HK1081191B - 3-(4-oxo-4h-chromen-2-yl)-(1h)-quinoline-4-one derivatives, method for preparing same and pharmaceutical compositions containing same - Google Patents

Description

3- (4-oxo-4H-benzopyran-2-yl) - (1H) -quinoline-4-one derivatives, process for their preparation and pharmaceutical compositions containing them

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The present invention relates to novel 3- (4-oxo-4H-chromen-2-yl) - (1H) -quinolin-4-one compounds, processes for their preparation, pharmaceutical compositions containing them and their use as anti-cancer agents.

Anticancer therapy requires continuous development of new antitumor drugs with the aim of obtaining drugs with stronger activity and better tolerance.

In addition to the compounds of the present invention being novel, they have very valuable antitumor properties.

On the one hand, they have a pro-apoptotic effect, which is independent of the expression of p53, pRb and Bc1-2, and a marked antiangiogenic effect, and on the other hand, they have a synergistic effect with many cytostatic therapeutics, without any additional haematotoxicity or in general any signs of intolerance.

Due to the above properties, the compounds of the present invention are highly effective and well tolerated adjuvants for chemotherapy, while the presently disclosed medicaments are capable of maintaining and prolonging the effects of such chemotherapy when the chemotherapy is discontinued for a variety of reasons, such as intolerance, completion of the course of treatment, discontinuation due to surgical reasons, and the like.

Due to the properties of the compounds of the invention, they can be advantageously combined with all cytotoxic treatments and radiotherapy currently used (without increasing their toxicity) and with various hormonal therapies directed against aggressive cancers (breast and prostate cancers).

More specifically, the present invention relates to compounds of formula (I):

wherein:

●R₁、R₂、R₃、R₄、R₆、R₈、R₉and R₁₀Which may be the same or different, each represents a group selected from: hydrogen, hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl, in which the alkoxy is linear or branched (C)₁-C₆) Arylalkoxy of alkoxy, in which the alkoxy is each straight-chain or branched (C)₁-C₆) Alkoxycarbonylalkoxy of alkoxy, and OR 'wherein R' represents an ionized OR ionizable group, e.g. phosphoric acid group-PO (OH)₂Sulfuric acid radical-SO₃H. Wherein the alkyl group is linear or branched (C)₁-C₆) Carboxyalkylcarbonyl of an alkyl group, wherein the alkyl groups are each straight-chain or branched (C)₁-C₆) Dialkylaminoalkylcarbonyls of alkyl groups or in which the alkyl group is straight-chain or branched (C)₁-C₆) The carboxyalkylaminocarbonyl group of the alkyl group,

●R₅represents a linear or branched chain selected from (C)₁-C₆) The radicals of alkyl, aryl and heteroaryl,

●R₇represents a group selected from hydrogen, hydroxyl, linear or branched (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl and (C)₃-C₇) Radicals of cycloalkyl radicals, or R₇Denotes a nitrogen-or oxygen-containing heterocycle, its optical isomers if present, its addition salts with a pharmaceutically acceptable acid and its hydrates and solvates.

Among the pharmaceutically acceptable acids, there may be mentioned, without any limitation, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid, oxalic acid, methanesulfonic acid, benzenesulfonic acid, camphoric acid.

Aryl is understood to be phenyl, biphenyl, naphthyl or tetrahydronaphthyl, each of which is optionally substituted by one or more identical or different atoms or groups selected from: halogen atom and straight or branched chain (C)₁-C₆) Alkyl, hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Polyhaloalkyl, amino (optionally substituted by one or two straight or branched chains (C)₁-C₆) Alkyl substituted), nitro and (C)₁-C₂) An alkylenedioxy group.

Heteroaryl is understood to be a 5-to 12-membered group which is either an aromatic monocyclic ring or a bicyclic ring in which at least one ring is aromatic in character, and which contains one, two or three heteroatoms selected from oxygen, nitrogen and sulfur, it being understood that: heteroaryl groups may be optionally substituted with one or more atoms or groups, which may be the same or different, selected from: halogen atom and straight or branched chain (C)₁-C₆) Alkyl, hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Polyhaloalkyl and amino (optionally substituted by one or two straight or branched chains (C)₁-C₆) Alkyl substituted). Among heteroaryl groups, mention may be made, without any limitation, of thienyl, pyridyl, furyl, pyrrolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolinyl, isoquinolinyl and pyrimidinyl groups.

Nitrogen-containing heterocycles are understood as being saturated or unsaturated 5-to 7-membered nitrogen-containing monocyclic radicals, which are optionally substituted by one or more groups selected from: hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl, aryl in which the alkyl moiety is linear or branched- (C)₁-C₆) Alkyl, and wherein the alkyl moiety is linear or branched and wherein the amino group is optionally substituted with one or two linear or branched (C)₁-C₆) Alkyl-substituted amino- (C)₁-C₆) An alkyl group.

Preferred nitrogen-containing heterocycles are optionally substituted piperidinyl and tetrahydropyridinyl.

