BENZOYL UREA DERIVATIVES HAVING ANTI-TUMOR ACTIVITY

Abstract: The invention relates to new benzoyl urea derivatives of the formula: It has been found that these compounds have anti-tumor activity. The compounds of this formula in which at least one of the phenyl groups contains an optionally esterified hydroxy group have been found to possess the strongest activity. The new compounds can be prepared according to methods known for the synthesis of analogous compounds.

Description: Benzoyl urea derivatives having anti-tumor activity. The invention relates to new benzoyl urea derivatives having anti-tumor activity, to the preparation of such compounds, and to compositions comprising at least one of the new compounds as the active substance. It is known from Unites States Patent Specification 3,748,356 that a group of l-benzoyl-3-phenyl-urea compounds has insecticidal properties. Said insecticidal activity is based on a mechanism which has not yet been fully cleared. What is clear is that the insects are killed in that said urea derivatives inhibit the chitin formation. Furthermore it it known from European Patent Application 0,025,363 that some N-benzoyl-N'-pyridyloxyphenyl urea compounds have an anti-tumor activity. It has now been found that l-benzoyl-3-phenyl -urea compounds containing at least one hydroxyl or mercapto group, or at least one group which can generate a hydroxyl or mercapto group have excellent anti-tumor activity. The new substituted benzoyl urea compounds according to the invention can be represented by the formula: EMI1.1 in which: : - X is an oxygen or sulfur-atom; - R1 is a hydrogen atom, a group (O-A)n, wherein A re presents a hydrogen atom, an alkanoyl stoup @@ving 1 to 6 carbon atoms, a carbamoyl group optionally substituted by alkyl containing 1 to 6 carbon atoms or by phuy, a benzylcarbonyl group optionally substituted by halogen, a benzoyl group, a benzyl group, an aryloxycarbonyl group optionally substituted by halogen, a phenyl group optionally substituted by halogen, or an alkoxycarbonyl alkyl group containing from 1 to 6 carbon atoms, or R1 is an alkylmercapto group, having 1 to 3 carbon atoms or a group -S-A, wherein A has the above qiven meaning: : R2 is a hydrogen or halogen atom, an alkyl group ha ving 1 to 6 carbon atoms optionally substituted by halo gen, or R2 is a nitro group; R3 is a hydrogen or halogen atom, a nitro group, a 1 to 6 carbon atoms containing alkyl group optionally sub stituted by halogen, an alkoxy group containing 1 to 6 carbon atoms optionally substituted by halogen, a cyclo alkoxy group, containing 5-10 carbon atoms, optionally substituted by alkyl groups having 1-3 carbon atoms, an alkanoyl group containing from 1 to 6 carbon atoms, a benzoyl group, a nitrile group, an amino group optional ly substituted by 1 or 2 alkyl groups having 1 to 3 car bon atoms, an alkoxycarbonyl group containing from 1 to 6 carbon atoms, or a piperidinocarbonyl group; ; R4 is a hydrogen atom, a group (O-A) or S-A, where in A has the above given meaning, or an alkylmercapto group wherein the alkyl group contains