JP3244325B2 - Pyrimidine ring fused cyclopentylidene derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a glutamic acid-bonded pyrimidine ring-fused cyclopentylidene derivative, a process for producing the same,
And its uses.

[0002]

2. Description of the Related Art Methotrexate (referred to as MTX)
Is the only drug currently in clinical use as an antifolate antitumor agent. In addition to being used alone in choriocarcinoma, osteosarcoma and acute lymphocytic leukemia, its application has been expanded through combination therapy with other antitumor agents and the introduction of new administration methods. MTX is a dihydrofolate reductase (D
HFR). However, DHFR is present in tumor tissues and normal host tissues and affects both of them. For this reason, MTX has strong side effects, which are limiting factors for clinical use. MTX
However, they generally have a low therapeutic effect on solid tumors. In addition, there is a drawback in that tumor cells acquire resistance after being used for a certain period of time, and drug efficacy is significantly reduced. These are M
It has reduced the clinical value of TX.

[0003] There has been active research in this area for over 40 years. In particular, a large number of antifolates have been synthesized by the structural change of the folate molecule, and pharmacological studies have been conducted. In general, the molecular structure of an antifolate substance is conveniently divided into four parts: a heterocyclic part, a bridge part, a benzoic acid part, and an amino acid part. As the heterocyclic moiety, a pteridine ring is a typical structure as seen in folic acid, MTX and the like, and in fact, many folate antagonists in this category are known. Relatively recently, various deazapteridine compounds such as 5-deazapteridine, 8-deazapteridine, 5,
8-Dideazapteridine or its dihydro or tetrahydro form has been studied as an antifolate and has been published in large numbers. Inhibition of folate-related enzymes and tumor cell growth of these compounds, as well as antitumor effects using tumor-bearing mice, have been measured, and information has been accumulated to date on the correlation between structure and activity or structure and toxicity. Was. These are, for example, Progre by AndreRosowsky
ss in Medicinal Chemistry, 26, 1-252 (1989), Elsevi
Comprehensive and detailed in erScience Publishers. The first of the structural features of these antifolates is in the heterocyclic portion, and most of them have a bicyclic heterocyclic structure in which two 6-membered rings are fused together like a pteridine ring. On the other hand, folate antagonists having a bicyclic heterocyclic structure formed by the condensation of a 5-membered ring and a 6-membered ring have not been known so far. Journal Medicin by LTWeinstock
al Chemistry, 13, 995 (1970) describes a compound in which a pteridine ring is converted to a purine ring, but it is reported that DHFR inhibition or antitumor activity was not observed. For this reason, it has been considered that pteridine or a closely related structure is required as a structural factor capable of exerting an antifolate action. However, recently, Miwa et al., Journal Medicinal Chemistry, 34,
555-560 (1991) describe that a benzoylglutamic acid derivative having a pyrrolo [2,3-d] pyrimidine ring has a folate antagonism and exhibits a higher inhibitory activity on the growth of human tumor cells than MTX.

[0004]

As described above, despite the recent academic progress in this field, there has not yet been any product that has been put into practical use as a pharmaceutical and provided to a clinical setting since MTX. Accordingly, the present invention has been developed to provide a selective toxicity to tumor tissue with this systemic compound and a therapeutic effect.
There is still a strong demand for providing a useful anticancer agent to a clinical setting. In view of the above situation, an object of the present invention is to provide a novel pyrimidine ring-fused cyclopentylidene derivative having excellent antitumor activity. It is another object of the present invention to provide a method for producing the compound and a pharmaceutical composition containing the compound as an active ingredient.

[0005]

Means for Solving the Problems The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, have found that a glutamic acid-bonded pyrimidine ring-fused cyclopentylidene derivative can be a novel and useful antitumor agent. Completed the invention. That is, the present invention provides a pyrimidine ring-fused cyclopentylidene derivative represented by the general formula (1) or a pharmacologically acceptable salt thereof.

[0006]

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(Wherein, R ¹ represents a hydroxyl group or an amino group; R ² represents a hydrogen atom, an amino group or a methyl group. X
Is a divalent linear group composed of 1 to 4 atoms and may have a substituent. Y represents a phenylene group, a thiendiyl group, a furandiyl group, a thiazolediyl group, an indolediyl group or an indolinediyl group. Also, the present invention provides a compound represented by the general formula (2):

[0008]

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Wherein R ¹ , R ² , X and Y are as defined above, or a reactive derivative at the carboxyl group thereof, and a compound represented by the general formula (3):

[0010]

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(Wherein R ³ and R ⁴ represent the same or different carboxyl-protecting groups) and are condensed with a compound represented by the general formula (4)

[0012]

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(Wherein R ¹ , R ² , R ³ , R ⁴ , X and Y are as defined above), and then the ester is converted to an acid or an alkali. And a method for producing a pyrimidine ring-fused cyclopentylidene derivative represented by the above general formula (1) or a pharmacologically acceptable salt thereof. Further, the present invention provides a compound represented by the general formula (5):

[0014]

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(Wherein R ¹ , R ² , X and Y are as defined above. —COOR ⁵ represents a carboxyl group which may be esterified with a protecting group) or a compound thereof. It provides salt. Further, the present invention provides an antitumor agent comprising a pyrimidine ring-fused cyclopentylidene derivative represented by the above general formula (1) or a pharmacologically acceptable salt thereof as an active ingredient.

In the above formula, when R ¹ is a hydroxyl group or an amino group, compounds (1), (2), (4) and (5) are compatible with keto-enol type or imine-enamine type. The isomers exist in equilibrium. In the present specification, for the sake of convenience of description, the enol type (hydroxyl type) or the enamine type (amine type) is described and named, but in any case, the compound of the present invention is not only enol type but also keto type (oxo). Body, i.e., amide form) or enamine form as well as imine form. In addition, in the present compound (1), in addition to the asymmetric center of the L-glutamic acid moiety, when a substituent is present on X, the asymmetric center may be present on the carbon atom at the base of the substituent, Except that the absolute configuration of the asymmetric carbon atom in the glutamic acid moiety is S (L-type), the absolute configuration of the other asymmetric center may be any of S, R or a mixture of RS. In this case, compound (1) is obtained as diastereoisomers or a mixture of diastereoisomers, and these components can be easily separated and purified by a conventional method such as fractional crystallization or chromatography, if necessary. I can do it. Any diastereoisomers that are separated fall within the scope of the invention. The compound (1) of the present invention is generally obtained as a powder, a crystalline solid or a crystal.

In the above formula, X is a divalent linear group consisting of 1 to 4 atoms which may have a substituent, and the atoms constituting the linear chain include carbon and nitrogen. , Oxygen, sulfur and the like, and examples of the substituent include a hydrocarbon group, a halogen atom, and a substituted or unsubstituted amino group.

In the above formula, the phenylene group represented by Y is, for example, a 1,4- or 1,3-phenylene group, and the thienediyl group and the frangylyl group are, for example, 2,2.
A thiophene group or a furan group having a bond at the 5 (or 5,2) -position or at the 3,5 (or 5,3) -position, and examples of the indolediyl group and the indolinediyl group include 1,1
An indole group having a bond at the 4-position, 1,5-position or 1,6-position is exemplified.

The carboxyl-protecting groups represented by R ³ , R ⁴ and R ⁵ include alkyl groups having 1 to 5 carbon atoms,
Examples include a benzyl group which may have a substituent, a phenyl group which may have a substituent, and a trisubstituted silyl group. Examples of the alkyl group herein include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a tertiary butyl group, and a pentyl group, and a benzyl group which may have a substituent. A phenyl group, a methoxyphenyl group, a nitrophenyl group or the like as a phenyl group which may further have a substituent, and a tert-butyldimethylsilyl group as a trisubstituted silyl group. And tert-butyldiphenylsilyl groups.

