorine.

As used herein, the term "aryl" means an aromatic group, monocyclic or bicyclic such as phenyl or naphthyl, and the like. The aryl group is preferably a monocyclic C₆ aryl (i.e. phenyl).

As used herein, the term "heteroaryl" means a mono- or bicyclic heteroaromatic group containing one or more heteroatom(s) preferably selected from N, O and S, such as, but not limited to, pyridyl, quinolinyl, furanyl, thienyl, oxadiazolyl, thiadiazolyl, thiazolyl, oxazolyl, pyrazolyl, triazolyl, tetrazolyl, isoxazolyl, isothiazolyl, isoquinolinyl, naphthyridinyl, imidazolyl, phenazinyl, phenothiazinyl, phthalazinyl, indolyl, pyridazinyl, quinazolinyl, quinolizinyl, quinoxalinyl, tetrahydroisoquinolinyl, pyrazinyl, indazolyl, indolinyl, pyrimidinyl, thiophenetyl, pyranyl, carbazolyl, chromanyl, cinnolinyl, acridinyl, benzimidazolyl, benzodioxanyl, benzodioxepinyl, benzodioxolyl, benzofuranyl, benzothiazolyl, benzobenzoxadiazolyl, benzoxazinyl, benzoxazolyl, benzomorpholinyl, benzoselenadiazolyl, benzothienyl, purinyl, and pteridinyl.

Thus, in one aspect of the invention, there is provided compounds of formula (I)

(I) as defined herein above.

In formula (I), n denotes the number of -CH₂- linking X to R , and is either 0 or 1. In one 5 embodiment n is 1 ; however, more preferably n is 0.

The groups R and R² are independently selected from hydrogen; branched or unbranched C₁-C₈ alkyl, C₂-C₈ alkenyl, or C₂-C₈ alkynyl; monocyclic or bicyclic, saturated or unsaturated C₃-C₈ carbocyclyl; and monocyclic or bicyclic, saturated or unsaturated C₁-C₇ heterocyclyl wherein the heteroatoms are independently selected from N, O and S; and wherein said alkyl, 10 alkenyl, alkynyl, carbocyclyl or heterocyclyl optionally are substituted with 1, 2, or 3 groups R . In one embodiment, R¹ and R² are independently selected from hydrogen and branched or unbranched C₁-C₈ alkyl, C₂-C₈ alkenyl and C₂-C₈ alkynyl, wherein said alkyl, alkenyl, or alkynyl are substituted with 1, 2, or 3 groups R^a. In particular, R¹ and R² may be independently selected from hydrogen and branched or unbranched C₁-C₄ alkyl, C₂-C₄ alkenyl and C₂-C₄ alkynyl, said ^5 alkyl, alkenyl, or alkynyl optionally being substituted with 1, 2, or 3 groups R .

In another embodiment, R¹ and R² are independently selected from hydrogen and branched or unbranched C₁-C₈ alkyl, said alkyl optionally being substituted with 1, 2, or 3 groups R^a. In particular, R and R² may be independently selected from hydrogen and branched or unbranched C_J-C₄ alkyl, e.g. methyl, said alkyl, alkenyl, or alkynyl optionally being substituted with 1, 2, or !0 3 groups R , e.g. 1 or 2 groups R , or 1 group R , or no group R .

1 0 1

In one embodiment, R is hydrogen, and R is as defined herein above. For example, R is hydrogen, and R² is as defined herein above, but is not hydrogen.

In one particular embodiment, R¹ is hydrogen, and R² is methyl.

In a compound of formula (I), R³ is selected from monocyclic or bicyclic C₆-C₁₀ aryl; and '5 monocyclic or bicyclic Ci -Cg heteroaryl, wherein the heteroatoms independently are selected from N, O and S; said aryl or heteroaryl optionally being substituted with 1, 2, 3, 4 or 5 groups R^b. In one embodiment, R³ is selected from monocyclic C₆ (i.e. phenyl); and monocyclic C₁-C₅ heteroaryl, wherein the heteroatoms independently are selected from N, O and S; said aryl or heteroaryl optionally being substituted with 1, 2, 3, 4 or 5 groups R .

In another embodiment, R is selected from monocyclic or bicyclic C₆-Ci_O aryl; said aryl 5 optionally being substituted with 1, 2, 3, 4 or 5 groups R .

For example, R may be phenyl; optionally substituted with 1, 2, 3, 4 or 5 groups R . In one embodiment, R³ is selected from (unsubstituted) phenyl, fluorophenyl, chlorophenyl, isopropylphenyl, difluoromethoxyphenyl, dimethylphenyl and methoxyphenyl. For example, R may be selected from phenyl, 2-fluorophenyl, 4-fluorophenyl, 4-chlorophenyl, 4- 10 isopropylphenyl, 4-(difluoromethoxy)phenyl, 2,6-dimethylphenyl, and 3 -methoxyphenyl.

In formula (I), the moiety Y is - is selected from -C(O)-; -S(O)-; and -S(O)₂-, but preferably is -C(O)-.

The moiety X is selected from -NR⁰-; -O-; and -S-; but preferably is -NR^C-.

As mentioned herein above, any alkyl, alkenyl, or alkynyl in either of R and R optionally 15 may be substituted with 1, 2, or 3 groups R^a. For example, the number of substituents R e.g. may be 1 or 2, or 1, and each R is independently selected from halogen; hydroxy; carbonyl; methoxy; halomethoxy; dihalomethoxy; and trihalomethoxy; e.g. from halogen. In one embodiment R^a is absent.

In R , any aryl or heteroaryl optionally is substituted with 1, 2, 3, 4 or 5 groups R . For !0 example, the number of groups R present on any given ring in a monocyclic or bicyclic moiety may be 1-4, or 1-3, e.g.l or 2.

Each R is independently selected from halogen, branched or unbranched C₁-C₄ alkyl, C₂- C₄ alkenyl or C₂-C₄ alkynyl; branched or unbranched C₁-C₄ alkyloxy, C₂-C₄ alkenyloxy or C₂-C₄ alkynyloxy; branched or unbranched C₁-C₄ alkylthio, C₂-C₄ alkenylthio or C₂-C₄ alkynylthio; '.5 wherein any alkyl, alkenyl, alkynyl, alkyloxy, alkenyloxy alkynyloxy, alkylthio, alkenylthio or alkynylthio group optionally is substituted with 1, 2 or 3 halogens.

In one embodiment, each R is independently selected from halogen, branched or unbranched Ci-C₄ alkyl; branched or unbranched C₁-C₄ alkyloxy; and branched or unbranched C₁-C₄ alkylthio; wherein any alkyl, alkyloxy or alkylthio group optionally is substituted with 1, 2 0 or 3 halogens. For example, each R may be independently selected from halogen, branched or unbranched C₁-C₄ alkyl; and branched or unbranched C₁-C₄ alkyloxy; wherein any alkyl or alkyloxy group optionally is substituted with 1, 2 or 3 halogens. In another embodiment, each R is independently selected from halogen, branched or unbranched C₁-C₄ alkyl, C₂-C₄ alkenyl or C₂-C₄ alkynyl; and branched or unbranched C₁-C₄ alkyloxy, C₂-C₄ alkenyloxy or C₂-C₄ alkynyloxy; wherein any alkyl, alkenyl, alkynyl, alkyloxy, alkenyloxy, or alkynyloxy group optionally is substituted with 1, 2 or 3 halogens. 5 In one embodiment, any C₁-C₄ alkyl moiety present in R is selected from C₁- C₃ alkyl.

Also, any C₂-C₄ alkenyl moiety in R may be selected from C₂-C₃ alkenyl. Any C₂-C₄ alkynyl moiety in R may be selected from C₂-C₃ alkynyl.

In one embodiment, each R is selected from halogen, e.g. fluoro, chloro and bromo, in particular fluoro and chloro.

LO In another embodiment of the invention, each R is independently selected from halomethyl; dihalomethyl and trihalomethyl.

In another embodiment, each R is selected from fluoro, chloro; methyl, isopropyl, and methoxy, the methyl, isopropyl and methoxy groups optionally being substituted by 1, 2 or 3 hhaallooggeennss.. FFoorr eexxaaimple, R may be selected from fluoro, chloro, methyl, isopropyl, methoxy, and difluoromethoxy.

The group R^c is selected from hydrogen and branched or unbranched C₁-C₄ alkyl, C₂-C₄ alkenyl or C₂-C₄ alkynyl. In one embodiment, R is selected from hydrogen and branched or unbranched C₁-C₄ alkyl, e.g. hydrogen and methyl. In one embodiment, R is hydrogen.

In one particular embodiment of the invention, there is provided compounds of formula (I), ',0 wherein R¹ and R² are independently selected from hydrogen, C₁-C₄ alkyl, C₂-C₄ alkenyl and C₂- C₄ alkynyl; R^a is halogen; X is -NR^C- and n is 0 (zero).