from 1 to 3 car bon atoms; R5 is a hydrogen atom, or represents from 1 to 3 halo gen atoms; - n+m has the value from 1 to 6, with the proviso that R1 and R4 re not simultaneously hydrogen, and the following compounds are disclaimed: : 1) l-(2, 6-difluorobenzoyl)-3-(2-hydroxy-4-chlorophenyl)urea 2) l-( 2, 6-difluorobenzoyl )-3-( 3-hydroxy-4-chlorophenyl )urea 3) 1-(2,6-difluorobenzoyl)-3-(4-hydroxyphenyl)urea 4) l-(2,6-difluorobenzoyl)-3-(2-hydroxy-4-trifluoromethylphe nyl ) urea 5) 1-(2-chlorobenzoyl)-3-(2-hydroxy-4-trifluoromethoxyphenyl)- urea 6) l-(2-chlorobenzoyl)-3-(2-hydroxy-4-trifluoromethylphenyl) urea. These disclaimed compounds are mentioned in the non-prepublished European patent application no. 83201263.7 (publication no. 0107214). It has been found in particular that the compounds of the above formula wherein at least one of the phenyl rings contains a hydroxyl or an esterified hydroxyl group have a very strong anti-tumor activity in the test-model mentioned hereinafter. The anti-tumor activity of the new compounds seems to be based on a new mechanism, which differs from that of known cytostatics. There are indications that the compounds according to the invention do not influence cell-cycle but are active on membrane-level. It has been found that the membrane-fluidity decreases under the influence of the new compounds. The new compounds according to the invention can be obtained in a manner known for the synthesis of analogous compounds. For example, the compounds according to the invention can be prepared by reacting a substituted aniline of the general formula EMI3.1 wherein R3, R4 and R5 have the above given meanings, with a compound of the general formula EMI4.1 wherein R1, R2, R3 and X have the above given meanings. The new compounds according to the invention can also be prepared by reacting a substituted benzamide of the formula EMI4.2 wherein R1, R2, R3 and X have the above given meanings, with a compound of the formula EMI4.3 wherein R3, R4, R5 and X have the above mentioned meaning. The above reactions are preferably carried out in the presence of an organic solvent, such as an aromatic hydrocarbon, an alkyl halide, a non-cylcic or cyclic dialkyl ether or acetonitrile, at a reaction temperature between 0 C and the boiling point of the solvent used. Although the above mentioned methods of preparing are the best suitable, the new compounds can also be prepared in a different manner, for example as described in Netherlands patent application 7105350, or according to the methods described in Netherlands patent applications 7806678 or 8005588. The compounds can be processed to compositions in any usual manner. A great advantage of the compounds according to the invention as compared with known cytostatics is that they have a very low toxicity. The anti-tumor activity was tested in an in vitro test. In this test the sensitivity of B16 melanoma cells to the new compounds according to the invention was determined. B16 melanoma cells grow as a "monolayer". The doubling time of said cells in a cell culture is 12-16 hours. "6 