Next, the method for producing the compound (1) of the present invention will be described. The production of the compound of the present invention can be carried out using an appropriate reaction generally performed. Examples are shown below, but the present invention is not limited to these. Compound (1) is
As shown in the following reaction formula, the carboxylic acid represented by the formula (2) or a reactive derivative thereof is condensed with a glutamic acid derivative represented by the formula (3) to obtain an intermediate represented by the formula (4). It can be obtained by removing the carboxyl protecting group by a method such as hydrolysis.

[0021]

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(Wherein R ¹ , R ² , R ³ , R ⁴ , X and
Y has the same meaning as described above. ) As the above-mentioned condensation method, a conventional synthesis technique for peptide formation can be applied. For example, a compound (2) may be added to a carboxylic acid activating reagent such as chlorocarbonates, organic acid anhydrides, diphenylphosphoryl azide, diethyl cyanophosphate, carbonyldiimidazole, chlorophosphate or carbodiimide in the presence of a base. Or act in the absence,
Then, the intermediate compound (4) can be obtained by reacting the compound (3). The amount of the activating reagent to be used for the compound (2) is 1 to 25 molar equivalents, preferably 1 to 25 molar equivalents.
Selected from 5 molar equivalents. Methyl chlorocarbonate or ethyl chlorocarbonate is used as the chlorocarbonate used as the activating reagent, and acetic anhydride, chloroacetic anhydride or the mixed acid anhydride method is used as the organic acid anhydride. Diphenyl chlorophosphate, diethyl chlorophosphate and the like can be used.
As the carbodiimides, dicyclohexylcarbodiimide is practically preferable. In addition, carbodiimides can be appropriately selected from carbodiimides such as diphenylcarbodiimide, 1,3-diparatolylcarbodiimide or 1-cyclohexyl-3 (2-morpholinoethyl) carbodiimide. I can do it.

The compound (4) can be obtained by reacting the compound (2) with a halogenating reagent to convert the compound into a carboxylic acid halide (acyl halide) and then reacting the compound (3) in the same manner as described above. . Examples of the halogenating reagent include thionyl chloride, oxalyl chloride, phosphorus trichloride, and corresponding fluorides, bromides, iodides, and triphenylphosphine-
Carbon tetrachloride, triphenylphosphine-bromine and the like can be used.

The reaction for obtaining the intermediate compound (4) is preferably carried out in the presence of a solvent.
Water, alcohols (methanol, ethanol, propanol, etc.), ethers (diethyl ether, tetrahydrofuran, dioxane, etc.), nitriles (acetonitrile, etc.), aromatic hydrocarbons (benzene, toluene, etc.), halogenated hydrocarbons (dichloromethane, Chloroform, carbon tetrachloride, etc.), aliphatic hydrocarbons (hexane,
Pentane, etc.), acetone, pyridine, dimethylformamide, dimethylsulfoxide and the like, and a mixture of these solvents as appropriate may be used. Intermediate compound
The reaction to obtain (4) is carried out in the presence or absence of a base, the base used is sodium methylate, sodium ethylate, lithium hydroxide, sodium hydroxide,
It is appropriately selected from inorganic bases such as potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate and calcium carbonate, and organic bases such as triethylamine, trimethylamine, pyridine and triethanolamine. The reaction temperature is generally in the range of −10 ° C. to the boiling point of the reaction solvent, preferably 0 to 50 ° C., and the reaction time is generally 0.5 hour to 4 hours.
Can be implemented in days.

Next, the ester intermediate (4) obtained in the above-mentioned condensation step is hydrolyzed to a free carboxylic acid to obtain the compound (1). The hydrolysis reaction is carried out in an aqueous solution or, in some cases, in a water-affinity organic solvent such as methanol, ethanol, tetrahydrofuran, dioxane, dimethylformamide, or dimethylsulfoxide, in an aqueous inorganic acid such as hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid, or in trifluoroacetic acid. Using organic acids such as trichloroacetic acid, p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, or aqueous caustic alkalis such as sodium hydroxide or potassium hydroxide, sodium carbonate or potassium carbonate, sodium hydrogencarbonate, etc. Using alkali metal carbonates, alkali hydrogen carbonates, metal alkoxides such as sodium methoxide or sodium ethoxide. The reaction temperature is, for example, the boiling point of the solvent used under ice cooling, preferably 10 to
The reaction can be performed at a temperature of 70 ° C. for a reaction time of, for example, 1 hour to about 2 days. If an alkali is used, the product will have the glutamic acid moiety formed as an acid salt or intermediate salt.

Next, a method for producing the starting compound (2) will be described. Among the compounds (2), a compound wherein R ¹ is an amino group can be produced, for example, by the following reaction step.

[0027]

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[0028]

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In the above process, R ¹ represents an amino group, R ² ,
R ⁵ , X and Y are as defined above. Z represents a halogen atom such as a chlorine atom, a bromine atom and an iodine atom,
R ⁶ represents a general protecting group such as a linear or branched alkyl group having 1 to 10 carbon atoms, an aryl group such as a phenyl group and a substituted phenyl group, and an arylalkyl group such as a benzyl group. R ⁷
Represents a residue obtained by removing the -SH group from an aliphatic or aromatic thiol.

Hereinafter, the above reaction steps will be described.
2-cyanocyclopenten-1-one (7) is obtained by reacting cyclopentenone with an aromatic thiol or aliphatic thiol such as tri-lower alkylaluminum and thiophenol, for example, isopropyl thiol, and then reacting with tosyl cyanide. Silica gel or BF ₃ −
By reacting with a Lewis acid such as ether, the compound can be synthesized smoothly and in good yield. In this case, 2-cyanocyclopenten-1-one (7) is converted to an intermediate 3-alkylthio-2-one.
Produced via cyanocyclopentane.

The compound (8) can be synthesized by reacting a cyclic carboxylic acid ester (6) having an ω-halogenoalkyl bond with 2-cyanocyclopenten-1-one (7). In this case, the reaction is performed in the presence of a tin hydride such as trialkyltin hydride or triphenyltin hydride or a trialkylgermanium hydride and a radical initiator such as azobisisobutyronitrile and benzoyl peroxide. Further, 2-cyanocyclopenten-1-one is used in the presence of a copper salt such as copper cyanide.
Compound (8) can also be obtained by reacting (7) with an organic zinc compound derived from zinc and a cyclic carboxylic acid ester (6) having an ω-halogenoalkyl bond.

As another method for synthesizing compound (8), there is a method using 3-toluenesulfonylcyclopentan-1-one as a starting material. That is, as shown in the following reaction formula,
Glycol is allowed to act on 3-toluenesulfonylcyclopentan-1-one to convert it to a dioxolane compound (14), and then the compound is added in the presence of lithium diisopropylamide.
(6) to effect alkylation. The alkylated product (15) is subjected to an acidic treatment to give a 3-position-substituted cyclopentene-1.
-ON (16) can be obtained. Next, when cooled to a reaction solution obtained from toluenethiol and an equivalent amount of trimethylaluminum, the compound (16) is added, 4-toluenesulfonyl cyanide is acted on the reaction solution, and the obtained reaction product is purified by silica gel column chromatography. By compound
(8) (T. Yoshida et al., Bull. Chem. S.
oc. Jpn., 55, 393 (1982).

