JPS6120558B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the general formula This invention relates to a novel 5-fluorouracil derivative having the following. In the above formula, R ¹ and R ² are the same or different and represent a hydrogen atom or a β-D-glucopianacyl group. However, cases where R ¹ and R ² are the same and represent a hydrogen atom are excluded. To explain further, in the general formula (), R ¹ and R ² are the same formula or one of R ¹ and R ² represents a group having the above general formula (),
The other represents a hydrogen atom. The 5-fluorouracil derivative having the general formula () obtained by the present invention is a novel compound, and is a compound useful as a medicine that exhibits excellent anticancer activity. Conventionally, O-glycoside compounds in which a heterocyclic base and a saccharide are bonded via an oxygen atom are unstable to acids and alkalis. There have been no cases where the body has been isolated. The present inventors conducted extensive research on 5-fluorouracil derivatives with the aim of developing anticancer drugs with fewer side effects, and as a result, the O-glycoside compound of 5-fluorouracil having the general formula () above was found to be excellent. The present invention was completed based on the discovery that it exhibits anticancer activity and low toxicity. Specific compounds having the general formula () obtained by the present invention are described below. (1) O ² -(β-D-glucopyranosyl)-5-
Fluorouracil (2) O ⁴ -(β-D-glucopyranocil)-5-
Fluorouracil (3) O ² , O ⁴ -bis(β-D-glucopyranosyl)-5-fluorouracil The novel compound () according to the present invention can be produced by the method shown below. In the above formula, R ³ is an aliphatic acyl group such as formyl, acetyl, n-propionyl, n-butyryl, isobutyryl, or an aromatic acyl group such as benzoyl, p-nitrobenzoyl, p-methylbenzoyl, p-methoxybenzoyl. ^R4 represents an acyl group used as a protective group for a hydroxyl group of a sugar such as a nitro group, a halogen atom such as chlorine or bromine, an alkoxy group such as methoxy, or an alkylene dioxy group such as methylenedioxy in the aromatic ring. M
is the condensation of sugars with heterocyclic bases such as monovalent silver, monovalent and divalent mercury such as HgCl and HgBr, divalent copper such as CuCl and CuBr, and alkali metals such as sodium and potassium. represents a metal ion used in the reaction to produce an O-glycoside compound, and X represents a halogen atom such as chlorine or bromine. The manufacturing method generally involves condensing a 5-fluorouracil metal salt compound with a halogen sugar, and then carrying out an appropriate combination of reactions in which the pyrimidine nucleus and the protecting group for the hydroxyl group of the sugar are removed from the resulting O-glycoside compound. , Each step will be explained in detail below according to the above-mentioned reaction formula. The first step is a step of producing an O-glycoside compound having the general formula (), and is a step of producing an O-glycoside compound having the general formula ().
- This is a step in which a fluorouracil metal salt is subjected to a condensation reaction with a halogenopyranose derivative having the general formula (). In carrying out the reaction of this step, the condensation reaction is carried out by bringing the compound having the general formula () into contact with the compound having the general formula () in the presence of a solvent. The solvent used is not particularly limited as long as it does not participate in this reaction, but aromatic hydrocarbons such as toluene and xylene, fatty acid dialkylamides such as dimethylformamide and dimethylacetamide, and dialkyl sulfoxides such as dimethyl sulfoxide are used. , phosphoric acid amides such as hexamethylphosphoramide, nitroalkanes such as nitromethane and nitriles such as acetonitrile are preferred. There is no particular limitation on the reaction temperature, but it is usually carried out at 0 to 150°C, and generally at 100 to 150°C when using a nonpolar solvent.
It is preferable to carry out the reaction at 150°C, or around room temperature when a polar solvent is used. The reaction time varies mainly depending on the reaction temperature and the solvent used, but it is about 5
The duration ranges from minutes to 30 hours. In this reaction, instead of the metal salt having the general formula (), in the presence of a solvent, the general formula (In the formula, R ⁴ has the same meaning as described above.) Mercury cyanide, mercury oxide,
While adding silver carbonate, silver oxide, silver perchlorate, cupric carbonate, cadmium carbonate, etc. and heating, the general formula
It can also be carried out by adding a halogenopyranose derivative having (). After the reaction is completed, the target compound () of this step is collected from the reaction mixture according to a conventional method. For example, after separating insoluble materials from the reaction mixture, the solvent is distilled off under reduced pressure, and the residue is extracted using an organic solvent such as ethyl acetate. It can be obtained by washing the extract with water and drying it, distilling off the solvent from the extract, adding an organic solvent such as ethanol to the residue, allowing it to stand, and collecting the precipitated crystals. The second step is a step of producing a compound having the general formula (), in which the protecting group for the hydroxyl group in the pyrimidine core portion is removed from the compound having the general formula () by a reduction reaction. In carrying out the reaction in this step, the reaction is carried out by bringing the compound having the general formula () into contact with a reducing agent in the presence of a solvent. The reducing agent used is not particularly limited as long as it is generally used for removing protective groups such as benzyl groups, but hydrogen and catalytic reduction catalysts such as palladium-carbon are suitable. The solvent to be used is not particularly limited as long as it does not participate in this reaction and dissolves the raw material compound (), but alcohols such as methanol and ethanol, and ethers such as tetrahydrofuran and dioxane are suitable. The reaction temperature is not particularly limited, but it is usually suitably carried out around room temperature. The reaction time is usually such that the reaction is carried out until one equivalent of hydrogen is absorbed. After the reaction is completed, the target compound () of this step is collected from the reaction mixture according to a conventional method. For example, it can be obtained by separating insoluble matter from the reaction mixture, distilling off the solvent under reduced pressure, adding an organic solvent such as ethanol to the residue, allowing it to stand, and collecting the precipitated crystals. The third step is the general formula of the target compound of the present invention.
Has (). This is a process for producing O ² -(β-D-glucopyranosyl)-5-fluorouracil derivatives, in which the acyl protecting group of the hydroxyl group of the sugar moiety is removed from the compound having the general formula (). In carrying out the reaction of this step, the reaction is carried out by bringing the compound having the general formula () into contact with a base such as ammonia or an alkali metal alkoxide in the presence of a solvent. Bases used include ammonia; sodium methoxide, sodium ethoxide or potassium tert.
-Butoxide and the like are preferred. The solvent to be used is not particularly limited as long as it does not participate in this reaction, but alcohols such as methanol, ethanol, and tert-butanol, and combinations of these alcohols and ethers such as tetrahydrofuran and dioxane, methylene chloride, chloroform, etc. A mixed solvent with an organic solvent such as a halogenated hydrocarbon or an aromatic hydrocarbon such as benzene, toluene, or xylene is suitable, and it is preferable to carry out the reaction under anhydrous conditions. There is no particular limitation on the reaction temperature, but it is usually −
The reaction is carried out at a temperature of 30 to 50°C, and preferably at a temperature of 0°C to room temperature. The reaction time varies mainly depending on the reaction temperature, but is usually about 2 to 20 hours. After completion of the reaction, the target compound () of the present invention is collected from the reaction mixture according to a conventional method. For example, it can be obtained by distilling off the solvent from the reaction mixture under reduced pressure, dissolving the residue in a small amount of anhydrous methanol, adding chloroform, and collecting the precipitate. The fourth step is a step of producing a compound having the general formula (), and is a step of removing the acyl protecting group of the hydroxyl group of the sugar moiety from the compound having the general formula (). The reaction conditions and post-treatment method for the reaction in this step are the same as in the case of the reaction in the third step described above. The fifth step is the general formula of the target compound of the present invention.
O ² -(β-D-glucopyranosyl) with ()
This is a separate process for producing -5-fluorouracil derivatives, in which the protecting group for the hydroxyl group in the pyrimidine core is removed from the compound having the general formula () by a reduction reaction. The reaction conditions and post-treatment method for the reaction in this step are the same as in the case of the reaction in the second step described above. The sixth step is a step of producing an O-glycoside compound having the general formula (), and is a step of producing an O-glycoside compound having the general formula ().
- This is a step in which a fluorouracil metal salt is subjected to a condensation reaction with a halogenopyranose derivative having the general formula (). The reaction conditions for the reaction in this step are the same as those in the first step described above, but instead of the metal salt having the general formula (), mercuric cyanide is added to 5-fluorouracil in the presence of a solvent. While adding mercury oxide, silver carbonate, silver oxide, silver perchlorate, cupric carbonate, cadmium carbonate, etc. and heating, the general formula ()
It can also be carried out by a method of adding a halogenopyranose derivative having the following. After the reaction is completed, the target compound () of this step is collected from the reaction mixture using a conventional method and a post-treatment method similar to that of the first step described above. The seventh step is the general formula of the target compound of the present invention.
A process for producing an O ² , O ⁴ -bis(β-D-glucopyranosyl)-5-fluorouracil derivative having (XI), in which the acyl protecting group of the hydroxyl group of the sugar moiety is removed from the compound having the general formula (). It is. The reaction conditions and post-treatment method for the reaction in this step are the same as in the case of the reaction in the third step described above. The eighth step is a step of producing an O-glycoside compound having the general formula (), in which a 5-fluorouracil metal salt having the general formula (XII) is subjected to a condensation reaction with a halogenopyranose derivative having the general formula (). It is. The reaction conditions for the reaction in this step are the same as those in the first step described above, but in the presence of a solvent instead of the metal salt having the general formula (XII). (In the formula, R ⁴ has the same meaning as described above.) Mercury cyanide, mercury oxide,
While adding silver carbonate, silver oxide, silver perchlorate, cupric carbonate, cadmium carbonate, etc. and heating, the general formula
It can also be carried out by adding a halogenopyranose derivative having (). After the reaction is completed, the target compound () of this step is collected from the reaction mixture using a conventional method and a post-treatment method similar to that of the first step described above. The ninth step is a step of producing a compound having the general formula (), and is a step of removing the hydroxyl group of the pyrimidine core from the compound having the general formula () by a reduction reaction. The reaction conditions and post-treatment method for the reaction in this step are the same as in the case of the reaction in the second step described above. The 10th step is the general formula of the target compound of the present invention.
This is a process for producing an O ⁴ -(β-D-glucopyranosyl)-5-fluorouracil derivative having (), in which the acyl protecting group of the hydroxyl group of the sugar moiety is removed from the compound having the general formula (XI). The reaction conditions and post-treatment method for the reaction in this step are the same as in the case of the reaction in the third step described above. The 11th step is a step of producing a compound having the general formula (), and is a step of removing the acyl protecting group of the hydroxyl group of the sugar moiety from the compound having the general formula (). The reaction conditions and post-treatment method in this step are the same as those in the third step described above. The twelfth step is the general formula of the target compound of the present invention.
In a separate process for producing the O ⁴ -(β-D-glucopyranosyl)-5-fluorouracil derivative having the formula (), the protecting group for the hydroxyl group in the pyrimidine core is removed from the compound having the general formula () by a reduction reaction. It is a process. The reaction conditions and post-treatment method in this step are the same as those in the second step described above. The target compound obtained in each of the above steps can be further purified, if necessary, by conventional methods such as recrystallization, column chromatography, or reprecipitation. All of the 5-fluorouracil derivatives of the present invention having the general formula () have shown remarkable anticancer activity in pharmacological tests.Next, the results of pharmacological tests regarding anticancer effects will be exemplified.

【table】

[Table] Therefore, the compound having the above general formula () is useful as an anticancer agent for various cancer diseases. The administration form includes, for example, parenteral administration by intravenous injection, tablets, capsules, granules, powders, etc.
Oral administration such as syrup can be given. The dosage varies depending on symptoms, age, body weight, etc., but the usual daily dose for adults is 5 to 50 mg/Kg for intravenous injection, and the daily dose for oral administration. The dose is 200 to 2000 mg and can be administered in 1 to 4 divided doses. Next, the present invention will be explained in more detail with reference to manufacturing examples. Production example 1 O ² -(β-D-tetra-O-acetylglucopyranosyl)-O ⁴ -benzyl-5-fluorouracil O ⁴ -benzyl-5-fluorouracil silver salt 1.95
g (6.1 mmole) was suspended in 100 ml of anhydrous xylene (dehydrated with calcium hydride and then distilled), heated to reflux, distilled off about 10 ml to remove water azeotropically, and left to cool. Add 2.51 g (6.1 mmole) of 2,3,4,6-tetraacetyl-α-D-glycopyranosyl bromide and reflux for 5 minutes. After the reaction was completed, the precipitated insoluble materials were removed and the solvent was distilled off under reduced pressure. The resulting caramel-like residue was dissolved in 30 ml of ethyl acetate, and dissolved once with 20 ml of 5% sodium bicarbonate solution.
After washing twice with 20 ml of water, the aqueous layer was further extracted with 15 ml of ethyl acetate, the organic layers were combined and dried over anhydrous magnesium sulfate, and the caramel obtained by distilling off the bath agent under reduced pressure was mixed with 15 ml of ethanol. Dissolve and leave to obtain white crystals. These crystals were further recrystallized with a mixed solvent of 15 ml of ethanol and 4 ml of benzene, and the melting point was
1.986 g of the target compound having a temperature of 140-141°C is obtained. Furthermore, after concentrating the mother liquor to dryness, it is subjected to column chromatography using silica gel, and the fraction eluted with chloroform is separated and purified and crystallized to obtain 274 mg of the above-mentioned target compound. Elemental analysis value C ₂₅ H ₂₇ O ₁₁ N ₂ F Calculated value C, 54.55; H, 4.94; N, 5.09; F, 3.45 Actual value C, 54.34; H, 5.08; N, 5.04; F, 3.30 Ultraviolet absorption spectrum λ ^{0.1N-HClinMe
OH} _nax nm
(ε): 264 (5200); λ ^MeOH _nax nm(ε): 264
(5100);λ ^{0.1N-NaOHinMeOH} _nax nm
(ε)：265
(5500). Production example 2 O ² -(β-D-tetra-O-acetylglucopyranosyl)-5-fluorouracil After dissolving 550 mg (1 mmole) of O ² -(β-D-tetra-O-acetylglucopyranosyl)-O ⁴ -benzyl-5-fluorouracil in 30 ml of anhydrous methanol and purging the inside of the reduction reaction vessel with dry nitrogen gas,
Carefully add 100 mg of 10% palladium-carbon and absorb 24 ml of hydrogen gas at normal pressure while stirring at room temperature. After the reaction was completed, the catalyst was removed by purging with nitrogen gas again, and the liquid was concentrated to dryness under reduced pressure. The resulting crystalline residue was heated and dissolved in 45 ml of absolute ethanol and left in the refrigerator overnight to precipitate. When you take the crystal,
373 mg of the target compound is obtained as white crystals. When the melting point of this crystal was measured, it melted at 140-142℃, 197
Color decomposes at ~212℃. Elemental analysis value as C ₁₈ H ₂₁ C ₁₁ N ₂ F Calculated value C, 46.96; H, 4.60; N, 6.09; F, 4.13 Actual value C, 47.23; H, 4.90; N, 5.73; F, 3.95 Ultraviolet absorption Spectrum λ ^MeOH _nax nm(ε):
266
(6200) Production example 3 O ² -(β-D-glucopyranocil)-5-fluorouracil O ² -(β-D-tetra-O-acetylglucopyranosyl)-5-fluorouracil 460mg
Dissolve (1 mmole) in 25 ml of anhydrous methanol, saturate with dry ammonia gas under ice cooling, and leave for 4 hours, then leave in the refrigerator overnight. After the reaction is complete,
The solvent was distilled off under reduced pressure, the mixture was dissolved in a small amount of anhydrous methanol, and 10 ml of chloroform was added to remove the precipitated powder, yielding 307 mg of the target compound.
When the melting point of this powder is measured, it partially melts at 103-109°C and foams and decomposes at 116-119°C. Elemental analysis value C ₁₀ H ₁₃ O ₇ N ₂ F・CH ₃ OH ・NH ₃・H ₂ O Calculated value C, 36.77; H, 6.17; N, 11.70; F, 5.29 Actual value C, 36.0.; H, 5.73; N, 12.07; F, 5.38. Ultraviolet absorption spectrum λ0.02M−Shosphate buffer (PH6.86) max
nm(ε): 268 (7000) Production example 4 O ² , O ⁴ -bis(β-D-tetra-O-acetylglucopyranosyl)-5-fluorouracil Add 2,3 to 50ml of well-dried acetonitrile.
Dissolve 4.11 g (10 mmole) of 4,6-tetraacetyl-α-D-glucopyranosyl bromide,
- Add 2.17 g (10 mmole) of fluorouracil silver salt and stir at room temperature for 30 hours. After the reaction, the precipitated insoluble matter was removed, and the liquid was concentrated to dryness under reduced pressure.
3.93 g of colorless caramel are obtained. This caramel was subjected to column chromatography using silica gel, separated and purified using chloroform containing 1% methanol as a developing solvent, and the resulting fraction was crystallized from ethanol, resulting in white crystals with a melting point of 183-190°C. 1.342 g of the target compound was obtained. Elemental analysis value as C ₃₂ H ₃₉ O ₂₀ N ₂ F Calculated value C, 48.61; H, 4.97; N, 3.54; F, 2.40 Actual value C, 46.66; H, 5.08; N, 3.74; F, 2.24 Ultraviolet absorption Spectrum λ ^H+inMeOH _nax
nm:
265;λ ^MeOH _nax nm(ε):265(6800);λ
^OH-inMeOH _nax nm:267. Production example 5 O ² , O ⁴ -bis(β-D-glucopyranosyl)
-5-Fluorouracil O ² , O ⁴ -bis(λ-D-tetra-O-acetylglucopyranosyl)-5-fluorouracil
790 mg (1 mmole) was suspended in 50 ml of anhydrous methanol, saturated with dry ammonia gas while cooling with ice water, sealed with a stopper, and stirred overnight at 5°C. After the reaction, the solvent was distilled off under reduced pressure, and anhydrous methanol was added and distilled off, which was repeated twice. Dissolved in 5 ml of anhydrous methanol and added 40 ml of chloroform to collect the precipitated white precipitate, washed with chloroform, On drying, 320 mg of the target compound are obtained as a white powder with a melting point of 110-113° C. (decomposition). Elemental analysis value C ₁₆ H ₂₃ O ₁₂ N ₂ F ・CH ₃ OH・2H ₂ O Calculated value C, 39.08; H, 5.98; N, 5.36; F, 3.64 Actual value C, 39.28; H, 5.64; N, 5.60 ; F, 3.57. Ultraviolet absorption spectrum λ0.02N−Phosphate buffer (PH6.86) max
nm(ε): 266 (5800) Production example 6 O ⁴ -(β-D-tetra-O-acetylglucopyranosyl)-O ² -benzyl-5-fluorouracil 2-benzyl-5-fluorouracil silver salt 4.91
g (15 mmole) was suspended in 220 ml of dry xylene and heated to reflux. 20 ml of xylene was distilled off to remove water, and 2,3,4,6-tetraacetyl-
α-D-glucopyranosyl bromide 6.17g
(15 mmole) and heated under reflux for 5 minutes. After the reaction, the precipitated gray-yellow insoluble matter was removed, the solvent was concentrated to dryness under reduced pressure, and the residue was dissolved in ethyl acetate 100%.
After washing once with 70 ml of 5% sodium bicarbonate solution and twice with 70 ml of water, the aqueous layer was sequentially extracted with 50 ml of ethyl acetate, the organic layers were combined and dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure. The caramel-like substance obtained by distillation is dissolved in ethanol, isopropyl ether is added, the precipitated crystals are collected and dried, and the melting point
Target compound as white crystals with 95-96℃
Obtain 5.795g. Elemental analysis value as C ₂₅ H ₂₇ O ₁₁ N ₂ F Calculated value C, 54.54; H, 4.94; N, 5.09; F, 3.26 Actual value C, 54.33; H, 4.92; N, 4.70; F, 3.26. Ultraviolet absorption spectrum λ ^{0.1N-HClinMe
OH} _nax nm
(ε): 270 (7200); λ ^MeOH _nax nm(ε): 270
(7100);λ ^{0.1N-MaOHinMeOH} _nax
nm(ε)：270
(8300). Production example 7 O ⁴ -(β-D-tetra-O-acetylglucopyranosyl)-5-fluorouracil 5.795 g (10.5 mmole) of O ⁴ -(β-D-tetra-O-acetylglucopyranosyl)-O ² -benzyl-5-fluorouracil was dissolved in 130 ml of distilled tetrahydrofuran, and the inside of the reduction reaction vessel was replaced with nitrogen gas. 10% palladium-carbon 526mg
was carefully added and stirred at room temperature and pressure until 254 ml of hydrogen gas was absorbed, then palladium-carbon 52
After replacing mg with nitrogen gas, add it and absorb an additional 14 ml of hydrogen gas. After the reaction, the catalyst was removed by purging with nitrogen gas, and the liquid was concentrated to dryness under reduced pressure. Absolute ethanol was added to the obtained crystalline residue to collect the crystals, and the target compound with a melting point of 154-155°C was obtained. Obtain 4.318g. Elemental analysis value: Calculated value as C ₁₈ H ₂₁ O ₁₁ N ₂ F: C, 46.96; H, 4.60; N, 6.09; F, 4.13. Actual measurements: C, 46.87; H, 4.67; N, 5.93; F, 3.94. Ultraviolet absorption spectrum λ ^MeOH _nax nm(ε):
288
(4200) Production example 8 O ⁴ -(β-D-glucopyranocil)-5-fluorouracil O ⁴ -(β-D-tetra-O-acetylglucopyranosyl)-5-fluorouracil 2.3g
(5 mmole) was dissolved in 125 ml of anhydrous methanol, saturated with dry ammonia gas under ice cooling, and then
Stir for an hour and leave in the refrigerator overnight. After the reaction is completed, the solvent is distilled off under reduced pressure, and then anhydrous ethanol is added and distilled off. The gel-like substance produced is taken and dried, and the melting point
1.40 g of the target compound are obtained as a white powder having a temperature of 114-115°C. Elemental analysis value C ₁₀ H ₁₃ O ₇ N ₂ F ・1/2H ₂ O Calculated value C, 39.87; H, 4.68; N, 9.30; F, 6.31. Actual measurements: C, 39.36; H, 5.02; N, 9.31; F, 6.13. Ultraviolet absorption spectrum λ0.02M−Phosphate
buffer (PH6.86) max nm(ε): 286
(4600) Production example 9 O ² -benzyl-O ⁴ -(β-D-glucopyranosyl)-5-fluorouracil O ⁴ -(β-D-tetra-) obtained in Production Example 6
Dissolve 550 mg of O-acetylglucopyranosyl)-O ² -benzyl-5-fluorouracil in 100 ml of anhydrous methanol, saturate the solution with dry ammonia gas at room temperature, seal the solution, and stir for 2 hours. After the reaction is completed, the solvent is distilled off under reduced pressure to obtain 410 mg of caramel. This was subjected to thin layer chromatography using a preparative silica gel plate, separated and purified using chloroform containing 10% methanol as a developing solvent, and 310 mg of the target compound was obtained as a caramel, giving a single spot by thin layer chromatography. get. Production example 10 O ⁴ -(β-D-glucopyranosyl)-5-fluorouracil Dissolve 310 mg of O ² -benzyl-O ⁴ -(β-D-glucopyranosyl)-5-fluorouracil in 20 ml of absolute methanol, and add 50 mg of 10% palladium-carbon.
and stir while passing hydrogen gas under room temperature and normal pressure. After 5 minutes, the catalyst was removed and the solvent was distilled off under reduced pressure. The resulting caramel was dissolved in ethanol and allowed to stand, and the precipitated crystals were collected to obtain the desired compound as a white powder. The physical properties of this product are Production Example 8
matches what was obtained.

Claims (1)

[Claims] 1. General formula (In the formula, R ¹ and R ² are the same or different and represent a hydrogen atom or a β-D-glucopyranosyl group. However, this excludes cases where R ¹ and R ² are the same and represent a hydrogen atom.) Fluorouracil derivatives.