Multi-well" tissue culture plates having a flat bottom 2 having an area of 6 cm proved to be best suited for carrying out the test. On day 0 a quantity of 5 x 10 B 16 melanoma cells is provided in each cell culture dish. On day 1 the compounds to be tested are vortexed in the presence of glass-beads. The compound to be tested is then added to the dishes comprising melanoma cells in the desired quantity. Incubat on takes place in a CO2- -incubator at 37 C for 20 hours. The inc:ubati n is termi- nated by removing the culture medium with therein . pound to be tested, after which the cells are washed twice and fresh medium is added. Forty-eight hours after the beginning of the incubation the quantity of cells in each dish is measured by means of a microcell Coulter Counter. The results thus obtained are expressed by indication the activity cf the compounds as 1+, 2+, 3+, 4+ or 5+, which has the following meaning: 1+ : cel growth is inhibited 1- 20% with respect to control. 2+ : cel growth is inhibited 20- 40% with respect to control. 3+ : cel growth is inhibited 40- 60% with respect to control. 4+ : cel growth is inhibited 60- 80% with respect to control. 5+ : cel growth is inhibited 80-100% with respect to control. Sensitivity of B16 melanoma cells. Various compounds according to the invention were tested in the above-described manner in a dosage of 500 /ug/ml in the form of an aqueous suspension. The results obtained are recorded in the table below: EMI7.1 EMI7.2 Compound <tb> <SEP> No. <SEP> X1 <SEP> X2 <SEP> R6 <SEP> R7 <SEP> M.P.( C) <SEP> Activity <tb> <SEP> 1 <SEP> O <SEP> O <SEP> 2,6-dichloro-3-hydroxyphenyl <SEP> 4-chlorophenyl <SEP> 201 <tb> <SEP> 2 <SEP> O <SEP> O <SEP> 2,6-dichloro-3-acetoxyphenyl <SEP> 4-chlorophenyl <SEP> 211 <tb> <SEP> 3 <SEP> O <SEP> O <SEP> 2,6-dichloro-3-benzyloxyphenyl <SEP> 4-chlorophenyl <SEP> 151 <SEP> 3+ <tb> <SEP> 4 <SEP> O <SEP> O <SEP> 2,6-dichloro-3-hydroxyphenyl <SEP> 4-trifluoromethylphenyl <SEP> 204 <SEP> 5+ <tb> <SEP> 5 <SEP> O <SEP> O <SEP> 2,6-dichloro-3-acetoxyphenyl <SEP> 4-trifluoromethylphenyl <SEP> 169 <SEP> 5+ <tb> <SEP> 6 <SEP> O <SEP> O <SEP> 2-chloro-6-hydroxyphenyl <SEP> 4-chlorophenyl <SEP> 240 <SEP> 4+ <tb> <SEP> 7 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-chlorophenyl <SEP> 174 <SEP> 5+ <tb> <SEP> 8 <SEP> O <SEP> O <SEP> 2-chloro-6-propionyloxyphenyl <SEP> 4-chlorophenyl <SEP> 159 <SEP> 4+ <tb> <SEP> 9 <SEP> O <SEP> O <SEP> 5-nitro-2-hydroxyphenyl <SEP> 4-chlorophenyl <SEP> 211 <SEP> 4+ <tb> <SEP> 10 <SEP> O <SEP> O <SEP> 2-hydroxyphenyl <SEP> 2-chloro-4-nitrophenyl <SEP> 221 <SEP> @@ <tb> <SEP> 11 <SEP> O <SEP> O <SEP> 2,6-dihydroxyphenyl <SEP> 4-chlorophenyl <SEP> 280 <SEP> @+ <tb> <SEP> 12 <SEP> O <SEP> O <SEP> 3,5-diiodo-2-hydroxyphenyl <SEP> 4-chlorophenyl <SEP> 187 <SEP> @ <tb> <SEP> 13 <SEP> O <SEP> O <SEP> 2,6-dihydroxyphenyl <SEP> 2-methyl-4-chlorophenyl <SEP> 201 <SEP> @@ <tb> <SEP> 14 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-(1,1,2,2-tetrafluoroethoxyphenyl <SEP> 105 <tb> <SEP> 15 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-methyloxyphenyl <SEP> 