[0033]

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(In the above reaction formula, R ⁵ , X, Y and Z are as defined above.) From 3-substituted alkyl-2-cyanocyclopentan-1-one (8) to lower alkyl enol ether (9) Conversion to diazomethane, TMS diazomethane or its analog, 2,2-dimethoxypropane and paratoluenesulfonic acid, or a reagent for introducing a protecting group to a common hydroxyl group such as orthoformate or trialkyloxonium fluoroborate Can be achieved by:

1-alkoxy-2-cyano-3-substituted alkyl-5-halogeno-1-cyclopentene compound (10)
Is obtained by reacting the above enol ether (9) with an N-halogenated amide (imide) compound in the presence of a solvent (for example, an ether-based solvent or benzene) which is not easily subjected to a radical reaction under heat or light irradiation. Halogenation at the allylic position occurs, and the compound (10) can be obtained. N-
As the halogenated amide (imide) compound, for example,
1,3-dibromo-5,5-dimethylhydantoin, N, N
-Dichlorourethane, N-chloro- or N-bromosuccinimide, N-bromocaprolactam, N-chloro-N-cyclohexylbenzenesulfonamide, tert-butyl hypochloride and the like can be used.

The corresponding thiol compound (11) can be obtained by allowing an aliphatic or aromatic thiol to act on the halogenated compound (10). Examples of the aliphatic or aromatic thiol include benzenethiol, substituted benzenethiol (substituents include an alkyl group, an alkoxy group, an amino group, and a halogen atom), and various alkylthiols (the alkyl group is an ethyl group, Butyl, octyl, isopropyl, tertiary butyl and the like). These aliphatic or aromatic thiols can also be used in this reaction as thiol alkali metal salts. In this case, when an aqueous solvent is used and a small amount of a phase transfer catalyst is present, a favorable result may be obtained in some cases.

1-alkoxy-2-cyano-3-substituted alkyl-5-alkylthio (or arylthio)-
1-cyclopentene compound (11) under normal pressure or in a sealed tube,
When guanidine or a salt thereof is allowed to act under heating, the desired 2,4-diamino-7-mercapto-6,7-dihydro-
5H-cyclopenta [d] pyrimidine compound (12, R ¹ = NH
₂ ) can be obtained. Guanidine is an enol ether
An excess (2 to 20-fold, preferably 5 to 10-fold) of (11) is used, and the solvent is a protic solvent such as methanol, ethanol, methoxyethanol, propanol,
Use of 2-methyl-2-propanol or the like is preferable, and the temperature is preferably 50 to 200 ° C, preferably 100 to 185 ° C, and the reaction time is preferably 8 to 70 hours. However, a carboxylic acid may be directly obtained under high-temperature, long-time reaction conditions. Next, the thioether (12) can be converted to the corresponding sulfoxide by treating it with a peroxide such as perbenzoic acid at room temperature or under ice cooling. The sulfoxide group is eliminated by heating to give the corresponding olefin compound (13). In this case, it is preferable to use a trialkyl phosphite or a tertiary base. Olefin compound (13) is a 6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene compound
(5) Besides 5-substituted-5H-cyclopenta [d] pyrimidine compound (5a) and 5-substituted-7H-cyclopenta [d]
Contains a pyrimidine compound (5b). These can be separated by ordinary separation means such as liquid chromatography. 6,7-dihydrocyclopenta [d] pyrimidine-5
-The above carboxylic acid compound (2) can be obtained by deprotecting the ester group of the ylidene compound (5). The carboxylic acid compound (2) can be obtained by isomerizing the compound (5a) and the compound (5b) in the process of deprotection with an acid or an alkali (or a base). As another method for synthesizing the carboxylic acid ester (5), the method shown in the following reaction formula can be mentioned.

[0038]

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(In the above reaction formula, R ¹ , R ² , R ⁵ , R ⁶ , R ⁷ , X,
Y and Z are as defined above. That is, a reaction solution obtained by allowing cyclopentenone to react with trimethylaluminum and an equivalent amount of toluenethiol is added to the reaction solution.
-The thiol adduct (17) can be obtained by the action of toluenesulfonyl cyanide. The compound (18) can be obtained by reacting the thiol adduct (17) with an alkylating reagent such as diazomethane to form an enol ether, and then reacting with hydrogen peroxide or perbenzoic acid. This is reacted with an alkyl halide (6) under basic conditions such as lithium diisopropylamide (LDA) to give a compound (19). The carboxylic acid ester (5) can be obtained by heating this in a sealed tube with guanidine or an acidic salt thereof in the presence of an organic solvent.

In general, a carboxylic acid (2) having a 6,7-dihydrocyclopenta [d] pyrimidine ring is prepared by converting a corresponding carboxylic acid ester (5) from a compound (4) according to a method for producing a compound (1). It can be obtained by hydrolysis in the presence of an acid or alkali.

Next, among the compounds (5), those in which R ¹ is a hydroxyl group can be produced by the following reaction steps.

[0042]

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[0043]

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In the above process, R ¹ represents a hydroxyl group, and R ² , R ⁵ ,
R ⁶ , R ⁷ , X, Y and Z are as defined above, and R ⁸ is a carboxyl-protecting group. 2-carboalkoxy-2-
The synthesis of cyclopenten-1-one (20) can be performed by a known method. That is, MAGuaciaro et al. Tetrahedrons,
47 , 4661 (1978).
-On-synthesis or H.Reich et al., J.Am.Chem.So
c., 97, 5434 (1975) and can be used for the purpose of the present invention by synthesizing from 2-carboethoxycyclopentanone.

The keto compound (21) is obtained by converting a cyclic carboxylic acid ester (6) having an ω-halogenoalkyl group into a 2-carboalkoxy-2-yl group in the presence of a trialkyl or triphenyltin hydride and azobisisobutyronitrile. It can be synthesized by reacting with cyclopenten-1-one (20). Conversion of the keto form, i.e., the cyclopentanone ring-containing carboxylic acid ester (21) to the enol ether (22) can be carried out by a general method such as diazomethane or an analog thereof, 2,2-dimethoxypropane and paratoluenesulfonic acid, or orthoformate. The reaction proceeds smoothly by the action of a reagent for introducing a protecting group into a hydroxyl group.

The 1-alkoxy-2-alkoxycarbonyl-3-substituted alkyl-5-halogeno-1-cyclopentene compound (23) is prepared by adding an N-halogenated amide (imide) compound to the above enol ether (22) by heat or heat. When acted under light irradiation, halogenation at the allylic position occurs and compound (23) can be obtained. As the N-halogenated amide (imide) compound, for example, 1,3-dibromo-5,5-
Dimethylhydantoin, N, N-dichlorourethane, N
-Chloro- or N-bromosuccinimide, N-bromocaprolactam, N-chloro-N-cyclohexylbenzenesulfonamide, tert-butyl hypochloride and the like can be used.

The corresponding thiol compound (24) can be obtained by allowing an aliphatic or aromatic thiol to act on the halogenated compound (23). Examples of the aliphatic or aromatic thiol include benzenethiol, substituted benzenethiol (substituents are groups such as alkyl, alkoxy, amino, and halogen), and various alkylthiols (the alkyl group is ethyl, butyl, octyl, isopropyl, and tertiary). Butyl and the like) can be used. These aliphatic or aromatic thiols can also be used in this reaction as thiol alkali metal salts.