In another embodiment of the invention, there is provided compounds of formula (I), wherein n is 0 (zero); R and R are independently selected from hydrogen and C₁-C₈ alkyl; R is a monocyclic C₆ aryl; substituted with 1, 2, 3, 4 or 5 groups each independently selected from '5 R ; Y is -C(O)-; X is -NR⁰-; and R is independently selected from halogen; halomethyl; dihalomethyl; and trihalomethyl.

In another embodiment of the invention, there is provided compounds of formula (I), wherein R¹ represents hydrogen; R² represents C₁-C₄ alkyl; X represents NR⁰; R° represents hydrogen; R³ represents a monocyclic C₆ aryl (phenyl), substituted with R ; R represents 0 halogen; and n represents 0 (zero). In another embodiment of the invention, there is provided compounds of formula (I), wherein R represents hydrogen; R represents C₁-C₄ alkyl; X represents NR ; R represents hydrogen; R³ represents a monocyclic C₆ aryl (phenyl), substituted with R ; R represents halomethyl; dihalomethyl or trihalomethyl; and n represents 0 (zero).

In any of the above embodiments, wherein any C_p alkyl, alkynyl or alkenyl group has a number p > 4 of carbon atoms, said alkyl, alkenyl or alkynyl group optionally includes a C_q carbocyclic portion of q of carbon atoms, whereby 3 < q < p.

In another embodiment of the invention, there is provided a compound of formula (I), which is:

4-(4-fluorophenylamino)-6-(methylcarbamoyl)quinoline-3 -carboxylic acid; or

4-(4-chlorophenylamino)-6-(methylcarbamoyl)quinoline-3-carboxylic acid;

4- [(4-isopropylphenyl)amino] -6-(methylcarbamoyl)quinoline-3 -carboxylic acid;

-[[4-(difluoromethoxy)phenyl]amino]-6-(methylcarbamoyl)quinoline-3-carboxylic acid;

-benzylamino-6-methylcarbamoyl-quinoline-3 -carboxylic acid;

-(2,6-dimethyl-phenylamino)-6-methylcarbamoyl-quinoline-3-carboxylic acid;

-(3 -methoxy-phenylamino)-6-methylcarbamoyl-quinoline-3 -carboxylic acid;

4-(2-fluoro-phenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid;

6-niethylcarbamoyl-4-phenylamino-quinoline-3-carboxylic acid, or a pharmaceutically acceptable salt thereof.

The compounds of the invention can be present as salts, which are also within the scope of this invention. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred.

For example, the inventive compounds can form acid addition salts, e.g. at the amino function. These may be formed, for example, with strong inorganic acids, such as mineral acids, for example sulfuric acid, phosphoric acid or a hydrohalic acid; strong organic carboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atoms which are unsubstituted or substituted, for example, by halogen, for example acetic acid, saturated or unsaturated dicarboxylic acids, for example oxalic, malonic, succinic, maleic, fumaric, phthalic or terephthalic acid, hydroxycarboxylic acids, for example ascorbic, glycolic, lactic, malic, tartaric or citric acid, amino acids, (for example aspartic or glutamic acid or lysine or arginine), or benzoic acid, or with organic sulfonic acids, such as (Ci-C₄) alkyl or arylsulfonic acids which are unsubstituted or substituted, for example by halogen, for example methyl- or p-toluene- sulfonic acid. Corresponding acid addition salts can also be formed having, if desired, an additionally present basic center.

The compounds of formula I having at least one acid group (for example COOH) can also form salts with bases. Suitable salts with bases are, for example, metal salts, such as alkali metal or alkaline earth metal salts, for example sodium, potassium or magnesium salts, or salts with ammonia or an organic amine, such as morpholine, thiomorpholine, piperidine, pyrrolidine, a mono-, di- or tri-lower alkylamine, for example ethyl, tert-butyl, diethyl, diisopropyl, triethyl, tributyl or dimethyl-propylamine, or a mono, di or trihydroxy lower alkylamine, for example mono-, di- or triethanolamine. Corresponding internal salts may furthermore be formed. Salts that are unsuitable for pharmaceutical uses but which can be employed, for example, for the

5 isolation or purification of free compounds of formula I or their pharmaceutically acceptable salts are also included.

The present invention also includes prodrugs. In fact, the esters of formula I display improved uptake in vivo and are hydrolyzed to their corresponding carboxylic acids in vivo. The term "prodrug" is intended to represent a compound bonded to a carrier, which prodrug is

.0 capable of releasing the active ingredient when the prodrug is administered to a mammalian subject. Release of the active ingredient occurs in vivo. Prodrugs of compounds of the invention include compounds wherein a hydroxyl, amino, carboxylic, or a similar group is modified. Examples of prodrugs include, but are not limited to, esters (e.g. acetate, formate, and benzoate derivatives), carbamates (e.g., N,N-dimethylaminocarbonyl of hydroxyl or amino functional

5 groups of the present invention), amides (e.g., trifluoroacetylamino, acetylamino, and the like), and the like.

The compounds of the invention may be administered as is or as an alternative prodrug, for example in the form of an in vivo hydrolysable ester or in vivo hydrolysable amide. An in vivo hydrolysable ester of a compound of the invention containing carboxy or hydroxyl group is, for

!0 example, a pharmaceutically acceptable ester which is hydrolysed in the human or animal body to produce the parent acid or alcohol. Suitable pharmaceutically acceptable esters for carboxy include Q-Cealkyloxymethyl esters (e.g., methoxymethyl) d-Cβalkanoyloxymethyl esters (e.g., pivaloyloxymethyl), phthalidyl esters, Ca-Cscycloalkyloxycarbonyloxy-Ci-C_δalkyl esters (e.g. 1- cyclohexylcarbonyloxyethyl), l,3-dioxolen-2-onylmethyl esters (e.g., 5 -methyl- 1,3 -dioxolen-2-

15 onylmethyl) and Cj-Cealkyloxycarbonyloxyethyl esters (e.g., 1-methoxycarbonyloxyethyl) and may be formed at any appropriate carboxy group in the compounds of the invention.

An in vivo hydrolysable ester of a compound of the invention containing a hydroxyl group includes inorganic esters such as phosphate esters and acyloxyalkyl ethers and related compounds which as a result of the in vivo hydrolysis of the ester breakdown to give the parent

0 hydroxy group. Examples of acyloxyalkyl ethers include acetoxymethoxy and 2,2- dimethylpropionyloxy-methoxy. A selection of in vivo hydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl and 7V-(N,iV-dialkylamino- ethyl)-7V-alkylcarbamoyl (to give carbamates), A^N-dialkylaminoacetyl and carboxyacetyl. Examples of substituents on benzoyl include morpholino and piperazino linked from a ring nitrogen atom via a methylene group to the 3- or 4- position of the benzoyl ring. A suitable value for an in vivo hydrolysable amide of a compound of the invention containing a carboxy group is, for example, an N-Ci-C₆alkyl or 7V,7V-diCj-C₆alkyl amide such as iV-methyl, iV-ethyl, JV-propyl, ΛyV-dimethyl, iV-ethyl-iV-methyl or N_^/V-diethyl amide. Upon administration of a compound of the invention, or an alternative prodrug thereof, the prodrug undergoes chemical conversion by metabolic or chemical processes to yield another compound, for example a salt and/or solvate thereof. Solvates of the compounds of the present invention include, for example hydrates. An administration of a therapeutic agent of the invention includes administration of a therapeutically effective amount of the agent of the invention. The term "therapeutically effective amount" as used herein refers to an amount of a therapeutic agent to treat or prevent a condition treatable by administration of a composition of the invention. That amount is the amount sufficient to exhibit a detectable therapeutic or preventative or ameliorative effect. The effect may include, for example, treatment or prevention of the conditions listed herein. The precise effective amount for a subject will depend upon the subject's size and general condition, the nature and extent of the condition being treated, recommendations of the treating physician, and the therapeutics or combination of therapeutics selected for administration. Thus, it is not useful to exactly specify an exact effective amount in advance. In the case of oral administration the dosage might, however, vary from about 0.01 mg to about 1000 mg per day of a compound of formula (I) or the corresponding amount of a pharmaceutically acceptable salt thereof.

The composition according to the invention may be prepared for any route of administration, e.g. oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, or intraperitoneal. The precise nature of the carrier or other material will depend on the route of administration. For parenteral administration, a parenterally acceptable aqueous solution is employed, which is pyrogen free and has requisite pH, isotonicity and stability. Those skilled in the art are well able to prepare suitable solutions and numerous methods are described in the literature.