Oxygen-containing heterocycles are understood as being saturated or unsaturated, oxygen atom-containing, 5-to 7-membered monocyclic radicals which are optionally substituted by one or more radicals selected from the group consisting of: hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl, aryl in which the alkyl moiety is linear or branched- (C)₁-C₆) Alkyl, and wherein the alkyl moiety is linear or branched and wherein the amino group is optionally substituted with one or two linear or branched (C)₁-C₆) Alkyl-substituted amino- (C)₁-C₆) An alkyl group.

An advantageous aspect of the present invention relates to wherein R₅A compound of formula (I) representing an aryl group.

Another advantageous aspect of the present invention relates to wherein R₇A compound of formula (I) representing a hydrogen atom.

Another advantageous aspect of the present invention relates to wherein R₇A compound of formula (I) representing an optionally substituted nitrogen-containing heterocycle.

Preferred compounds of formula (I) are those in which R is₅Represents phenyl and R₇Represents a hydrogen atom or a substituted 1, 2, 3, 6-tetrahydro-4-pyridyl group.

Among the preferred compounds of the invention, mention may be made more particularly of:

-3- (5-hydroxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-quinolin-4-one,

-3- [5, 7-dimethoxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one,

-3- (5, 7-dihydroxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one,

-and 3- [5, 7-dihydroxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one.

The invention also relates to a process for the preparation of a compound of formula (I), which process is characterized in that:

reacting a compound of formula (II):

wherein R is₁、R₂、R₃、R₄And R₅As defined in formula (I),

with diethyl ethoxymethylenemalonate to produce a compound of formula (III):

wherein R is₁、R₂、R₃、R₄And R₅As defined above, and Et represents ethyl,

cyclizing the compound of formula (III) under acid conditions to produce a compound of formula (IV):

wherein R is₁、R₂、R₃、R₄And R₅And Et is as defined above, and Et,

hydrolyzing the compound of formula (IV) to produce a compound of formula (V):

wherein R is₁、R₂、R₃、R₄And R₅As defined above, the above-mentioned,

the compound of formula (V) is reacted with thionyl chloride to convert it to an acid chloride, which is then reacted with a compound of formula (VI):

wherein R is₇、R₈、R₉And R₁₀As defined in formula (I),

to produce a compound of formula (VII):

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉And R₁₀As defined above, the above-mentioned,

reacting a compound of formula (VII) with a base to produce a compound of formula (VIII):

wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉And R₁₀As defined above, andindicating that the resulting compound is in the form of a keto-enol mixture depending on the molecule involved,

the compound of formula (VIII) is then subjected to acidic conditions to produce the compound of formula (I), which is purified, if necessary, according to conventional purification techniques, separated into its optical isomers, if necessary, according to conventional separation techniques, and converted, if necessary, into an addition salt with a pharmaceutically acceptable acid.

Wherein one or more substituents R₁-R₄And R₆-R₁₀Compounds of formula (I) which represent hydroxy groups may also be obtained by cleavage of the corresponding substituents which represent linear or branched (C)₁-C₆) Alkoxy, and compounds of formula (I).

Wherein one or more substituents R₁-R₄And R₆-R₁₀Represents alkoxycarbonylalkoxy, arylCompounds of formula (I) which are arylalkoxy OR OR' groups may also be prepared starting from compounds of formula (I) in which the corresponding substituents represent hydroxy groups.

In addition to the fact that they are novel compounds, the compounds of the present invention have very valuable anti-tumor properties, which makes them useful for the treatment of cancer.

Among the types of cancer that can be treated by the compounds of the invention, there may be mentioned, without any limitation, adenocarcinomas, carcinomas, sarcomas, gliomas and leukemias.

They can also be used in combination with another anticancer drug such as paclitaxel, tamoxifen and its derivatives, cisplatin and its analogs, irinotecan and its metabolites, various alkylating agents, and in therapy, the main examples of which are cyclophosphamide, etoposide, vinca alkaloids, doxorubicin and other anthracyclines and nitrosoureas.

The invention also relates to pharmaceutical compositions containing, as active principle, at least one compound of formula (I) and one or more suitable inert non-toxic excipients. Among the pharmaceutical compositions of the present invention, mention may be made in particular of those suitable for oral, parenteral (intravenous, intramuscular or subcutaneous) or nasal administration, tablets or dragees, sublingual tablets, capsules, lozenges, suppositories, creams, ointments, dermal gels, injectable preparations, drinkable suspensions and the like.

Useful doses may be adapted to the nature and severity of the disease, the route of administration, the age and weight of the patient and any associated treatment, and are administered in the range of 0.5mg to 2g once or in several portions every 24 hours.

The following examples illustrate the invention without any limitation.

The starting materials used are known products or are prepared according to known methods.

The structures of the compounds described in the examples were determined according to the usual spectrometric techniques (infrared spectroscopy, NMR, mass spectrometry).

Example 1: 3- (7-methoxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Step A: n, N-Diphenylaminomethylene malonic acid diethyl ester

10mmol of diethyl ethoxymethylenemalonate was added to 10mmol of diphenylamine, and the mixture was heated at 140 ℃ and 150 ℃ for 5 hours. After returning to ambient temperature, the solid formed was washed with 100ml of diethyl ether and recrystallized from hexane to give the expected product in the form of a brown solid.