172 <SEP> 4+ <tb> <SEP> 16 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-piperidinocarbonylphenyl <SEP> 172 <SEP> 5+ <tb> <SEP> 17 <SEP> O <SEP> O <SEP> 2-benzoyloxyphenyl <SEP> 4-chlorophenyl <SEP> 183 <SEP> 4@ <tb> <SEP> 18 <SEP> O <SEP> O <SEP> 3,5-dichloro-4-hydroxyphenyl <SEP> 4-chlorophenyl <SEP> 244 <SEP> 4+ <tb> <SEP> 19 <SEP> O <SEP> O <SEP> 3,5-dichloro-4-acetoxyphenyl <SEP> 4-chlorophenyl <SEP> 227 <SEP> @ <tb> EMI8.1 Compound <tb> <SEP> No. <SEP> X1 <SEP> X2 <SEP> R6 <SEP> R7 <SEP> M.P.( C) <SEP> Activity <tb> <SEP> 20 <SEP> O <SEP> O <SEP> 5-chloro-2-hydroxyphenyl <SEP> 2-chloro-4-nitrophenyl <SEP> 228 <SEP> 3+ <tb> <SEP> 21 <SEP> O <SEP> O <SEP> 2,6-dichloro-4-hydroxyphenyl <SEP> 3,4-dichlorophenyl <SEP> 260 <SEP> 4+ <tb> <SEP> 22 <SEP> O <SEP> O <SEP> 2,6-dibenzyloxyphenyl <SEP> 2-methyl-4-chlorophenyl <SEP> 206 <SEP> 3+ <tb> <SEP> 23 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-benzoylphenyl <SEP> 155 <SEP> 5+ <tb> <SEP> 24 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 4-chloro-2-acetoxyphenyl <SEP> 150 <SEP> 5+ <tb> <SEP> 25 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 4-trifluoromethyl-2-acetoxyphenyl <SEP> 162 <SEP> 5+ <tb> <SEP> 26 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 3-trifluoromethyl-4-hydroxyphenyl <SEP> 194 <SEP> 5+ <tb> <SEP> 27 <SEP> O <SEP> O <SEP> 2-chlorophenyl <SEP> 3-trifluoromethyl-4-hydroxyphenyl <SEP> 209 <SEP> 3+ <tb> <SEP> 28 <SEP> O <SEP> O <SEP> 2-chlorophenyl <SEP> 2-trifluoromethoxy-4-hydroxyphenyl <SEP> 156 <SEP> 5+ <tb> <SEP> 29 <SEP> O <SEP> O <SEP> 2-chlorophenyl <SEP> 4-trifluoromethyl-2-acetoxyphenyl <SEP> 178 <SEP> 3+ <tb> <SEP> 30 <SEP> O <SEP> O <SEP> 2-chlorophenyl <SEP> 4-trifluoromethoxy-2-acetoxyphenyl <SEP> 170 <SEP> 3+ <tb> <SEP> 31 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 3-chloro-4-(4-chlorophenoxy)phenyl <SEP> 189 <SEP> 3+ <tb> <SEP> 32 <SEP> O <SEP> O <SEP> 2,6-dichlorophenyl <SEP> 2-methylcarbamoyloxyphenyl <SEP> 186 <SEP> 4+ <tb> <SEP> 33 <SEP> O <SEP> O <SEP> 2,6-dichlorophenyl <SEP> 3-phenylcarbamoyloxyphenyl <SEP> 166 <SEP> 5+ <tb> <SEP> 34 <SEP> O <SEP> O <SEP> 2-chlorophenyl <SEP> 3-chloro-4-(4-chlorobenzylcarbonyloxy)phenyl <SEP> 184 <SEP> 3+ <tb> <SEP> 35 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 3-trifluoromethyl-4-ethoxycarbonylmethoxy- <SEP> 128 <SEP> 4+ <tb> <SEP> phenyl <tb> <SEP> 36 <SEP> O <SEP> O <SEP> 2-methylphenyl <SEP> 4-hydroxyphenyl <SEP> 214 <SEP> 4+ <tb> <SEP> 37 <SEP> O <SEP> O <SEP> 2-chlorophenyl <SEP> 3,5-di-tert. <SEP> butyl-4-hydroxyphenyl <SEP> 80 <SEP> 5+ <tb> <SEP> 38 <SEP> O <SEP> O <SEP> 2-chloro-6-hydroxyphenyl <SEP> 4-chloro-3-hydroxyphenyl <SEP> 198 <SEP> 4+ <tb> <SEP> 39 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-chloro-3-acetoxyphenyl <SEP> 181 <SEP> 5+ <tb> <SEP> 40 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-hydroxyphenyl <SEP> 140 <SEP> 4+ <tb> <SEP> 41 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 5-methyl-2-hydroxyphenyl <SEP> 185 <SEP> 5+ <tb> EMI9.1 Compound <tb> <SEP> No. <SEP> X1 <SEP> X2 <SEP> R6 <SEP> R7 <SEP> M.P.( C) <SEP> Activity <tb> <SEP> 42 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-nitro-2-hydroxyphenyl <SEP> 181 <SEP> 5+ <tb> <SEP> 43 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 5-methyl-2-acetoxyphenyl <SEP> 190 <SEP> 5+ <tb> <SEP> 44 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 4-n-butoxycarbonyl-3-hydroxyphenyl <SEP> 159 <SEP> 4+ <tb> <SEP> 45 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 3,5-dibromo-2-hydroxyphenyl <SEP> 185 <SEP> 5+ <tb> <SEP> 46 <SEP> O <SEP> O <SEP> 2-chloro-6-acetoxyphenyl <SEP> 3-acetyl-4-hydroxyphenyl <SEP> 181 <SEP> 5+ <tb> <SEP> 47 <SEP> O <SEP> O <SEP> 3,5-di-tert. <SEP> butyl-4-hydroxy- <SEP> 4-acetoxyphenyl <SEP> 205 <SEP> 4+ <tb> <SEP> phenyl <tb> <SEP> 48 <SEP> O <SEP> O <SEP> 3-chloro-4-acetoxyphenyl <SEP> 4-hydroxyphenyl <SEP> 250 <SEP> 4+ <tb> <SEP> 49 <SEP> O <SEP> O <SEP> 2-acetoxyphenyl <SEP> 3,5-dichloro-4-hydroxyphenyl <SEP> 201 <SEP> 5+ <tb> <SEP> 50 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 2-mercaptophenyl <SEP> 187 <SEP> 5+ <tb> <SEP> 51 <SEP> O <SEP> S <SEP> phenyl <SEP> 2-acetoxyphenyl <SEP> 138 <SEP> 4+ <tb> <SEP> 52 <SEP> S <SEP> S <SEP> 2-chloro-6-methylthiophenyl <SEP> 4-chlorophenyl <SEP> 170 <SEP> 5+ <tb> <SEP> 53 <SEP> O <SEP> S <SEP> 2,4-dichlorophenyl <SEP> 3-hydroxyphenyl <SEP> 155 <SEP> 5+ <tb> <SEP> 54 <SEP> O <SEP> O <SEP> 2-benzyloxyphenyl <SEP> 4-chlorophenyl <SEP> 185 <SEP> 2+ <tb> <SEP> 55 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 3,5-dichloro-4-(4-chlorobenzoyl)phenyl <SEP> 237 <SEP> 3+ <tb> <SEP> 56 <SEP> O <SEP> O <SEP> 3-chloro-4-acetoxyphenyl <SEP> 3-trifluoromethyl-4-hydroxyphenyl <SEP> 225 <SEP> 3+ <tb> <SEP> 57 <SEP> O <SEP> O <SEP> 2,6-difluorophenyl <SEP> 4-(4-chlorophenoxycarbonyloxy)phenyl <SEP> 220 <SEP> 2+ <tb> The anti-tumor activity of the compounds is confirmed by preliminary results obtained in an in vivo test in mice. The preparation of the compounds is illustrated in the following examples: EXAMPLE 1 To a stirred solution of 2.67 g of 2-hydroxy-3,5 -dibromoaniline in 50 ml. of dry ether was added slowly 2.39 g of 2-acetoxy-6-chlorobenzoylisocyanate. After stirring overnight the precipitate was collected and washed with ether. Yield: 1.88 g of 1-(2-acetoxy-6-chloroben zoyl)-3-( 2-hydroxy-3, 5-dibromophenyl)urea (compound no. 45 as listed above) having a melting point of 185"C. The acetoxy group of the obtained compound was removed by boiling the compound in a solution of ethanol/ HC1 (2N) for a few hours. In this manner the corresponding compound carrying a free hydroxyl group is obtained. Example II A solution of 4.5 g of 2-benzyloxybenzamide and 3.07 g of 4-chlorophenylisocyanate in 50 ml of dry xylene was refluxed during 16 hours. After cooling the precipitate was collected and washed with ether. Compound no. 54, i.e. l-(2-benzyloxybenzoyl)-3-(4-chlorophenyl)urea was obtained in a yield of 6.9 g; m.p. 185"C. The benzyl group was removed by heating the obtained compound in acetic acid/HBr at 55-600C for a few hours. The bove listed compounds all have been prepared according to tht methods of the above described examples.

Claims: Claims 1. Compounds having anti-tunor activity of the formula EMI11.1 wherein - X is an oxygen or sulfur atom; R1 is a hydrogen atom, a group (O¯A)n, wherein A represents a hydrogen atom, an alkanoyl group havinq 1 to 6 carbon atoms, a carbamoyl group optionally substi tuted by alkyl containing 1 to 6 carbon atoms or by phe nyl, a benzylcarbonyl group optionally substituted by halogen, a benzoyl group, a benzyl group, an aryloxycar bonyl group optionally substituted by halogen, a phenyl group optionally substituted by halogen, or an alkoxy carbonylalkyl group containing from 1 to 6 carbon atoms or R1 is an alkylmercapto group having 1 to 3 carbon atoms or a group -S-A, wherein A has the above given meaning; ; R2 is a hydrogen or halogen atom, an alkyl group ha ving 1 to 6 carbon atoms optionally substituted by halo gen, or R2 is a nitro group; R3 is a hydrogen or halogen atom, a nitro group, a 1 to 6 carbon atoms containing alkyl group optionally sub stituted by halogen, an alkoxy group containing 1 to 6 carbon atoms optionally substituted by halogen, a cyclo alkoxy group, containing 5-10 carbon atoms, optionally substituted by alkyl groups having 1-3 carbon atoms, an alkanoyl group containing from 1 to 6 carbon atoms, a benzoyl group, a nitrile group, an amino group optional ly substit°.ted by 1 or 2 alkyl groups having i to 3 car bon atoms, an alkoxycarbonyl group containing from 1 to 6 carbon atoms, or a piperic;nocarbonyl group; ; - R4 is a hydrogen atom, a group (O-A) or S-A, where in A has the above given meaning, or an alkylmercapto group wherein the alkyl group contains from 1 to 3 car bon atoms; - R5 is a hydrogen atom, or represents from 1 to 3 halo gen atoms; - n+m has the value from 1 to 6, with the proviso that R1 and R4 are not simultaneously hydrogen, and the following compounds are disclaimed: : 1) l-(2, 6-difluorobenzoyl)-3- (2-hydroxy-4-chlorophenyl)urea 2) 1-(2,6-difluorobenzoyl)-3-(3-hydroxy-4-chlorophenyl)urea 3) 1-(2, 6-difluorobenzoyl )-3- (4-hydroxyphenyl)urea 4) l-(2,6-difluorobenzoyl )-3-( 2-hydroxy-4-trifluoromethyl- phenyl ) urea 5) l-(2-chlorobenzoyl)-3-( 2-hydroxy-4-trifluoromethoxyphenyl)- urea 6) l-( 2-chlorobenzoyl )-3-( 2-hydroxy-4-trifluoromethylphenyl)- urea. 2. Compounds according to claim 1, wherein R2, R3, R5 and X have the meanings given in claim 1 and R1 and R4 represent from 1 to 6 hydroxyl groups or esterified hydroxyl groups. 3. Compounds according to claim 2, wherein esterified hydroxyl groups are acetoxy groups. 4. A composition having anti-tumor activity, characterized in that it contains at least one compound according to claim 1 as active component. 5. A method of preparing compounds as claimed in claim 1, characterized in that a) a compound of the formula EMI12.1 is reacted with a compound of the formula EMI13.1 or b) a compound of the formula EMI13.2 is reacted with a compound of the formula EMI13.3 in which formulae R1 to R5 and X have the meanings given in claim 1. 6. A method of combating tumors in living beings, characterized in that a composition as claimed in claim 4 is administered to the living being.

Inventors: BROUWER, MARIUS S., VAN HES, ROELOF
Applicants: DUPHAR INTERNATIONAL RESEARCH B.V
Application year: 1984
Main classification: A61K31
IPC codes: A61K31, C07C239, C07C325, C07D295, C07C275, C07C67, C07C335, A61P35