The pharmaceutically acceptable excipients described herein, for example, vehicles, adjuvants, carriers or diluents, are well-known to those who are skilled in the art and are readily available to the public. The pharmaceutically acceptable carrier may be one that is chemically inert to the active compounds and that has no detrimental side effects or toxicity under the conditions of use. Examples of pharmaceutical formulations can be found in Remington: The Science and Practice of Pharmacy. A. R. Gennaro, Editor. Lippincott, Williams and Wilkins, 20th edition (2000). All stereoisomers of the compounds of the instant invention are contemplated, either in admixture or in pure or substantially pure form. The compounds of the present invention can have asymmetric centers at any of the carbon atoms including any one of the R substituents. Consequently, compounds of formula I can exist in enantiomeric or diasteromeric forms or in

5 mixtures thereof. The processes for preparation can utilize racemates, enantiomers or diasteromers as starting materials. When diastereomeric or enantiomeric products are prepared, they can be separated by conventional methods, which for example is chromatographic or fractional crystallization.

The effectiveness of the compounds of the invention in preventing or treating disease may

[O be improved by administering the compounds in combination with another agent that is effective for those purposes, such as, but not limited to, another antiangiogenic compounds inhibiting VEGF, VEGFR tyrosine kinase, integrin inhibitors, phototherapies, antibodies against VEGF, or one or more conventional therapeutic agents such as, alkylating agents, folic acid antagonists, anti-metabolites of nucleic acid metabolism, pyrimidine analogs, 5-fluorouracil, purine

5 nucleosides. Such other agents may be present in the composition being administered or may be administered separately. Also, the compounds of the invention are suitably administered serially or in combination with radiological treatments, whether involving irradiation or administration of radioactive substances.

The term antiangiogenic as used herein by itself or as a part of another definition refers to a

!0 compound with the ability to inhibit angiogenesis, which is the growth of new blood vessels into a solid tumor.

The number of mechanisms for antiangiogenic agents is diverse and may include, but not limited to, compounds that inhibit cell proliferation, inhibit cell migration of endothelial cells, activate immune system, downregulate angiogenesis stimulators, stimulate angiogenesis

!5 inhibitor formation, inhibits binding of angiogenesis stimulators, inhibit basement membrane degradation, induce apoptosis of endothelial cells, inhibit survival of endothelial cells, inhibit cell adhesion and inhibit survival of endothelial cells.

The number of compounds or monoclonal antibodies that are antiangiogenic may include, but is not limited to, Avastin® (bevacizumab) carboxyamidotriazole (5-Amino-l-((3,5-dichloro-

0 4-(4-chlorobenzoyl)phenyl)methyl)- 1 H- 1 ,2,3 -triazole-4-carboxamide), TNP-470

((3R,4S,5S,6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-2-butenyl)-oxiranyl]-l-oxaspiro [2,5] oct-6-yl(chloroacetyl) carbamate), CM-IOl (a bacterial polysaccharide exotoxin produced by group B Streptococcus (GBS), also referred to as GBS toxin), Germanin® (also known as suramin, CAS number 145-63-1), SU5416 (semaxinib, (3Z)-3-[(3,5-dimethyl-lH-pyrrol-2- yl)methylidene]-l,3-dihydro-2H-indol-2-one), TSP (thrombospondins, a group of secreted proteins with antiangiogenic abilities), angiostatic steroids and heparin in combination, matrix metalloproteinase inhibitors, Angiostatin™, Macugen® (pegaptanib sodium injection), Endostatin™, 2-methoxyestradiol, Tecogalan sodium (DS-4152, a bacterial polysaccharide), 5 prolactin (or luteotropic hormone (LTΗ), a peptide hormone), linomide (LS-2616, [N-methyl-N- phenyl-l,2-dihydro-4-hydroxy-l-methyl-2-oxo-quinoline-3-carboxamide]) and the like.

The term VEGF (vascular endothelial growth factor) as used herein refers to a sub-family of growth factors, which are platelet-derived growth factor family of cystine-knot growth factors. They are important signaling proteins involved in angiogenesis, as well as vasculogenesis (de 10 novo formation of the embryonic circulatory system).

The term VEGFR tyrosine kinase as used herein refers to the tyrosine kinase receptors that the members of the VEGF family bind to.

The term integrin as used herein by itself or as a part of another definition refers to a family of transmembrane glycoproteins consisting of noncovalent heterodimers. The integrins consist of .5 at least three identified families where each family contains a common beta-subunit combined with one or more distinct alpha-subunits. These receptors participate in cell-matrix and cell-cell adhesion in many physiologically important processes, including oncogenic transformation.

The compounds according to formula (I) will be useful for treating various diseases such as cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic !0 myocardium, atherosclerosis, macular edema and psoriasis. The treatment may be preventive, palliative or curative.

The compounds of the invention provides a method of treating a mammal suffering from a disease or disorder related to VEGFR tyrosine kinase or integrin activity, comprising administering to said mammal in need thereof, a therapeutically effective amount of a compound 15 of formula (I). The said mammal can be a human.

The compounds of the present invention may be used or administered in combination with one or more additional drugs useful in the treatment of hyperproliferative diseases, e.g. antiangiogenic agents, including both compounds and monoclonal antibodies, and a cytostatic agent. The components may be in the same formulation or in separate formulations for 0 administration simultaneously or sequentially. The compounds of the present invention may also be used or administered in combination with other treatment such as irradiation for the treatment of cancer.

Examples of cytotstatic agents for use as indicated herein above are DNA alkylating compounds, topoisomerase I inhibitors, topoisomerase II inhibitors, compounds interfering with RNA and DNA synthesis, compounds polymerising the cytoskeleton, and compounds depolymerising the cytoskeleton.

The invention is illustrated by the following non-limiting Examples.

EXAMPLES Example 1: 4-(4-fluorophenylamino)-6-(methylcarbamoyl)quinoline-3-carboxylic acid.

(a) Preparation of intermediary compound diethyl 2-((4-bromophenylamino)methylene)- malonate:

4-Bromoaniline (10 g) and diethoxymethylene malonate (12.6 g) were heated at 150°C for 3 hours in a sealed tube. The reaction mixture was then cooled and diluted with n-hexane when the solid product precipitated out. This solid was filtered, washed several times with n-hexane and dried under vacuum to afford 17.8 g of 2-[(4-bromo-phenylamino)methylene]-malonic acid diethyl ester. ¹R NMR (300 MHz, CDCl₃) δ 11.03 (d, IH, J = 13 Hz, -NH- ), 8.48 (d, IH, J =13 Hz, -CH=C), 7.49 (m, 2Η, aromatic), 7.10-7.01 (m, 2H, aromatic), 4.42-4.22 (m, 4H, -CH₂-CH₃), 1.45-1.26 (m, 6H, -CH₂-CH₅); LC-MS (rn/z) 343.9 (M+l).

(b) Preparation of intermediary compound 6-bromo-4-chloroquinoline-3-carboxylic acid ethyl ester:

2-[(4-Bromophenylamino)methylene]malonic acid diethyl ester (5 g) was heated with POCl₃ (phosphoryl chloride, 31.5 mL) at 150⁰C in a sealed tube for about 6 hours. The excess POCl₃ was removed in vacuo and the reaction mixture was diluted with dichloromethane. The dichloromethane extract was washed with aqueous sodium hydroxide solution (10 %), dried over sodium sulphate and purified by column chromatography (Silica gel, hexane/ethyl acetate 80:20) to give 2.3 g of 6-bromo-4-chloroquinoline-3-carboxylic acid ethyl ester. H NMR (300 MHz, CDCl₃) δ 9.22 (s, IH, aromatic), 8.60 (d, IH, J = 2.1 Hz, aromatic), 8.04 (d, IH, J = 9 Hz, aromatic), 7.95-7.85 (m, IH, aromatic), 4.53 (q, 2H, J = 7 Hz, -CH₂-), 1.50 (t, 3H, J = 7 Hz, - CH₅); LC-MS (m/z) 315.8 (M+ 1).

(c) Preparation of intermediary compound ethyl 6-bromo-4-(4-fluorophenylamino)- quinoline-3 -carboxylate:

/7-Fluoroaniline (0.106 g) and 6-bromo-4-chloroquinoline-3-carboxylic acid ethyl ester (0.3 g, 0.95 mmol) were mixed in dioxane and irradiated in a microwave reactor at 150°C for 30 minutes. The reaction mixture was diluted with petroleum ether. The solid product obtained was filtered and dried to give 0.33 g of ethyl 6-bromo-4-(4-fluorophenyl-amino)quinoline-3- carboxylate. LC-MS (m/z) 389.4 (M+l).