Next, a thiol compound (24) was prepared using 1N hydrochloric acid.
Is converted again into a keto form, that is, a cyclopentanone ring-containing carboxylic acid ester (25), and then reacted with guanidine under normal pressure or in a sealed tube under heating to close the ring to give 2-amino-4-.
Hydroxy-6,7-dihydro-5H-cyclopenta [d] pyrimidine compound (26) can be obtained. Guanidine converts its hydrochloride or other salt to potassium t-
It can be obtained using butoxide or other alkoxides. Guanidine is used in excess (2 to 20 times, preferably 5 to 10 times by mole) with respect to the cyclopentanone ring-containing carboxylic acid ester (25), and the solvent is a protic solvent such as methanol, ethanol, methoxyethanol, propanol, or the like. The use of 2-methyl-2-propanol or the like is preferable, and the temperature is 10 to 200 ° C, preferably 12 to 1 ° C.
Preferred conditions are 85 ° C. and a reaction time of 8 to 70 hours. However, a carboxylic acid may be directly obtained under high-temperature, long-time reaction conditions. Next, 2-amino-4-
Hydroxy-6,7-dihydro-5H-cyclopenta [d] pyrimidine compound (26) can be converted to the corresponding sulfoxide by treating it with a peroxide such as perbenzoic acid at room temperature or under ice-cooling. The sulfoxide group is eliminated by heating to give the corresponding olefin compound (27). In this case, when a trialkyl phosphite or a tertiary base is present, a favorable result may be obtained. Olefin compound (27)
6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene compound (5), 5-substituted-5H-cyclopenta [d] pyrimidine compound (5a) and 5-substituted-7H-cyclopenta [d] pyrimidine compound (5b) is contained. These can be separated from each other by a general separation operation such as liquid chromatography. The carboxylic acid compound (2) described above can be obtained by deprotecting the ester group of the 6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene compound (5).

Next, in the compound (5), R ¹ is an amino group, X
There -NH- or -NR ⁹ - (R ⁹ represents a divalent functional group forming a condensed ring with an acyl group, an alkyl group or Y)
If it contains (X = X'-NR ⁹ - or -X'-NH- (X 'represents an alkylene group)), compound (5) can be prepared by the reaction steps shown below.

[0050]

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[0051]

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In the above process, R ¹ represents an amino group, and R ² ,
R ⁵ , R ⁶ , R ⁷ , R ⁹ , X ′, Y and Z are as defined above,
R ¹⁰ represents an acyl group as a protecting group for a hydroxyl group. By reacting the ω-halogenoalkyl compound (28) with 2-cyanocyclopenten-1-one (7), 3- (acyloxyalkyl) -2-cyanocyclopentan-1-one (2
9) can be synthesized. In this case, the reaction can be carried out according to the method for synthesizing compound (8). 3- (acyloxyalkyl) -2
The conversion of -cyanocyclopentan-1-one (29) to the corresponding enol ether (30) can be carried out according to the above-mentioned reaction for deriving compound (9) from compound (8). Below, 1
-Alkoxy-2-cyano-3- (acyloxyalkyl) -5-halogeno-1-cyclopentene compound (31),
The corresponding alkylthio compound (32), 2,4-diamino-7
-Alkylthio-6,7-dihydrocyclopenta [d] pyrimidine compound (33) can be carried out according to the above-mentioned synthesis reaction of compounds (10), (11) and (12) and the synthesis conditions. When R ¹ is an amino group and R ² is a hydrogen atom or a methyl group, formamidine or acetamidine may be used instead of guanidine.

In the next step, 2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidenealkyl compound (34) was prepared from compound (12) to compound (13) and compound (13). 5) can be implemented according to the method of obtaining When R ² is a hydrogen atom or a methyl group, it can be achieved by a similar synthetic reaction. In the compound (33), the acyl group may be eliminated during the reaction to directly obtain the alkyl alcohol (34),
When the acyl group is not eliminated, an appropriate alcohol treatment (34) can be obtained by appropriate treatment based on the properties of the acyl group. Then, a halogenating reagent is allowed to act on this to form an alkyl halide (35), and then a monocyclic or bicyclic aromatic carboxylic acid having an acylamino (R ⁹ = acyl group) or alkylamino (R ⁹ = alkyl group) bond The corresponding compound (5) (X = -X'-NR
⁹ -, R ⁹ = acyl or alkyl group) can be obtained. In the case of a bicyclic heterocycle, a structure in which R ⁹ is fused to Y 2 may be adopted. In this case, R ⁹ represents a divalent functional group (for example, an ethylene group, a vinylene group and the like can be mentioned).
When R ⁹ is a protecting group for an amino group, that is, an acyl group, the target compound (X = −X′-NH—) can be obtained by deprotecting R ^9.
Can be led to. As described above, the ester compound (5), which is an important intermediate, can be obtained.

The compounds of the present invention can form pharmacologically acceptable salts with organic or inorganic acids. Examples of acids suitable for salt formation include hydrochloric, sulfuric, phosphoric, acetic, citric, malonic, salicylic, malic, fumaric, maleic, succinic, ascorbic, methanesulfonic, and the like. it can. Organic or inorganic acid salts can be prepared in a conventional manner by adding an equivalent amount of an acid to the free base form. Free bases can be regenerated by treating salts of organic and inorganic acids with a base. Sodium hydroxide, potassium carbonate, ammonia, aqueous solutions of sodium bicarbonate and the like are suitable for this purpose. The compounds of the present invention can also produce pharmaceutically acceptable carboxylate salts by reacting a suitable base with one or more free carboxyl groups. Suitable bases also include alkali metal hydroxides or carbonates, for example sodium hydroxide, potassium hydroxide and the corresponding carbonates, as well as ammonia, nitrogen bases such as alkylamines such as triethylamine and the like.

Specific examples of the compound (1) of the present invention include: N- {4- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl〕
Benzoyl} -L-glutamic acid N- {5- [3- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] -2-thenoyl} -L-glutamic acid N- {4- [3- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] -benzoyl} -L-glutamic acidN- {5- [2- ( 2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl]
-2-Thenoyl} -L-glutamic acid N- {5- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-ethyl-ethyl] -2 -Thenoyl} -L-glutamic acid N- {4- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-ethyl-ethyl] benzoyl} -L -Glutamic acidN- {5- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-methyl-ethyl] -2-thenoyl} -L-glutamic acid N- {4- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-methyl-ethyl] benzoyl} -L-glutamic acid N- {5 -[3- (2,4-diamino-6,7-dihydrocyclopen [D] pyrimidine-5-ylidene) -1-methyl-propyl] -2-thenoyl} -L-glutamic acidN- {4- [3- (2,4-diamino-6,7-dihydrocyclopenta [d ] Pyrimidine-5-ylidene) -1-methyl-propyl] benzoyl} -L-glutamic acidN- {4- [2- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl] benzoyl} -L-glutamic acidN- {5- [2- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl] -2-thenoyl} -L-glutamic acidN- {4- [N '-[2- (2,4-diamino-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl] -N'-methyl] aminobenzoyl} -L
-Glutamic acid N- {4- [3- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-]
Fluoropropyl] benzoyl} -L-glutamic acidN- {4- [3- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] benzoyl} -L-glutamic acidN- {5- [3- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] -2-thenoyl} -L-glutamic acid N- {1- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine -5-ylidene) ethyl] indol-5-ylcarbonyl} -L-glutamic acidN- {1- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) Ethyl] -2,3-dihydroindol-5-ylcarbonyl} -L-glutamic acidN- {5- [2- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-ethyl-ethyl] -2-thenoyl} -L-glutamic acidN- {4- [2- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-ethyl-ethyl] benzoyl} -L-glutamic acidN- {5- [2- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-methyl-ethyl] -2-thenoyl} -L-glutamic acidN- {4- [2- (2-amino-4-oxo-6,7-
Dihydrocyclopenta [d] pyrimidine-5-ylidene) -1-methyl-ethyl] benzoyl} -L-glutamic acid.

