DIHYDROBENZOPYRAN, THIOCHROMAN, TETRAHYDROQUINOLEINE AND TETRAHYDRONAPHTHALENE DERIVATIVES AND THEIR USE IN ANTICANCER THERAPY The present invention relates to new compounds, particularly dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives, to pharmaceutical compositions comprising thereof and to their use for treatment of diseases, particularly proliferative disorder such as cancer.

BACKGROUND OF THE INVENTION

Proliferative disorder such as cancer is becoming a more and more important cause of death amongst humans. More than 80% of all anti-cancer drugs are directed towards the apoptosis pathway of tumor cells and are cytotoxic upon activating said pathway. They are called antineoplastic agents, neoplastic referring to cancer cells. The very nature of antineoplastic agents makes them harmful to healthy constantly dividing cells and tissues, as well as the cancerous cells. A large number of cancer cells such as glioblastomas (brain cancers), brain metastases, melanomas, pancreatic cancers, lung cancers of the NSCLC-type, refractory prostate cancers (HRPC), breast cancers such as triple negatives and other types are naturally resistant to apoptosis and cannot be treated by the many known drugs and chemotherapeutics.

The present invention therefore investigated the potential of new compounds, to have a cytotoxic and/or a cytostatic effect on apoptosis resistant tumor cells or cancer cells, especially in the context of brain cancers including gliomas.

There is a continuous need in the art for improving the efficacy of antiproliferative treatments in humans by providing suitable combinations of new drugs with conventional antineoplastic agents. SUMMARY OF THE INVENTION

We have now found new compounds particularly dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives that provide a solution to the above stated problem.

We have unexpectedly found that the compounds of the invention appear to have important anti-cancer effects, while not impairing normal cell biology of non-tumor or non-cancer cells.

The new compounds are represented by general formula (I):

wherein

X represents Oxygen, Sulphur or an element selected from the group consisting of ( -NR ₅ , -CR5R6 or -SO2); wherein R ₅ and R ₆ which can be identical or different, denotes each independently one element selected from the group consisting of (hydrogen, a d-12-alkyl optionally mono- or polysubstituted with a halogen atom or a C3-8-cycloalkyl optionally mono- or polysubstituted with a halogen atom) assuming that if Z represents Oxygen or Sulphur, R ₅ cannot be hydrogen;

Y represents Oxygen or Sulphur;

Z represents Oxygen or Sulphur or NH; if Z represents NH, then R ₄ is different from H.

R-i , R ₂ and R ₄ represent a hydrogen atom, a C-i-12-alkyl optionally mono- or polysubstituted with a halogen atom or a C3-8-cycloalkyl group optionally mono- or polysubstituted with a halogen atom;

Ri and R ₂ optionally form together a hydrocarbon ring of 3 to 6 atoms, one carbon atom optionally being replaced by an oxygen atom.

R ₃ represents an element selected from the group consisting of (hydrogen, halogen, C-i-12-alkyl optionally mono- or polysubstituted with a halogen atom, C3-8-cycloalkyl optionally mono- or polysubstituted with a halogen atom, hydroxy, C-i-6-alkoxy, C1-6- alkoxy-C-i-6-alkyl, nitro, amino, aminomethyl, cyano, cyanomethyl, perhalomethyl, C-i- 6-monoalkyl- or dialkylamino, sulfamoyl, C-i-6-alkylthio, C-i-6-alkylsulfonyl, C1-6- alkylsulfinyl, formyl, C-i-6-alkylcarbonylamino, R ₇ arylthio, R ₇ arylsulfinyl, R ₇ arylsulfonyl, C-i-6-alkoxycarbonyl, Ci-6-alkoxycarbonyl-Ci-6-alkyl, carbamoyl, carbamoylmethyl, C-i- 6-monoalkyl- or dialkylaminocarbonyl, C-i-6-monoalkyl- or dialkylaminothiocarbonyl, ureido, C-i-6-monoalkyl- or dialkylaminocarbonylamino, thioureido, C-i-6-monoalkyl- or dialkylaminothiocarbonylamino, Ci-6-monoalkyl- or dialkylaminosulfonyl, carboxy, carboxy-Ci-6-alkyl, acyl, R ₇ aryl, R ₇ arylalkyl, R ₇ aryloxy), where R ₇ denotes one or several elements selected from the group consisting of (hydrogen, C-i-6-alkyl, halogen, hydroxy or C-i-6-alkoxy).

"C-i-6-alkyl" as used herein, alone or in combination, refers to a straight or branched, saturated hydrocarbon chain having 1 to 6 carbon atoms such as methyl, propyl, butyl, isopentyl, hexyl, 1 -methylbutyl, 1 ,2-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl and the like.

"C-i-12-alkyl" as used herein, alone or in combination, refers to a straight or branched, saturated hydrocarbon chain having 1 to 12 carbon atoms.

"C3-8-cycloalkyl" as used herein refers to a radical of a saturated cyclic hydrocarbon chain having 3 to 8 carbon atoms such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

"C-i-6-alkoxy" as used herein, alone or in combination, refers to a straight or branched monovalent substituent comprising a d-6-alkyl group linked through an ether oxygen having its free valence bond from the ether oxygen and having 1 to 6 carbon atoms such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, pentoxy, tert- butoxy and the like.

"Ci-6-alkoxy-Ci-6-alkyl" as used herein refers to a group of 2-12 carbon atoms interrupted by an oxygen atom such as -CH ₂ -0-CH ₃ , -CH2CH2O-CH3, -CH ₂ -0- CH2CH3, -CH ₂ -0-CH(CH ₃ ) ₂ , -CH ₂ CH ₂ -0-CH(CH ₃ )2 , -CH(CH ₃ )CH ₂ -0-CH ₃ and the like.

"halogen" means fluorine, chlorine, bromine or iodine. "perhalomethyl" means trifluoromethyl, trichloromethyl, tribromomethyl or triiodomethyl.

"C-i-6-monoalkylamino" as used herein refers to an amino group wherein one of the hydrogen atoms is substituted with a straight or branched, saturated hydrocarbon chain having 1 to 6 carbon atoms such as methylamino, ethylamino, propylamino, isopropylamino, butylamino, tert-butylamino, isopentylamino, hexylamino and the like. "Ci-6-dialkylamino" as used herein refers to an amino group wherein the two hydrogen atoms independently are substituted with a straight or branched, saturated hydrocarbon chain having 1 to 6 carbon atoms such as dimethylamino, N-ethyl-N- methylamino, N-methyl-N-isopropylamino, N-butyl-N-methylamino, dihexylamino and the like.

"C-i-6-alkylthio" as used herein, alone or in combination, refers to a straight or branched monovalent substituent comprising a d-6-alkyl group linked through a divalent sulfur atom having its free valence bond from the sulfur atom and having 1 to 6 carbon atoms such as methylthio, ethylthio, propylthio, isopropylthio, butylthio, pentylthio, 3-methylpentylthio and the like.

"C-i-6-alkylsulfonyl" as used herein refers to a monovalent substituent comprising a C-i-6-alkyl group linked through a sulfonyl group (-S(=0)2-) such as methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, pentylsulfonyl, 2- methylpentylsulfonyl and the like.

"C-i-6-alkylsulfinyl" as used herein refers to a monovalent substituent comprising a C-i-6-alkyl group linked through a sulfinyl group (-S(=0)-) such as methylsulfinyl, ethylsulfinyl, propylsulfinyl, isopropylsulfinyl, tert-butylsulfinyl, pentylsulfinyl, 2- ethylbutylsulfinyl and the like.

"acyl" as used herein refers to a monovalent substituent comprising a

C-i-6-alkyl group linked through a carbonyl group such as acetyl, propionyl, butyryl, isobutyryl, pivaloyl, valeryl and the like.

"C-i-6-alkylcarbonylamino" as used herein refers to an amino group wherein one of the hydrogen atoms is substituted with an acyl group such as acetamido, propionamido, isopropylcarbonylamino, 2-ethylbutylcarbonylamino and the like. "aryl" as used herein refers to phenyl, 1 -naphthyl, or 2-naphthyl.

"arylthio" as used herein, alone or in combination, refers to an aryl group linked through a divalent sulfur atom having its free valence bond from the sulfur atom, the aryl group is substituted or not by one or several elements of R ₇ such as phenylthio, 1 -naphthylthio, 2-methylphenylthio, 3-methoxyphenylthio and the like.

"arylsulfinyl" as used herein, alone or in combination, refers to an aryl group linked through a sulfinyl group (-S(=0)-), the aryl group is substituted or not by one or several elements of R ₇ such as phenylsulfinyl, 2-methylphenylsulfinyl, 3-chloro-1 - naphthylsulfinyl and the like.

"arylsulfonyl" as used herein, alone or in combination, refers to an aryl group linked through a sulfonyl group (-S(=0)2-), the aryl group is substituted or not by one or several elements of R ₇ such as phenylsulfonyl, 2-methylphenylsulfonyl, 4- iodophenylsulfonyl, 2-naphthylsulfonyl and the like.

"C-i-6-alkoxycarbonyl" as used herein refers to a monovalent substituent comprising a C-i-6-alkoxy group linked through a carbonyl group such as methoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl, 2-methylpentoxycarbonyl and the like.

"Ci-6-monoalkylaminocarbonyl" as used herein refers to a monovalent substituent comprising a d-6-monoalkylamino group linked through a carbonyl group such as methylaminocarbonyl, isopropylaminocarbonyl, butylaminocarbonyl, 2- methylbutylaminocarbonyl and the like.

"C-i-6-dialkylaminocarbonyl" as used herein refers to a monovalent substituent comprising a C-i-6-dialkylamino group linked through a carbonyl group such as dimethylaminocarbonyl, diethylaminocarbonyl, N-methyl-N-isopropylaminocarbonyl, N-methyl-N-butylaminocarbonyl, N-propyl-N-2-methylbutylaminocarbonyl and the like.

"Ci-6-monoalkylaminothiocarbonyl" as used herein refers to a monovalent substituent comprising a C-i-6-monoalkylamino group linked through a thiocarbonyl group such as methylaminothiocarbonyl, isopropylaminothiocarbonyl, butylaminothiocarbonyl, 3- methylpentylaminothiocarbonyl, 1 ,2-dimethylbutylaminothiocarbonyl and the like.

"Ci-6-dialkylaminothiocarbonyl" as used herein refers to a monovalent substituent comprising a d-6-dialkylamino group linked through a thiocarbonyl group such as dimethylaminothiocarbonyl, diethylaminothiocarbonyl N-methyl-N- isopropylaminothiocarbonyl, N-methyl-N-butylaminothiocarbonyl N-tert-butyl-N- hexylaminothiocarbonyl and the like.

"Ci-6-monoalkylaminocarbonylamino" as used herein refers to an amino group wherein one of the hydrogen atoms is substituted with a C-i-6- monoalkylaminocarbonyl group such as methylaminocarbonylamino, ethylaminocarbonylamino, propylaminocarbonylamino, 3- methylbutylaminocarbonylamino, 1 ,2-dimethylbutylaminocarbonylamino and the like.

"Ci-6-dialkylaminocarbonylamino" as used herein refers to an amino group wherein one of the hydrogen atoms is substituted with a C-i-6-dialkylaminocarbonyl group such as dimethylaminocarbonylamino, diethylaminocarbonylamino, N-methyl-N- ethylaminocarbonylamino, N-methyl-N-isopropylaminocarbonylamino, N-propyl-N- pentylaminocarbonylamino and the like.

"Ci-6-monoalkylaminothiocarbonylamino" as used herein refers to an amino group wherein one of the hydrogen atoms is substituted with a Ci-6-monoalkylaminothiocarbonyl group such as methylaminothiocarbonylamino, ethylaminothiocarbonylamino, propylaminothiocarbonylamino, 3- methylpentylaminothiocarbonylamino and the like. "Ci-6-dialkylaminothiocarbonylamino" as used herein refers to an amino group wherein one of the hydrogen atoms is substituted with a C-i-6- dialkylaminothiocarbonyl group such as dimethylaminothiocarbonylamino, diethylaminothiocarbonylamino, N-methyl-N-ethylaminothiocarbonylamino, N-methyl- N-propylaminothiocarbonylamino , N-isopropyl-N-hexylaminothiocarbonylamino, N- 3-methylpentyl-N-pentylaminothiocarbonylamino and the like.

"Ci-6-monoalkylaminosulfonyl" as used herein refers to a monovalent substituent comprising a C-i-6-monoalkylamino group linked through a sulfonyl group such as methylaminosulfonyl, ethylaminosulfonyl, propylaminosulfonyl, hexylaminosulfonyl, tert-butylaminosulfonyl, 1 ,2-dimethylbutylaminosulfonyl and the like.

"C-i-6-dialkylaminosulfonyl" as used herein refers to a monovalent substituent comprising a d-6-dialkylamino group linked through a sulfonyl group such as dimethylaminosulfonyl, diethylaminosulfonyl, N-methyl-N-ethylaminosulfonyl N- methyl-N-propylaminosulfonyl, N-hexyl-N-3-methylbutylaminosulfonyl and the like.

"Carbamoyl " as used herein refers to NH ₂ CO-.

"sulfamoyl" as used herein refers to NH2-SO2-. "ureido" as used herein means -NH-CO-NH ₂ . "thioureido" as used herein means -NH-CS-NH ₂ . "arylalkyl" as used herein refers to a straight or branched saturated carbon chain containing from 1 to 6 carbons substituted with an aromatic carbohydride, the aryl group is substituted or not by one or several elements of R ₇ . "aryloxy" as used herein refers to phenoxy, 1 -naphthyloxy or 2-naphthyloxy, the aryl group is substituted or not by one or several elements of R ₇ .

"R ₇ aryl as used herein refers to aryl substituted by R _7. "Spiro compounds" as used herein refers to the compounds of Formula (I) wherein Ri and R ₂ form together a hydrocarbon ring of 3 to 6 atoms providing polycyclic compounds such as spiro[2/-/-1 -benzopyran-2, 1 '-cyclobutane] derivatives, spiro[2/-/- 1 -benzopyran-2,1 '-cyclopentane] derivatives and the like. "Spiro carbon" as used herein refers to the quaternary carbon atom connecting the two rings.

"dihydrobenzopyran derivatives" as used herein refers to the compounds of Formula (I) wherein -X- is represented by -0- (oxygen),

"thiochroman derivatives" as used herein refers to the compounds of Formula (I) wherein -X- is represented by -S- (sulphur) or -SO2-,

"tetrahydroquinoleine derivatives as used herein refers to the compounds of Formula (I) wherein -X- is represented by -NR ₅ -, and

"tetrahydronaphthalene derivatives" as used herein refers to the compounds of Formula (I) wherein -X- is represented by -CR5R6-.

This invention also refers to all optical isomers of the compounds represented by formula (I), particularly to all optically active isomers of dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives, and their mixtures including racemic mixtures thereof. When one has an asymetrical carbon atom in the compound of general formula (I), the invention also refers to pure optical isomers than to racemic mixture.

The invention refers also to tautomeric forms of the compounds represented by formula (I), particularly to tautomeric forms of dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives and to pharmacologically acceptable salts of said compounds.

By pharmacologically acceptable salts of the derivatives, one means pharmaceutically acceptable acid addition salts, pharmaceutically acceptable metal salts or optionally alkylated ammonium salts.

Preferred compounds of the Invention are:

R/S-/V-2-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran- 4-yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran- 4-yl)urea

R/S-/V-4-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran- 4-yl)urea

R/S-/V-2-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran- 4-yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran- 4-yl)urea

R/S-/V-4-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran- 4-yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-ie f-butoxycarbonylaminothiochroman-4-yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-ie f-butoxycarbonylaminothiochroman-4-yl)urea

R/S-/V-4-chlorophenyl-/V'-(6-ie f-butoxycarbonylaminothiochroman-4-yl)urea

R/S-/V-2-chlorophenyl-/V'-(6-ie f-butoxycarbonylaminothiochroman-4-yl)urea

R/S-/V-4-cyanophenyl-/V'-(6-ie f-butoxycarbonylaminothiochroman-4-yl)urea

R/S-/V-2-cyanophenyl-/V'-(6-ie f-butoxycarbonylaminothiochroman-4-yl)urea R/S-/V-4-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- yl)urea

R/S-/V-2-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- yl)urea

R/S-/V-4-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- yl)urea

R/S-/V-2-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2/-/- 1 -benzopyran-4-yl)urea

R/S-/V-4-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2H- 1 -benzopyran-4-yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2/-/- 1 -benzopyran-4-yl)urea

R/S-/V-4-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2H- 1 -benzopyran-4-yl)urea

R/S-/V-4-fluorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-yl)urea

R/S-/V-3-fluorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-yl)urea

R/S-/V-3-bromophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2/-/- 1 -benzopyran-4-yl)urea

R/S-/V-3,5-difluorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl- 2/-/-1 -benzopyran-4-yl)urea

R/S-/V-4-nitrophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2H-1 - benzopyran-4-yl)urea

R/S-/V-4-aminophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2H- 1 -benzopyran-4-yl)urea R/S-/V-3-nitrophenyl-/V'-(6-terf-^

benzopyran-4-yl)urea

R/S-/V-3-aminophenyl-/V'-(6-fe f-butoxycarbonylamino-3,4-dihydro-2,2-dimeth 1 -benzopyran-4-yl)urea

R/S-A/-4-tnfluoromethylphenyl-A/'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

R/S-/V-3-trifluoromethylphenyl-/V'-(6-ferf-butoxycarbonylami no-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

R/S-/V-3,5-dichlorophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl- 2/-/-1 -benzopyran-4-yl)urea

R/S-N-2-nitrophenyl-/V'-(6-te t-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-yl)urea

R/S-A/-2-trifluoromethylphenyl-A/'-(6-teri-butoxycarbonylami no-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

R/S-/V-3,5-di(trifluoromethyl)phenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2 dimethyl-2/-/-1 -benzopyran-4-yl)urea

R/S-A/-2-chloro-4-trifluoromethylphenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro- 2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

R/S-A/-2-chloro-5-trifluoromethylphenyl-/V'-(6-ie t-butoxycarbonylamino-3,4-dihydro^ 2,2-dimethyl-2H-1 -benzopyran-4-yl)urea

R/S-/V-2-chloro-6-trifluoromethylphenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro- 2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

R/S-/V-4-chloro-3-trifluoromethylphenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro 2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

R/S-A/-4-bromophenyl-/V'-(6-feri-butoxycarbonylamino-3,4-dih ydro-2,2-dimethyl-2H- 1 -benzopyran-4-yl)urea

R/S-/V-2-chlorophenyl-/V'-(6-ferf-butoxycarbonylamino-3,4-di hydro-2,2-dimethyl-2H- 1 -benzopyran-4-yl)urea

R/S-/V-2-cyanophenyl-/V'-(6-ferf-butoxycarbonylamino-3,4-dih ydro-2,2-dimethyl-2/-/- 1 -benzopyran-4-yl)urea

R/S-A/-4-chloro-2-trifluoromethylphenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro- 2,2-dimethyl-2H-1 -benzopyran-4-yl)urea R/S-/V-4-sulfamoylphenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimeth 2/-/-1 -benzopyran-4-yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethylthiochroman-4- yl)urea

R/S-/V-4-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethylthiochroman-4- yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethylthiochroman-4- yl)urea

R/S-/V-4-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethylthiochroman-4- yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethyl-1 , 1 - dioxothiochroman-4-yl)urea

R/S-/V-4-cyanophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethyl-1 , 1 - dioxothiochroman-4-yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethyl-1 , 1 - dioxothiochroman-4-yl)urea

R/S-/V-4-chlorophenyl-/V'-(6-ie f-butoxycarbonylamino-2,2-dimethyl-1 , 1 - dioxothiochroman-4-yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-dih ydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclopentan]-4-yl)urea

R/S-/V-4-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-dih ydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclopentan]-4-yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-di hydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclopentan]-4-yl)urea

R/S-/V-4-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-di hydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclopentan]-4-yl)urea

R/S-/V-3-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-dih ydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclobutan]-4-yl)urea

R/S-/V-4-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-dih ydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclobutan]-4-yl)urea

R/S-/V-3-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-di hydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclobutan]-4-yl)urea R/S-/V-4-chlorophenyl-/V 6-tert-butoxycarbonylamino-3,4-dihydrospiro[2H-1 - benzopyran-2, 1 '-cyclobutan]-4-yl)urea

Most preferred compounds are:

R/S-/V-3-cyanophenyl-/V -(6-tert-butoxycarbanoylamino-3,4-dihydro-2,2-dimethyl-2/-/- 1 -benzopyran-4-yl)urea,

R/S-/V-4-trifluoromethylphenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea,

R/S-/V-3-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-dih ydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclopentan]-4-yl)urea and

R/S-/V-3-cyanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4-dih ydrospiro[2/-/-1 - benzopyran-2, 1 '-cyclobutan]-4-yl)urea.

The present invention also refers to compounds particularly dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives or their combination for use as a medicament.

In another aspect the invention relates to compounds particularly to dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives of the general formula (I) or a pharmaceutically acceptable acid addition salt thereof for use as a therapeutically acceptable substance, preferably for use as a therapeutically acceptable substance in the treatment or prevention of proliferative disorder, particularly cancer and most particularly brain cancer. Preferably, the compounds of the present invention are used in the treatment or prevention of brain cancer such as for example gliomas, primitive neuroectodermal tumors (PNETs) and brain metastases.

The compounds, particularly dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives of formula (I) and combinations thereof can be formulated in compositions such as tablets, capsules or elixirs for oral administration, in sterile solutions or suspensions. Accordingly, the invention also refers to a pharmaceutical composition comprising an effective amount of compounds particularly dihydrobenzopyran, thiochroman, tetrahydroquinoleine or tetrahydronaphthalene derivatives or a combination thereof with a pharmaceutical acceptable carrier.

The pharmaceutical composition according to the invention may be administered as a dose in a range from about 0.05 to 1000, preferably from about 0.1 to 500 and especially in the range from 50 to 200 mg per day.

The invention also relates to a process for the manufacture of a medicament, particularly to be used in the treatment of proliferative disorder, which process comprising bringing a dihydrobenzopyran, thiochroman, tetrahydroquinoleine or tetrahydronaphthalene derivative of formula (I) or a pharmaceutically acceptable salt thereof into a galenic dosage form.

In still another aspect, the present invention relates to a method for preparing the above-mentioned compounds particularly the dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives.

The method comprises the following steps:

Reagents: i: X'-C(RiR ₂ )-CH ₂ -COOH or C(RiR ₂ )=CH-COOH; ii: H ₂ S0 ₄ or SOCI ₂ , then AICI ₃ ; iii: Fe, NH CI; iv: R ₅ -NCO or R5-OCOCI or (R ₅ OCO) ₂ 0; iv: mCPBA ; vi: NH ₂ OH.HCI, NaHC0 ₃ ; vii: R -X\ NaH; viii: Raney Ni, H ₂ ; ix: R ₃ -C ₆ H ₄ -NCY

Scheme 1

Reagents: i: Ac ₂ 0, HOAc; ii: CH3COCI, AICI ₃ ; iii: RiR ₂ C=0, pyrrolidine, CH ₃ OH;

iv: HCI, EtOH.

Scheme 2

To synthesize the final compounds, the 6-amino-substituted compound obtained after step (iv) of scheme 2 is then engaged in the same sequence of reactions as described for the corresponding 6-amino-substituted compound obtained after step (iii) of scheme 1 .

Reagents: i: SOCI ₂ , then AICI ₃ ; ii: Fe, NH ₄ CI.

Scheme 3

To synthesize the final compounds, the 6-amino-substituted compound obtained after step (ii) of scheme 3 is then engaged in the same sequence of reactions as described for the corresponding 6-amino-substituted compound obtained after step (iii) of scheme 1 .

The starting materials are either known compounds or compounds, which may be prepared in analogy with the preparation of known compounds or in analogy with known methods.

DESCRIPTION OF THE INVENTION

The method of preparing the compound and particularly dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives of formula (I) is further illustrated in the following examples, which, however, are not to be construed as limiting.

Example 1 : synthesis of R/S-/V-2-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihvdro-2/-/-1 -benzopyran-4-yl)urea 3-( 4-nitrophenoxy)propionic acid A solution of 4-nitrophenol (7 g) in KOH 10% (70 mL) was supplemented with β- bromopropionic acid (7.6 g) and heated at reflux for 5 hours. After cooling, the mixture was acidified by means of 6N HCI and then extracted with ethyl acetate (3 x 100 mL). The organic layers were collected and extracted with a saturated solution of sodium hydrogenocarbonate (3 x 100 mL). The aqueous layers were mixed together and acidified by means of 6N HCI. The resulting precipitate was collected by filtration, washed with water and dried (yield: 40%); melting point: 1 17-1 18°C.

6-nitro-3, 4-dihydro-2H- 1 -benzopyran-4-one

A solution of 3-(4-nitrophenoxy)propionic acid (4 g) in concentrated sulphuric acid (40 mL) was heated at 50°C for 5 h. The reaction mixture was poured on ice (400 g) and, after complete melting of the ice, extracted with ethyl acetate (3 x 100 mL). The organic layers were collected, dried over anhydrous magnesium sulphate, filtered, and the filtrate was concentrated to dryness. The residue was dissolved in ethyl acetate (25 mL) and the solution was supplemented with hexane (75 mL) under stirring. The resulting precipitate was collected by filtration, washed with hexane and dried (yield: 85%; melting point: 176-178°C.

6-amino-3, 4-dihydro-2H- 1 -benzopyran-4-one

A solution of 6-nitro-3,4-dihydro-2/-/-1 -benzopyran-4-one (3 g) in a 3: 1 mixture of ethanol and water (60 mL) was heated at 80°C. Ammonium chloride (1 .5 g) and iron powder (6 g) were then added and the reaction mixture was heated at 80°C for 15 min. The warm reaction mixture was filtered and the insoluble material was washed with warm ethanol (50 mL). The filtrate was treated with charcoal and, after filtration, was concentrated to dryness. The residue was taken off with water, collected by filtration, washed with water and dried (yield: 70%); melting point: 140-142°C.