(d) Preparation of intermediary compound ethyl 4-(4-fiuorophenylamino)-6- (methylcarbamoyl)quinoline-3 -carboxylate :

Ethyl 6-bromo-4-(4-fluorophenyl-amino)quinoline-3-carboxylate (0.3 g) was added to tetrahydrofuran followed by trørø-di(μ-acetato)-bis[o-(di-ø-tolylphosphino)- benzyl]dipalladium(II) (Herrmann's palladacycle, 0.038 mmol), tri tertiarybutyl phosphonium hexafluoborate) ([(/-Bu)₃PH]BF₄, 0.0385 mmol), molybdenum hexacarbonyl (Mo(CO)₆, 1.54 mmol), methylamine (4.6 mmol, 2N in tetrahydrofuran) and l,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 7.7 mmol). The reaction mixture was irradiated at 130°C for 5 minutes in a microwave reactor. The reaction mixture was concentrated and then purified on column (silica gel, dichloromethane/methanol 98:2) to give 0.39 g of ethyl 4-(4-fluorophenylamino)-6- (methylcarbamoyltøuinoline-S-carboxylate as a solid. ¹H NMR (300 MHz, CDCl₃) δ 9.88(s, IH, -CONH-), 8.89(s, IH, aromatic), 8.72(s, IH, aromatic), 8.59(d, 1H,J=4 Hz, aromatic), 8.15(d,lH, J= 8.7 Hz, aromatic), 7.98(d, IH, J=8.7 Hz, aromatic), 7.16(m, 4H, aromatic), 3.98(g, 2H, J=7 Hz, -CH₂-), 2.80(s, 3H₅-NCH₃), 1.16(t, 2Η, J=7 Hz, -CH₃); LC-MS (m/z) 368.1 (M+l). (e) Ethyl 4-(4-fluorophenylamino)-6-(methylcarbamoyl)quinoline-3-carboxylate (0.03 g) was stirred with lithium hydroxide (0.128 g) in a mixture of 6 mL of methanol/tetrahydrofuran /water (2:2:2,) overnight. The reaction mixture was concentrated and the aqueous layer was washed with ethyl acetate. The aqueous layers were collected and acidified with aqueous hydrochloric acid and the precipitate formed was filtered and dried to give 0.022 g of 4-(4- fluorophenylamino)-6-(methylcarbamoyl)quinoline-3-carboxylic acid as a yellow solid. ¹H NMR (300 MHz, CD₃OD) δ 12.47(bs, IH₅-COOH), 9.12(s, IH, aromatic), 8.46(s, IH, aromatic), 8.23(s, IH, aromatic), 8.07(d, IH, J=8.4 Hz, aromatic), 7.92(d, IH, J=8.4 Hz, aromatic), 7.15(m, 4H, aromatic), 2.17(s, 3H, -NCH₃) ; LC-MS (m/z) 340.2 (M+l).

Example 2: 4-(4-chlorophenylamino)-6-(methylcarbamoyl)quinoIine-3-carboxylic acid.

(a) Preparation of intermediary compound ethyl 6-bromo-4-(4-chlorophenylamino)- quinoline-3 -carboxylate :

/7-Chloroaniline (0.2 g) and 6-bromo-4-chloroquinoline-3-carboxylic acid ethyl ester (0.5 g) were mixed in dioxane and irradiated in a microwave reactor at 150°C for 30 minutes. The reaction mixture was diluted with petroleum ether. The solid product obtained was filtered and dried to give 0.565 g of ethyl 6-bromo-4-(4-chlorophenylamino)-quinoline-3-carboxylate. LC- MS (m/z) 406.7 (M+l). (b) Preparation of intermediary compound ethyl 4-(4-chlorophenylamino)-6- (methylcarbamoyl)quinoline-3 -carboxylate :

Ethyl 6-bromo-4-(4-chlorophenylamino)-quinoline-3-carboxylate (0.52 g) was added to tetrahydrofuran followed by /rαo>s'-di(μ-acetato)-bis[o-(di-o- tolylphosphino)benzyl]dipalladium(II) (Herrmann's palladacycle, 0.06 g), [(t-Bu)₃PH]BF₄ (tri tertiarybutyl phosphonium hexafluoborate) (0.074 mg), molybdenum hexacarbonyl (Mo(CO)₆, 0.677 mg), methylamine (7.6 mmol, 2N in tetrahydrofuran) and l,8-diazabicyclo[5.4.0]undec-7- ene (DBU, 1.95 g). The reaction mixture was irradiated at 130°C for 5 minutes in a microwave reactor. The reaction mixture was concentrated and then purified on column (silica gel, dichloromethane/methanol 98:2) to give 0.25 g of ethyl 4-(4-chlorophenylamino)-6-(methyl- carbamoyl)quinoline-3-carboxylate. ¹H NMR (300 MHz, CDCl₃) δ 9.83(s, IH, -CONH-), 8.92(s, IH, aromatic), 8.75(s, IH, aromatic), 8.61(d, IH, J= 4.5Hz, aromatic), 8.17(d, IH, J=8.7Hz, aromatic), 8.01(d, IH, J=8.7 Hz, aromatic), 7.35(d, 2H, J= 8.7 Hz, aromatic), 7.09(d, 2H, J= 8.7 Hz, aromatic), 3.98(q, 2H, J= 7Hz, -CH₂-), 2.81(s, 3Η, N-CH₃), 1.41(t, J= 7Ηz, -CH₃)LC-MS im/z) 384.1 (M+l).

(c) Ethyl 4-(4-chlorophenylamino)-6-(methylcarbamoyl)quinoline-3-carboxylate (0.025 g) was stirred with lithium hydroxide (10 mg) in a mixture of 3 mL of methanol/ tetrahydrofuran/water (2:2:2,) overnight. The reaction mixture was concentrated and the aqueous layer was washed with ethyl acetate. The aqueous layers were collected and acidified with aqueous hydrochloric acid and the precipitate formed was filtered and dried to give 0.02 g of 4- [(4-chloro-phenyl)amino]-6-(methylcarbamoyl)-quinoline-3-carboxylic acid as a solid. Η NMR (300 MHz, MeOD) δ 11.8 (broad s, IH₅-COOH), 9.05 (s, 1Η, aromatic), 8.68(m, 2Η, aromatic), 8.66 (d, IH, J=4.5 Hz, aromatic), 8.24 (d, IH, J=9 Hz, aromatic), 8.02 (d, IH, J=9 Hz, aromatic), 7.42 (d, 2H, J=8.4 Hz, aromatic), 7.23 (d, 2H, J=8.4 Hz, aromatic), 2.77 (s, 3H, -NCH₃) ; LC- MS (m/z) 356.4 (M+l). Example 3: 4-[(4-Isopropylphenyl)amino]-6-(methylcarbamoyl)quinoline-3-carboxylic acid

(a) Preparation of intermediary compound ethyl 6-bromo-4-[(4-isopropylphenyl)- amino] quinoline-3 -carboxylate :

4-Isopropyl aniline (0.195 g, 1.4 mmol) and 6-bromo-4-chloroquinoline-3-carboxylic acid ethyl ester (1.0 g, 3.2 mmol) were dissolved in dioxane (7 mL) and irradiated in a microwave reactor at 150°C for 30 minutes. The reaction mixture was diluted with petroleum ether, and the precipitate formed filtered off and dried. This gave 0.5 g (85 % yield) of ethyl 6-bromo-4-[(4- isopropylphenyl)-amino]quinoline-3-carboxylate. LC-MS (m/z) 413.3 (M+l). ¹H NMR (300 MHz, CDCl₃) δ 9.15 (d, IH, J= 3 Hz), 8.06 (d, IH, J= 9 Hz), 7.86 (d, IH, J= 9 Hz), 7.79 (s, IH), 7.49 (d, 2H, J= 8.4 Hz), 7.35 (d, 2H, J= 8.4 Hz), 4.46 (dd, 2H, J= 7 Hz), 3.10 (m, IH), 1.45 (t, 3H, J= 7 Hz), 1.35 (d, 6H, J= 5 Hz).

(b) Preparation of intermediary compound ethyl 6-bromo-4-[(4-isopropylphenyl)- amino]quinoline-3-carboxylate:

Ethyl 6-bromo-4-[(4-isopropylphenyl)amino]quinoline-3-carboxylate (0.45 g, 1.08 mmol) was added to tetrahydrofuran followed by trans-di(μ-acetato)bis[o-(di-o-tolylphosphino)- benzyl]dipalladium(II) (50 mg, 0.054 mmol), tri-tert-butylphosphonium tetrafluoroborate (0.063 g, 0.2 mmol), molybdenum hexacarbonyl (0.575 g, 2.17 mmol), methyl amine (2 N in tetrahydrofuran, 3.2 mL, 6.5 mmol) and l,8-diazabicyclo[5.4.0]undec-7-ene (1.7 mL, 10 mmol). The reaction mixture was irradiated at 130 °C for 5 minutes in a microwave reactor. The reaction mixture was concentrated and the crude product was purified on column (flash chromatography, silica gel, dichloromethane/methanol 98:2). This gave 0.33 g (79% yield) of ethyl 6-bromo-4- [(4-isopropylphenyl)amino]quinoline-3-carboxylate. LC-MS (m/z) 392.1 (M+l). ¹H NMR (MeOD) δ 9.11 (s, IH), 8.23 (s, IH), 8.03 (d, IH, J= 4.5 Hz), 7.91 (d, IH, J= 8 Hz), 7.27 (d, 2H, J= 8.5 Hz), 7.10 (d, 2H, J= 8.5 Hz), 4.41 (dd, 2H, J= 7 Hz), 2.96 (m, IH), 2.82 (s, 3H), 1.43 (triplett, 3H, J= 7 Hz), 1.29 (d, J= 7 Hz).