[0056]

The following pharmacological experimental examples are provided to describe the effects of the compounds of the present invention. Experimental Example 1 Inhibitory activity of dihydrofolate reductase (DHFR) was basically determined by the method of DKMisra et al. (Nature, 18
9, 39-42 (1961). DHFR was prepared from mouse leukemia cell P388 cells. Preparation method is JMWhi
teley et al. (Arch.Biochem.Biophys., 150, 15-22
(1972)). The method is potassium phosphate buffer (pH
7.5, 75 mM), mercaptoethanol (7.5 mM), NADPH
(0.25 mM) is added to the purified DHFR solution to make a total volume of 0.75 ml. To this, 0.015 ml of a serial dilution of the compound of the present invention is added, and preheating is performed at 37 ° C. for 5 minutes.
Next, 0.015 ml of a 25 μM solution of dihydrofolate is added, and the reaction is carried out at 37 ° C. for 5 minutes. The decrease in absorbance at 339 nm per minute was measured to determine the amount of tetrahydrofolic acid produced. Based on comparison with a control solution (without compound), the
The 50% DHFR inhibitory concentration (IC ₅₀ ) is calculated. Table 1 shows the results.

[0057]

[Table 1]

Experimental Example 2 Tumor Cell Growth Inhibitory Activity As tumor cells, mouse leukemia cell P388 cell, mouse leukemia cell P388 / MTXr E-2 cell, mouse colon cancer cell Colon26 cell, and human nasopharyngeal empty cell KB cell were used. Each of the above cancer cell solutions prepared in RPMI-1640 medium (10% fetal bovine serum) is put into a 96-well microplate in a fixed amount using a pipette. These were placed in a carbon dioxide incubator containing 5% CO ₂ and 37
Incubate at ℃ for 1 day. After dissolving the compound of the present invention in DMSO,
A culture solution is added to prepare a serial dilution. After adding a certain amount of these solutions to each well containing the cultured cells, the cells are cultured at 37 ° C. for 3 days. Viable cells are measured by the MTT method to determine the 50% cell growth inhibitory concentration (IC ₅₀ ). The measurement method was based on the method by MC Alley et al. (Cancer Research, 48, 589-601 (1988)). Table 2 shows the results.

[0059]

[Table 2]

According to the above experimental examples, the compound of the present invention has a dihydrofolate reductase (DHFR) inhibitory effect, and has mouse leukemia cell P388, mouse leukemia cell P388 / MTXr E-2, mouse colon cancer cell Colon26 cell, and human nasopharynx. It shows clear growth inhibition in cancer cells and KB cells. The compound of the present invention can be administered orally or parenterally as it is or in the form of a pharmaceutically acceptable carboxylate or acid addition salt. The dose varies depending on the severity of the disease; the weight, age, sex, and sensitivity of the patient; the administration method (route), the administration interval (schedule); the type of active ingredient; However, the usual daily dose is 1 mg to 300 mg / man, preferably 10 mg to 150 mg / ma for a subject weighing 60 kg (1.62 m ² / man).
n and continuous administration once a day or intermittent administration 1 to 3 times a week.

Forms suitable for oral therapeutic administration of the compound of the present invention include tablets, buccals, capsules, suspensions, syrups, troches and the like. The ratio (%) of active compound to excipient in each preparation is approximately 0.
It is convenient to have a weight of 5-50%. The amount of such active compounds is such that a suitable dosage will be obtained. Tablets, capsules, troches and the like may be added with binders such as tragacanth gum, gum arabic, starch and gelatin; excipients such as dicalcium phosphate, various starches, alginic acid and stearic acid, disintegrants and lubricants. Tablets, capsules and the like may be coated with a coating agent such as methylcellulose, shellac, sugar and the like. Forms suitable for parenteral therapeutic administration of the compounds of the present invention include injections, dispersions, and emulsions. Solutions of the active compounds can be prepared in water by mixing with a surfactant such as hydroxypropylcellulose. Injectables include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of injection solutions. The carrier can be a solvent including water, ethanol, glycerol, polyols, mixtures thereof, and vegetable oils. Dispersions can be prepared in water and in oils containing glycerol, liquid polyethylene glycols and mixtures thereof. For use in preparing the injection solution-a preferred method for preparing a sterile powder is a method in which the sterilized and filtered solution is concentrated under reduced pressure or freeze-dried to obtain a desired active ingredient.

[0062]

Next, the present invention will be described more specifically with reference to Production Examples and Examples, but the present invention is not limited to the following Production Examples and Examples. In the structural formulas in the examples, Me represents a methyl group, Et represents an ethyl group, and t-Bu represents a tert-butyl group.

Production Example 1 Production of 2-cyano-2-cyclopenten-1-one

[0064]

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P-Toluenethiol (13.7 g, 0.11 mol)
The anhydrous methylene chloride solution of was stirred under ice cooling in a nitrogen stream,
A hexane solution of trimethylaluminum (7.9 g, 0.11 mol) was added thereto, followed by stirring for about 20 minutes. The reaction solution was cooled in a dry ice-acetone bath, and a methylene chloride solution of 2-cyclopentenone (8.41 g, 0.10 mol) was added dropwise thereto.
The mixture was further stirred for 30 minutes. After the reaction solution was diluted with anhydrous tetrahydrofuran, a solution of p-toluenesulfonyl cyanide (22.9 g, 0.12 mol) in tetrahydrofuran was added at −50 ° C. or lower, and the mixture was stirred for about 30 minutes, and further stirred for about 60 minutes in an ice bath. did. After about 40 ml of methanol was added dropwise, diethyl ether was added, and this was washed with aqueous hydrochloric acid, aqueous sodium hydrogen carbonate, and brine, then dehydrated and concentrated, and the concentrated solution was left standing or a small amount of n-hexane. : Ethyl acetate = 1: 1 A mixed solution was added and allowed to stand, and a crystal powder of 2-cyano-3- (4-methylphenyl) thio-1-cyclopentanone was deposited. Yield 22g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.82 to 2.00 (m, 1
H), 2.37 (s, 3H), 2.30-2.60 (m, 3H), 3.04 (d, J = 9.6Hz, 1H),
3.68 to 3.78 (m, 1H), 7.20 (d, J = 8.4 Hz, 2H), 7.42 (d, J = 8.4H
z, 2H) The compound (10 g, 43 mmol) obtained in the previous operation was dissolved in tetrahydrofuran, adsorbed on a silica gel column, and then subjected to chromatography using a mixed solution of n-hexane and ethyl acetate as a developing solution to obtain the desired compound. The containing fraction was concentrated to give the target compound as a colorless to yellow oil. Yield 2.9 g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 2.55 to 2.58 (m, 2
H), 2.90 to 2.94 ((m, 2H), 8.32 (t, J = 2.8 Hz, 1H) IR (neat) ν (cm ^-1 ): 2238,1728,1607 Production Example 2 4-vinylbenzoic acid tertiary Production of butyl ester

[0068]