Melting point: 146 ℃ and 148 ℃.

MS(EI，m/z)：339.9(M^+.)。

And B: 4-oxo-1-phenyl-1H-1, 4-dihydroquinoline-3-carboxylic acid ethyl ester

To 10mmol of the compound obtained in the previous step, 13.3g of polyphosphoric acid was added. The mixture, which gradually became liquid, was then heated at 150-160 ℃ for 45 minutes and then cooled to 90 ℃. After hydrolysis, the mixture was neutralized with 10% NaOH solution and isolated to give the desired product.

IR (NaCl plate, cm)^-1)：1733(υC＝O)，1610(υC＝C)，1645(υC＝O)，690(υC-H(ar))。

MS(EI，m/z)：293.3(M^+.)。

And C: 4-oxo-1-phenyl-1H-1, 4-dihydroquinoline-3-carboxylic acid

To 10mmol of the compound obtained in the previous step dissolved in methanol was added 38mL of 2M NaOH solution. The reaction mixture was then heated under methanol reflux for 10 hours, and then the solvent was removed in vacuo. Water was added to the resulting residue, and the mixture was neutralized with 4M HCl solution. The resulting grey solid was washed with water and then dried to give the desired product.

Melting point: 210 ℃ and 213 ℃.

IR(KBr，cm^-1): 3320(υ OH (acid)), 1733(υ C ═ O), 1610(υ C ═ C), 1645(υ C ═ O), 690(υ C-h (ar)).

MS(EI，m/z)：265.3(M^+.)。

Step D: 4-oxo-1-phenyl-1H-1, 4-dihydroquinoline-3-carboxylic acid (2-acetyl-5-methoxy) phenyl ester

10mmol of the compound obtained in the preceding step are added to 20mmol of thionyl chloride dissolved in dichloroethane. The reaction mixture was heated at reflux of the solvent for 2 hours, then concentrated in vacuo, and the excess thionyl chloride was distilled off under vacuum with dichloroethane entrainment and repeated several times.

The acid chloride thus obtained (white solid) was added in small portions to 6.7mmol of commercially available 2-hydroxy-4-methoxyacetophenone dissolved in pyridine. After stirring for 12 hours at ambient temperature under an inert atmosphere, the reaction mixture is passed through a column chromatography on silica gel (eluent: CH)₂Cl₂MeOH: 95/5) to give the expected product as a yellow powder.

Melting point: 137-139 ℃.

IR(KBr，cm^-1)：2865(OCH₃υ CH), 1740(υ C ═ O), 1655(υ C ═ O), 1590-.

MS (electrospray, m/z): 413.4 (M)^+.)。

Step E: 3- [3- (2-hydroxy-4-methoxyphenyl-1, 3-dioxopropan-1-yl) -1H-1-phenyl-1, 4-dihydroquinolin-4-one

12mmol of potassium tert-butoxide are slowly added to 10mmol of the compound obtained in the preceding step dissolved in a mixture of dimethylformamide and tetrahydrofuran (35/75) under an inert atmosphere at ambient temperature. The reaction mixture was stirred for 2 hours and poured into 55mL of 0 ℃ aqueous solution containing 1.3mL of 10% hydrochloric acid. The resulting precipitate was filtered off, washed with copious amounts of water and then dried. The resulting solid was subjected to silica gel column chromatography (eluent: CH)₂Cl₂) Purification to give the expected product in the form of a keto-enol mixture.

Melting point: 228 ℃ and 230 ℃.

Step F: 3- (7-methoxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one

To the compound obtained in the previous step (10 mmol) dissolved in 25mL of glacial acetic acid was slowly added 25mL of an acetic acid solution containing 20% sulfuric acid. A yellow precipitate formed. After 2 hours and 30 minutes at ambient temperature, the mixture was poured into ice water (4 ℃). Insoluble material was filtered off and washed with copious amounts of water to give the expected product as a white powder.

Melting point: 297 deg.C.

IR(cm^-1)：2825(υOCH₃)，1750(υC＝O)，1675(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 396.1 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.634.633.39

Measured value: 72.464.573.27

Example 2: 3- (7-hydroxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-quinolin-4-one

10mmol of the compound from example 1 are added to 500mmol of phenol dissolved in 214mL of hydroiodic acid (57% aqueous solution) under inert atmosphere protected from light with stirring. The heterogeneous reaction mixture was then heated at 160 ℃ for 15 hours. The initially yellow solution turned orange. After returning to ambient temperature, the solution is poured onto ice and the resulting precipitate is washed with water and then with diethyl ether to remove residual phenol, after recrystallization, the expected product is obtained in the form of a yellow powder.

Melting point: 295 ℃ and 300 ℃ (acetone).

IR(cm^-1)：3280(υOH)，1770(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 381.1 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.174.293.51

Measured value: 72.374.373.55

Example 3: 3- (6-methoxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Following the procedure described in example 1, but substituting 2-hydroxy-5-methoxyacetophenone for 2-hydroxy-4-methoxyacetophenone in step D, the expected product was obtained.