(c) Ethyl 4-[(4-isopropylphenyl)amino]-6-(methylcarbamoyl)-quinoline-3-carboxylate (30 mg, 0.77 mmol) was stirred with lithium hydroxide monohydrate (128 mg, 3.0 mmol) in a mixture of methanol, tetrahydrofuran and water (6 mL, 2:2:2) overnight. The reaction mixture was concentrated and the aqueous layer washed with ethyl acetate. The aqueous layers were collected, acidified with aqueous hydrochloric acid and the precipitate formed was filtered off and dried to afford 22 mg (80 %) of 4-(4-isopropyl-phenylamino)-6-methylcarbamoyl-quinoline- 3-carboxylic acid as a yellow solid. LC-MS (m/z) 364.2 (M+1).1H NMR (MeOD) δ 12.08 (broad s, IH), 8.99 (s, IH), 8.82 (s, IH), 8.35 (d, IH, J= 8.7 Hz), 8.14 (d, IH, J= 8.7 Hz), 7.35 (m, 4H), 3.54 (m, IH), 2.50 (s, 3H), 1.23 (d, 6H, J= 5.1 Hz).

Example 4: 4-[[4-(Difluoromethoxy)phenyl]amino]-6-(methylcarbamoyl)quinoline-3- carboxylic acid.

(a) Preparation of intermediary compound ethyl-6-bromo-4-[[4-(difluoromethoxy)- phenyl]amino]quinoline-3-carboxylate:

6-bromo-4-chloroquinoline-3-carboxylic acid ethyl ester (0.30 g, 0.95 mmol) and 4- difluoromethoxyaniline (0.23 g, 1.43 mmol) was mixed in dioxane (4 mL) and heated under microwave conditions at 150°Cfor 20 minutes. The reaction mixture was cooled, concentrated in vacuo and suspended in dichloromethane. The reaction mixture was filtered, the residue evaporated and triturated with diethyl ether. The yellow crystals were washed twice with cold diethyl ether and dried in vacuo to afford ethyl-6-bromo-4-[[4-(difluoromethoxy)phenyl] amino] - quinoline-3-carboxylate in 90% purity (monitored by LC-MS). Yield: 250 mg (60 %). LC-MS (m/z) 437 (M+l).

(b) Preparation of intermediary compound ethyl 4-[[4-(difluoromethoxy)phenyl]amino]-6- (methylcarbamoyl)quinoline-3 -carboxylate :

Ethyl 6-bromo-4-[[4-(difluoromethoxy)phenyl]amino]quinoline-3-carboxylate (0.100 g, 0,21 mmol) , /rø«s-di(i-acetato)-bis[ø-(di-ø-tolylphosphino)benzyl]dipalladium(II) (Herrmann's palladacycle, 10.0 mg, 0.01 mmol), tri-tert-butylphosphonium tetrafluoroborate ([(^-Bu)₃PH]BF₄, 11.5 mg, 0.04 mmol) and molybdenum hexacarbonyl (111 mg, 0,42 mmol) were dissolved in tetrahydrofuran (2 mL), followed by and methylamine in tetrahydrofuran (2N, 0.7 mL, 1.4 mmol) and l,8-diazabicyclo[5.4.0]-undec-7-ene (32 mg, 0,21 mmol). The mixture irradiated under microwave conditions at 120 ⁰C for 5 minutes. The reaction mixture was evaporated and the crude product purified by column (flash chromatography on silica gel, dichloromethane/ methanol 97.5 : 2.5). This gave 25 mg (26 %) of ethyl 4-[[4-(difluoromethoxy)-phenyi]amino]-6- (methylcarbamoyl)quinoline-3-carboxylate in 95% purity as a yellow solid. LC-MS (m/z) 416 (M+l). ¹NMR (CDCl₃) δ 9.53 (s, IH), 8.46-8.38 (m, 2H), 8.20 (s, IH), 7.49 (s, 4H), 7.05-6.68 (triplett, IH, J= 73.1 Hz), 4.74 (dd, 2H, J= 7.2 Hz), 3.16 (d, 3H, J= 5.07 Hz), 1.74 (triplett, 3H, J= 7.2 Hz).

(c) Lithium hydroxide monohydrate (3.6 mg, 0.09 mmol) was added to a solution of ethyl 4-[[4-(difluoromethoxy)phenyl]amino]-6-(methylcarbamoyl)quinoline-3-carboxylate (15 mg, 0.036 mmol) in a mixture of tetrahydrofuran and water (1.5 mL, 8:2 mL) at room temperature. The reaction mixture was stirred overnight, acidified with aqueous hydrochloric acid (2N) and concentrated in vacuo. The yellow solid was suspended in water, filtered, washed twice with water and dried to give 5 mg (36 % yield) of 4-[[4-(difluoromethoxy)phenyl]amino]-6- (methylcarbamoyl)quinoline-3-carboxylic acid with a purity of 95%. LC-MS (m/z) 388 (M+l). NMR (CD₃OD) δ 9.19 (s, IH), 8.24-8.17 (m, 2H), 7.94 (d, IH, J= 8.6 Hz), 7.46 (d, 2H, J= 8.7 Hz), 7.32 (d, 2H, J= 8.7 Hz), 6.95 (triplett, IH, J= 73.5 Hz), 2.83 (s, 3H).

Example 5. 4-Ben2ylamino-6-methylcarbamoyl-quinoline-3-carboxylic acid:

(a) Preparation of intermediary compound iV-methyl-4-nitrobenzamide:

To a solution of aqueous methylamine (1.09 g, 35.1 mmol) and sodium hydroxide (1.2 g, 29.7 mmol) in water (27.5 mL) was added a solution of 4-nitrobenzoyl chloride (5.0 g, 27.0 mmol) in 2-butanone (6.5 mL) in such a way that the temperature of the reaction mixture not increased above 40 ⁰C. After completion of addition, water (20 mL) was added and the reaction mixture was stirred well for 3 hours. The solid precipitate was filtered, washed with cold water (2 x10 mL) and cold methanol (2 ^χ5 mL) to afford 3.5 (72 % yield) of Λ^-methyl-4-nitro- benzamide as a brown solid. ¹H-NMR (300 MHz, DMSO-d₆) δ 8.78 (bs, IH), 8.31 (d, J= 8.7 Hz, 2H), 8.06 (d, J= 8.7 Hz, 2H), 2.82 (d, J= 4.5 Hz, 3H). LC-MS (m/z, ⁰A): 180.5 (M+l, 99), 202.5 (M+Na).

(b) Preparation of intermediary compound 4-amino-iV-methylbenzamide:

To a solution of 7V-methyl-4-nitrobenzamide (23.0 g, 127.8 mmol) in ethanol (460 rnL) was added palladium on carbon (Pd/C, 10%, 50% moistened, 15.0 g). The mixture was hydrogenated under 2.0 atmospheric pressure at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered through Celite and thoroughly washed with ethanol. The filtrate was evaporated and the residue washed with diisopropyl ether to give 16.0 g (84 % yield) of 4-amino-N-methylbenzamide as an off-white solid. ¹H-NMR (300 MHz, DMSOd₆) δ 7.94 (bs, IH), 7.54 (d, J= 8.7 Hz, 2H), 6.52 (d, J= 8.7 Hz, 2H), 5.57 (s, 2H), 2.72 (d, J= 4.5 Hz, 3H). LC-MS 172.5 (M+Na).