6-tert-butoxycarbonylamino-3, 4-dihydro-2H- 1-benzopyran-4-one The solution of 6-amino-3,4-dihydro-2/-/-1 -benzopyran-4-one (2.5 g) in a 1 : 1 mixture of THF and water (60 ml_) was supplemented with potassium carbonate (4 g) and di- fe/f-butyl dicarbonate (3.2 g), then the reaction mixture was heated at 40°C for 15 h. After cooling, the organic layer was separated and preserved. The aqueous layer was extracted with ethyl acetate (2 x 50 ml_) and the organic layers were collected, dried over magnesium sulphate and filtered. The filtrate was concentrated to dryness and the residue was dissolved in ethyl acetate (20 ml_). The solution was supplemented with hexane (60 ml_) under stirring and the resulting precipitate was collected by filtration, washed with hexane and dried (yield: 90%); melting point: 146- 148°C.

6-tert-butoxycarbonylamino-3,4-dihydro-2H-1-benzopyran-4- hydroxyimine

The solution of 6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran-4-one (2.5 g) in ethanol (30 ml_) was supplemented with sodium hydrogenocarbonate (2 g) and hydroxylamine hydrochloride (1 g). The reaction mixture was stirred at room temperature during 8 h, then poured on ice. The resulting precipitate was collected by filtration, washed with water and dried (yield: 95%); melting point: 194.5-195.5°C.

R/S-4-amino-6-tert-butoxycarbonylamino-3,4-dihydro-2H-1-b enzopyran The solution of 6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 -benzopyran-4- hydroxyimine (2.5 g) in ethanol (25 ml_) was supplemented with Raney nickel (3 g). The suspension was introduced in a sealed vessel and submitted to hydrogen gas under pressure (10 bars) during 4 h. The catalyst was eliminated by filtration over celite and the filtrate was evaporated to dryness. The residue was dissolved in methanol (10 ml_) and the resulting solution was supplemented with water (80 ml_). The precipitate was collected by filtration, washed with water and dried (yield: 70%); melting point: 109-1 1 1 °C. R/S-N-2-chlorophenyl-N'-( 6-tert-butoxycarbonylamino-3, 4-dihydro-2H-1-benzopyran- 4-yl)urea To the solution of R/S-4-amino-6-ie f-butoxycarbonylamino-3,4-dihydro-2/-/-1 - benzopyran (4.0 mmol) in dichloromethane (5 ml_) was added under stirring 2- chlorophenyl isocyanate (4.4 mmol). The reaction mixture was stirred for 30 min and then evaporated to dryness under reduced pressure. The residue was dissolved in the minimum volume of ethyl acetate and then supplemented with two volumes hexane. The resulting precipitate was collected by filtration, washed with hexane and dried (yield: 80%); melting point: 156-159°C.

Example 2: synthesis of R/S-/V-3-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihvdro-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ferf-butoxycarbonylarnino-3,4- dihydro-2/-/-1 -benzopyran (1 eq.) and 3-chlorophenyl isocyanate (1.1 eq.); melting point: 133-135°C.

Example 3: synthesis of R/S-/V-4-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihvdro-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ferf-butoxycarbonylarnino-3,4- dihydro-2/-/-1 -benzopyran (1 eq.) and 4-chlorophenyl isocyanate (1.1 eq.); melting point: 216-218°C.

Example 4: synthesis of R/S-/V-2-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihvdro-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ferf-butoxycarbonylamino-3,4- dihydro-2/-/-1 -benzopyran (1 eq.) and 2-cyanophenyl isocyanate (1.1 eq.); melting point: 224-227°C.

Example 5: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihydro-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2/-/-1 -benzopyran (1 eq.) and 3-cyanophenyl isocyanate (1.1 eq.); melting point: 212-214°C.

Example 6: synthesis of R/S-/V-4-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihydro-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2/-/-1 -benzopyran (1 eq.) and 4-cyanophenyl isocyanate (1.1 eq.); melting point: 204-206°C.

Example 7: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-feff-butoxycarbonylamino- thiochroman-4-yl)urea

3-( 4-nitrophenylsulfanyl)propionic acid

The solution of 4-nitrothiophenol (10 g) in a 10% aqueous solution of KOH (100 mL) was supplemented with β-bromopropionic acid (1 1 g), and the solution was refluxed for 5 h. After cooling, the reaction mixture was extracted with ethyl acetate (10 mL) and the aqueous layer was separated and treated with 6N HCI until pH 1 . The resulting precipitate was collected by filtration, washed with water and dried (yield: 94%); melting point: 129-130°C.

6-nitrothiochroman-4-one

The solution of 3-(4-nitrophenylsulfanyl)propionic acid (6 g) in thionyl chloride (60 mL) was refluxed for 4 h. The reaction mixture was evaporated to dryness and the residue was dissolved in anhydrous dichloromethane (60 mL). The solution was supplemented with aluminium chloride (5 g) and the reaction mixture was stirred for 4 h at room temperature. After careful addition of water (60 mL) to the mixture, the organic layer was collected. Then, the aqueous layer was extracted twice with dichloromethane (2 x 50 mL) and the three combined organic layers were dried over anhydrous magnesium sulphate, filtered and evaporated to dryness under reduced pressure. The residue was dissolved in ethyl acetate (25 mL) and the resulting solution was mixed with hexane (75 mL). The precipitate was collected by filtration, washed with hexane and dried (yield: 75%); melting point: 168-170°C.

6-aminothiochroman-4-one

The solution of 6-nitrothiochroman-4-one (3 g) in a 3:1 mixture of ethanol and water (60 mL) was heated at 80°C and then supplemented with ammonium chloride (1 .5 g) and iron (6 g). After 15 min, the hot reaction mixture was filtered and the insoluble material was washed with hot ethanol (50 mL). The filtrate was treated with charcoal, filtered and then concentrated to dryness. The residue was suspended in water, collected by filtration, washed with water and dried (yield: 86%); melting point: 1 19-121 °C.

6-tert-butoxycarbonylaminothiochroman-4-one The solution of 6-aminothiochroman-4-one (2.5 g) in a 1 : 1 mixture of THF and water was supplemented with potassium carbonate (4 g) and di-fe/f-butyl dicarbonate (3.2 g) and the reaction mixture was heated at 40°C for 15 h. After cooling, the organic phase was collected and the aqueous phase was extracted twice with ethyl acetate (2 x 50 ml_). The three organic phases were combined and dried over anhydrous magnesium sulphate, then filtered and evaporated to dryness. The residue was dissolved in ethyl acetate (20 ml_) and mixed with hexane (60 ml_). The resulting precipitate was collected by filtration, washed with hexane and dried (yield: 96%); melting point: 138-140°C.

6-tert-butoxycarbonylaminothiochroman-4-hydroxyimine

The solution of 6-ie f-butoxycarbonylaminothiochroman-4-one (2.5 g) in ethanol (30 ml_) was supplemented with sodium hydrogenocarbonate (2 g) and hydroxylamine hydrochloride (1 g). The reaction mixture was stirred at room temperature for 8 h and then poured on ice. The resulting precipitate was collected by filtration, washed with water and dried (yield: 95%); melting point: 192-194°C.

R/S-4-amino-6-tert-butoxycarbonylaminothiochroman

To the solution of 6-ie f-butoxycarbonylaminothiochroman-4-hydroxyimine (2.5 g) in ethanol (25 ml_) was added Raney Nickel (3 g) and the resulting suspension was submitted to hydrogen gas (10 bars) in a sealed vessel at room temperature during 24 h. The catalyst was then removed by filtration over celite and the filtrate was evaporated to dryness. The residue was dissolved in methanol (10 ml_) and mixed with water (80 ml_). The resulting precipitate was collected by filtration, washed with water and dried (yield: 70%). The crude product was used in the next step without further purification.

R/S-N-3-cyanophenyl-N'-(6-teii-butoxycarbonylaminothiochr oman-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 3-cyanophenyl isocyanate (1.1 eq.); melting point: 185-187°C.

Example 8: synthesis of R/S-/V-3-chlorophenyl-/V'-(6-te f-butoxycarbonylamino- thiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 3-chlorophenyl isocyanate (1 .1 eq.); melting point: 189-191 °C.

Example 9: synthesis of R/S-/V-4-chlorophenyl-/V'-(6-te f-butoxycarbonylamino- thiochroman-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 4-chlorophenyl isocyanate (1 .1 eq.); melting point: 125-128°C.

Example 10: synthesis of R/S-/V-2-chlorophenyl-/V'-(6-te f-butoxycarbonylamino- thiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 2-chlorophenyl isocyanate (1 .1 eq.); melting point: 208-210°C. Example 1 1 : synthesis of R/S-/V-4-cvanophenyl-/V'-(6-feff-butoxycarbonylamino- thiochroman-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 4-cyanophenyl isocyanate (1.1 eq.); melting point: 126-130°C.

Example 12: synthesis of R/S-/V-2-cvanophenyl-/V'-(6-feff-butoxycarbonylamino- thiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 2-cyanophenyl isocyanate (1.1 eq.); melting point: 189-191 °C.

Example 13: synthesis of R/S-/V-4-cvanophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 - dioxo-thiochroman-4-yl)urea

6-tert-butoxycarbonylamino- 1, 1-dioxo-thiochroman-4-one The solution of 6-ie f-butoxycarbonylaminothiochroman-4-one (2.5 g) in dichloromethane (30 mL) was cooled at 0°C and then supplemented with mCPBA (10 g). After 10 min, water (30 mL) was added and the reaction mixture was alcalized by means of a 10% aqueous solution of NaOH. The organic phase was collected and the aqueous phase was extracted twice with ethyl acetate (2 x 50 mL). The three organic phases were combined, dried over anhydrous magnesium sulphate, filtered and evaporated to dryness. The residue was dissolved in ethyl acetate (25 mL) and mixed with hexane (75 mL). The resulting precipitate was collected by filtration, washed with hexane and dried (yield: 83%); melting point: 179- 182°C.

6-tert-butoxycarbonylamino- 1, 1-dioxo-thiochroman-4-hydroxyimine

The title compound was obtained by following the experimental conditions described in example 7 for the synthesis of 6-fe/f-butoxycarbonylamino-thiochroman-4- hydroxyimine starting from 6-ie f-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- one; melting point: 163-165°C.

R/S-4-amino-6-tert-butoxycarbonylamino-1, 1 -dioxo-thiochroman The title compound was obtained by following the experimental conditions described in example 7 for the synthesis of R/S-4-amino-6-ie f-butoxycarbonylamino- thiochroman starting from 6-ie f-butoxycarbonylamino-1 , 1 -dioxo-thiochroman-4- hydroxyimine.

R/S-N-4-cyanophenyl-N'-( 6-tert-butoxycarbonylamino- 1, 1 -dioxo-thiochroman-4- y I) urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-1 , 1 - dioxo-thiochroman (1 eq.) and 4-cyanophenyl isocyanate (1 .1 eq.); melting point: 160-162°C.

Example 14: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 - dioxo-thiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-1 , 1 - dioxo-thiochroman (1 eq.) and 3-cyanophenyl isocyanate (1 .1 eq.); melting point: 150-180°C.

Example 15: synthesis of R/S-/V-2-cvanophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 - dioxo-thiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-1 , 1 - dioxo-thiochroman (1 eq.) and 2-cyanophenyl isocyanate (1 .1 eq.); melting point: 187-189°C.

Example 16: synthesis of R/S-/V-4-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 - dioxo-thiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-1 , 1 - dioxo-thiochroman (1 eq.) and 4-chlorophenyl isocyanate (1.1 eq.); melting point: 145-160°C.

Example 17: synthesis of R/S-/V-3-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 - dioxo-thiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-1 , 1 - dioxo-thiochroman (1 eq.) and 3-chlorophenyl isocyanate (1.1 eq.); melting point: 130-150°C.

Example 18: synthesis of R/S-/V-2-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-1 , 1 - dioxo-thiochroman-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-1 , 1 - dioxo-thiochroman (1 eq.) and 2-chlorophenyl isocyanate (1.1 eq.); melting point: 135-155°C.

Example 19: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihvdro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea N-( 4-methoxyphenyl)acetamide

To the solution of sodium acetate (10.24 g) in acetic acid (41 ml_) was added 4- methoxyaniline (50 g) and the reaction mixture was cooled at 0°C. Then acetic anhydride (45.05 ml_) was added dropwise under stirring. After 30 min, the reaction mixture was supplemented under stirring with water (100 ml_) and the crystalline precipitate of the title compound was collected by filtration. The product was crystallized in hot water and, after cooling, collected by filtration, washed with water and dried (yield: 92%): melting point: 126-127 °C.

N-(3-acetyl-4-hydroxyphenyl)acetamide

To the suspension of /V-(4-methoxyphenyl)acetamide (20 g) in carbon disulfide (50 ml_) and acetyl chloride (25 ml_) was added aluminum chloride (55 g) portionwise. The reaction mixture was heated at 80-90°C for 90 min, then evaporated under reduced pressure. The residue was suspended in water and ice and the insoluble material was collected by filtration. The product was dissolved in a 5% aqueous solution of sodium hydroxide. The solution was treated with charcoal, filtered and acidified with 12N HCI. The precipitate was collected by filtration, washed with water and dried (yield: 90%); melting point: 162-164°C. 6-acetamido-3, 4-dihydro-2, 2-dimethyl-2H- 1-benzopyran-4-one

The solution of /V-(3-acetyl-4-hydroxyphenyl)acetamide (25 g) in the mixture of pyrrolidine (15 mL), acetone (32 mL) and methanol (575 mL) was stirred at room temperature overnight. Then, the reaction mixture was evaporated under reduced pressure and the residue was suspended in water. The suspension was adjusted to pH 1 by means of 12N HCI and the insoluble material was then collected by filtration. The precipitate was dissolved in methanol and the resulting solution was treated with charcoal, filtered. The filtrate was mixed with water, collected by filtration, washed with water and dried (yield: 91 %); melting point: 164-165°C.

6-amino-3, 4-dihydro-2, 2-dimethyl-2H-1-benzopyran-4-one The solution of 6-acetamido-3,4-dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-one (20 g) in a 5N HCI ethanolic solution (250 mL) was refluxed for 3 h. The reaction mixture was mixed with water (500 mL) and the title compound was precipitated by the addition of a 40% aqueous solution of sodium hydroxide until basic pH. The insoluble material was collected by filtration, washed with water and dried (yield: 95%); melting point: 148-149 °C.

6-tert-butoxycarbonylamino-3, 4-dihydro-2, 2-dimethyl-2H-1-benzopyran-4-one The solution of 6-amino-3,4-dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-one (15 g) in a 1 :1 mixture of THF and water (385 mL) was supplemented with potassium carbonate (21 .9 g) and di-fe/f-butyl dicarbonate (20.1 g). The reaction mixture was stirred at room temperature for 4 days. The organic layer was collected and the aqueous phase was extracted with ethyl acetate (3 x 100 mL). The organic phases were combined, dried over anhydrous magnesium sulphate, filtered and concentrated to dryness under reduced pressure. The residue was dissolved in methanol and the title compound precipitated by addition of two volumes of water. The precipitate was collected by filtration, washed with water and dried (yield: 95%); melting point: 168- 170°C.

6-tert-butoxycarbonylamino-3, 4-dihydro-2, 2-dimethyl-2H-1 -benzopyran-4- hydroxy/ mine

The title compound was obtained by following the experimental conditions described in example 7 for the synthesis of 6-fe/f-butoxycarbonylamino-thiochroman-4- hydroxyimine starting from 6-tert-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2H- 1 -benzopyran-4-one (yield: 94%); melting point: 162-163°C.

R/S-4-amino-6-tert-butoxycarbonylamino-3, 4-dihydro-2, 2-dimethyl-2H-1 -benzopyran The title compound was obtained by following the experimental conditions described in example 7 for the synthesis of R/S-4-amino-6-ie f-butoxycarbonylamino- thiochroman starting from 6-tert-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl-2/-/- 1 -benzopyran-4-hydroxyimine (yield: 88%); melting point: 144-146°C.

R/S-N-3-cyanophenyl-N'-(6-tert-butoxycarbonylamino-3,4-di hydro

1 -benzopyran-4-yl) urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3-cyanophenyl isocyanate (1.1 eq.); melting point: 191 -193°C.

Example 20: R/S-/V-4-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-cyanophenyl isocyanate (1.1 eq.); melting point: 231 -233°C.

Example 21 : R/S-/V-3-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3-chlorophenyl isocyanate (1.1 eq.); melting point: 1 13°C (decomp.).

Example 22: R/S-/V-4-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-chlorophenyl isocyanate (1.1 eq.); melting point: 1 13°C (decomp.).

Example 23: R/S-/V-4-fluorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-fluorophenyl isocyanate (1.1 eq.); melting point: 208-210°C.

Example 24: R/S-/V-3-fluorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ferf-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3-fluorophenyl isocyanate (1.1 eq.); melting point: 1 15-125°C.

Example 25: R/S-/V-3-bromophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3-bromophenyl isocyanate (1 .1 eq.); melting point: 210-21 1 °C. Example 26: R/S-/V-3,5-difluorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro- 2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3,5-difluorophenyl isocyanate (1 .1 eq.); melting point: 120-155°C.

Example 27: R/S-/V-4-nitrophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-nitrophenyl isocyanate (1.1 eq.); melting point: 233-234°C.

Example 28: R/S-/V-4-aminophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea The solution of R/S-/V-4-nitrophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea (500 mg) in a 3: 1 mixture of ethanol and water (10 ml_) was heated at 80°C and then supplemented with ammonium chloride (250 mg) and iron (1 g). After 15 min, the hot reaction mixture was filtered and the insoluble material was washed with hot ethanol (50 ml_). The filtrate was treated with charcoal, filtered and then concentrated to dryness. The residue was suspended in water, collected by filtration, washed with water and dried (yield: 80%); melting point: 170-180°C.

Example 29: R/S-/V-3-nitrophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3-nitrophenyl isocyanate (1.1 eq.); melting point: 120-155°C.

Example 30: R/S-/V-3-aminophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 28 starting from R/S-/V-3-nitrophenyl-/V'-(6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea; melting point: 130-140°C.

Example 31 : R/S-/V-4-trifluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 186-188°C. Example 32: R/S-/V-3-tnfluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihvdro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 140-155°C. Example 33: R/S-/V-3,5-dichlorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro- 2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3,5-dichlorophenyl isocyanate (1 .1 eq.); melting point: 142-155°C.

Example 34: R/S-/V-2-nitrophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 2-nitrophenyl isocyanate (1.1 eq.); melting point: 218-219°C.

Example 35: R/S-/V-2-trifluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 2-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 239-240°C. Example 36: R/S-/V-3,5-di(trifluoromethyl)phenyl-/V'-(6-te f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 3,5-di(trifluoromethyl)phenyl isocyanate (1 .1 eq.); melting point: 186-190°C.

Example 37: R/S-/V-2-chloro-4-trifluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino- 3,4-dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 2-chloro-4-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 234-235°C.

Example 38: R/S-/V-2-chloro-5-trifluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino- 3,4-dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 2-chloro-5-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 233-234°C. Example 39: R/S-/V-2-chloro-6-trifluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino- 3,4-dihydro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 2-chloro-6-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 243-244°C. Example 40: R/S-/V-4-chloro-3-tnfluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino- 3,4-dihvdro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ferf-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-chloro-3-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 213-214°C.

Example 41 : R/S-/V-4-bromophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-bromophenyl isocyanate (1 .1 eq.); melting point: 181 -184°C.

Example 42: R/S-/V-2-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 2-chlorophenyl isocyanate (1.1 eq.); melting point: 192-194°C. Example 43: R/S-/V-2-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro-2,2- dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 2-cyanophenyl isocyanate (1.1 eq.); melting point: 206-208°C. Example 44: R/S-/V-4-chloro-2-tnfluoromethylphenyl-/V'-(6-te f-butoxycarbonylamino- 3,4-dihvdro-2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-ie f-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran (1 eq.) and 4-chloro-2-trifluoromethylphenyl isocyanate (1 .1 eq.); melting point: 217-219°C.

Example 45: R/S-/V-4-sulfamoylphenyl-/V'-(6-te f-butoxycarbonylamino-3,4-dihvdro- 2,2-dimethyl-2/-/-1 -benzopyran-4-yl)urea

To the solution of R/S-4-amino-6-ie f-butoxycarbonylamino-3,4-dihydro-2,2-dimethyl- 2/-/-1 -benzopyran (0.1 g) in dichloromethane (2 mL) was added under stirring 4- (chlorosulfonyl)phenyl isocyanate (0.08 g). The reaction mixture was stirred for 30 min and then evaporated to dryness under reduced pressure. The residue was dissolved in dioxane (2 mL) and added dropwise to a 10% solution of ammonium hydroxide. After 2 h at room temperature, the excess of ammonium hydroxide was removed by evaporation under reduced pressure and the solution was acidified by means of HCI 6N. The resulting precipitate was collected by filtration, washed with water and dried (yield: 55%); melting point: 180-190°C.

Example 46: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethylthiochroman-4-yl)urea

3- ( 4-acetamidophenylsulfanyl)propionic acid

4- acetamidothiophenol (5 g), 3,3-dimethylacrylic acid (3,3 g) and piperidine (1 ,5 mL) were introduced in a sealed PAAR apparatus and heated at 1 10°C for 24 h. After cooling, the residue was dissolved in a 10% aqueous solution of NaOH (100 mL) and the solution was washed using ethyl acetate (10 mL). The aqueous layer was acidified to pH 1 using HCI 6N and the precipitate was collected by filtration, washed with water and dried (yield: 79%); melting point: 126-128°C. 6-acetamido-2,2-dimethylthiochroman-4-one 3-(4-acetamidophenylsulfanyl)propionic acid (6 g) and polyphosphoric acid (10 g) were heated at 60°C for 1 h. The mixture was mixed with water (200 mL); the resulting precipitate was filtered, suspended in a 10% aqueous solution of NaOH (100 mL) and extracted with ethyl acetate (3 x 100 mL). The organic phases were combined, dried over anhydrous magnesium sulphate, filtered and concentrated to dryness under reduced pressure. The residue was dissolved in the minimum volume of ethyl acetate and then supplemented with two volumes of hexane. The resulting precipitate was collected by filtration, washed with hexane and dried (yield: 53%); melting point: 182-185°C.

6-amino-2,2-dimethylthiochroman-4-one

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-amino-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-one starting from 6-acetamido-2,2-dimethylthiochroman-4-one; melting point: 144-146°C.

6-tert-butoxycarbonylamino-2,2-dimethylthiochroman-4-one

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-tert-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-one starting from 6-amino-2,2-dimethylthiochroman-4- one; melting point: 171 -173°C.

6-teii-butoxycarbonylamino-2,2-dimethylthiochroman-4-hydr oxyimine The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-tert-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2H-1 -benzopyran-4-hydroxyimine starting from 6-tert-butoxycarbonylamino- 2,2-dimethylthiochroman-4-one; melting point: 135.5-137.5°C.

R/S-4-amino-6-tert-butoxycarbonylamino-2,2-dimethylthioch roman

The title compound was obtained by following the experimental conditions described in example 7 for the synthesis of R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman starting from 6-tert-butoxycarbonylamino-2,2- dimethylthiochroman-4-hydroxyimine. The crude product was used in the next step without further purification.

R/S-N-3-cyanophenyl-N'-(6-teii-butoxycarbonylamino-2,2-di methylthiochroman y I) urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-2,2- dimethylthiochroman (1 eq.) and 3-cyanophenyl isocyanate (1.1 eq.); melting point: 125-130°C.

Example 47: synthesis of R/S-/V-4-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethylthiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 4-cyanophenyl isocyanate (1.1 eq.); melting point: 213-215°C. Example 48: synthesis of R/S-/V-3-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethylthiochroman-4-yl)urea

The title compound was obtained by following the expenmental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 3-chlorophenyl isocyanate (1 .1 eq.); melting point: 208-210.5°C.

Example 49: synthesis of R/S-/V-4-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethylthiochroman-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman (1 eq.) and 4-chlorophenyl isocyanate (1 .1 eq.); melting point: 218-219.5°C.

Example 50: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman-4-yl)urea

6-tert-butoxycarbonylamino -2, 2-di methyl- 1, 1-dioxothiochroman-4-one

The title compound was obtained by following the experimental conditions described in example 13 for the synthesis of 6-tert-butoxycarbonylamino-1 , 1 - dioxothiochroman-4-one starting from 6-tert-butoxycarbonylamino-2,2- dimethylthiochroman-4-one; melting point: 177-181 °C.

6-tert-butoxycarbonylamino-2, 2-di methyl- 1, 1-dioxothiochroman-4-hydroxyimine

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-tert-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-hydroxyimine starting from 6-tert-butoxycarbonylamino- 2,2-dimethyl-1 , 1 -dioxothiochroman-4-one; melting point: 196-198°C.

R/S-4-amino-6-tert-butoxycarbonylamino-2, 2-dimethyl- 1, 1 -dioxothiochroman

The title compound was obtained by following the experimental conditions described in example 7 for the synthesis of R/S-4-amino-6-fe/f- butoxycarbonylaminothiochroman starting from 6-tert-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman-4-hydroxyimine; melting point: 91 -94°C.

R/S-N-3-cyanophenyl-N'-( 6-tert-butoxycarbonylamino-2, 2-dimethyl- 1, 1- dioxothiochroman-4-yl) urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman (1 eq.) and 3-cyanophenyl isocyanate (1 .1 eq.); melting point: 218-219.5°C.

Example 51 : synthesis of R/S-/V-4-cvanophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman (1 eq.) and 4-cyanophenyl isocyanate (1 .1 eq.); melting point: 165-175°C.

Example 52: synthesis of R/S-/V-3-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman-4-yl)urea The title compound was obtained by following the expenmental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman (1 eq.) and 3-chlorophenyl isocyanate (1 .1 eq.); melting point: 209-210.5°C.

Example 53: synthesis of R/S-/V-4-chlorophenyl-/V'-(6-te f-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-2,2- dimethyl-1 , 1 -dioxothiochroman (1 eq.) and 4-chlorophenyl isocyanate (1 .1 eq.); melting point: 150-155°C.

Example 54: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentan1-4-yl)urea

6-acetamidospiro[2H-1-benzopyran-2, 1 '-cyclopentan]-4(3H)-one

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-acetamido-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-one starting from /V-(3-acetyl-4-hydroxyphenyl)acetamide (5 g) and cyclopentanone (4.5 ml_) (yield: 86%); melting point: 121 -124°C.

6-aminospiro[2H-1 -benzopyran-2, 1 '-cyclopentan]-4(3H)-one

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-amino-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-one starting from 6-acetamidospiro[2/-/-1 -benzopyran-2, 1 '- cyclopentan]-4(3/-/)-one (yield: 91 %); melting point: 126-128°C. 6-tert-butoxycarbonylaminospiro[2H-1-benzopyran-2, 1 '-cyclopentan]-4(3H)-one

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-tert-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-one starting from 6-aminospiro[2/-/-1 -benzopyran-2, 1 '- cyclopentan]-4(3/-/)-one (yield: 73%); melting point: 164-166°C.

6-tert-butoxycarbonylaminospiro[2H-1-benzopyran-2, 1 '-cyclopentan]-4(3H)- hydroxyimine

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-tert-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2H-1 -benzopyran-4-hydroxyimine starting from 6-tert- butoxycarbonylaminospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentan]-4(3/-/)-one (yield: 81 %); melting point: 166-168°C.

R/S-4-amino-6-tert-butoxycarbonylamino-3, 4-dihydrospiro[2H-1 -benzopyran-2, 1 '- cyclopentane]

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran starting from 6-tert- butoxycarbonylaminospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentan]-4(3H)-hydroxyimine (yield: 93%); melting point: 160.5-162.5°C.

R/S-N-3-cyanophenyl-N'-( 6-tert-butoxycarbonylamino-3, 4-dihydrospiro[2H- 1- benzopyran-2, 1 '-cyclopentan]-4-yl)urea The title compound was obtained by following the expenmental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2,1 '-cyclopentane] (1 eq.) and 3-cyanophenyl isocyanate (1.1 eq.); melting point: 130-150°C.

Example 55: synthesis of R/S-/V-4-cvanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentan1-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentane] (1 eq.) and 4-cyanophenyl isocyanate (1.1 eq.); melting point: 197-199°C.

Example 56: synthesis of R/S-/V-3-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentan1-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentane] (1 eq.) and 3-chlorophenyl isocyanate (1.1 eq.); melting point: 1 19-142°C.

Example 57: synthesis of R/S-/V-4-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentan1-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclopentane] (1 eq.) and 4-chlorophenyl isocyanate (1.1 eq.); melting point: 159-165°C. Example 58: synthesis of R/S-/V-3-cvanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclobutan1-4-yl)urea

6-acetamidospiro[2H-1-benzopyran-2, 1 '-cyclobutan]-4(3H)-one

The title compound was obtained by following the expenmental conditions described in example 19 for the synthesis of 6-acetamido-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-one starting from /V-(3-acetyl-4-hydroxyphenyl)acetamide (3 g) and cyclobutanone (2.4 ml_) (yield: 70%); melting point: 192-195°C.

6-aminospiro[2H-1 -benzopyran-2, 1 '-cyclobutan]-4(3H)-one

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-amino-3,4-dihydro-2,2-dimethyl-2/-/-1 - benzopyran-4-one starting from 6-acetamidospiro[2/-/-1 -benzopyran-2, 1 '-cyclobutan]- 4(3H)-one (yield: 96%); melting point: 160-162.5°C.

6-tert-butoxycarbonylaminospiro[2H-1 -benzopyran-2, 1 '-cyclobutan]-4(3H)-one

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-tert-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2/-/-1 -benzopyran-4-one starting from 6-aminospiro[2/-/-1 -benzopyran-2, 1 '- cyclobutan]-4(3/-/)-one (yield: 87%); melting point: 161 -162.5°C.

6-tert-butoxycarbonylaminospiro[2H-1 -benzopyran-2, 1 '-cyclobutan]-4(3H)- hydroxyimine

The title compound was obtained by following the experimental conditions described in example 19 for the synthesis of 6-tert-butoxycarbonylamino-3,4-dihydro-2,2- dimethyl-2H-1 -benzopyran-4-hydroxyimine starting from 6-tert- butoxycarbonylaminospiro[2H-1 -benzopyran-2, 1 '-cyclobutan]-4(3H)- 90%); melting point: 197-199°C.

R/S-4-amino-6-tert-butoxycarbonylaminospiro[2H-1-benzopyr an-2, 1 '-cyclobutane]

The title compound was obtained by following the expenmental conditions described in example 19 for the synthesis of R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydro-2,2-dimethyl-2/-/-1 -benzopyran starting from 6-tert- butoxycarbonylaminospiro[2/-/-1 -benzopyran-2, 1 '-cyclobutan]-4(3/-/)-hydroxyimine. The crude product was used in the next step without further purification.

R/S-N-3-cyanophenyl-N'-( 6-tert-butoxycarbonylamino-3, 4-dihydrospiro[2H- 1- benzopyran-2, 1 '-cyclobutan]-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2,1 '-cyclobutane] (1 eq.) and 3-cyanophenyl isocyanate (1.1 eq.); melting point: 123-152°C.

Example 59: synthesis of R/S-/V-4-cvanophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclobutan1-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclobutane] (1 eq.) and 4-cyanophenyl isocyanate (1 .1 eq.); melting point: 222-225°C.

Example 60: synthesis of R/S-/V-3-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cvclobutan]-4-yl)urea The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2,1 '-cyclobutane] (1 eq.) and 3-chlorophenyl isocyanate (1.1 eq.); melting point: 1 17-155°C.

Example 61 : synthesis of R/S-/V-4-chlorophenyl-/V'-(6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclobutan1-4-yl)urea

The title compound was obtained by following the experimental conditions described in example 1 (last step) starting from R/S-4-amino-6-tert-butoxycarbonylamino-3,4- dihydrospiro[2/-/-1 -benzopyran-2, 1 '-cyclobutane] (1 eq.) and 4-chlorophenyl isocyanate (1.1 eq.); melting point: 214-216°C.

BRIEF DESCRIPTION OF FIGURES

Figure 1 a,b,c depict illustrative phase contrast pictures (the videomiscroscopy related pictures) obtained in an in vitro cellular imaging approach on human U373 glioblastoma cells untreated or treated with compounds according to the invention. Cellular imaging: pictures (time= Oh, 24h, 48h, 72h) of each U373 cell line left untreated or treated with each compound according to the invention at their IC50 growth inhibitory concentration.

Fig 1 a illustrates the videomiscroscopy related pictures of U373 glioblastoma cells left untreated (control) or treated with compound of example 19 (71 1 ), compound of example 24 (732), compound of example 29 (736) (class I) at their IC50 growth inhibitory concentration (see table 1 ). Fig 1 b illustrates the videomiscroscopy related pictures of U373 glioblastoma cells left untreated (control) or treated with compound of example 32 (738), compound of example 33 (739) (class II) at their IC50 growth inhibitory concentration (see table 1 ). Fig 1 c illustrates the videomiscroscopy related pictures of U373 glioblastoma cells left untreated (control) or treated with compound of example 25 (733), compound of example 16 (713), compound of example 22 (714) (class III) at their IC50 growth inhibitory concentration (see table 1 ).

PHARMACOLOGICAL METHOD

The compounds and particularly dihydrobenzopyran, thiochroman, tetrahydroquinoleine and tetrahydronaphthalene derivatives of the invention were evaluated for their anti-proliferative effect (by means of the colorimetric MTT assay), in the following human glioblastoma cell-lines U373, T98G and Hs683 and in the mouse glioma cell line GL261 . Normal brain cells (primocultures from newborn mice) were also used in order to determine the bioselectivity (B ^{N C} ) of the compounds of interest, i.e. the ratio between the concentrations that inhibits 50% of the growth of brain cancer (C) cells (glioma cells) and the concentration that inhibits 50% of the growth of normal (N) brain cells (primocultures of astrocytes). The ideal situation to be reached is a bioselectivity of B ^{N C} > 10.

The inventors have unexpectedly found that the compounds of the invention appear to have important anti-cancer effects at a given concentration, while not impairing normal cell biology of non-tumor or non-cancer cells at said concentration.

In addition, the inventors could establish that the compounds of the invention act through a non-apoptosis-related mechanism thanks to a unique experimental approach used by the inventors, i.e. computer-assisted phase-contrast microscopy (quantitative videomicroscopy), making them good candidates as anti-cancer drugs for treating apoptosis-resistant tumor or cancer cells and for overcoming problems linked to the known anti-cancer drugs. In vitro characterization of the biological effects of the compounds according to the invention A/ Effect on overall cell growth

MTT tests were performed in order to rapidly, i.e. within 5 days, measure the effect of compounds of this invention on the overall cell growth. The test measured the number of metabolically active living cells that were able to transform the yellow product 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (herein referred as MTT) into the blue product formazan dye by mitochondrial reduction. The amount of formazan obtained at the end of the experiment, measured by means of a spectrophotometer, is directly proportional to the number of living cells. Optical density determination thus enabled a quantitative measurement of the effect of the investigated compounds as compared to the control condition (untreated cells) and/or to other reference compounds.

Three human cancer cell lines were used in the following MTT tests.

To perform the assay, cells were allowed to grow in 96-well micro-wells with a flat bottom with an amount of 100 μΙ of cell suspension per well with 1 ,000 to 8,000 cells/well depending on the cell type used. Each cell line was seeded in its appropriate culture medium.

The detailed experimental procedure was the following: after a 24-hour period of incubation at 37°C, the culture medium was replaced by 100 μΙ of fresh medium in which the tested compound was previously dissolved, at the following molar concentrations: 10 ^"8 M, 5.10 ^"8 M, 10 ^"7 M, 5.10 ^"7 M, 10 ^"6 M, 5.10 ^"6 M, 10 ^"5 M, 5.10 ^"5 M and 10 ^"4 M. Each experiment was performed in sextuplicate (6 times).

After 72 hours of incubation at 37°C with (experimental conditions) or without (control condition) the compound to be tested, the medium was replaced by 100 μΙ MTT dissolved in RPMI (1640 without phenol red) at a concentration of 0.5 mg/ml. The micro-wells were subsequently incubated during 3 hours and a half at 37° C and centrifuged at 1300 rpm during 10 minutes. MTT was removed and formazan crystals formed were dissolved in 100 μΙ DMSO. The micro-wells were shaken for 5 minutes and read on a spectrophotometer at wavelengths of 570 nm (maximal formazan absorbance).

For each experimental condition, the mean optical density was calculated, allowing the determination of the percentage of remaining living cells in comparison to the control.

Table 1 shows the IC50 (representing the range of concentration of the compound tested that resulted in a 50% inhibition of overall tumour cells growth) for each compound in each cell line investigated.

The origin of all the cell lines we used, along with their biological characteristics, and the validation procedures of the MTT colorimetric assay as employed here are fully detailed in Van Quaquebeke et al., J Med Chem 48:849-856, 2005; Ingrassia et al., J Med Chem 52: 1 100-1 1 14, 2009; Mathieu et al., J Cell Mol Med, Feb. 20, 2009; Maes W et al., Neuro-Oncol 1 1 :529-542, 2009.

Table 1 : IC ₅₀ (μΜ) values: Concentration of compound needed in order to inhibit cell population growth by 50%

ICso (μΜ) In Vitro Growth Inhibitory Concentra tions (MTT Colorimetri c Assay)

Compounds Determine d in three Human G Noma Cell Example n° Lines

Mean ±

U373 T98G Hs683

SEM

19 2 3 2 2 ± 1

36 4 5 3 4± 1

29 1 7 16 8±4

33 6 12 9 9±2

26 6 14 7 9±3

54 4 17 7 9±4

58 4 20 7 10±5

31 10 15 11 12 ± 1

47 18 9 8 12 ±3

32 9 21 8 13±4

34 12 22 13 16±3

21 18 20 12 17 ±3

20 10 26 18 18±5

41 22 21 21 21 ± 1

40 21 23 22 22 ± 1

24 20 26 19 22 ±2

25 22 26 18 22 ±2

38 15 38 12 22 ±8

27 24 26 20 23 ±2

56 25 24 21 23 ± 1

1 26 24 23 24 ± 1

55 28 18 26 24 ±3

22 22 26 26 25 ± 1

2 19 29 28 25 ±3

Table 2: Bioselectivity determined by dividing the IC ₅₀ obtained with glioma cell lines by the IC ₅₀ obtained with normal astrocytes. Bioselectivity was determined only for those compounds whose mean + SEM values on the three human glioma cell lines were < 20 μΜ.

Compounds

In vitro anti-cancer activity and bioselectivity

(Mean IC ₅₀ MTT values + SEM (μΜ))

Glioma

Astrocytes

Example n°

Human Brain Bioselectivity (I / II)

Mouse normal brain

cancer cells

cells

(I - 72h)

(II - 72h)

19 2 ± 1 >20 > 10

36 4± 1 -20 -5

29 8±4 <40 < 5

33 9±2 -40 < 5

26 9±3 -45 -5

31 12 ± 1 > 120 > 10

32 13±4 -20 <2

21 17 ±3 >200 > 12

20 18±5 >200 > 10 B/ Impairment of cell proliferation and cell migration triggered by the compound of the present invention in cancer cells As the MTT test is based on the mitochondria functions, we also investigated the effects of the compounds according to the invention on cell proliferation, migration and morphology by means of a cellular imaging approach (Debeir et al., Cytometry 60:29-40, 2004; Debeir et al., IEEE Trans Med Imaging 24:697-71 1 , 2005) in the human U373 glioma cell line, which is apoptosis-resistant but autophagy-sensitive (Lefranc F et al., Neurosurgery 2008).

Cellular imaging relied on the use of computer-assisted phase-contrast microscopy making possible to film the behaviour of living cells in culture dishes for several days.

Cells were seeded in a 25-cm ² flask at a low density, treated or not with the compounds of the invention (at their IC50 concentration determined previously by MTT tests) and filmed thereafter for a period of 72h. The experiment was conducted in triplicate.

The behaviour of the cells, in terms of morphology, growth and death were thus investigated. The effect on the overall growth was measured by counting the number of cells on the first (Oh) and the last image (72h) of each film. The global growth ratio (GGR) was then deduced by dividing the number of cells on the last image by the number of cells on the first image. The ratio GGRtreated ceiis / GGR _CO ntroi ceiis, was further calculated thereby obtaining a value that describes the effect of compounds of the present invention on the overall cell growth.

The methodology is fully described and validated in Debeir et al., 2008, Exp Cell Res 314:2985-2998 and Mathieu et al., 2009, J Cell Mol Med, Feb 20, 2009.

The recordings (not shown) and data obtained clearly show that the compounds according to the invention impair cell morphology and proliferation of human cancer cells. Illustrative pictures (time= Oh, 24h, 48h and 72h) of glioblastoma left untreated or treated with compounds according to the invention (IC50 values calculated by means of the MTT colorimetric assay; see Table 1 ) are provided in Figure 1. (Figure 1 ).

Thanks to the quantitative videomicroscopy (QVM) approach, we classified the compounds into 4 distinctive classes on the basis of their cytostatic versus cytotoxic effects (Table 3).

• Class I : actually cytostatic for 3 days (< 10 % of cell death at the 3 ^rd day)

• Class II: cytostatic but becomes cytotoxic with < 10% cell death on the 2 ^nd day and > 20% of cell death on the 3 ^rd day

· Class III: cytostatic but rapidly becomes cytotoxic with < 10% cell death on the 1 ^st day, > 20% of cell death on the 2 ^nd day and > 50% of cell death on the 3 ^rd day

• Class IV: cytotoxic (> 50% of cell death on the first day)

In Fig. 1 a, compounds of example 19 (5 μΜ), example 24 (20 μΜ) and example 29 (5 μΜ) belong to QVM Class I because they display cytostatic effects on human U373 GBM cells during the whole duration of the experiment. Cytostatic effects mean that < 10 % of cell death was recorded at the end of the experiment, i.e. at the end of the 3 ^rd day. QVM analyses enable the global growth ratio (GGR) to be calculated in each experiment. The GGR index is obtained by dividing the number of cells in the last image by the number of cells in the first image of the experiment, knowing that 1080 images are taken during the 72 h-period of observation. The ratio _{GG R} treated ceiis _; QQR ^C ontroi ceiis _{wgs furthei} - calculated thereby obtaining a value that describes the effect of compounds on the overall cell growth. Thus, a GGR index of 0.2 means that they remain only 20% of treated cells compare to the control cells at the end of the experiment.

In Fig. 1 b, compounds of example 32 (10 μΜ) and example 33 (10 μΜ) belong to the QVM Class II because they display cytostatic effects on human U373 GBM cells but this effect becomes cytotoxic on the third day of experiment with < 10% cell death on the 2nd day and > 20% of cell death on the 3rd day.

In Fig. 1 c, compounds of example 25 (25 μΜ), example 21 (20 μΜ), example 22 (25 μΜ) belong to the QVM Class III because they are cytostatic but they become rapidly cytotoxic with < 10% cell death on the 1 st day, > 20% of cell death on the 2nd day and > 50% of cell death on the 3rd day.

Table 3. Quantitative videomicroscopy classification on the basis of the compound cytotoxicity

Example n° QVM Class

19 I

36 I

29 I

33 II

26 I

31 I

32 II

34 I

21 III

20 I

24 I

25 III

38 II

27 IV

22 III

37 II

23 I

39 III 40 III

41 III

43 I

44 II

5

10

15

20

25