Melting point: 265 deg.C.

IR(cm^-1)：2830(υOCH₃)，1740(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 396.1 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.634.633.39

Measured value: 72.744.463.36

Example 4: 3- (5-methoxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Following the procedure described in example 1, but substituting 2-hydroxy-6-methoxyacetophenone for 2-hydroxy-4-methoxyacetophenone in step D, the expected product was obtained.

Melting point: 271 ℃.

IR(cm^-1)：2830(υOCH₃)，1744(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 3961 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.634.633.39

Measured value: 72.464.573.27

Example 5: 3- (5, 7-dimethoxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Step A: 2, 4-dimethoxy-6-hydroxyacetophenone:

adding 14.5mmol of K₂CO₃Added in one portion to 10mmol of radiculoacetophenone monohydrate dissolved in acetone. 20mmol of dimethyl sulfate are then added in 30 minutes under an inert atmosphere and the reaction mixture is heated under acetone reflux for 12 hours. After returning to ambient temperature, the mixture was poured into water to give a white suspension which was then filtered. The resulting white powder was washed and then recrystallized from methanol to give the desired product.

Melting point: 80-81 deg.C (methanol).

And B: 3- (5, 7-dimethoxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one:

the expected product was obtained according to the procedure described in example 1, but replacing 2-hydroxy-4-methoxyacetophenone in step D by the compound obtained in step a above.

Melting point: 282 deg.C.

IR(cm^-1)：2825(υOCH₃)，1744(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 425.45 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated value 70.424.773.16

Measured value: 70.334.763.26

Example 6: 3- (5-hydroxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-quinolin-4-one

The expected product is obtained according to the procedure described in example 2, starting from the compound of example 4.

Melting point: > 300 deg.C (acetone).

IR(cm^-1)：3200(υOH)，1770(υC＝O)，1635(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 381.1 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.174.293.51

Measured value: 72.334.403.65

Example 7: 3- (5-hydroxy-7-methoxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-quinolin-4-one

Under the inert atmosphere and in the dark, 11mmol of BBr are added₃(1M solution in dichloromethane) was added to a 10mmol suspension of the compound of example 5 in dichloromethane over 15 minutes, resulting in the formation of a yellow precipitate. The reaction mixture was stirred vigorously at ambient temperature for 6 hours and then cooledTo 0 ℃. Ethanol was then added and the solution was concentrated under vacuum. The resulting residue was then poured into an aqueous alcohol solution (50%) and the mixture was stirred vigorously for 10 minutes. The precipitate obtained is filtered off, washed with water and then with diethyl ether and, after recrystallization, the expected product is obtained as a beige powder.

Melting point: 295 ℃ and 296 ℃ (acetone).

IR(cm^-1)：3224(υOH)，1780(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 411.41 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.994.163.40

Measured value: 72.704.103.45

Example 8: 3- (4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-quinoline-4-one

Following the procedure described in example 1, but substituting acetophenone for 2-hydroxy-4-methoxyacetophenone in step D, the expected product was obtained.

Melting point: 327 ℃ and 328 ℃ (acetone).

MS (electrospray, m/z): 365.4 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 78.894.143.83

Measured value: 78.604.103.60

Example 9: 3- [5, 7-dimethoxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Step A: 4- (2, 4-dimethoxy-6-hydroxy-5-methylcarbonylphenyl) -1-methyl-1, 2, 5, 6-tetrahydropyridine

To 10mmol of the compound obtained in step A of example 5 dissolved in glacial acetic acid was added 11.5mmol of 1-methylpiperidin-4-one, slowly so as not to exceed 25 ℃. When the addition was complete, a stream of hydrogen chloride gas was bubbled for 1 hour 40 minutes, and then the reaction mixture was heated at a temperature of 95 to 100 ℃ for 5 hours. The acetic acid was removed by vacuum distillation, and the residual oil was then treated with water and extracted with diethyl ether. The aqueous phase was made alkaline by adding 40% NaOH solution. The resulting precipitate is filtered off, washed with copious amounts of water and recrystallized from petroleum ether to give the expected product.

Melting point: 143 ℃ and 144 ℃.

IR(KBr，cm^-1)：3400-3200(υOH)，2843(υOCH₃)，1680(υC＝O)，1655(υC＝C)。

MS(EI，m/z)：291(M^+.)。

And B: 3- [5, 7-dimethoxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridinyl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected compound was obtained according to the procedure described in example 1, but replacing 2-hydroxy-4-methylacetophenone in step D with the compound obtained in step A above.

Melting point: 248-250 deg.C (acetone).

IR(cm^-1)：2835(υOCH₃)，1755(υC＝O)，1675(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 520.6 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 73.285.105.22

Measured value: 73.835.425.38

Example 10: 3- (5, 7-dihydroxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product is obtained starting from the compound of example 5 according to the procedure described for example 2.

Melting point: 365-.

IR(cm^-1)：3224(υOH)，1780(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 397.4 (M)^+..)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.543.803.52

Measured value: 72.204.013.33

Example 11: [2- (4-oxo-1-phenyl-1, 4-dihydroquinolin-3-yl) -4H-1-benzopyran-7-yloxy ] acetic acid ethyl ester

To 10mmol of the compound from example 2 suspended in acetone 20mmol of potassium carbonate and then 20mmol of ethyl bromoacetate are slowly added. The mixture was heated at reflux of the solvent for 2 hours 30 minutes and then, after returning to ambient temperature, poured into water. The resulting insoluble material was filtered off and washed with copious amounts of water to give the desired product as a white solid.

Melting point: 330 deg.C (acetone).

IR(cm^-1)：1744(υC＝O)，1680(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 467.48 (M)^+.)。

Elemental microanalysis

％C ％H ％N

Calculated values: 71.944.533.00

Measured value: 71.704.773.40

Example 12: 3- [5, 7-dimethoxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-methyl-1H-1, 4-dihydroquinolin-4-one

Step A: N-Phenylaminomethylene malonic acid diethyl ester

The expected product was obtained according to the procedure described in example 1, step a, but replacing the diphenylamine with aniline.

Melting point: 46-48 deg.C (hexane).

MS(EI，m/z)：263(M^+.)。

And B: N-methyl-N-phenylaminomethylene malonic acid diethyl ester

To 10mmol of the compound obtained in the previous step dissolved in tetrahydrofuran, 12mmol of 95% NaH are slowly (in small portions) added under an inert atmosphere, followed by the dropwise addition of 30mmol of methyl iodide. The reaction mixture was then stirred at ambient temperature under an inert atmosphere for 12 hours. 1ml of methanol was added to neutralize the excess sodium hydride. The solution was then concentrated in vacuo and water was then added to the resulting residual oil. After extraction with dichloromethane, the combined organic phases are dried, filtered and concentrated under reduced pressure to give the expected product in the form of a colourless oil.

MS(EI，m/z)：276(M^+.)。

And C: 3- [5, 7-dimethoxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-methyl-1H-1, 4-dihydroquinolin-4-one

The expected product was obtained according to the procedure described in example 1, steps B to F, but replacing in step B the compound obtained in step a of example 1 with the compound obtained in step B above and replacing in step D the 2-hydroxy-4-methoxyacetophenone with the compound obtained in step a of example 9.

Melting point: 289 deg.C (acetone).

IR(cm^-1)：2835(υOCH₃)，1755(υC＝O)，1675(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 458.5 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 70.735.726.11

Measured value: 70.255.485.78

Example 13: 3- [5, 7-dimethoxy-8- [1- (4-fluorobenzyl) -1, 2, 5, 6-tetrahydropyridin-4-yl ] -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Step A: 1- (4-fluorobenzyl) piperidin-4-one:

to 10mmol of piperidin-4-one hydrochloride monohydrate and 20mmol of triethylamine dissolved in dichloromethane was slowly added 10mmol of 4-fluorobenzyl chloride; the reaction mixture was then heated at reflux of the solvent for 48 hours while stirring vigorously. After returning to ambient temperature, water was added, the phases were separated, the organic phase was dried, filtered and concentrated in vacuo to give the expected product as an orange oil.

MS(EI，m/z)：207.2(M^+.)。

And B: 4- (3-acetyl-4, 6-dimethoxy-2-hydroxyphenyl) -1- (4-fluorobenzyl) -1, 2, 5, 6-tetrahydropyridine

To 10mmol of the compound obtained in step A of example 5 dissolved in glacial acetic acid, 11mmol of the compound obtained in step A above were added, slowly so as not to exceed a temperature of 25 ℃. When the addition was complete, a stream of hydrogen chloride gas was bubbled through the solution for 2 hours, and then the reaction mixture was heated at 95-100 ℃ for 5 hours. Acetic acid was removed by vacuum distillation and then water was added to the resulting residual oil. After extraction with diethyl ether, the aqueous phase was made basic with 40% NaOH solution. The resulting precipitate was filtered off, washed with copious amounts of water and recrystallized from a mixture of ether/ethyl acetate (90/10) to give the expected product in the form of a beige solid.

Melting point: 147 ℃ and 150 ℃.

IR(cm^-1)：3400-3200(υOH)，2871(OCH₃υ CH), 1633(υ C ═ O), 1655(υ C ═ C), 1350-.

MS(EI，m/z)：385.4(M^+.)。

And C: 3- [5, 7-dimethoxy-8- [1- (4-fluorobenzyl) -1, 2, 5, 6-tetrahydropyridin-4-yl ] -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product was obtained following the procedure described in example 1, but replacing 2-hydroxy-4-methoxyacetophenone in step D by the compound obtained in step B above.

Melting point: 218 deg.C and 219 deg.C (acetone).

IR(cm^-1)：2835(υOCH₃)，1755(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 614.66 (M)^+.)。

Example 14: 3- [5, 7-dihydroxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Dispersing 10mmol of the compound of example 9 in pyridinium chloride; the mixture was then heated in a sealed tube at 180 ℃ for 12 hours. After cooling to 100 ℃, the reaction mixture was poured into water and adjusted to pH 7-8(pH starting value 1) using 10% sodium bicarbonate solution. Insoluble material was separated by filtration and washed with water to give the desired product.

Melting point: is > 250 ℃.

IR(cm^-1)：3330(υOH)，1785(υC＝O)，1665(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 492.5 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 72.945.125.87

Measured value: 73.164.915.69

Example 15: 3- [5, 7-dimethoxy-8- (1-benzyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product is obtained according to the procedure described in steps B to C of example 13, but replacing in step B the compound obtained in step A of example 13 by 1-benzylpiperidin-4-one.

Melting point: 248-249 deg.C (acetone).

IR(cm^-1)：2830(OCH₃υ CH), 1765(υ C ═ O), 1655(υ C ═ O), 1570-.

MS (electrospray, m/z): 596.9 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 76.495.414.69

Measured value: 76.065.034.93

Example 16: 3- [5, 7-dihydroxy-8- (1-benzyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product is obtained according to the procedure described for example 14, starting from the compound of example 15.

Melting point: 230 ℃ and 231 ℃ (acetone).

IR(cm^-1)：3340(υCH)，1752(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 568.6 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 76.044.964.93

Measured value: 76.395.415.22

Example 17: 3- [5, 7-dihydroxy-8- [1- (4-fluorobenzyl) -1, 2, 5, 6-tetrahydropyridin-4-yl ] -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product is obtained according to the procedure described for example 14, starting from the compound of example 13.

Melting point: 197 ℃ and 198 ℃ (acetone).

IR(cm^-1)：3245(υCH)，1775(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 586.6 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 73.714.644.78

Measured value: 73.204.284.34

Example 18: 3- [5, 7-dimethoxy-8- [1- (4-methoxybenzyl) -1, 2, 5, 6-tetrahydropyridin-4-yl ] -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product was prepared according to the procedure described for example 13, but substituting 4-methoxybenzyl chloride for 4-fluorobenzyl chloride in step A.

Melting point: 232 ℃ and 235 ℃ (acetone).

IR(cm^-1)：2845(υOCH₃)，1750(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 626.7 (M)^+.)。

Example 19: 3- [5, 7-dimethoxy-8- (1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

To a suspension of 10mmol of the compound from example 18 in glacial acetic acid under inert atmosphere was added 0.9mg Pd/C (10% by weight). The reaction mixture was heated at 70 ℃ and stirred under an atmosphere of hydrogen at atmospheric pressure for 5 hours. The reaction mixture was then filtered through celite, then washed with methanol. The solvent was removed by distillation in vacuo. To the resulting residue was added water adjusted to pH 8-9. By CH₂Cl₂Extraction of the/MeOH (90/10) mixture gave the expected product as a white solid.

Melting point: 251 ℃ and 253 ℃ (diethyl ether/acetone).

IR(cm^-1)：3387(υN-H)，2880(υOCH₃)，1780(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 506.5 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 73.505.175.53

Measured value: 73.125.584.98

Example 20: 3- [5, 7-dimethoxy-8- (1-isopropyl-1, 2, 5, 6-tetrahydropyridinyl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product is obtained according to the procedure described in steps B to C of example 13, but replacing in step B the compound obtained in step A of example 13 by N-isopropylpiperidin-4-one.

Melting point: 248-249 deg.C (acetone).

IR(cm^-1)：2850(υOCH₃)，1755(υC＝O)，1650(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 548.6 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 74.435.885.11

Measured value: 73.785.235.78

Example 21: 3- [7- (4-Bromobenzyloxy) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-quinolin-4-one

The expected product is obtained according to the procedure described for example 11, starting from the compound of example 2 but replacing ethyl bromoacetate by 4-bromobenzyloxy chloride.

IR(cm^-1)：1744(υC＝O)，1655(υC＝O)，1570-1590(υC＝C)。

MS (electrospray, m/z): 549.4 (M)^+.)。

Elemental microanalysis:

％C ％H ％N

calculated values: 67.653.662.54

Measured value: 66.954.212.44

Example 22: 3- [5, 7-dimethoxy-8- [1- (2-dimethylaminoethyl) -1, 2, 5, 6-tetrahydropyridin-4-yl ] -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

Following the procedure described for example 13, but substituting 2-chloro-N, N-dimethylethylamine for 4-fluorobenzyl chloride in step A, the expected product was obtained.

Example 23: 3- [5, 7-dihydroxy-8- [1- (2-dimethylaminoethyl) -1, 2, 5, 6-tetrahydropyridin-4-yl ] -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one

The expected product is obtained according to the procedure described in example 2, starting from the compound of example 22.

Pharmacological study of the Compounds of the invention

Example 24: in vitro cytotoxicity Studies of Compounds of the invention

Eleven cell lines from carcinomas of different origin and location (lung, breast, prostate, colon, blood, bladder, skin, ovary, brain) were cultured to study various compounds compared to a reference substance.

Cells were incubated with different concentrations of the compounds of the invention for 96 hours.

In vitro cytotoxic activity was measured by Carmichael in Cancer Res. -1987; 47(4): MTT [3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide ] assay as described in 936-942.

Activity is termed IC₅₀Indicating, that is to say, the concentration which inhibits 50% of the proliferation of the tumor cells.

In this model, the compounds of the invention have intrinsic cytotoxic activity against one or more tumor cell lines.

For example, the compound of example 9 has cytotoxic activity against 8 of the 11 cell lines tested (depending on the cell line used, IC)₅₀Values of 2 to 10. mu.M).

IC of compound of example 14 for 5 of the 11 cell lines tested₅₀The value is 0.05 to 0.25. mu.M.

Example 25: in vitro synergistic effects of the compounds of the invention with known anticancer agents

Three cell lines sensitive to three anticancer drugs were used: breast cancer cells associated with paclitaxel treatment, lung cancer cells associated with Cisplatin (CDDP) treatment, and colon cancer cells associated with the treatment of SN38, a metabolite of CPT-11 (irinotecan).

Tumor cells were incubated with 5 different concentrations of each compound of the present invention and 5 concentrations of each anti-cancer agent for 96 hours.

The in vitro cytotoxic activity was determined by the MTT assay described in example 24.

According to published Trends Pharmaceutical sci-1983; 4: chou and Talabay, 450 for data analysis.

The compounds of the present invention show a synergistic effect with the different anticancer agents tested, that is to say they enhance the cytotoxic activity of the anticancer agents administered simultaneously.

For example, the compounds of examples 14 and 16 show synergistic effects with paclitaxel and cisplatin.

Example 26: apoptosis of Compounds of the invention

Apoptosis is a natural mechanism that allows the body to remove its own abnormal cells such as cancer cells.

The proapoptotic effect of the compounds of the invention was studied in the prostate cancer line (LN CaP). Placing the cells in IC₅₀Culturing at the concentration for 8-96 hours.

Then according to Sgonc in Trends Genetics-1994; 10: 41 the TUNEL assay was performed.

The compounds of the invention are capable of causing apoptosis, with the time to maximum intensity varying from compound to compound.

The compounds of the present invention differ from the previously described agents in their ability to cause earlier apoptosis. For example, the compound of example 6 resulted in a very high degree of apoptosis early (8 hours).

Example 27: pharmaceutical composition

Preparation of 1000 tablets each containing 10mg of active ingredient

Compound … … … … … … … … … … … … … … … … … … … … … … 10g of example 9

Hydroxypropyl cellulose … … … … … … … … … … … … … … … … … … … … … … … … 2g

… … … … … … … … … … … … … … … … … … … … … … … … … … 10g of wheat starch

Lactose … … … … … … … … … … … … … … … … … … … … … … … … … … … … 100g

… … … … … … … … … … … … … … … … … … … … … … … … … … 3g of magnesium stearate

Talc … … … … … … … … … … … … … … … … … … … … … … … … … … … … 3g

Claims (13)

1. A compound of formula (I), an optical isomer thereof, if present, an addition salt thereof with a pharmaceutically acceptable acid, or a hydrate or solvate thereof,

wherein:

●R₁、R₂、R₃、R₄、R₆、R₈、R₉and R₁₀May be the same or different and each represents a group selected fromThe following groups: hydrogen, hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl, in which the alkoxy is linear or branched (C)₁-C₆) Arylalkoxy of alkoxy, in which the alkoxy is each straight-chain or branched (C)₁-C₆) Alkoxycarbonylalkoxy of alkoxy, and OR ', wherein R' represents a group selected from: -PO (OH)₂、-SO₃H. Wherein the alkyl group is linear or branched (C)₁-C₆) Carboxyalkylcarbonyl of an alkyl group, wherein the alkyl groups are each straight-chain or branched (C)₁-C₆) Dialkylaminoalkylcarbonyls of alkyl groups and wherein the alkyl group is straight or branched (C)₁-C₆) The carboxyalkylaminocarbonyl group of the alkyl group,

●R₅represents a linear or branched chain selected from (C)₁-C₆) The radicals of alkyl, aryl and heteroaryl,

●R₇represents a group selected from hydrogen, hydroxyl, linear or branched (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl and (C)₃-C₇) Radicals of cycloalkyl radicals, or R₇Represents a nitrogen-or oxygen-containing heterocyclic ring,

aryl represents phenyl, biphenyl, naphthyl or tetrahydronaphthyl, each of which is optionally substituted by one or more identical or different atoms or groups selected from: halogen atom and straight or branched chain (C)₁-C₆) Alkyl, hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Polyhaloalkyl, optionally substituted by one or two straight or branched chains (C)₁-C₆) Alkyl-substituted amino, nitro and (C)₁-C₂) An alkylenedioxy group, a hydroxyl group,

heteroaryl denotes a 5-to 12-membered group which is either an aromatic monocyclic ring or a bicyclic ring in which at least one ring is aromatic in character, and which contains one, two or three heteroatoms selected from oxygen, nitrogen and sulfur, and in which: heteroaryl groups may be optionally substituted with one or more atoms or groups, which may be the same or different, selected from: halogen atom and straight or branched chain (C)₁-C₆) Alkyl, hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Polyhaloalkyl and optionally substituted by one or two straight or branched chains (C)₁-C₆) An amino group substituted with an alkyl group,

nitrogen-containing heterocyclic ring means a saturated or unsaturated 5-to 7-membered monocyclic group containing nitrogen atoms, which is optionally substituted by one or more groups selected from: hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl, aryl in which the alkyl moiety is linear or branched- (C)₁-C₆) Alkyl, and wherein the alkyl moiety is linear or branched and wherein the amino group is optionally substituted with one or two linear or branched (C)₁-C₆) Alkyl-substituted amino- (C)₁-C₆) An alkyl group, a carboxyl group,

oxygen-containing heterocycle represents a saturated or unsaturated 5-to 7-membered monocyclic group containing oxygen atoms, which is optionally substituted by one or more groups selected from: hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl, aryl in which the alkyl moiety is linear or branched- (C)₁-C₆) Alkyl, and wherein the alkyl moiety is linear or branched and wherein the amino group is optionally substituted with one or two linear or branched (C)₁-C₆) Alkyl-substituted amino- (C)₁-C₆) An alkyl group.

2. A compound of formula (I) according to claim 1, wherein R₁、R₂、R₃、R₄、R₆、R₈、R₉And R₁₀Which may be the same or different, each represents a group selected from: hydrogen, hydroxy, straight or branched chain (C)₁-C₆) Alkoxy, straight or branched chain (C)₁-C₆) Alkyl, in which the alkoxy is linear or branched (C)₁-C₆) Arylalkoxy of alkoxy and wherein the alkoxy is each straight-chain or branched (C)₁-C₆) Alkoxycarbonylalkoxy group of alkoxy group.

3. A compound of formula (I) according to any one of claims 1-2, wherein R₅Represents an aryl group.

4. A compound of formula (I) according to any one of claims 1-2, wherein R₇Represents a hydrogen atom.

5. A compound of formula (I) according to any one of claims 1-2, wherein R₇Represents an optionally substituted nitrogen-containing heterocycle.

6. A compound of formula (I) according to any one of claims 1-2, wherein R₅Represents phenyl and R₇Represents a hydrogen atom or a substituted 1, 2, 3, 6-tetrahydro-4-pyridyl group.

7. A compound of formula (I) according to claim 1 which is 3- (5-hydroxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-quinolin-4-one.

8. A compound of formula (I) according to claim 1 which is 3- [5, 7-dimethoxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one.

9. A compound of formula (I) according to claim 1, which is 3- (5, 7-dihydroxy-4-oxo-4H-1-benzopyran-2-yl) -1-phenyl-1H-1, 4-dihydroquinolin-4-one.

10. A compound of formula (I) according to claim 1 which is 3- [5, 7-dihydroxy-8- (1-methyl-1, 2, 5, 6-tetrahydropyridin-4-yl) -4-oxo-4H-1-benzopyran-2-yl ] -1-phenyl-1H-1, 4-dihydroquinolin-4-one.

11. Preparation of a compound according to claim 1 wherein R₆A process for the preparation of a compound of formula (I) which is hydrogen, characterized in that:

reacting a compound of formula (II):

wherein R is₁、R₂、R₃、R₄And R₅As defined in claim 1, the first and second,

with diethyl ethoxymethylenemalonate to produce a compound of formula (III):

wherein R is₁、R₂、R₃、R₄And R₅As defined in claim 1, and Et represents ethyl,

cyclizing the compound of formula (III) under acid conditions to produce a compound of formula (IV):

wherein R is₁、R₂、R₃、R₄And R₅As defined in claim 1, and Et represents ethyl,

hydrolyzing the compound of formula (IV) to produce a compound of formula (IV):

wherein R is₁、R₂、R₃、R₄And R₅As defined in claim 1, the first and second,

the compound of formula (V) is reacted with thionyl chloride to convert it to an acid chloride, which is then reacted with a compound of formula (VI):

wherein R is₇、R₈、R₉And R₁₀As defined in claim 1, the first and second,

to produce a compound of formula (VII):

wherein R is₁、R₂、R₃、R₄、R₅、R₇、R₈、R₉And R₁₀As defined in claim 1, the first and second,

reacting a compound of formula (VII) with a base to produce a compound of formula (VIII):

wherein R is₁、R₂、R₃、R₄、R₅、R₇、R₈、R₉And R₁₀As defined in claim 1, andindicating that the resulting compound is in the form of a keto-enol mixture depending on the molecule involved,

the compound of formula (VIII) is then subjected to acidic conditions to produce the compound of formula (I), which is purified, if necessary, according to conventional purification techniques, separated into its optical isomers, if necessary, according to conventional separation techniques, and converted, if necessary, into an addition salt with a pharmaceutically acceptable acid.

12. A pharmaceutical composition comprising as active ingredient a compound according to any one of claims 1 to 10 together with one or more inert non-toxic and pharmaceutically acceptable carriers.

13. Use of a compound of formula I according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 12 for the preparation of an anti-cancer medicament.