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Methyltriphenylphosphonium bromide (18.2 g, 50.9 mmol) was suspended in anhydrous tetrahydrofuran, and dried under a stream of nitrogen in a dry ice-acetone mixture.
A 1.6 M butyllithium hexane solution (49.7 mmol) was added, and after stirring for a while, the mixture was stirred under ice cooling for about 30 minutes. In addition, terephthalaldehyde acid tertiary butyl ester (1
(0 g, 48.5 mmol) in anhydrous tetrahydrofuran and stirred for about 40 minutes. The resulting precipitate is removed by filtration, the filtrate is concentrated to dryness, and the obtained residue is subjected to silica gel column chromatography (developing solution n-hexane: ethyl acetate = 20:
Purification in 1) gave the desired compound. Yield 8.47 g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.59 (s, 9H), 5.3
6 (dd, J = 0.8Hz, 10.8Hz, 1H), 5.84 (dd, J = 0.8Hz, 17.6Hz, 1
H), 6.74 (dd, J = 10.8Hz, 17.6Hz, 1H), 7.43 (d, J = 8.0Hz, 2
H), 7.94 (d, J = 8.0 Hz, 2H) IR (neat) ν (cm ⁻¹ ): 2978,1710,1292,1166,916 Production Example 3 Tertiary butyl 4- (2-hydroxyethyl) benzoate Ester production

[0071]

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To a solution of 4-vinylbenzoic acid tertiary butyl ester (8.47 g, 42.1 mmol) in anhydrous tetrahydrofuran was added 9-borabicyclo [3.3.1] nonane (9-BBN, 5.65 g, 46.3 mmol) in a stream of nitrogen under anhydrous nitrogen. A tetrahydrofuran solution was added, and the mixture was stirred at room temperature for 3 hours. After 15 ml of water and then 17 ml of 3N NaOH were added to the reaction solution, 30% aqueous hydrogen peroxide was added dropwise at 50 ° C. or lower. After stirring for 1 hour as it was, the reaction solution was poured into water and extracted with ethyl acetate. The extract was washed with brine, dehydrated and evaporated, and the obtained residue was purified by silica gel column chromatography (developing solution n-hexane: ethyl acetate = 1: 1) to obtain the desired compound. Yield 8.71 g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.41 (t, J = 6.6H
z), 1.59 (s, 9H), 2.92 (t, J = 6.6Hz, 2H), 3.88 (q, J = 6.6Hz, 2
H), 7.28 (d, J = 8.0Hz, 2H), 7.93 (d, J = 8.0Hz, 2H) IR (neat) ν (cm ^-1 ): 3150-3700,2978,1713,1294,11
67 Production Example 4 Production of tert-butyl 4- (2-bromoethyl) benzoate

[0074]

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4- (2-hydroxyethyl) benzoic acid
To a methylene chloride solution of tertiary butyl ester (8.7 g, 39.1 mmol) and triethylamine (5.9 g, 58.7 mmol) was added dropwise a solution of methanesulfonyl chloride (5.8 g, 50.8 mmol) in methylene chloride under cooling with dry ice-acetone. Then, the mixture was stirred for 30 minutes under ice cooling. 0.5N aqueous sodium hydrogen sulfite was added to the reaction solution, and the mixture was stirred, and then the organic layer was separated. This was washed with brine, dried, and the solvent was distilled off to obtain 11.7 g of methanesulfonic acid ester. This was dissolved in 2-butanone, lithium bromide (5.1 g, 58.4 mmol) was added, and the mixture was stirred at 65 ° C for 12 hours in a nitrogen stream. After leaving at room temperature, the resulting precipitate was filtered off, the filtrate was concentrated to dryness, and the residue was purified by silica gel column chromatography (developing solution n-hexane: ethyl acetate = 4: 1) to obtain the desired compound. 9.8 g of colorless to pale brown crystals were obtained.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.59 (s, 9H), 3.2
1 (t, J = 7.2Hz, 2H), 3.58 (t, J = 7.2Hz, 2H), 7.26 (d, J = 8.0Hz,
2H), 7.94 (d, J = 8.0 Hz, 2H) Production Example 5 4- [2- (2-cyano-1-cyclopentanone-3-
Production of tert-butyl benzoate)

[0077]

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4- (2-Bromoethyl) benzoic acid tert-butyl ester (3.21 g, 11.3 mmol) was dissolved in anhydrous benzene, and the solution obtained in Production Example 1 was refluxed under heating and reflux.
-Cyano-2-cyclopenten-1-one (2.41 g, 2
Anhydrous benzene solution of 2.5 mmol) and an anhydrous benzene solution of azobisisobutyronitrile (AIBN) (0.05 g, catalytic amount) and tri-n-butyltin hydride (3.62 g, 12.4 mmol) were simultaneously slowly added dropwise. After heating under reflux for about 20 minutes, the solvent was distilled off, and the residue was purified by silica gel column chromatography to obtain the desired product. Yield 0.43g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.59 (s, 9H), 1.7
8 to 1.96 (m, 1H), 2.02 to 2.22 (m, 1H), 2.24 to 2.58 (m, 4
H), 2.76 to 2.94 (m, 3H), 7.26 (d, J = 8.2 Hz, 2H), 7.93 (d, J = 8.
2Hz, 2H) IR (neat) ν (cm ⁻¹ ): 2977,2933,2246,1760,1713 Mass (FAB) M + H ⁺ = 314 Production Example 6 4- [2- (2-cyano-3-methoxy-2) -Cyclopenten-1-yl) ethyl] benzoic acid tertiary butyl ester

[0080]

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4- [2- (2-cyano-1-cyclopentanone-3-yl) ethyl] benzoic acid tert-butyl ester (0.43 g, 1.37 mmol) and N, N-diisopropylethylamine (0.21 g, 1.64 mmol) was dissolved in a mixture of methanol and acetonitrile (1: 5), and a 10% by weight solution of trimethylsilyldiazomethane in hexane 3.
2 g was added and the mixture was stirred at room temperature for 2 hours. After adding a small amount of acetic acid and stirring, the solvent was distilled off. The residue was dissolved in diethyl ether, washed with dilute hydrochloric acid, dilute alkali, and brine, dehydrated and concentrated to dryness, and the residue was purified by silica gel column chromatography to obtain the desired product. Yield 0.23
g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.52 to 1.72 (m, 2
H), 1.59 ((s, 9H), 2.02 to 2.18 (m, 2H), 2.46 to 2.52 (m, 2
H), 2.62 to 2.82 (m, 2H), 2.82 to 2.94 (m, 1H), 4.05 (s, 3H),
7.24 (d, J = 8.2 Hz, 2H), 7.90 (d, J = 8.2 Hz, 2H) IR (neat) ν (cm ⁻¹ ): 2977,2933,2203,1710,1632 Mass (FAB) M + H ⁺ = 328 Production Example 7 4- [3- (2-cyano-1-cyclopentanone-3-
Production of tertiary butyl ester

[0083]

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Using tert-butyl 4- (3-bromopropyl) benzoate (1.5 g, 5 mmol) as a starting material, the corresponding target compound was obtained in the same manner as in Production Example 5. Yield 0.43g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.40 to 1.66 (m, 2
H), 1.59 (s, 9H), 1.70-1.92 (m, 3H), 2.20-2.56 (m, 4H),
2.60 ~ 2.80 (m, 2H), 2.81 (d, J = 11.6Hz, 1H), 7.22 (d, J = 8.0H
z, 2H), 7.91 (d, J = 8.0 Hz, 2H) IR (neat) ν (cm ⁻¹ ): 2958,2932,2248,1760,1710 Mass (EI) M ⁺ = 327 Production Example 8 4- [ Production of 3- (2-cyano-3-methoxy-2-cyclopenten-1-yl) propyl] benzoic acid tertiary butyl ester

[0086]

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Using the compound (0.41 g) obtained in Production Example 7 as a starting material, the reaction was carried out in the same manner as in Production Example 6 to obtain the desired product. Yield 0.31 g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 1.30 to 1.41 (m, 1
H), 1.41 to 1.55 (m, 1H), 1.59 (s, 9H), 1.55 to 1.82 (m, 3H),
2.02 to 2.12 (m, 1H), 2.42 to 2.48 (m, 2H), 2.61 to 2.76 (m, 2
H), 2.82 -2.91 (m, 1H), 4.03 (s, 3H), 7.22 (d, J = 8.0Hz, 2
H), 7.90 (d, J = 8.0 Hz, 2H) IR (neat) ν (cm ⁻¹ ): 2978,2935, 2203, 1712, 1632 Mass (EI) M ⁺ = 341 Production Example 9 4- [3- Production of (4-phenylthio-2-cyano-3-methoxy-2-cyclopenten-1-yl) propyl] benzoic acid tertiary butyl ester

[0089]

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The tert-butyl 4- [3- (2-cyano-3-methoxy-2-cyclopenten-1-yl) propyl] benzoate obtained in Production Example 8 (7.4
g, 21.7 mmol) and N, N-dichlorourethane (2.8 ml,
23.9 mmol) was dissolved in benzene, and the mixture was stirred at room temperature for 1 hour under ultraviolet irradiation with a high-pressure mercury lamp. After adding a small amount of aqueous sodium bisulfite solution and stirring, the solvent was distilled off. Add ether and benzenethiol to the residue (11 ml, 10
7 mmol), an aqueous solution of sodium hydroxide (4.3 g, 108 mmol), and catalytic amounts of tetrabutylammonium iodide and benzyltriethylammonium bromide were added and stirred.
The reaction mixture was extracted with ethyl acetate, the extract was dried, and the residue obtained by distilling off the solvent was purified by silica gel chromatography (developing solution n-hexane: ethyl acetate = 10: 1) to obtain the target compound. Obtained. Yield 6.8 g.

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.93 to 7.88 (m, 2
H), 7.39 to 7.16 (m, 7H), 4.18 (s, 3H), 4.02 (ddd, J = 0.8, 6.
4,9.2Hz, 1H × 0.7), 3.94 (dt, J = 4.0,16.8Hz, 1H × 0.3), 2.
77-2.48 (m, 5H), 1.59 (s, 9H), 1.68-1.47 (m, 4H) Production Example 10 4- [3- (2,4-diamino-6,7-dihydro-7-phenylthio-5H -Cyclopenta [d] pyrimidine-5
-Yl) propyl] benzoic acid tertiary butyl ester

[0092]

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Tertiary butyl 4- [3- (4-phenylthio-2-cyano-3-methoxy-2-cyclopenten-1-yl) propyl] benzoate obtained in Production Example 9 (6.8 g, 15 mmol) And guanidine carbonate (6.0g,
33 mmol) was mixed with 2-methyl-2-propanol, placed in a pressure-resistant reactor under a nitrogen atmosphere, and stirred at 150 ± 3 ° C. for 19 hours.
Heated for hours. The reaction solution was allowed to cool, the precipitate was filtered off, the filtrate was concentrated, and the residue was separated and purified by silica gel column chromatography (developing solution: chloroform: methanol = 10: 1) to obtain 1.2 g (2.5 mmol) of the desired product. Was done.

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.90 (d, J = 5 Hz, 2
H), 7.40 (d, J = 4Hz, 2H), 7.29-7.15 (m, 5H), 5.18 (brs, 2H),
4.88 (brs, 2H × 0.5), 4.83 (brs, 2H × 0.5), 4.41 (dd, J = 6.3,
7.8Hz, 1H × 0.5), 4.35 (dd, J = 4.0,8.8Hz, 1H × 0.5), 2.95
~ 2.80 (m, 1H), 2.67 ~ 2.56 (m, 3H), 2.40 ~ 2.15 (m, 1H),
1.65 to 1.52 (m, 4H), 1.59 (s, 9H) Production Example 11 4- [2- (4-phenylthio-2-cyano-3-methoxy-2-cyclopenten-1-yl) ethyl] benzoic acid tertiary Production of butyl ester

[0095]

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The 4- [2- (2-cyano-3-methoxy-2-cyclopenten-1-yl) ethyl] benzoic acid tert-butyl ester obtained in Production Example 6 (3.5 g,
10.7 mmol) and N, N-dichlorourethane (1.8 ml, 15.4 mm
ol) was dissolved in benzene, and the mixture was stirred at room temperature for 30 minutes under irradiation with ultraviolet light using a high-pressure mercury lamp. After adding a small amount of aqueous sodium bisulfite solution and stirring, the solvent was distilled off. Ether and benzenethiol (5.5 ml, 54 mmol) in the residue
And an aqueous solution of sodium hydroxide (2.1 g, 53 mmol), and catalytic amounts of tetrabutylammonium iodide and benzyltriethylammonium bromide were added and stirred. The reaction mixture was extracted with ethyl acetate, the extract was dried, and the residue obtained by distilling off the solvent was purified by silica gel chromatography (developing solution n-hexane: ethyl acetate = 10: 1) to obtain the target compound. 2.6 g were obtained.

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.89 (d, J = 8.0H
z, 2H), 7.43 ~ 7.29 (m, 5H), 7.16 (d, J = 8.0Hz, 2H), 4.19 (s,
3H), 4.04 (ddd, J = 1.0,6.2,9.2Hz, 1H × 0.8), 3.79 (ddd, J =
1.8,2.6,8.6Hz, 1H × 0.2), 2.81 ~ 2.48 (m, 3H), 2.06 ~ 1.8
6 (m, 2H), 1.67 to 1.55 (m, 2H), 1.59 (s, 9H) Production Example 12 4- [2- (2,4-diamino-6,7-dihydro-7-phenylthio-5H-cyclopentane [D] pyrimidine-5
-Yl) ethyl] benzoic acid tertiary butyl ester

[0098]

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Using the tert-butyl 4- [2- (4-phenylthio-2-cyano-3-methoxy-2-cyclopenten-1-yl) ethyl] benzoate obtained in Production Example 11 as a raw material, Synthesized in the same manner as in Example 10.

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.92 to 7.88 (m, 2
H), 7.49-7.40 (m, 2H), 7.34-7.16 (m, 5H), 5.31 (brs, 2
H), 4.92 (brs, 2H), 4.45 (t, J = 8Hz, 1H × 0.5), 4.39 (dd, J = 4,
8Hz, 1H × 0.5), 2.97 ~ 2.28 (m, 5H), 1.70 ~ 1.55 (m, 2H),
1.59 (s, 9H) Example 1 Production of 4- [3- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] benzoic acid

[0101]

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The 4- [3- (2,4-
Diamino-6,7-dihydro-7-phenylthio-5H-
Cyclopenta [d] pyrimidin-5-yl) propyl]
Benzoic acid To a dichloromethane solution of tertiary butyl ester was added 3-chloroperbenzoic acid (650 mg,
3.7 mmol) was added slowly. After stirring for 15 minutes under ice cooling, an aqueous solution of sodium thiosulfate was added and stirred. After the reaction solution was extracted with dichloromethane, it was dehydrated and the solvent was distilled off. To the residue was added a toluene solution (50 ml) of trimethyl phosphite (2.6 ml, 22 mmol), and the mixture was refluxed for 30 minutes. The reaction solution was concentrated as it was, and columnar chromatography was used to remove phosphorus and sulfur-derived contaminants. To this was added a 1N hydrochloric acid-acetic acid solution (10 ml), and the mixture was stirred for 3 hours. The reaction solution was concentrated, and the residue was dried at 145 ° C. for 3 hours to obtain 400 mg of the desired product.

TLC (Merk Art. 5715) CHCl ₃ : CH ₃ OH: CH ₃ CO
_{2 H = 10: 1: 1} , Rf = 0.2 Example 2 N-{4-[3- (2,4 - diamino -6,7 - dihydrocyclopenta [d] pyrimidin-5-ylidene) propyl] benzoyl} -Production of L-glutamic acid diethyl ester

[0104]

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The 4- [3- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] benzoic acid (400 mg) obtained in Example 1 was added to N,
It was dissolved in N-dimethylformamide (4 ml), and glutamic acid diethyl ester hydrochloride (523 mg, 2.2 m
mol), diphenyl azide phosphate (0.5 ml, 2.3 mmol),
Triethylamine (0.8 ml, 5.7 mmol) was added. After stirring overnight at room temperature as it is, the solvent was distilled off and thin layer chromatography (developing solution: chloroform: methanol = 1: 1)
0: 1) to give 256 mg of the desired product.

¹ H-NMR (CDCl ₃ ) δ (ppm): 7.73 (d, J = 8 Hz, 2
H), 7.25 (d, J = 8Hz, 2H), 7.13 (d, J = 2Hz, 1H), 5.22 (tt, J = 2,7
Hz, 1H), 5.16 (brs, 2H), 5.05 (brs, 2H), 4.78 (dt, J = 5,8Hz, 1
H), 4.21 (dq, J = 1,7Hz, 2H), 4.09 (dq, J = 2,7Hz, 2H), 2.71〜
2.79 (m, 2H), 2.37 to 2.64 (m, 6H), 2.25 to 2.34 (m, 1H), 2.1
2 (dt, J = 8,16Hz, 1H), 1.28 (t, J = 7Hz, 3H), 1.20 (t, J = 7Hz, 3
H) Example 3 Preparation of N- {4- [3- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] benzoyl} -L-glutamic acid

[0107]

Embedded image

The N- {4- [3- (2,2) obtained in Example 2
4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) propyl] benzoyl] -L-
1.5 ml of a 1 N aqueous solution of sodium hydroxide for a tetrahydrofuran aqueous solution of glutamic acid diethyl ester
Was added and stirred at room temperature overnight. Crystals were precipitated by adding 1N hydrochloric acid to the reaction solution. This was collected by filtration and dried to obtain 140 mg of the desired product.

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.49 (d, J = 8H
z, 1H), 7.78 (d, J = 8Hz, 2H), 7.32 (d, J = 8Hz, 2H), 6.25 (brs, 2H
H), 6.01 (s, 2H), 5.51 (t, J = 8Hz, 1H), 4.35 (dt, J = 12,4Hz, 1
H), 2.74 (t, J = 8 Hz, 2H), 2.52 to 2.56 (m, 2H), 2.30 to 2.45 (m,
6H), 2.05 (dt, J = 12, 8 Hz, 1H), 1.92 (dt, J = 24, 8 Hz, 1H) TLC (Merk Art. 5715) CHCl ₃ : CH ₃ OH: CH ₃ CO ₂ H = 10: 3:
2, Rf = 0.34 Example 4 Production of 4- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl] benzoic acid

[0110]

Embedded image

4- [2- (2,4-) obtained in Production Example 12
Diamino-6,7-dihydro-7-phenylthio-5H-
Cyclopenta [d] pyrimidin-5-yl) ethyl] benzoic acid tertiary butyl ester was used as a starting material to synthesize the target compound in the same manner as in Example 1. TLC (Merk Art.5715) CHCl ₃ : CH ₃ OH: CH ₃ CO ₂ H = 10: 1:
1, Rf = 0.2 Example 5 N- {4- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl]
Production of benzoyl} -L-glutamic acid diethyl ester

[0112]

Embedded image

Using 4- [2- (2,4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl] benzoic acid obtained in Example 4 as a raw material, It was synthesized in the same manner as described above. TLC (Merk Art. 5715) CH ₂ Cl ₂ : C ₂ H ₅ OH = 10: 1, Rf = 0.5 Example 6 N- {4- [2- (2,4-diamino-6,7-dihydrocyclopentane) [D] pyrimidine-5-ylidene) ethyl]
Production of benzoyl｝ -L-glutamic acid

[0114]

Embedded image

The N- {4- [2- (2,2) obtained in Example 5
Example 3 Using 4-diamino-6,7-dihydrocyclopenta [d] pyrimidine-5-ylidene) ethyl] benzoyl} -L-glutamic acid diethyl ester as a raw material, Example 3
It was synthesized in the same manner as described above.

¹ H-NMR (DMSO-d ₆ ) δ (ppm): 8.45 (d, J = 8H
z, 1H), 7.79 (d, J = 8Hz, 2H), 7.34 (d, J = 8Hz, 2H), 6.24 (brs, 2
H), 5.99 (brs, 2H), 5.70 (t, J = 8Hz, 1H), 4.37 (dd, J = 8,12Hz,
1H), 3.47 (t, J = 8Hz, 2H), 2.35-2.65 (4H), 2.31 (t, J = 12Hz,
2H), 2.04 (dt, J = 8,12Hz, 1H), 1.92 (dt, J = 8,12Hz, 1H) MS (FAB) m / e: 426 (MH ⁺ )

────────────────────────────────────────────────── ─── Continued on the front page (51) Int.Cl. ⁷ Identification code FI C07D 409/12 C07D 409/12 417/12 417/12 (72) Inventor Kyosuke Kito 1-10-10 Tokodaidai, Tsukuba, Ibaraki 8 (58) Fields investigated (Int. Cl. ⁷ , DB name) C07D 239/70 A61K 31/517 C07D 403/12 C07D 405/12 C07D 409/12 C07D 417/12 CA (STN) REGISTRY (STN)

Claims (5)

(57) [Claims] 1. A pyrimidine ring-fused cyclopentylidene derivative represented by the general formula (1) or a pharmacologically acceptable salt thereof. Embedded image (Wherein, R ¹ represents a hydroxyl group or an amino group; R ² represents a hydrogen atom, an amino group or a methyl group; X is a divalent linear group composed of 1 to 4 atoms, and a substituent Y represents a phenylene group, a thiendiyl group, a furanyl group, a thiazolediyl group, an indolediyl group or an indolinediyl group.) 2. X is a divalent linear group consisting of 1 to 4 atoms selected from carbon, nitrogen, oxygen or sulfur, and is selected from a hydrocarbon group, a halogen atom and a substituted or unsubstituted amino group. The pyrimidine ring-fused cyclopentylidene or a pharmacologically acceptable salt thereof according to claim 1, which is a group which may have a selected substituent. 3. A compound of the general formula (2) (Wherein R ¹ , R ² , X and Y are as defined above) or a reactive derivative at the carboxyl group thereof, and a compound represented by the general formula (3): (Wherein R ³ and R ⁴ represent the same or different carboxyl-protecting groups) and are condensed to give a compound of the general formula (4) (Wherein, R ¹ , R ² , R ³ , R ⁴ , X and Y are as defined above), and a carboxylic acid ester represented by the formula:
2. The method for producing a pyrimidine ring-fused cyclopentylidene derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein the ester is hydrolyzed in the presence of an acid or an alkali. 4. A compound of the general formula (5) (In the formula, R ¹ , R ² , X and Y have the same meanings as those in claim 1. —COOR ⁵ represents a carboxyl group which may be esterified with a protecting group.) Or its salt. 5. An antitumor agent comprising the pyrimidine ring-fused cyclopentylidene derivative according to claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient.