JPH0210154B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to 7,8-dehydroprostaglandin A, a method for producing the same, and an anticancer agent containing the same as an active ingredient. More specifically, the present invention provides novel 7,8-dehydroprostaglandin A compounds such as 7,8-dehydroprostaglandin A ₁ having excellent anticancer effects, a method for producing the same, and an anticancer agent containing the same as an active ingredient. Regarding. Prostaglandin is a compound that has unique biological activities such as inhibiting platelet aggregation and lowering blood pressure, 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 (EJCorey5 , J. Amer.
See Chem.Soc., 95, 6831 (1973). ). The present invention has synthesized novel 7,8-dehydroprostaglandin A having a double bond at the 7-position as a derivative of prostaglandin A, and investigated the pharmacological effects of such a compound. The present invention was achieved by discovering that novel 7,8-dehydroprostaglandin A is useful as an anticancer agent. That is, the present invention is based on the following formula [] [In the formula, R represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a monoequivalent cation, and A represents -CH ₂ -
Represents CH ₂ −. ] 7,8-dehydroprostaglandin A represented by the following, a method for producing the same, and an anticancer agent containing the same as an active ingredient. In the 7,8-dehydroprostaglandin A of the above formula [], R is a hydrogen atom, and has 1 carbon atom.
~10 alkyl groups or one equivalent of cation.
Here, examples of the alkyl group having 1 to 10 carbon atoms include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl,
n-pentyl, n-hexyl, n-heptyl, n
-octyl, n-nonyl, n-decyl and the like. Among these, methyl and ethyl are particularly preferred. One equivalent cation is, for example, an alkali metal cation such as Na ⁺ or K ⁺ ; 1/2
Divalent or trivalent metal cations such as Ca ²⁺ , 1/2Mg ²⁺ , 1/2Zn ²⁺ , 1/3Al ³⁺ ; NH ₄ ⁺ , tetramethylammonium, monomethylammonium, dimethylammonium, trimethylammonium,
Examples include ammonium cations such as benzylammonium, phenethyl ammonium, monoethanol ammonium, and piperidinium cations. Among these, sodium ions and ammonium ions are particularly preferred. In the above formula [], A represents _-CH2 - _CH2- . Also shown in the above formula []

[Formula] represents the case where cis (7Z) or trans (7E) or cis and trans coexist in any ratio with respect to the oxo group (C=0) at the 9-position. Specific examples of such 7,8-dehydroprostaglandeins A include (1) (7E)-7,8-dehydroprostaglandein A ₁ , (2) (7Z)-7,8-dehydroprostaglandein A ₁ , (3) (7E)-7,8-dehydroprostaglandein A ₂ , (4) (7Z)-7,8-dehydroprostaglandein A ₂ , (5) (7E)-7,8- Dehydroprostaglandein A ₁ methyl ester, (6) (7Z)-7,8-dehydroprostaglandein A ₁ methyl ester, (7) (7E)-7,8-dehydroprostaglandein A ₂ methyl ester, ( 8) (7Z)-7,8-dehydroprostaglandein A ₂ methyl ester, (9) 5,6-dehydro form of the compound of (3), (10) 5,6-dehydro form of the compound of (4) , (11) Sodium salt of the compound of (1), (12) Sodium salt of the compound of (2), (13) Sodium salt of the compound of (3), (14) Sodium salt of the compound of (4), ( 15) Ammonium salt of the compound of (1), (16) Ammonium salt of the compound of (2), (17) Ammonium salt of the compound of (3), (18) Ammonium salt of the compound of (9), etc. It will be done. The 7,8-dehydroprostaglandin A of the present invention has the following formula [] [In the formula, R' represents a hydrogen atom or an alkyl group having 1 to 10 carbon atoms, and A represents _-CH2 - _CH2- . ] It is produced by subjecting 7,8-dehydroprostaglandin E represented by the following to a dehydration reaction, and then subjecting it to a salt-forming reaction as necessary. In the above formula [], R' represents a hydrogen atom or an alkyl group having 1 to 10 carbon atoms. Examples of the alkyl group having 1 to 10 carbon atoms include the same alkyl groups as described above. A and display

[Formula] is also the same as described above. The 7,8-dehydroprostaglandin E of the above formula [] is expressed by the following formula: 7-Hydroxyprostaglandin E class As expressed by It will be done. Examples of dehydrating agents used when subjecting 7,8-dehydroprostaglandin E of the above formula [] to a dehydration reaction include inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, and phosphoric acid; acetic acid; , propionic acid, oxalic acid, citric acid, maleic acid, and other organic carboxylic acids; methanesulfonic acid, ethanesulfonic acid,
Examples include organic sulfonic acids such as benzenesulfonic acid and p-toluenesulfonic acid, among which inorganic acids are preferred. The amount of such dehydrating agent used is 7,8
-Dehydroprostaglandin E is preferably used in an amount of 0.5 to 50 equivalents, particularly preferably 1 to 10 equivalents. Reaction solvents include ethers such as tetrahydrofuran, dioxane, dimethoxyethane, and diethyl ether; alcohols such as methanol and ethanol; and dimethyl sulfoxide, dimethylformamide, hexamethylphosphoric triamide, acetonitrile, water, etc., singly or in any combination thereof. Used in combination. The reaction temperature is preferably in the range from 0 to 80°C, particularly preferably from 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 7,8-dehydroprostaglandin E of the above formula [] as a raw material compound, 7,8-dehydroprostaglandin E of the above formula [] whose hydroxyl groups at positions 11 and 15 are protected with, for example, t-butyldimethylsilyl group, is used. Using 8-dehydroprostaglandein E, it is dehydrated with a dehydrating agent such as hydrofluoric acid to deprotect its 11th and 15th positions, and then the dehydration reaction is continued. The target compound can also be obtained by 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 thus obtained has a carboxyl group at the 1-position, 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 consists of carboxylic acid and approximately the same amount of sodium hydroxide,
Basic compounds such as potassium hydroxide and sodium carbonate, or ammonia, trimethylamine,
This is done by neutralizing with monoethanolamine, morpholine, etc. using a conventional method. In this way, 7,8-dehydroprostaglandin A of the above formula [] is produced. The 7,8-dehydroprostaglandin A of the present invention particularly reaches L1210 leukemia cells and exhibits a strong anticancer effect at extremely low concentrations, making it extremely useful as an anticancer agent. The 7,8-dehydroprostaglandin A of the present invention can be administered orally, rectally, subcutaneously, intramuscularly,
It can be administered parenterally, such as intravenously. For oral administration, solid or liquid preparations can be provided. Examples of solid preparations include tablets, pills,
There are powders, granules, etc. In such solid formulations, one or more of the 7,8-dehydroprostaglandin A's may be combined with, for example, sodium bicarbonate, calcium carbonate, potato starch,
It is mixed with sucrose, mannitol, carboxymethyl cellulose, etc. 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 solvents and suspending agents include propylene glycol, polyethylene glycol, olive oil, and injectable organic esters such as ethyl oleate. Such preparations also contain preservatives, emulsifiers,
It can contain a dispersant, a stabilizer, etc. 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 7,8-dehydroprostaglandin A of the present invention can also be used by forming an inclusion compound with α, β or γ-cyclodextrin or methylated cyclodextrin. The dosage of 7,8-dehydroprostaglandin A 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-body weight/day. can be administered in amounts. Such a dose may be administered once or in several divided doses, for example 2 to 6 doses. As detailed above, the present invention provides 7,8-dehydroprostaglandin A which is extremely useful for the treatment of cancer patients, a method for producing the same, and a cancer drug containing the same as an active ingredient. The present invention will be explained in more detail below with reference to Examples. Example 1 (7E)-7,8-dehydroprostaglandin
Dissolve 160mg of E1 in 3ml of tetrahydrofuran,
2 ml of 0.5N hydrochloric acid was added and stirred at room temperature for 4 days. Saturated brine was added and the mixture was extracted with ethyl acetate. The organic layers were combined, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration and concentration, the mixture was separated and purified using silica gel TLC to obtain 95 mg (7E)-7,8-dehydroprostaglandin A ₁ (yield 63%). TLC: Rf0.58 (developing solvent; hexane: acetone =
1:2) IR (CHCl ₃ solvent): 3650-2400, 1698, 1646, 967
cm ^-1 NMR: δCDCl ₃ 0.88 (t, 3H, J = 6.0Hz), 1.0~
2.0 (m, 14H), 2.0~2.8 (m, 4H), 3.4~4.6
(m, 4H), 5.1~5.9 (m, 2H), 6.34 (dd, 1H,
J = 6.0, 2.4Hz), 6.61 (dd, 1H, J = 7.8, 1.5
Hz), 7.36 (dd, 1H, J=6.0, 2.7Hz) Example 2 (7E)-7,8-dehydroprostaglandin
22 mg of E1,11,15-bis-t-butyldimethylsilyl ether was dissolved in a solution of 1 ml of acetonitrile and 50 µ of 47% hydrofluoric acid and stirred at room temperature for 20 minutes. After adding a saturated aqueous sodium hydrogen carbonate solution, the pH was adjusted to 1 using oxalic acid. Extraction was performed with ethyl acetate, and the organic layers were combined, washed with saturated brine, and dried over anhydrous magnesium sulfate. After filtration and concentration, the mixture was separated and purified by silica gel TLC to obtain 4.7 mg (yield 37%) of (7E)-7,8-dehydroprostaglandin A ₁ . Example 3 Measurement of anticancer activity Cancer cells were 10%
The cells were grown in RPMI 1640 medium containing serum. 7,8-dehydroprostaglandin A ₁
was dissolved in 99.5% ethanol and added to the medium to a final concentration of 0.1% ethanol or less before use. The medium containing 7,8-dehydroprostaglandin A ₁ 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 and grown for 4 days. The number of viable cells was determined by trypan blue staining. The results are shown in the table.

[Table] Example 4 Manufacture of tablets Tablets each having the following composition were manufactured. (7E)-7,8-dehydroprostaglandein A ₁ 10mg Lactose 250mg Dial starch 70mg Polyvinylpyrrolidone 10mg Magnesium stearate 5mg (7E)-7,8-dehydroprostaglandein A ₁ obtained in Example 1, lactose and ginger starch were mixed, evenly moistened 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. Example 5 Synthesis of (7E)-7,8-dehydro PGA ₁ methyl ester and 12-ep(7E)-7,8-dehydro PGA ₁ methyl ester; (7E)-7,8 dehydro PGA ₁ methyl ester
1.0 g of a mixture of 11,15-bis(t-butyldimethylsilyl)ether and 15-Epi-Ent-(7E)-7,8-dehydroPGE ₁ methyl 11,15-bis(t-butyldimethylsilyl)ether. The mixture was suspended in a 2:1:1 solution of acetic acid:tetrahydrofuran:water, and stirred at 50°C for 14 hours and at 60°C for 3 hours. After concentration, water and sodium carbonate were added to neutralize, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and then dried over anhydrous magnesium sulfate. After filtration and concentration, silica gel column chromatography (200
g, hexane:ethyl acetate=1:1), 12
-Epi-(7E)-7,8-dehydroPGA ₁ methyl ester 184 mg (31% yield) and (7E)-7,8
-dehydroPGA ₁ methyl ester 184 mg (yield 31
%) was obtained. Spectral data of 12-Epi-(7E)-7,8-dehydroPGA ₁ methyl ester TLC: Rf = 0.57 (hexane: ethyl acetate = 1:
3) IR: (liquid film) 3560, 1738, 1700, 1648, 1577cm ^-1 NMR: δ ^CDCl3 0.90 (brt, 3H) 1.0-2.0 (m, 14H),
2.0~2.7 (m, 5H), 3.66 (S, 3H), 3.75~4.45
(m, 2H), 5.45 (dd, 1H, J=15.5, 6.5Hz),
5.71 (dd, 1H, J=15.5, 6.0Hz), 6.30 (dd,
1H, J=6.0, 2.0Hz), 6.63(brt, 1H, J=8.0
Hz), 7.41 (dd, 1H, J = 6.0, 3.0Hz) Spectral data of (7E)-7,8-dehydroPGA ₁ methyl ester TLC: Rf = 0.51 (hexane: ethyl acetate = 1:
3) IR: (liquid film) 3450, 1739, 1701, 1648, 1578 cm ^-1 NMR: δ ^CDCl3 0.90 (brt, 3H) 1.0-2.0 (m, 14H),
2.0~2.8 (m, 5H), 3.66 (S, 3H), 3.7~4.5
(m, 2H), 5.46 (dd, 1H, J=15.5, 6.5Hz),
5.71 (dd, 1H, J=15.5, 6.0Hz), 6.34 (dd,
1H, J=6.0, 2.0Hz), 6.63(brt, 1H, J=8.0
Hz), 7.41 = (dd, 1H, J = 6.0, 3.0Hz) Example 6 Toxicity Data (1) Measurement of LD ₅₀ value The test drug was administered (intravenous injection) to mice, and the LD ₅₀ value was observed.

[Table] (2) Continuous administration test When the test drug was administered intraperitoneally to male ICR mice (6 weeks old) for 10 days, 25.0mg/Kg/day was administered.
20% mortality at 50.0mg/Kg/Day, 100% at 50.0mg/Kg/Day
Died. Test drug;

Claims (1)

[Claims] 1. The following formula [] [In the formula, R represents a hydrogen atom, an alkyl group having 1 to 10 carbon atoms, or a monoequivalent cation, and A represents -CH ₂ -
Represents CH ₂ −. ] 7,8 dehydroprostaglandin A represented by: 2 The following formula [] [In the formula, R' represents a hydrogen atom or an alkyl group having 1 to 10 carbon atoms, and A represents _-CH2 - _CH2- . ] The following formula [] is characterized in that 7,8-dehydroprostaglandin E represented by is subjected to a dehydration reaction, and then subjected to a salt production reaction as necessary. [In the formula, the definitions of A and R are the same as above. ] A method for producing 7,8-dehydroprostaglandin A represented by: 3. The method for producing 7,8-dehydroprostaglandin A according to claim 2, wherein the dehydration reaction is carried out using an inorganic acid. 4. An anticancer agent containing 7,8-dehydroprostaglandin A represented by the above formula [] as an active ingredient.