JPH024215B2 - - Google Patents

Description

[Detailed description of the invention]

<Industrial Application Field> The present invention relates to 4-substituted-5-alkylidene-2-cyclopentenones and a method for producing the same. More specifically, the present invention relates to novel 4-substituted-5-alkylidene-2-cyclopentenones having excellent anticancer activity and a method for producing the same. <Prior Art> Prostaglandin is a compound that has unique biological activities such as platelet aggregation inhibiting action and blood pressure lowering action, and is a useful natural product that has recently been used as a peripheral circulation treatment drug in the medical field.
Among prostaglandins, prostaglandin A is known to have a double bond in its cyclopentane ring. For example, prostaglandin A ₂ is expected to be a drug with a blood pressure lowering effect (EJCorey , J.Amer.Chem.
See Soc., 95, 6831 (1973). ). The present inventors synthesized a novel 7,8-dehydroprostaglandin A as a derivative of prostaglandin A and investigated the pharmacological effects of this compound, and found that it had excellent anticancer activity. Ta. The present invention was achieved as a result of extensive and intensive research based on this knowledge. <Object of the present invention> An object of the present invention is to provide a prostaglandin A derivative having excellent anticancer activity and relatively low prostaglandin-like physiological activity, and a method for producing the same. <Structure of the invention> The 4-substituted-5-alkylidene-2-cyclopentenone of the present invention has the following formula [] [In the formula, the symbol 〓 represents a single bond or a triple bond, and the symbol 〓 represents a single bond or a double bond. R ¹ represents an alkyl group having 3 to 5 carbon atoms substituted with -COOR ⁴ (where R ⁴ represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a monoequivalent cation). R ² represents an alkyl group having 1 to 7 carbon atoms or a cycloalkyl group having 5 carbon atoms. R ³ represents a hydrogen atom, a hydroxyl group or a t-butyldimethylsilyloxy group. however

[Formula] is

In the case of [expression]

【formula】 but

Except for the case of [formula]. ] It is expressed as . In the above formula [], the symbol 〓 represents a single bond or a triple bond. The symbol 〓 represents a single bond or a double bond. R ¹ represents a substituted alkyl group having 3 to 5 carbon atoms. Here, as the alkyl group having 3 to 5 carbon atoms,
n-propyl, iso-propyl, n-butyl, sec
-butyl, tert-butyl, n-pentyl and other linear or branched ones. Examples of the substituent for such a substituted alkyl group having 3 to 5 carbon atoms include -COOR ⁴ (where R ⁴ represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a cation of one equivalent). can. Examples of R ⁴ in -COOR ⁴ include a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, and a monoequivalent cation, and examples of the alkyl group having 1 to 10 carbon atoms include methyl, ethyl, n-propyl. , iso
-propyl, n-butyl, sec-butyl, tert-
Straight chain or branched ones such as butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl and n-decyl can be mentioned. As a cation equivalent, for example,
Ammonium cations such as NH ₄ ⁺ , tetramethylammonium, monomethylammonium, dimethylammonium, trimethylammonium, benzylammonium, phenethyl ammonium, morpholinium cation, monoethanolammonium, piberidinium cation; Na ⁺ , K ⁺ etc. Alkali metal cation; 1/2Ca ²⁺ , 1/
Examples include divalent or trivalent metal cations such ^as 2Mg ²⁺ , 1/2Zn ^{2+ ,} and 1/3Al 3+ . In the above formula [], R ² represents an unsubstituted alkyl group having 1 to 7 carbon atoms or an unsubstituted cycloalkyl group having 5 carbon atoms. Here, the number of unsubstituted carbon atoms is 1
-7 alkyl groups include, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl,
sec-butyl, t-butyl, n-pentyl, n-
Examples include hexyl, 2-methyl-1-hexyl, 2-methyl-2-hexyl, n-heptyl and the like. When R ² represents an unsubstituted cycloalkyl group having 5 carbon atoms, an example of the cycloalkyl group having 5 carbon atoms is cyclopentyl. In the above formula [], R ³ represents a hydrogen atom, a hydroxyl group, or a protected hydroxyl group (a group in which the hydrogen atom of a hydroxyl group is substituted with a protective group). As the protecting group for the protected hydroxyl group, tri(C ₁ -C ₇ ) hydrocarbon-
Mention may be made of silyl groups. As a tri(C ₁ -C ₇ ) hydrocarbon silyl group,
For example, trimethylsilyl, triethylsilyl, t
-tri(C ₁ -C ₄ ) such as butyldimethylsilyl group
Preferred examples include alkylsilyl and the like. The 4-substituted-5-alkylidene-2-cyclopentenones of the present invention are particularly suitable for the following formula ['] [In the formula, R ² and R ³ are as defined above, and R ⁴ represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a cation of one equivalent. ] Compounds represented by these are preferred. The above formula [′]
In the formula, R ⁴ represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a cation of one equivalent. These specific examples are the same as those described above. Also R ² ,
R ³ and the indications 〓 and 〓 are also the same as described above. Specific examples of such 4-substituted-5-alkylidene-2-cyclopentenones include (1) 4-butyl-5-(6-carboxyhexylidene)-2-cyclopentenone (2) 4-octyl -5-(6-carboxyhexylidene)-2-(cyclopentenone(3) 4-((1E)-1-octenyl)-5-(6-carboxyhexylidene)-2-cyclopentenone( 4) 4-Butyl-5-(6-carboxy-2-hexynylidene)-2-cyclopentenone (5) 4-Butyl-5-[2-(3-carboxypropylthio)ethylidene] 2-cyclopentenone ( 6) Methyl ester of (1) to (5) (7) Ethyl ester of (1) to (5) (8) Sodium salt of (1) to (5) (9) Ammonium of (1) to (5) Salt (10) 4-(3-hydroxy-5-methyl-(E)-
Examples include 1-nonenyl)-5-(6-carboxyhexylidene)-2-cyclopentenone. The 4-substituted-5-alkylidene-2-cyclopentenones of the present invention have the following formula [] [In the formula, the expressions 〓, 〓, R ¹ , R ² and R ³ are the same as defined above. ] 4-substituted-5-alkylidene-3- represented by
It is produced by subjecting hydroxycyclopentanones to a dehydration reaction, and then subjecting them to deprotection, hydrolysis, and salt-forming reactions as necessary. In the above formula [], R ¹ is substituted with 3 to 3 carbon atoms
5 represents an alkyl group, and the same ones as those listed in the above formula [] are exemplified. R ² represents an unsubstituted alkyl group having 1 to 7 carbon atoms or an unsubstituted cycloalkyl group having 5 carbon atoms, and the same alkyl groups as mentioned above are exemplified. The indications 〓 and 〓 are also the same as described above. 4-substituted-5-alkylidene- of the above formula []
3-Hydroxycyclopentanones are produced by a synthetic method represented by the following synthetic route. That is, it can be obtained by reacting the corresponding 5-hydroxymethylcyclopentanone derivative with methanesulfonyl chloride in the presence of a basic compound, and then removing the protective group. 4-substituted-5-alkylidene- of the above formula []
Examples of dehydrating agents used when subjecting 3-hydroxycyclopentanones to dehydration reactions include inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, and phosphoric acid; acetic acid, propionic acid, oxalic acid, and citric acid. , organic carboxylic acids such as maleic acid; methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid,
Examples include organic sulfonic acids such as p-toluenesulfonic acid, among which inorganic and organic carboxylic acids are preferred. The amount of such dehydrating agent used is 4-substituted-5
-Alkylidene-3-hydroxycyclopentanones preferably use 0.5 to 100 equivalents, particularly preferably 1 to 50 equivalents. Reaction solvents include ethers such as tetrahydrofuran, dioxane, dimethoxyethane, and diethyl ether; alcohols such as methanol and ethanol; dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, acetonitrile, water, etc., or any combination thereof. Use in combination. The reaction temperature is preferably in the range of 0 to 80°C, particularly preferably in the range of 10 to 50°C. The reaction time varies depending on the raw material compound, dehydrating agent, and reaction solvent used, but is usually carried out in a range of 10 minutes to 10 days, preferably in a range of 20 minutes to 5 days. In the dehydration reaction, instead of using 4-substituted-5-alkylidene-3-hydroxycyclopentanones of the above formula [] as the raw material compound, the hydroxyl group at the 3-position was protected with, for example, a t-butyldimethylsilyl group. 4-substituted-5-alkylidene-3
-Using hydroxycyclopentanones, the 3-position is deprotected by performing a deprotection reaction with a deprotection aid such as acetic acid, followed by a dehydration reaction to obtain the target compound. You can also do that. After the dehydration reaction, the target compound can be isolated and purified by means such as silica gel column chromatography, silica gel thin layer chromatography, florisil column chromatography, and high performance liquid chromatography. When the target compound has a protecting group in its molecule, it can be subjected to a deprotection reaction. When the protecting group is a group that forms an acetal bond together with the oxygen atom of the hydroxyl group, the protective group can be removed using acetic acid, a pyridinium salt of p-toluenesulfonic acid, or a cation exchange resin as a catalyst, such as water or tetrahydrofuran. , ethyl ether, dioxane, acetone, acetonitrile, etc. as the reaction solvent. The reaction is usually carried out at a temperature range of -78°C to +30°C for about 10 minutes to 3 days. In addition, when the protecting group is a tri(C ₁ -C ₇ ) hydrocarbon-silyl group, the same method is used in the reaction solvent as described above in the presence of, for example, acetic acid, tetrabutylammonium fluoride, cesium fluoride, etc. carried out for a similar period of time at temperature. In addition, when the protecting group is an acyl group, for example, an aqueous solution of caustic soda, caustic potash, calcium hydroxide or a water-
Hydrolysis can be carried out in an alcohol mixed solution, or in a methanol or ethanol solution containing sodium methoxide, potassium methoxide, or sodium ethoxide. If the target compound has an ester group, it can be subjected to hydrolysis, for example, using an enzyme such as lipase in water or a solvent containing water at -10°C to +
It is carried out at a temperature range of 60°C for about 10 minutes to 24 hours. When the target compound has a carboxyl group in its molecule, it can then be further subjected to a salt-forming reaction if necessary to obtain the corresponding carboxylate. The salt-forming reaction is known per se, and is carried out in a conventional manner using a basic compound such as sodium hydroxide, potassium hydroxide, sodium carbonate, etc., or ammonia, trimethylamine, monoethanolamine, morpholine, etc., in an approximately equal amount to a carboxylic acid. This is done by causing a sum reaction. In this way, 4-substituted-5-alkylidene-2-cyclopentenones of the above formula [] are produced. <Effects of the Invention> 4-Substituted-5-alkylindene-2- of the present invention
Cyclopentenones exhibit strong anticancer effects at extremely low concentrations, especially against L1210 leukemia cells.
It is extremely useful as an anticancer agent. It is also useful as a compound with antiviral activity. The 4-substituted-5-alkylidene-2-cyclopentenones of the present invention can be administered orally or parenterally, such as intrarectally, subcutaneously, intramuscularly, intravenously. For oral administration, solid or liquid preparations can be provided. Examples of solid preparations include tablets, pills, powders, and granules. In such solid formulations, one or more
-Substituted-5-alkylidene-2-cyclopentenones are mixed with, for example, sodium bicarbonate, calcium carbonate, potato starch, sucrose, mannitol, carboxymethylcellulose, and the like. Preparation operations are carried out according to conventional methods. The solid preparation may also contain lubricants such as calcium stearate, magnesium stearate, and glycerin, sweeteners, stabilizers, and preservatives. Liquid preparations for oral administration include, for example, emulsions, solutions, suspensions, syrups, and elixirs. Also wetting agents, suspension aids,
Sweeteners, flavoring agents, aromatics, stabilizers, etc. can be included. Liquid preparations can also be placed in capsules made of absorbable material such as gelatin. For rectal administration, conventional suppositories such as gelatin soft capsules are used. For preparations for parenteral administration other than rectal administration,
Examples include preparations for subcutaneous, intramuscular, and intravenous injection in the form of sterile aqueous or nonaqueous solutions, suspensions, and emulsions. Examples of non-aqueous solutions and suspensions include propylene glycol, polyethylene glycol, olive oil, and oleic acid. Such preparations also contain preservatives,
Emulsifiers, dispersants, stabilizers, etc. can be contained. Furthermore, these injectable preparations can be sterilized by appropriate treatment such as filtration through a bacteria retention filter, addition of a sterilizing agent, or irradiation. The 4-substituted-5-alkylidene-2-cyclopentenones of the present invention can also be used by forming an inclusion compound with α, β or γ-cyclodextrin. The dosage of the 4-substituted-5-alkylidene-2-cyclopentenones of the present invention varies depending on the condition, age, sex, body weight, administration route, etc. of the subject receiving administration, but is usually about 1 μg to 100 mg/Kg. -Can be administered in amounts per body weight/day. Such dosage is 1 per day
It can also be administered once or several times, for example, in 2 to 6 divided doses. As detailed above, the present invention provides 4-substituted-5-alkylidene-2-cyclopentenones that are extremely useful in the treatment of cancer patients, a method for producing the same, and an anticancer agent containing the same as an active ingredient. Ru. The present invention will be explained in more detail below with reference to Examples. Example 1 4-Butyl-5-(6-methoxycarbonylhexylidene)-3-hydroxycyclopentanone (81 mg, 0.27 mmol) was added to a mixed solvent of acetic acid (2 ml), tetrahydrofuran (1 ml), and water (1 ml). The mixture was dissolved and heated and stirred at 60°C for 7 hours and at 90°C for 50 hours.
After the reaction was completed, the reaction solution was concentrated under reduced pressure, neutralized with an aqueous sodium hydrogen carbonate solution, and then extracted with ethyl acetate.
The separated organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration and concentration, it was purified by silica gel column chromatography (10 g of silica gel; hexane: ethyl acetate = 6:1),
4-Butyl-5-(6-methoxycarbonylhexylidene)-2-cyclopentenone (Compound A)
(61 mg, 0.22 mmol, 81%) was obtained. IR (liquid film; cm ^-1 ); 1739, 1703, 1656, 1580 NMR (CDCl ₃ ; δ (ppm)); 0.89 (3H, t), 1.0 to 2.0 (12H, m), 2.0 to 2.6
(4H, m), 3.3-3.8 (1H, m), 3.67 (3H, S),
6.35 (1H, dd, J=6.0, 2.0Hz), 6.56 (1H,
t), 7.59 (1H, dd, J = 6.0, 3.0Hz) Example 2 4-octyl-5-(6-carboxyhexylidene) 3-hydroxycyclopentanone (169 mg,
0.5 mmol) was dissolved in tetrahydrofuran (3 ml), 2 ml of 0.5N hydrochloric acid was added, and the mixture was stirred at room temperature for 4 days.
Saturated brine was added and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration and concentration, separation and purification using preparative cylinder thin layer chromatography resulted in 4-octyl-5-
(6-carboxyhexylidene)-2-cyclopentenone (112 mg, 0.35 mmol, 70%) was obtained. IR (CHCl ₃ solution; cm ^-1 ); 3650-2400, 1700, 1645, 1580 NMR (CDCl ₃ δ (ppm)); 0.89 (3H, t), 1.0-2.0 (20H, m), 2.0-2.6
(4H, m), 3.3-3.8 (1H, m), 6.33 (1H, dd,
J=6,2Hz), 6.60 (1H, t), 7.53 (1H, dd,
J=6.3Hz), 11.5 (1H) Example 3 4-Butyl-5-(6-methoxycarbonyl-
2-Hexynylidene)-3-hydroxycyclopentanone (29 mg, 0.1 mmol) was dissolved in acetic acid (1 ml), tetrahydrofuran (0.5 ml), and water (0.5 ml) in the same manner as in Example 1, and heated at 50°C for 18 hours. Stirred.
After treatment and separation in the same manner as in Example 1, 4-butyl-
5-(6-methoxycarbonyl-2-hexynylidene)-2-cyclopentenone (13 mg,
0.047 mmol, 47%) was obtained. IR (liquid film; cm ^-1 ); 2190, 1730, 1680, 1620, 1565 NMR (CDCl ₃ ; δ (ppm)); 0.88 (3H, t), 1.0-2.2 (8H, m), 2.2-2.8
(4H, m), 3.3-3.8 (1H, m), 3.67 (3H, s),
6.40 (1H, dd, J=6.0, 2.0Hz), 6.55 (1H),
7.53 (1H, dd, J = 6.0, 3.0Hz) Example 4 3-t-Butydimethylsilyloxy-4-(3
-t-Butyldimethylsilyloxy-3-cyclopentyl-1-propenyl)-5-(6-methoxycarbonyl-2-hexynylidene)cyclopentanone (48 mg, 81 μmol), acetic acid (1.5 ml), tetrahydrofuran (1 ml), water ( 1 ml) of the mixed solvent was dissolved and stirred at 40°C for 35 hours. [A separate synthetic sample shows that 3-hydroxy-4-(3-hydroxy-3-cyclopentyl-1-propenyl)-5-(6-methoxycarbonyl-2-hexynylidene)cyclopentanone is produced in the early stage of the reaction. This was confirmed by the agreement with thin layer chromatography. ] After the reaction was completed, the reaction solution was concentrated under reduced pressure, neutralized by adding sodium hydrogen carbonate, and extracted with ethyl acetate. The separated organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After filtration and concentration, silica gel column chromatography (silica gel 3
g; Hexane:ethyl acetate=3:1) to purify 4-(3-hydroxy-3-cyclopentyl-1-propenyl)-5-(6-methoxycarbonyl-2-hexynylidene)-2-cyclopene Tenon (compound B) (19 mg, 55 μmol, 69%) was obtained. IR (liquid film; cm ^-1 ) 3450, 2190, 1728, 1683, 1620, 1565, 743 NMR (CDCl ₃ , δ (ppm)); 0.9 to 2.3 (m, 11H), 2.3 to 2.8 (m, 5H) , 3.70
(s, 3H), 3.45-4.35 (m, 2H), 5.5-5.9 (m,
2H), 6.3-6.7 (m, 2H), 7.4-7.7 (m, 1H) Example 5 7(E)-7,8-dehydro-17(S), 20-dimethylprostagra in the same manner as Example 4 Jin E ₁
7(E)-7,8-dehydro-17(S),20-dimethyl prostaglandin from methyl ester-11,15-bis-t-butyldimethylsilyl ether
A ₁ methyl ester (compound C) was obtained. nmr (δppm in CDCl ₃ ); 0.8 to 1.0 (6H, m), 1.0 to 2.0 (15H, m), 2.0 to
2.6 (4H, m), 3.65 (H, s), 4.15 (m, 3H),
5.4 (1H, dd, J = 15.6Hz), 5.75 (1H, dd, J
= 15.6Hz), 6.35 (1H, dd, J = 6.2Hz), 6.6
(1H, t, J=6Hz), 7.4 (1H, dd, J=6,
2Hz) Example 6 Measurement of anticancer activity Cancer cells were 10%, calf serum (fetal calf
The cells were grown in RPMI 1640 medium containing serum. 4-Butyl-5-(6-methoxycarbonylhexylidene)-2-cyclopentenone is dissolved in 99.5% ethanol and added to the medium to a final concentration of 0.1% ethanol or less before use. did. The medium containing 4-butyl-5-(6-methoxycarbonylhexylidene)-2-cyclopentenone (Compound A) was Mysopore filtered. 0.1% ethanol was used as a control. L1210 cancer cells were inoculated into the medium at a concentration of 1×10 ⁵ cells/ml.
It was grown for 4 days. The number of viable cells was determined by trypan blue staining. The results are shown in the table.

[Table] Therefore, the inhibitory concentration of the above compound for inhibiting the proliferation of L1210 cancer cells is IC ₅₀ =0.3 μg/ml. Similarly, the IC ₅₀ of compounds B and C is 0.3, respectively.
It showed a strong inhibitory activity of 0.2 μg/ml. Example 7 Manufacture of tablets Tablets each having the following composition were manufactured. 4-Butyl 5-(6-methoxycarbonylhexylidene)-2-cyclopentenone 10mg Lactose 250mg Potato starch 70mg Polyvinylpyrrolidone 10mg Magnesium stearate 5mg 4-Butyl-5-(6-methoxy obtained in Example 1) Carbonylhexylidene)-2-cyclopentenone, lactose, and ginger starch were mixed, evenly wetted with a 20% ethanol solution of polyvinylpyrrolidone, and then passed through a sieve. The granules thus obtained were mixed with magnesium stearate and compressed into tablets.

Claims (1)

[Claims] 1. The following formula [] [In the formula, the symbol 〓 represents a single bond or a triple bond, and the symbol 〓 represents a single bond or a double bond. R ¹ represents an alkyl group having 3 to 5 carbon atoms substituted with -COOR ⁴ (where R ⁴ represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a monoequivalent cation). R ² represents an alkyl group having 1 to 7 carbon atoms or a cycloalkyl group having 5 carbon atoms. R ³ represents a hydrogen atom, a hydroxyl group or a t-butyldimethylsilyloxy group. However, this excludes cases where [expression] is [expression] and [expression] is [expression]. ] 4-substituted-5-alkylidene-2- represented by
Cyclopentenones. 2 The following formula [] [In the formula, the symbol 〓 represents a single bond or a triple bond, and the symbol 〓 represents a single bond or a double bond. R ¹ represents an alkyl group having 3 to 5 carbon atoms substituted with -COOR ⁴ (where R ⁴ represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a monoequivalent cation). R ² represents an alkyl group having 1 to 7 carbon atoms or a cycloalkyl group having 5 carbon atoms. R ³ represents a hydrogen atom, a hydroxyl group or a t-butyldimethylsilyloxy group. However, this excludes cases where [expression] is [expression] and [expression] is [expression]. ] 4-substituted-5-alkylidene-2- represented by
The following formula [] is characterized in that cigaropentenones are subjected to a dehydration reaction, and then subjected to deprotection, hydrolysis, and salt-forming reactions as necessary. [In the formula, the expressions 〓, 〓, R ¹ , R ² and R ³ are the same as defined above. However, this excludes cases where [expression] is [expression] and [expression] is [expression]. ] 4-substituted-5-alkylidene-2- represented by
Method for producing cyclopentenones. 3. 4-Substituted-5- according to claim 2, wherein the dehydration reaction is carried out with an organic carboxylic acid or an inorganic acid.
A method for producing alkylidene-2-cyclopentenones.