(c) Preparation of intermediary compound 2-[(4-Methylcarbamoyl-phenylamino)- methylenejmalonic acid diethyl ester:

To a solution of 4-amino-iV-methyl-benzamide (7.5 g, 50 mmol) in anhydrous toluene (15 mL) was added diethoxymethylene malonate (11.88 g, 55.0 mmol) at room temperature. The reaction mixture was stirred at 100 ⁰C for 3 hours, cooled to 40 ⁰C and petroleum ether (200 mL) was added and the reaction mixture was subsequently stirred for 30 minutes. The solid formed, was collected by filtration, washed with diisopropylether and dried in vacuo to afford 15.0 g (94 % yield) of 2-[(4-methylcarbamoylphenylamino)methylene]malonic acid diethyl ester as a white solid. ¹H-NMR (300 MHz, CDCl₃) δ 11.09 (d, J= 13.5 Hz, IH), 8.54 (d, J= 13.5 Hz, 2H), 7.81 (d, J= 8.4 Hz, 2H), 7.18 (d, J= 8.7 Hz, 2H), 6.20 (bs, IH), 4.36-4.24 (m, 4H), 3.03 (d, J= 4.8 Hz, 3H), 1.41-1.33 (m, 6H). LC-MS (m/z, ⁰A): 319.6 (M-I, 99.7). (d) Preparation of intermediary compound 4-hydroxy-6-methylcarbamoylquinoline-3- carboxylic acid ethyl ester:

A mixture of 2-[(4-methylcarbamoylphenylamino)methylene]malonic acid diethyl ester (6.0 g, 18.8 mmol) and "Dowtherm A" (synthetic organic heat transfer fluid consisting of an eutectic mixture of biphenyl (C₁₂H₁₀) and diphenyl oxide (C₁₂H₁₀O), 60 mL) was heated to 200⁰C for 30 minutes. The temperature was raised to 24O⁰C and stirred for another 30 minutes. The reaction mixture was allowed to cool to room temperature and petroleum ether (50 mL) was added. A solid (0.6 g) precipitated out, was filtered and washed with dichloromethane (2 ^χ5 mL). The combined filtrates were concentrated in vacuo, the residue obtained mixed with "Dowtherm A" (60 mL) and heated to 240 ⁰C for 30 minutes. The reaction mixture was cooled to room temperature and treated with petroleum ether (50 mL). The formed solid precipitate was filtered and washed with dichloromethane (2 x10 mL) to afford 2.6 g (51 % yield) of 4-hydroxy-6- methylcarbamoylquinoline-3-carboxylic acid ethyl ester as a pale blue solid. IH-NMR (300 MHz, DMSOd₆) δ 12.46 (bs, IH), 8.68 (d, J= 4.5 Hz, IH), 8.65 (d, J= 1.8 Hz, 1H),8.58 (s, IH), 8.11 (dd, J= 9.9, 1.8 Hz, IH), 7.66 (d, J= 8.7 Hz, IH), 4.23 (q, J= 6.9 Hz, 2H), 2.81 (d, J = 4.5 Hz, 3H), 1.29 (triplet, J= 6.9 Hz, 3H). LC-MS (m/z, %): 275.2 (M+l, 98.7).

(e) Preparation of intermediary compound 4-chloro-6-methylcarbamoyl-quinoline-3- carboxylic acid ethyl ester:

4-Hydroxy-6-methylcarbamoyl-quinoline-3-carboxylic acid ethyl ester (1.0 g, 3.65 mmol) was added to phosphorus oxychloride (16.75 g, 109.2 mmol) at room temperature. The stirred mixture was heated at 100 ⁰C for 3 hours. The reaction mixture was cooled and concentrated in vacuo. The residue was dissolved in the tetrahydrofuran (10 mL) and the reaction mixture was neutralized (pH 7) by addition of triethylamine. The reaction mixture was diluted with dichloromethane (4 ^χ20 mL) and concentrated in vacuo. The crude residue was purified on column (flash chromatography on silica gel, chloroform/methanol 98:2) to give 0.8 g (75 % yield) of 4-chloro-6-methylcarbamoylquinoline-3-carboxylic acid ethyl ester as an off-white solid. ¹H-NMR (300 MHz, CDCl₃) δ 9.27 (s, IH), 8.79 (s, IH), 8.26-8.19 (m, 2H), 8.58 (s, IH), 6.48 (bs, IH), 4.53 (dd, J= 7.2 Hz, 2H), 3.13 (d, J= 4.8 Hz, 3H), 1.49 (triplett, J= 7.2 Hz, 3H). LC-MS (m/z, %): 293.1 (M+l, 98), 315 (M+Na).

(f) Preparation of intermediary compound 4-benzylamino-6-methylcarbamoylquinoline-3- carboxylic acid ethyl ester:

A 20 mL microwave vial was charged with 4-chloro-6-methylcarbamoylquinoline-3- carboxylic acid ethyl ester (150 mg, 0.514 mmol), benzylamine (0.082 g, 0.765 mmol), triethylamine (0.011 mL, 0.077 mmol) and dry AζiV-dimethylamine (10 mL). The vial was capped and the mixture irradiated with microwaves for 30 minutes at 160°C. The reaction mixture was cooled to room temperature and diluted with dichloromethane. The organic layer was washed with water (10 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was washed with ice-cold diisopropyl ether to give 90 mg (48 %) of 4-benzylamino- δ-methylcarbamoylquinoline-S-carboxylic acid ethyl ester. ¹H-NMR (300 MHz, CDCl₃) δ 9.87 (bs, IH), 9.19 (s, IH), 8.56 (d, J= 1.8 Hz, IH), 8.05- 7.96 (m, 2H), 7.52-7.37 (m, 5H), 5.62 (bs, IH), 5.05 (d, J= 6.0 Hz, 2H), 4.42 (q, J= 7.2 Hz, 2H), 2.9 (d, J= 4.8 Hz, 3H), 1.44 (t, J= 7.2 Hz, 3H). LC-MS (m/z, %): 364 (M+l, 98).

(g) To a solution of 4-benzylamino-6-methylcarbamoyl-quinoline-3-carboxylic acid ethyl ester (130 mg, 0.358 mmol) in a mixture of tetrahydrofuran (6 mL), methanol (6 mL) and water (6 mL) was added lithium hydroxide monohydrate (60 mg, 1.43 mmol) in one portion at room temperature. The clear solution was then stirred for 3 hours, during which time the reaction progress was monitored with thin layer chromatography. After completion of the reaction, the solvents were evaporated under reduced pressure. The highly viscous residue was treated with ice-cold aqueous hydrochloric acid (1.5 N) until the pH of the reaction mixture reached 1-2. The colourless solid precipitated out was filtered and washed with cold dichloromethane (3 mL) followed by cold diisopropyl ether (5 mL) to give 80 mg (75 % yield) of 4-benzylamino-6- methyl-carbamoylquinoline-3-carboxylic acid as an off-white solid. ¹H-NMR (300 MHz, CDCl₃) δ 11.31 (bs, IH), 9.04 (s, IH), 8.87 (d, J= 4.5 Hz, IH), 8.38 (d, J= 8.7 Hz, IH), 8.06 (d, J= 8.7 Hz, IH), 7.51-7.36 (m, 5H), 5.42 (bs, IH), 2.85 (d, J= 4.5 Hz, 3H). LC-MS (m/z, %): 336.2 (M+l, 98).

Example 6: 4-(2,6-Dimethyl-phenylamino)-6-methylcarbamoyl-quinoline-3-carboxyIic acid.

(a) Preparation of intermediary compound 4-(2,6-dimethyl-phenylamino)-6-methyl- carbamoyl-quinoline-3-carboxylic acid ethyl ester:

A 20 mL microwave vial was charged with 4-chloro-6-methylcarbamoylquinoline-3- carboxylic acid ethyl ester (400 mg, 1.370 mmol), 2,6-dimethylaniline (0.25 g, 2.066 mmol), triethylamine (0.028 mL, 2.06 mmol) and dry N,N-dimethylformamide (10 mL). The vial was capped and the mixture was irradiated with microwaves for 30 minutes at 160 °C. The reaction mixture was cooled to room temperature and diluted with dichloromethane. The organic layer was washed with water (10 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was washed with ice-cold diisopropyl ether to give 320 mg (62 % yield) of 4-(2,6- dimethylphenylamino)-6-methylcarbamoylquinoline-3 -carboxylic acid ethyl ester. H-NMR (300 MHz, CDCl₃) δ 10.98 (bs, IH), 9.28 (s, IH), 8.07 (dd, J= 8.7, 2.1 Hz, IH), 7.96 (d, J= 8.7 Hz, IH), 7.45 (d, J= 1.8 Hz, IH), 7.35-7.26 (m, 2H), 5.18 (d, J= 3.9 Hz, IH), 4.49 (dd, J= 7.2 Hz, 2H), 2.82 (d, J= 4.8 Hz, 3H), 2.18 (s, 6H), 1.50 (t, J= 7.2 Hz, 3H). LC-MS (m/z, %): 275.2 (M+l, 98.7).

(b) To a solution of 4-(2,6-dimethyl-phenylamino)-6-methylcarbamoylquinoline-3- carboxylic acid ethyl ester (0.36 g, 0.954 mmol) in a mixture of tetrahydrofuran (6 mL), methanol (6 mL) and water (6 mL), was added lithium hydroxide monohydrate (0.16 g, 3.81 mmol) in one portion at room temperature. The clear solution was stirred for 3 hours, during which time the reaction progress was monitored with thin layer chromatography. After completion of the reaction, the solvents were evaporated under reduced pressure. The highly viscous residue was treated with ice-cold aqueous hydrochloric acid (1.5 N) until the pH of the reaction mixture reached 1-2. The colourless solid precipitated out was filtered and washed with cold dichloromethane (3 mL) followed by cold diisopropyl ether (5 mL) to give 0.19 g (57 % yield) of 4-(2,6-dimethyl-phenylamino)-6-methylcarbamoyl-quinoline-3-carboxylic acid as off- white solid. ¹H-NMR (300 MHz, CDCl₃) δ 12.37 (bs, IH), 9.15 (s, IH), 8.56 (s, IH), 8.33 (d, J= 8.7 Hz, IH), 8.07(d, J= 8.7 Hz, IH), 7.36-7.26 (m, 3H), 2.71 (d, J= 3.6 Hz, 3H), 2.13 (s, 6H). LC-MS im/z, %): 350.1 (M+l, 97.6).

Example 7: 4-(3-Methoxy-phenylamino)-6-methyIcarbamoyl-quinoIine-3-carboxylic acid.

(a) Preparation of intermediary compound 4-(3-methoxy-phenylamino)-6-methyl- carbamoyl-quinoline-3-carboxylic acid ethyl ester.

A 20 mL microwave vial was charged with 4-chloro-6-methylcarbamoylquinoline-3- carboxylic acid ethyl ester (150 mg, 0.514 mmol), w-anisidine (0.095 g, 0.771 mmol), triethylamine (0.010 mL, 0,51 mmol) and dry iV,JV-dimethylformamide (10 mL). The vial was capped and the mixture was irradiated with microwaves for 30 minutes at 160°C. The reaction mixture was cooled to room temperature and diluted with dichloromethane. The organic layer was washed with water (10 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was washed with ice-cold diisopropyl ether to give 85 mg (43 % yield) of 4-(3- methoxyphenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid ethyl ester. ¹H-NMR (300 MHz, CDCl₃) δ 10.65 (bs, IH), 9.30 (s, IH), 8.07-7.99 (m, 2H), 7.95 (s, IH), 7.30-7.25 (m, IH), 6.83-6.80 (m, IH), 6.72-6.70 (m, IH), 5.54 (bs, IH), 4.47 (dd, J= 7.2 Hz, 2H), 3.78 (s, 3H), 2.88 (d, J= 4.8 Hz, 3H), 1.48 (triplet., J= 7.2 Hz, 3H).

(b) To a solution of 4-(3-methoxyphenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid ethyl ester (115 mg, 0.303 mmol) in a mixture of tetrahydrofuran (6 niL), methanol (6 mL) and water (6 mL) was added lithium hydroxide monohydrate (51 mg, 1.2 mmol) in one portion at room temperature. The clear solution was then stirred for 3 hours, during which time the reaction was monitored with thin layer chromatography. After completion of the reaction, the solvents were evaporated under reduced pressure. The highly viscous residue was treated with ice-cold aqueous hydrochloric acid (1.5 N) until the pH of the reaction mixture reached 1-2. The colourless solid precipitated out was filtered and washed with cold dichloromethane (3 mL) followed by cold diisopropyl ether (5 mL) to give 67 mg (63 % yield) of 4-(3-methoxy- phenylamino)-6-methylcarbamoyl-quinoline-3-carboxylic acid as an off-white solid. ¹H-NMR (300 MHz, DMSOd₆) δ 11.85 (bs, IH), 9.11 (s, IH), 8.82 (s, IH), 8.73 (d, J= 4.2 Hz, IH), 8.34 (d, J= 8.7 Hz, IH), 8.09 (d, J= 8.7 Hz, IH), 7.35 (t, J= 8.1 Hz, IH), 6.99 (s, IH), 6.91 (d, J= 8.1 Hz, 2H), 3.74 (s, 3H), 2.78 (d, J= 4 .5 Hz, 3H). LC-MS (m/z, %): 352.1 (M+l, 98.1).

Example 8: 4-(2-Fluoro-phenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid.

(a) Preparation of intermediary compound 4-(2-fluorophenylamino)-6-methylcarbamoyl- quinoline-3-carboxylic acid ethyl ester:

A 20 mL microwave vial was charged with 4-chloro-6-methylcarbamoylquinoline-3- carboxylic acid ethyl ester (360 mg, 1.233 mmol), 2-fluoroaniline (0.206 g, 1.853 mmol), triethylamine (0.025 mL, 1.85 mmol) and dry N^V-dimethylformamide (10 mL). The vial was capped and the mixture was irradiated with microwaves for 30 minutes at 160 °C. The reaction mixture was cooled to room temperature and diluted with dichloromethane. The organic layer was washed with water (10 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was washed with ice-cold diisopropyl ether to give 95 mg (21 % yield) of 4-(2- fluorophenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid ethyl ester. H-NMR (300 MHz, CDCl₃) δ: 10.54 (bs, IH), 9.32 (s, IH), 8.01 (d, J= 10.8 Hz, 3H), 7.28-7.09 (m, 4H), 5.68 (bs, IH), 4.48 (dd, J= 7.2 Hz, 2H), 2.89 (d, J= 4.5 Hz, 3H), 1.49 (t, J= 7.2 Hz, 3H).

(b) To a solution of 4-(2-fluorophenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid ethyl ester (115 mg, 0.313 mmol) in a mixture of tetrahydrofuran (6 mL), methanol (6 mL) and water (6 mL) was added lithium hydroxide monohydrate (53 mg, 1.3 mmol) in one portion at room temperature. The clear solution was stirred for 3 hours, during which time the reaction was monitored with thin layer chromatography. After completion of the reaction, the solvents were evaporated under reduced pressure. The highly viscous residue was treated with ice-cold aqueous hydrochloric acid (1.5 N) until the pH of the reaction mixture reached 1-2. The colourless solid precipitated out was filtered and washed with cold dichloromethane (3 mL) followed by cold diisopropyl ether (5 mL) to give 70 mg (66 % yield) of 4-(2-fluoro- phenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid as a pale yellow solid. H-NMR (300 MHz, CDCl₃) δ 12.09 (bs, IH), 9.32 (s, IH), 9.09 (s, IH), 8.84 (s, 2H), 8.38 (d, J= 8.7 Hz, IH), 8.15 (d, J= 8.7 Hz, IH), 7.48-7.38 (m, 3H), 7.30-7.24 (m, IH), 2.76 (d, J= 4.2 Hz, 3H). LC-MS (m/z, %): 340.1 (M+l, 98.9).

Example 9: 6-Methylcarbamoyl-4-phenylamino-quinoline-3-carboxyIic acid.

(a) Preparation of intermediary compound 6-methylcarbamoyl-4-phenylaminoquinoline-3- carboxylic acid ethyl ester:

A 20 mL microwave vial was charged with compound 4-chloro-6-methylcarbamoyl- quinoline-3-carboxylic acid ethyl ester (150 mg, 0.514 mmol), aniline (0.072 g, 0.774 mmol), triethylamine (0.011 mL, 0.077 mmol) and dry ΛζjV-dimethylfbrmamide (10 mL). The vial was capped and the mixture was irradiated with microwaves for 30 minutes at 160 °C. The reaction

5 mixture was cooled to room temperature and diluted with dichloromethane. The organic layer was washed with water (10 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was washed with ice-cold diisopropyl ether to give 90 mg (50 % yield) of 6- methylcarbamoyl-4-phenylaminoquinoline-3-carboxylic acid ethyl ester. ¹H-NMR (300 MHz, CDCl₃) δ 10.71 (bs, IH), 9.30 (s, IH), 8.06-7.99 (m, 2H), 7.86 (d, J= 1.5 Hz, IH), 7.44-7.38 (m,

10 2H), 7.31-7.27 (m, IH), 7.17 (d, J= 7.8 Hz, 2H), 5.47 (bs, IH), 4.47 (q, J= 7.2 Hz, 2H), 2.85 (d, J= 4.8 Hz, 3H), 1.48 (t, J= 7.2 Hz, 3H).

(b) To a solution of 6-methylcarbamoyl-4-phenylaminoquinoline-3-carboxylic acid ethyl ester (120 mg, 0.344 mmol) in a mixture of tetrahydrofuran (6 mL), methanol (6 mL) and water (6 mL) was added lithium hydroxide monohydrate (60 mg, 1.43 mmol) in one portion at room

5 temperature. The clear solution was then stirred for 3 hours, during which time the reaction was monitored with thin layer chromatography. After completion of the reaction, the solvents were evaporated under reduced pressure. The highly viscous residue was treated with ice-cold aqueous hydrochloric acid (1.5 N) until the pH of the reaction mixture reached 1-2. The colourless solid precipitated out was filtered and washed with cold dichloromethane (3 mL) followed by cold i0 diisopropyl ether (5 mL) to give 63 mg (57 % yield) of 6-methylcarbamoyl-4-phenylamino- quinoline-3-carboxylic acid ¹H-NMR (300 MHz, DMSO-d₆) δ 11.95 (bs, IH), 9.11 (s, IH), 8.58- 8.57 (m, 2H), 8.23 (d, J= 8.4 Hz, IH), 8.01 (d, J= 8.7 Hz, 1H),7.42 (triplet, J= 7.8 Hz, 2H), 7.29-7.27 (m, 3H), 2.74 (d, J= 4.5 Hz, 3H). LC-MS (m/z, %): 322.1 (M+l, 96.4).

:5 BIOLOGICAL ASSAYS

Chemotaxis assay

Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammation. Microporous membrane inserts are widely used for cell migration and invasion assays. The most widely 0 accepted of which is the Boyden Chamber assay. The Boyden chamber assay, is based on a chamber of two medium-filled compartments separated by a microporous membrane. Cells are placed in the upper compartment and are allowed to migrate through the pores of the membrane into the lower compartment, in which chemotactic agents are present. After an appropriate incubation time, the membrane between the two compartments is fixed and stained, and the number of cells that have migrated to the lower side of the membrane is determined. Therefore, the Boyden chamber-based cell migration assay has also been called chemotaxis assay. The compounds were tested for their capacity of influencing chemotaxis, using porcine aorta endothelial (PAE) cells expressing VEGFR2 and VEGFR3 (PAE/VEGFR-2 and PAE/VEGFR- 3) with VEGF-A and VEGF-C, respectively, as chemotactic agent. The method used a modified Boyden chamber assay. The migration of the PAE cells expressing VEGFR2 and VEGFR3 receptors toward VEGF-A and VEGF-C respectively used as chemo-attractant was studied though micropore polycarbonate filter and was scored in the absence of serum. The assay was performed in the presence of compounds at 10 μM concentrations and the quantified measure of the inhibiton activity of compounds is expressed in %.

Cell adhesion assay:

The cell adhesion assay is a measure of the ability of a compound to inhibit adhesion to fibronectin. Therefore a high % inhibition value indicates that the compounds inhibit adhesion to flbronectine and are therefore integrin antagonists for α5β 1.

Day 1 : Flat-bottomed microwell plates (Maxisorb, Nunc) are coated with 20 μg/ml of fibronectin (FN) and incubated over night at room temperature in a humified box.

Day 2: The excess of coating agent (fibronectin) is removed by washing three times in PBS. After washing, 50 μl 2 % PVP (polyvinylpyrrolidone) is added as a blocking agent and the plates are incubated one hour at room temperature. The blocking agent is then removed by washing 3 times in PBS. In the meantime HT1080 human fibrosarcoma cells (2 x 10 cells/ml) are pre-incubated 20 minutes at room temperature with substances to a final concentration of 100 μM (maximum DMSO concentration is 0.1%). The pre-incubated cells are added to the plates and incubated 20 minutes at 37 °C. After incubation the plates are washed 5 times in pre-warmed EMEM medium and the cells that had adhered to fibronectin is fixated in 4 % paraformaldehyde for 30 minutes in the humified box. To the fixated cells 0.5 % toluidine blue is added and the plates are incubated over night in the humified chamber.

Day 3: The plates are washed three times in deionised water and then 100 μl 2 % SDS is added to release the dye from the cells. After 20 minutes the plates are ready to be analysed by an ELISA-reader at 630 rim.

In Table 1, data from both the chemotaxis assay and the cell adhesion assay are shown. Thus, "Chemotaxis VEGFR-2 % inhibition of cell migration" and "Chemotaxis VEGFR-3 % inhibition of cell migration" show the percentage inhibition of PAE cells expressing VEGFR-2 or 3 in the presence of 10 μM of the indicated inventive compound. Data under "Cell adhesion assay % inhibition to fibronectin" show the percentage of inhibition of the compound at lOOμM.

Claims

1. A compound of formula (I)

(I) wherein: n is 0 (zero) or 1 ;

1 0

R and R are independently selected from hydrogen; branched or unbranched C]-C₈ alkyl, C₂-C₈ alkenyl, or C₂-C₈ alkynyl; monocyclic or bicyclic, saturated or unsaturated C₃-C₈ carbocyclyl; and monocyclic or bicyclic, saturated or unsaturated Ci-C₇ heterocyclyl wherein the heteroatoms are independently selected from N, O and S; said alkyl, alkenyl, alkynyl, carbocyclyl or heterocyclyl optionally being substituted with 1, 2, or 3 groups R^a'

R is selected from monocyclic or bicyclic C₆-C₁₀ aryl; and monocyclic or bicyclic C₁-C₉ heteroaryl, wherein the heteroatoms independently are selected from N, O and S; said aryl or heteroaryl optionally being substituted with 1, 2, 3, 4 or 5 groups R ;

Y is selected from -C(O)-; -S(O)-; and -S(O)₂-;

X is selected from -NR⁰-; -0-; and -S-; each R^a is independently selected from halogen; hydroxy; carbonyl; methoxy; halomethoxy; dihalomethoxy; and trihalomethoxy; each R is independently selected from halogen, branched or unbranched C₁-C₄ alkyl, C₂- C₄ alkenyl or C₂- C₄ alkynyl; branched or unbranched C₁-C₄ alkyloxy, C₂-C₄ alkenyloxy or C₂-C₄ alkynyloxy; branched or unbranched Cj-C₄ alkylthio, C₂-C₄ alkenylthio or C₂-C₄ alkynylthio; said alkyl, alkenyl, alkynyl, alkyloxy, alkenyloxy alkynyloxy, alkylthio, alkenylthio or alkynylthio group optionally being substituted with 1, 2 or 3 halogens;

R° is selected from hydrogen and branched or unbranched C₁-C₄ alkyl, C₂-C₄ alkenyl or C₂-C₄ alkynyl; or a pharmaceutically acceptable salt thereof.

2. A compound according to claim 1, wherein n is 0.

3. A compound according to claim 1 or claim 2, wherein R and R are independently selected from hydrogen and branched or unbranched C₁-C₈ alkyl, C₂-C₈ alkenyl and C₂-C₈ alkynyl, said alkyl, alkenyl, or alkynyl optionally being substituted with 1, 2, or 3 groups R .

4. A compound according to claim 3, wherein R and R are independently selected from hydrogen and branched or unbranched Cj-C₈ alkyl, said alkyl optionally being substituted with 1, 2, or 3 groups R .

5. A compound according to claim 3, wherein R is hydrogen and R is selected from C₁-C₄ alkyl.

6. A compound according to any one of claims 1 to 5, wherein R³ is phenyl, optionally substituted with 1, 2, 3, 4 or 5 groups R .

7. A compound according to any one of claims 1 to 6, wherein Y is -C(O)-.

8. A compound according to any one of claims 1 to 7, wherein X is -NR⁰-.

9. A compound according to any one of claims 1 to 8, wherein each R^a is halogen.

10. A compound according to any of claims 1 to 9, wherein each R is independently selected from halogen; and Ci-C₄ alkyl; and C₁-C₄ alkyloxy; wherein any alkyl or alkyloxy optionally is substituted with 1, 2 or 3 halogens.

11. A compound according to any one of claims 1 to 10, wherein R is hydrogen.

12. A compound according to claim 1 which is:

4-(4-fluorophenylamino)-6-(methylcarbamoyl)quinoline-3-carboxylic acid; 4-(4-chlorophenylamino)-6-(methylcarbamoyl)quinoline-3-carboxylic acid; 4-[(4-isopropylphenyl)amino]-6-(methylcarbamoyl)quinoline-3-carboxylic acid; 4-[[4-(difluoromethoxy)phenyl]amino]-6-(methylcarbamoyl)quinoline-3-carboxylic acid; 4-benzylamino-6-methylcarbamoyl-quinoline-3 -carboxylic acid; 4-(2,6-dimethyl-phenylamino)-6-methylcarbamoyl-quinoline-3-carboxylic acid; 4-(3-methoxy-phenylamino)-6-methylcarbamoyl-quinoline-3-carboxylic acid;

4-(2-fluoro-phenylamino)-6-methylcarbamoylquinoline-3-carboxylic acid; 6-methylcarbamoyl-4-phenylamino-quinoline-3 -carboxylic acid, or a pharmaceutically acceptable salt thereof.

13. A compound according to any one of claims 1 to 12 for use in therapy.

14. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any of claims 1 to 12, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable excipient.

15. A compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disorder selected from cancer, diabetic retinopathy, age- related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema and psoriasis.

16. The use of a compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, in the manufacturing of a medicament for use in the treatment of a disorder selected from cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema and psoriasis.

17. A method of treating a mammal suffering from cancer, diabetic retinopathy, age-related macular degeneration, inflammation, stroke, ischemic myocardium, atherosclerosis, macular edema or psoriasis, comprising administering to said mammal in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof.