DK155331B - 5-FLUORURACIL DERIVATIVES OR ACID ADDITION SALTS THEREOF USED AS INTERMEDIATES IN THE PREPARATION OF ANTITUMORALLY ACTIVE 5-FLOURURACIL COMPOUNDS - Google Patents

Description

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The invention relates to novel 5-fluorouracil derivatives or acid addition salts thereof for use as intermediates in the preparation of novel 5-fluorouracil compounds of the general formula (I) as set forth in the preamble of the claim. These compounds and their beneficial biological activity are described in more detail in Danish Patent Application No. 4781/77.

Certain 5-fluorouracil derivatives are known as antitumor agents. Eg. 5-fluorouracil itself (hereinafter referred to as "5-FU") and N 2 - (2-tetrahydrofuryl) -5-fluorouracil (hereinafter referred to as "FT-207") have been reported to show good antitumor activity . [R. Duschinsky et al., J. Amer. Chem. Soc. 79, 4559 (1957) and S.A. Hiller et al., Doc. Akad. Nauk. (USSR), 176, 332 (1967)]. However, there is a need for an agent that has as good or better antitumor activity than 5-FU and 0FT-207, but with fewer side effects and low toxicity.

This is achieved with the 5-fluorouracil derivatives of general formula (I) which can be prepared by a compound of formula: OR8 wherein R7 is benzyl optionally having a nitro or halogen substituent in the phenyl ring, or 8-halo-ethyl , and R8 represents a metal-containing ion, is reacted with a 2

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compound of the formula: R9 - X (III) wherein X is halogen and R9 is one of the glycoside residues R'0.CH2 r4'oco H / - ° \ HA- \ / h \ or / H Λ yoR1 ι / ττ l \ 9Rt Ψ vr

0RT5 * æf ^ t 0R, fsssaT3 «H

H R3 'H R3 wherein R' means an aliphatic acyl group having 2-4 carbon atoms or an aromatic acyl group having 7-11 carbon atoms, R3 means -OR 'where R' has the foregoing meaning, or benzyloxycarbonylamino, optionally has a nitro or halogen substituent in the phenyl ring, or β-haloethoxycarbonylamino, and R 4 'means straight or branched alkyl of 1-8 carbon atoms, cycloalkyl of 5-7 ring carbon atoms, aralkyl of 7-12 carbon atoms or aryl of 6-10 carbon atoms wherein the aromatic ring in the aralkyl or aryl group optionally has a substituent selected from alkyl of 1-4 carbon atoms, alkoxy of 1-4 carbon atoms and halogen to form a compound of formula: f, 3

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wherein R7 and R9 are as defined above and the compound of formula (IV) is optionally subjected to reduction to remove the hydroxy protecting group R7 and, if desired, to reduction of an amino protecting benzyloxycarbonyl or β-haloethoxycarbonyl.

Q I

nyl group in RJ and / or hydrolysis to remove the hydroxy protecting groups R 'and the carboxyl protecting group R4' to form a compound of formula (I), and if desired the compound of formula (I) formed is transferred into an acid addition salt thereof.

In the glycoside residues R9 in the reactant of formula (III), R 'is usually the same acyl group as represented by R in formula (I), R 1 when a protected amino group, usually the same protecting amino group as defined by R 3 in formula (I), and R 4 is generally the same group as represented by R 4 in formula (I).

When R 7 in formulas (II) and (IV) means a benzyl group having a nitro or halogen substituent in the phenylrin gene, it is e.g. o-nitrobenzyl, m-nitrobenzyl, p-nitrobenzyl, o-chlorobenzyl, m-chlorobenzyl, p-chlorobenzyl, m-fluorobenzyl or p-fluorobenzyl; and when it means a ph-haloethyl group, it is e.g. 2,2,2-trichloroethyl, 2,2,2-tribromethyl, 2,2-dichlorobenzyl or 2,2-dibromo-ethyl.

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When the group R means an aliphatic acyl group of 2-4 carbon atoms, it is e.g. acetyl, n-propionyl, n-butyryl or isobutyryl, and when it represents an aromatic acyl group of 7-11 carbon atoms, it is e.g. benzoyl, p-nitrobenzoyl, p-methylbenzoyl, p-methoxybenzoyl or naphthyl.

When an amino group is protected by a benzyl-oxycarbonyl group having a nitro or halogen substituent in the phenyl ring, the protecting group is e.g. o-nitrobenzyloxycarbonyl, m-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, o-chlorobenzyloxycarbonyl, m-chlorobenzyloxycarbonyl, p-chlorobenzyloxycarbonyl, o-bromobenzyloxycarbonyl, o-fluorobenzyloxycarbonyl, m-nitrobenzyloxycarbonyl, o-fluorobenzyloxycarbonyl, and when it is an amino group protected with a β-haloethoxycarbonyl group, the protecting group is e.g.

2,2,2-trichloroethoxycarbonyl, 2,2,2-tribromoethoxycarbonyl, 2,2-dibromoethoxycarbonyl or 2,2-dichloroethoxycarbonyl.

R 2 represents hydrogen, a straight or branched alkyl group having 1-8 carbon atoms, e.g. methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-pentyl, isopentyl, n-hexyl, sec.-hexyl, n-heptyl, o-ethyl or 2-ethylhexyl, a cycloalkyl group having 5 7 ring carbon atoms, e.g. cyclopentyl, cyclohexyl or cycloheptyl, an aralkyl group having 7-12 carbon atoms, the aromatic ring of which may be unsubstituted or having a substituent selected from alkyl of 1-4 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl or isobutyl), alkoxy of 1-4 carbon atoms (e.g. methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy or isobutoxy) and halogen (e.g., chlorine, bromine or fluoro), e.g. ben- 5

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zyl or phenethyl, or an aryl group of 6-10 carbon atoms whose aromatic ring may be unsubstituted or have a substituent selected from alkyl of 1-4 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl or isobutyl), alkoxy of 1-4 carbon atoms (e.g., methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy or isobutoxy) and halogen (e.g., chlorine, bromine or fluoro), e.g. phenyl.

The compounds of the above formula (IV) and their hydrolysis products, and such reducing products thereof, wherein only the optional protecting amino group

-5 I

R 2 is reduced, constituting the intermediates of the invention, which are characterized in that they have the general formula: OR 2 / γ wherein R 2 and R 7 are as previously defined, or are acid addition salts of such compounds wherein R 1 is amino.

Preferred examples of such compounds which can be prepared by the process described are given below. The numbers given to the connections in the following list are used to identify these connections in the following.

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1. 1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -β-D-glucopyranuronic acid.

2. Methyl 1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -β-D-glu-copyranuronate.

3. Ethyl 1- O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -S-6-glu-copyranuronate.

4. n-Propyl-1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -O-D-glucopyranuronate.

5. Isopropyl-1- (5-fluoro-1H-2-oxopyroidin-4-yl) -6-D-glucopyranuronate.

6. n-Butyl-1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -β-D-glucopyranuronate.

7. Isobutyl-1- (5-fluoro-1H-2-oxopyrimidin-4-yl) - (J-D-glucopyranuronate.

8. n-Pentyl-1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -β-D-glucopyranuronate.

9. n-Hexyl-1- (5-fluoro-1H-2-oxopyrimidin-4-yl) -3-D-glucopyranuronate.

10. 04- (8-D-gluopyranosyl) -5-fluorouracil.

11. O4- (2-Amino-2-deoxy-8-D-glucopyranosyl) -5-fluorouracil. 1 04- (3,4,6-tri-O-acetyl-2-amino-2-deoxy-8-D-glucopyranosyl) -5-fluorouracil.

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13. Isopentyl-1- (5-fluoro-1H-oxopyrimidin-4-yl) -B-D-glucopyranuronate.

14. Cyclohexyl-1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -3- D -glucopyranuronate.

15. O4- (2,3,4,6-tetra-O-acetyl- (5-D-glucopyranosyl) -5-fluorouracil.

In carrying out the process described, a compound of general formula (II) is reacted with a compound of general formula (III) to give a compound of formula (IV). This compound of formula (IV) is then subjected in any order to reduction to removal of the hydroxy protecting group and, if desired, reduction to removal of an amino protecting benzyloxycarbonyl or β-haloethoxycarbonyl group in R 3 'and / or hydrolysis to removal of the hydroxy protecting groups R 'and the carboxyl protecting group R4' to give the desired 5-fluorouracil derivative of formula (I).

In the formula OR8 1 J (D) 8

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optionally substituted benzyl or β-haloethyl ethyl as previously mentioned and R® a metal-containing ion. The metal-containing ion may be any monovalent ion containing a metal which may itself be monovalent or may be polyvalent. When the metal is monovalent, the metal-containing ion preferably consists of metal alone, e.g. a silver ion, a mercury (I) ion, Hg +, or an alkali metal ion, e.g. a sodium or potassium ion. Where the metal is polyvalent, the metal-containing ion contains the metal and one or more anionic ligands such that the metal-containing ion is itself monovalent; Examples include HgCl + and HgBr +. If the metal ion is polyvalent, the compound of formula (II) may also contain more than one 5-fluorouracil derivative residue, and the metal-containing ion is then considered to be composed of the metal and all but one of the residues mentioned.

Instead of adding the metal salt of formula (II) to the reaction mixture itself, the metal salt can be screened in situ from the corresponding free 5-fluorouracil derivative.

Eg. For example, the reaction can be carried out by adding e.g. mercury (II) cyanide to a solution of the corresponding free 5-fluorouracil derivative and then add the compound of formula (III).

The compound of formula (III) is an α-glycosyl halide, wherein the halogen atom is preferably chlorine or bromine. When this α-glycosyl halide reacts with the compound of formula (II), it gives the corresponding β-glycoside of the 5-fluorouracil derivative.

The reaction between compounds (II) and (III) is preferably carried out in the presence of a solvent. Any solvent can be used, provided it has no 9

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detrimental effect on the reaction. Examples of suitable solvents are aromatic hydrocarbons, e.g. toluene or xylene, dialkylamides of aliphatic acids, e.g. Ν, Ν-dimethylformamide or Ν, Ν-dimethylacetamide, dialkyl sulfoxides, e.g. dimethyl sulfoxide, phosphoric acid amides, e.g. hexamethylphosphoric acid diamide, nitroalkanes, e.g. nitromethane, and nitriles, e.g. of acetonitrile-Roll. Although there is no particular restriction on the reaction temperature, the reaction is generally carried out. o equality at a temperature of 0-150 C; where a non-polar solvent is used, the preferred reaction temperature is 100-150 ° C; where a polar solvent is used, it is preferred at room temperature.

The time required for the reaction will depend mainly on the reaction temperature and the solvent used, but will generally range from 5 minutes to 20 hours.

Upon completion of the reaction, the resulting compound of formula (IV) can be separated from the reaction mixture by conventional methods. Eg. after filtration of insoluble substances from the reaction mixture, the solvent can be removed by distillation under reduced pressure and then the residue is extracted with an organic solvent such as chloroform. The extract can then be washed with water and dried, after which the solvent is removed by distillation from the extract. An organic liquid such as ethanol can be added to the residue thus obtained and the mixture is allowed to stand in the cold, e.g. in a refrigerator to give the desired compound (IV) as crystals.

The compounds of general formula (IV) are useful as intermediates in the synthesis of antitumor agents with the 10

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of general formula (I) and, as indicated above, can be readily converted to such anti-tumor agents by reduction and optionally hydrolysis which may be carried out in any order.

The hydroxy protecting group in the pyrimidine moiety and / or any amino protecting group in the glycoside moiety can be removed by reduction by contacting the compound of formula (IV) with a reducing agent, preferably in the presence of a solvent. For the best results, the nature of the reducing agent used must be selected taking into account the nature of the protecting group to be removed. Where the hydroxy protecting group is a substituted or unsubstituted benzyl group, or the amino protecting group is a substituted or unsubstituted benzyloxycarbonyl group, the reducing agent is preferably hydrogen and a catalyst such as palladium on coal. Where the hydroxy protecting group is a β-haloethyl group or the amino protecting group is a β-haloethoxycarbonyl group, the reducing agent is preferably metallic zinc with an acid (eg acetic acid) or an alcohol (eg methanol or ethanol). The nature of the solvent used is not critical, as long as it has no adverse effect on the reaction. Suitable solvents include alcohols such as methanol or ethanol, and ethers such as tetrahydrofuran and dioxane. The reaction temperature is also not critical, and for convenience it is therefore preferred to carry out the reaction at about room temperature. The time required for the reaction will vary mainly depending on the nature of the reducing agent, but will generally be 5-60 minutes.

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Upon completion of the reaction, the product can be recovered by conventional methods. Eg. For example, after filtering off insoluble matter from the reaction mixture, the solvent can be removed by distillation under reduced pressure, and an organic liquid such as ethanol can be added to the residue, whereupon the mixture is allowed to obtain the desired product in crystalline form.

The hydrolysis reaction, which can be carried out before or after the reduction, can be carried out by contacting the compound of formula (IV) or the product obtained by reducing the compound of formula (IV) with a base in the presence of a solvent to remove acyl acid. the group represented by R '. The base used is preferably ammonia or an alkali metal alkoxide, e.g. sodium methoxide, sodium ethoxide or potassium t-butoxide.

Any solvent which does not damage the reaction may be used, although it is preferred to use an alcohol such as methanol, ethanol or t-butanol, or a mixture of one or more of these alcohols with one or more other organic solvents such as ethers ( e.g., tetrahydrofuran or dioxane) or halogenated hydrocarbons (e.g., methylene chloride or chloroform).

The reaction is preferably carried out under anhydrous conditions. There is no particular limit to the reaction temperature, and temperatures in the range of -30 to + 50 ° C, and most preferably from 0 ° C to about room temperature, give good results. The reaction time will vary, mainly depending on the reaction temperature, but will generally be 2-20 hours.

Upon completion of the reaction, the product can be recovered from the reaction mixture by conventional means.

Eg. For example, the solvent can be evaporated from the reaction mixture

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The residue is reduced under reduced pressure and the residue is dissolved in a small amount of anhydrous methanol, after which the addition of chloroform will result in crystals which can be filtered.

The hydrolysis of compound (IV) or the product obtained by reducing compound (IV) wherein R 4 'is an alkyl group, a cycloalkyl group, an aralkyl group or an aryl group, to the compound of formula (I) wherein R 4 is hydrogen is preferably carried out by about pH 11 using ammonium hydroxide in the presence of a solvent. The nature of the solvent is not limited, but does not adversely affect the reaction, and in general it is preferred to use a mixture of an alcohol (e.g. methanol or ethanol) with water. Also, there is no particular restriction on the reaction temperature, and in general it is preferred to carry out the reaction at a temperature between -10 and + 50 ° C, and for convenience, the reaction is preferably carried out at about room temperature. The time required for the reaction will depend mainly on the reaction temperature, but will generally be 3-24 hours.

Upon completion of the reaction, the product can be recovered by conventional means. Eg. For example, the solvent can be removed from the reaction mixture by distillation under reduced pressure and the residue then purified by fractionation by preparative thin layer chromatography on silica gel. The obtained fraction containing the desired compound can be extracted with an organic solvent such as methanol and the solvent then removed from the extract by distillation. Addition of an organic liquid such as ethanol to the residue and standing the mixture in the cold, e.g. in a refrigerator, give 13

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is the desired product in the form of crystals.

It is preferable to perform the hydrolysis of the acyl group (R *) prior to the reduction.

5-fluorouracil derivatives of formula (I) wherein R 3 represents an amino group can be converted to pharmaceutically acceptable acid addition salts by conventional means. Examples of suitable salts include the acid addition salts of mineral acids, e.g. hydrochloric acid, hydrobromic acid, sulfuric or phosphoric acid, or of organic acids, e.g. oxalic, succinic, maleic or citric.

The invention is further elucidated by the following preparation which illustrates the preparation of a starting material for the intermediates of the invention and by the following examples which illustrate the preparation of the intermediates of the invention and their use in the preparation of the antitumorally active compounds of formula (I).

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ISLAND

several other pesticide-active compounds and, if desired, one or more tolerant pesticide-acceptable thinners or carriers, surfactants and common excipients as previously described. Examples of other active compounds which may be included or used in conjunction with the herbicidal compositions of the invention include herbicides for e.g. to increase the spectrum ie the amount of the weed species that is being controlled, e.g. alachlor 10 15 20 25 1 35 15

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EXAMPLE 1

Methyl 1-O- (2-benzyloxy-5-fluoropyrimidin-4-yl) -2,3,4-tri-O-acetyl-g-D-glucopyranuronate ¢ 61) 5

cmcg T h ifi

/ 1V

in OCOCSi3> CHsCOO ^ / l 00C.Cfj3 7.9 g (24.2 ramol) of the silver salt of O ml of the solvent was removed by distillation using a water trap to remove water in the suspension. The remaining mixture was then allowed to cool. To the suspension was then added 9.1 g (24.2 mmol) of methyl 2,3,4-tri-O-acetyl-1-bromo-deoxy-α-D-glucopyranuroate, and the mixture was heated at reflux for 10 minutes. 5 minutes. The mixture was then allowed to cool and insoluble matter was filtered off. The solvent was removed by distillation under reduced pressure and the residue dissolved in chloroform. The chloroform solution was washed successively with 5 wt% aqueous sodium hydrogen carbonate solution and 20 wt% aqueous sodium of 16

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the chloroform phases were distilled off the solvent to give a caramel-like residue. Ethanol was added to this residue, and the mixture was allowed to stand overnight at 4 ° C to obtain 7.3 g of the desired methyl-10-acetyl-BD-glucopyranuronate in the form of crystals, which melted at 120 ° C. 121 C.

Element Analysis;

Calcd for C24H25O11N2F: C 53.73 - H 4.70 - N 5.22 - F 3.54 Found: C 53.94 - H 4.56 - N 5.19 - F 3.48.

Ultraviolet Absorption Spectrum: xmax (° 1N HCl, CH 3 OH) 271 nm (ε, 8000) * max (CH OH) 271 nm (ε, 7300) * max (° f NaN, CH 3 OH) 270 nm (e, 9000).

EXAMPLE 2

Methyl 1-Q- (2-benzyloxy-5-fluoropyrimidin-4-yl) -g-D-glucopyranuronate.

CH3O.CO4

Jkj (oh)

1 OH

17

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To 6.06 g (11.3 mmol) of methyl 1-O- (2-benzyloxy-5-fluoro-pyrimidin-4-yl) -2,3,4-tri-O-acetyl-8-D-glucopyranuronate (prepared as described in Example 1), 70 ml of a mixture of anhydrous methanol and methylene chloride were added in a 3: 2 volume ratio. After cooling the mixture with ice water, 6.6 ml of 1N methanolic sodium methoxide solution was added and the mixture was immediately stirred. The mixture was then stirred for an additional 40 πύο nuts at 0 C. After completion of the reaction, 2.5 ml of 1N hydrochloric acid was added to neutralize the mixture and the solvent was removed by distillation. The residue was dissolved in chloroform and the chloroform solution was then washed with water and dried over anhydrous magnesium sulfate. The solvent was removed from the dried solution by distillation to give 4.46 g of a caramel-like residue. This residue was subjected to column chromatography on 100 g of silica gel using chloroform containing 2.5% by volume of methanol. The eluate was collected and the solvent removed by distillation. Ethanol was added to the residue and then allowed to stand. The precipitated crystals were recovered by filtration to give 1.78 g of the desired methyl-1- (2-benzyl-oxy-5-fluoropyrimidin-4-yl) - (JD-glucopyranuronate in the form of white crystals melting at 159-160 C. An additional 540 mg of secondary crystals were obtained from the mother liquor.

Element Analysis:

Calcd for C 18 H 19 ° 8 N 2 F: C 52.68 - H 4.67 - N 6.83 - F 4.63 Found: C 53.05 - H 4.50 - N 6.93 - F 4.41.

m 18

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Ultraviolet absorption spectrum:

XmaxiOfiN HCl) 269 nm (ε, 7100)

Λmax (H °) 270 nm (ε, 7300) * max (° 1 NaOH) 269 nm (ε, 7700).

EXAMPLE 3

Methyl-l-O- (5-fluoro-lH-2-oxopyrimidine-4-yl) -B-D-glucopyranuronate.

EH3MQ γΝ

V-J

kJ / 1

· * CH

2.03 g (4.95 mmol) of methyl 1-O- (2-benzyloxy-5-fluoropyrimidin-4-yl) - β-D-glucopyranuronate were dissolved in 100 ml of anhydrous methanol. The atmosphere in the reaction vessel containing the solution was washed with nitrogen and then 400 mg of 10 wt% palladium on charcoal was added. The atmosphere was again replaced with fresh nitrogen gas and then hydrogen gas was introduced with stirring at atmospheric pressure for 15 minutes. After completion of the catalytic reduction, insoluble matter was filtered off and the solvent was then removed by distillation under reduced pressure. Ethanol was added to the precipitated crystals and the mixture was then filtered to give 19

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1.529 g of the desired methyl-1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -β-D-glucopyranuronate (Compound 2) were obtained as white crystals, melting at 122 124 ° C.

Element Analysis:

Calcd. For C 11 H 13 O 8 N 2 F · 0.5 H 2 O: C 40.13 - H 4.29 - N 8.51 - F 5.77 Found: C 39.97 - H 4.64 - N 8.44 - F 5.47.

Ultraviolet Absorption Spectrum: (H9O) 288 nm (ε, 5700).

The crystals obtained were recrystallized from a mixture of anhydrous methanol and ethanol in the volume ratio of 1: 1 to give white needles melting at 127-128 ° C.

EXAMPLE 4 1-O- (2-Benxyloxy-5-fluoropyrimidin-4-yl) -β-D-glucopyranuronic acid.

WH20 h \ By COOH 1 r l-oo (CH 1 /.

'QH

i 20

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1.025 g (2.5 mmol) of methyl-O- (2-benxyloxy-5-fluoropyrimidin-4-yl) - (3-D-glucopyranuronate (obtained as described in Example 2)) was dissolved in 23 ml of a mixture of methanol, water and concentrated ammonia water in a volume ratio of 3: 3.9: 0.1 The solution was then stirred at room temperature for 24 hours, then the solvent was removed by distillation under reduced pressure to give 1.2 g of a caramel-like residue. This residue was subjected to preparative thin layer chromatography on silica gel using chloroform containing 35% by volume of methanol as a developing solvent, a fraction containing the desired compound was extracted with methanol and the solvent was evaporated from the extract. to stand in the refrigerator overnight. The precipitated crystals were collected by filtration to give 724.5 mg of the desired product as white crystals which melted at 129-130 ° C.

Element Analysis:

Calcd. For C

Ultraviolet Absorption Spectrum: xmax (PH 4.01) 269 nm (ε, 8800)

Xmax (H2 °) 269 nm (ε, 8400)

Λmax (pH 9.18) 269 nm (ε, 8500).

EXAMPLE 5

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21 1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) -g-D-glucopyranuronic acid.

H

° YK

COOH I F

A

MH. / • \! /

1 OH

362.6 mg of 1-O- (2-benzyloxy-5-fluoropyrimidin-4-yl) -phi-D-glucopyranuronic acid were dissolved in 20 ml of anhydrous methanol. The atmosphere in the reaction vessel was replaced with nitrogen gas and then 72.5 mg of 10 wt% palladium on charcoal was added to the solution. The mixture was stirred at room temperature while introducing hydrogen gas into it. After completion of the reaction, the catalyst was filtered off from the reaction mixture and the solvent was removed by distillation under reduced pressure to give a crystalline powder. This powder was suspended in a small volume of ethanol, recovered by filtration and washed to give 254 mg of the desired 1-O- (5-fluoro-1H-2-oxopyrimidin-4-yl) - (JD-gluco -pyranuronic acid (compound 1) as a white powder.

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Elementarily see:

Calcd for CyoHnOg + F: C 39.2 - H 3.62 - N 9.15 - F 6.20

Found C 38.95 - H 3.51 - N 9.01 - F 6.31.

Ultraviolet Absorption Spectrum: ^ max®2®) 288 nm.

EXAMPLE 6

Methyl-l-O- (5-fluoro-lH-2-oxopyrimidine-4-yl) -2,3,4-tri-O-aeefcyl-B-D-glucopyranuronate.

IL · X.

CH3O.CO I f 1-ti. DOCCH3 1.2 g methyl-1- (2-benzyloxy-5-fluoropyrimidin-4-yl) -2,3,4-tri-O-acetyl-8-D-glucopyranuronate (prepared as described in Example 1) was dissolved in 120 ml of anhydrous methanol. The atmosphere in the reaction vessel was replaced with nitrogen and then 240 mg of 10 wt% of palladium-on-charcoal was added to the solution. The atmosphere in the reaction vessel was again replaced by fresh nitrogen gas, and 23

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the mixture was then stirred for 15 minutes while introducing hydrogen gas under atmospheric pressure. After completion of the reaction, insolubles were removed from the reaction mixture and then the solvent was removed by distillation under reduced pressure.

Ethanol was added to the residue and the precipitated crystals were recovered by filtration to give 891 mg of the desired methyl-1- (5-fluoro-1H-2-oxopyrimidin-4-yl) -2,3, 4-Tri-O-Acetyl - $ - D-glucopyranuroate in the form of white crystals melting at 115-117 ° C. Additional 106 mg of secondary crystals were obtained from the mother liquor.

Element Analysis:

Calcd for C 17 H 19 O 11 N 2 F 2 H 2 O: C 43.97 - H 4.56 - N 6.03 - F 4.10 Found: C 44.70 - H 4.55 - N 5.89 - F 4.26.

Ultraviolet absorption spectrum:

Xmax (H2 O) 288 nm (£ 4800).

EXAMPLE 7 24

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Isopropyl-O- (2-benzyloxy-5-fluoro-pyrimidin-4-yl) -2,3,4-tri-O-acetyl-B-D-glucopyranuronate.

"> VAF

P ^ CHOXQ I

} 00¾ 3.27 g (10 mmol) of the silver salt of O 2 -benzyl-5-fluorouracil was suspended in 150 ml of dry xylene. To remove the water, about 40 ml of the solvent was distilled off under atmospheric pressure and then the remaining suspension was allowed to cool. To the suspension was added 4.25 g of isopropyl-2,3,4-tri-O-acetyl-1-bromo-1-deoxy-α-D-glucopyranuronate and the mixture was heated under reflux for 5 minutes. After completion of the reaction, the mixture was allowed to cool and insoluble matter was then filtered off. The solvent was removed by distillation under reduced pressure leaving a residue which was dissolved in 100 ml of chloroform. The chloroform solution was washed successively with 50 ml of 5% aqueous solution of sodium bicarbonate and 50 ml of 20% aqueous solution of sodium chloride. The aqueous wash solutions were extracted again with 50 ml of chloro-25

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form, and all the chloroform phases were combined and dried. The solvent was removed by distillation to give a colorless caramel-like residue. This residue was dissolved in ethanol and the solution was allowed to stand overnight in the refrigerator. The precipitated crystals were collected by filtration to give 4.97 g of the desired isopropyl-1- (2-benzyloxy-5-fluoro-pyrimidin-4-yl) -2,3,4-tr acetyl-pD-glucopyran uronate in the form of white crystals melting at 138-139 ° C.

Element Analysis:

Calcd for C 26 H 29 ° 11 N 2 F: C 55.32 - H 5.18 - N 5.00 - F 3.37 Found: C 55.08 - H 5.21 - N 4.93 - F 3.12.

Ultraviolet Absorption Spectrum: * max (50% ° 1N HCl 50% Ch 3 H) 270 nm (ε, 10500)

Xmax (50% CH 3 OH, 50% H 2 O) 270 nm (e, 10300)

Xmax (50% O, 1N NaOH, 50% CH 3 OH) 270 nm (ε, 10800).

EXAMPLE 8 26

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Isopropyl-1-0- (2-benzyloxy-5-fluoro-pyrimidin-4-yl) -β-Ρ-qlucopyranuronat ♦

C8H5CH2O

L X

F

tCH3l2CHO.CC

Q ''

! OH

100 ml of anhydrous methanol was charged into a three-neck flask followed by 0.366 g of metallic sodium. The mixture was cooled to -30 while moisture was kept out. To the methanolic solution of sodium methoxide was added 4.50 g of isopropyl-1- (2-benzyloxy-5-fluoro-pyrimiclin-4-yl) -2,3,4-tri-O-acetyl-BD-glucopyranuronate, and the mixture was stirred for 6 hours. After completion of the reaction, the reaction mixture was neutralized by the addition of acetic acid and then evaporated to dryness. The residue was dissolved in 100 ml of ethyl acetate and the ethyl acetate solution was washed twice with a 2.5% by weight aqueous solution of sodium bicarbonate and with saturated aqueous sodium chloride solution. The organic phase was dried over magnesium sulfate and then dissolved

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The residue was removed by distillation under reduced pressure leaving a residue which was recrystallized from methanol to give the desired isopropyl-1- (2-benzyloxy-5-fluoropyrimidin-4-yl) -pD-glucopy o ranuronate, m.p. 68-69 ° C.

Element Analysis:

Calcd. For C20 H23 ° 8N2 + 1 / 3H2 ° C 54.05 - H 5.37 - N 6.30 - F 4.28 Found: C 54.06 - H 5.45 - N 6.24 - F 4, 04th

Ultraviolet absorption spectrum:

Λmax (CH 3 OH) 270 nm (ε, 7300).

EXAMPLE 9

Isopropyl-L-O- (5-fluoro-lH-2-oxopyrimidine-4-yl) -β-ρ-glucopyranuronate.

i ·

"Y * Y

H Jk <

ICHohCHD-CO

iH 'ΜΗ J /

OiMaeJr 1 OH

28

DK 155331 B

0.46 g of isopropyl-1-Q- (2-benzyloxy-5-fluoropyrimidin-4-yl) -β-D-glucopyranuronic acid was dissolved in 40 ml of anhydrous methanol. The atmosphere in the reaction vessel was replaced with nitrogen gas and then 52.5 mg of 10 wt% palladium on charcoal followed by hydrogen gas under atmospheric pressure was added. After absorbing 23 ml of hydrogen gas, the catalyst was removed from the reaction mixture by filtration and the solvent was then distilled off under reduced pressure. After adding anhydrous methanol to the reaction mixture, crystals were precipitated which were collected by filtration to give the desired isopropyl-1-O- (5-fluoro-1H-oxopyrimidin-4-yl) -D-D-glucopyranuronate (compound 5) , mp 151-152 0

Co.

Element Analysis:

Calculated for C C ^HxgOgMgFo S / BHgQ: C 42.21 - H 5.29 - N 7.57 - P 5.14 Found: C 42.25 - H 5.08 - N 7.63 - F 4.71

Ultraviolet absorption spectrum:

Xmax (pH 6.86 phosphate buffer) 288 nm (e, 5200) EXAMPLES 10-17

Compounds 3, 4, 6, 7, 8, 9, 13 and 14 (identified in the above list) were prepared using the procedure described in Examples 7, 8 and 9, except that the isopropyl-2 used in Example 7 was used. 3,4-Tri-O-acetyl-1-bromo-1-deoxy-α-D-glucopyranuronate was replaced with the corresponding halogen saccharide to give a compound of the following formula: 29

DK 155331 B

VK

R% CO F

) - We • (oh in mJ (

OH

1 wherein RXiS has the meaning given in the following Table 6. The properties of the compounds prepared are listed in the following Tables 6 and 7.

30

DK 155331 B

—I ΓΠ ^^ T p

“S 'I j I

g: S i c I i | ϊ a o [3 <H I! I: 'P <P - I 5 +3 S3, · —s, - \ j r ~ \ j y'-N / N /' "M I VM i M ft o ooo.o oo [o;

ω -P O 'o o o o. O o S P

ft cd cTi I V S- CO Φ V- - O 5 CQ B. <S- UK i Lf \ 4 L 'I ^ I: i P-l I v j' w 'I ^ —S% f J s ^ i S w' -p m I \ + f.

ω o co co I od co co co I cg f cu; η ί I co -CO CO co oo rø I O) o .; O ft CM CM CM CM CM CM | CM. | • h 1 is i

i> vo {I I I

os oo I - 'ii -p vo i III i S 5 I [j * I i _I' I _j ysio 111 v "i: i '; i ^ i ^ 9 ω IK \ [K \ I 4" I (C \ J CM I CM I CM! "S +» - pv I <r I <r- vr v- 1 · \ I ~ 6:71 * Η ^ ί ^ i |. | | Iiji I 1 ii Yes,; Φ0 O (O 4- o CO VD vo ^ £; S ίί O f CM f .fOl I 4- j K \ CM I CM j CM! · Røftwl ^ r IV {v I v v- {v- s v- j * ji 1 oi [1 1! CM IK \ i fC \ i

: (^ B B i ^ S

K \ ΓΟ, praise ΚΛ O o / \ I

KM B B M B ^ · I! . ”· I I

B · O O O o. B U \ w ί I

O CMlKN I v Ό Ιλ J \ cm cm _ l ^ BIO:, c ^ - l \ B · OJ CM O I CM Car B O B B CM B cvr _ O I i / i O O B O B Vi

v-y w O w O! IN

I! I 1 · Y I

. ii SJ ^ Ϊ, q cq o KV 4 "VD O- CO [2 ^ PH p O <D S3 j ^ ___________ I .....— '•. □ I — i CM ΡΛ <t LTN SP fi— CQ · i — J i — 1 rH i — ir — jr — ξ i—! IJ — i B s .__; _._________ 1- 31

DK 155331 B

κ \ (A ca la I fA O O OJ

O- LA OJ fe i LO lD LA LO

Pr. »» · · J 0 '*' · * LA lA LA lA -4 4 "-3 * '4

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γ * ί {t-j — t g · f - e el o · | l I

pq 1-4 LA LA LA LA LA LA LA LA I LA I

OJ 4 Oj CA fA O 4 I CM j ΓΑ fe LO O .OJ (AjOjOJ!

ο · β · 9 4 o i ° I 0 I

V oj ia. 4 lo 4 ζ- co 4 4 4-.-4- 4 'j. J -4 j 4-

cr \ I 4- LA o fA i fe LA I EN

4 v CA O EN AND LO LA

Ccj · »· ofcfc oj® IA LA 4- LA 4 4- 4- 4-

__ I

LO IN O IN AND LA LO 4 * j ^ O f LA NA tA Ά CA CO 'EN i vo £ 3 t «·! 0 - '' S0 · i · Γ- 4: CO (N «CN J AND CN I LO LO i LO I - ^ ____ I_j i '.. j:

S LA CA OJ IN LA i O EN! CM

Γη IN OJ LO LA IN O I 4 CO

Jp H · · * S o c S w I 0 I * I

S4 4 IA LA LA LAIlD LO -LA

ω _________ ________j

CM „fe CA j fe LA i ΕΝ I LO

LO OJ - AND OJ IN f LP O OJ

O · o f 0 1 0 ·! c I 0 0 0 I

fe .CM I ΓΑ j 4 i LO 1 4 "'-N Cp I

I 4- 4- - j 4 ^ _I 4 j 4 j fe 4 4- j o 00 I O I 0.1

; (M OJ I CM I Ϊ CM CM i - CM

w. ixi W .1 fcd. free in K

K \ la lK S O I ia la O la,

i \ \ \ CM o «CM

i, oj. lo Lo rd O fe- W -r- rrt_l * · 6 »o I o ·, I · .2 P Cq- & j · 'pa' I Qq I (34 'I J =) · I pq i (4

oj cm Pli OJ i oj i CM CM [CVI

0 O CO OD CO CO j CO CO! CO. CO · O O O o O i o o o P ^ LA IN (A CA I fe fe IA! Fe i

£ fe fe fe fej OJ CM i OJ I OJ

S W K tø ixi, 1 td tø K tø 4

OJ "4” 4 j LA LA LO j LO

fe fe fe fe fe i fe fe I fe OOO O. O j O j O I O -

i 1 I i j I

β! Ilt »ri 0) ® S ^ f I st

w * P. n B

O4- (2,3,4,6-tetra-O-acetyl-B-D-flucopyranosyl) -O2-benzyl-5-fluorouracil.

EXAMPLE 18

DK 155331B

32 \

AcOCH2 7

Al i

Sfc / OAeHf / OÅc

Ac = Acetyl 4.91 g (15 mmol) of the silver salt of O 2 -benzyl-5-fluorouracil was suspended in 220 ml of dry xylene. 20 ml of xylene was then distilled off to remove all water. Then, 6.17 g (15 mmol) of 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide were added to the suspension and the mixture was heated under reflux for 5 hours. After the reaction was complete, a greyish-yellow, insoluble material was filtered off and the solution was concentrated under reduced pressure and dried. The residue was dissolved in 100 ml of ethyl acetate, and the solution was washed once with 70 ml of a 5% by weight aqueous solution of sodium bicarbonate and twice with 70 ml of water. The aqueous washing solutions were extracted several times with 50 ml of ethyl acetate and the combined ethyl acetate

DK 155331 B

tat solutions were dried over anhydrous magnesium sulfate, then the solvent was distilled off under reduced pressure to give a caramel-like substance which was then dissolved in ethanol. Addition of isopropyl alcohol to the ethanol solution yielded the precipitate of crystals which were filtered off and filtered off. g of the desired 04- (2,3,4,6-tetra-O-acetyl-8-D-glucopyranosyl) -C1-benzyl-5-fluorouracil in the form of white crystals melting at 95-96 ° C .

Element Analysis:

Calcd for C 25 H 27 O 11 N 2 F: C 54.54 - H 4.94 - N 5.09 - P 3.26 Found: C 54.33 - H 4.92 - N 4.70 - F 3.26

Ultraviolet Absorption Spectrum: * max (° 1N HCl 1 CH 3 OH) 270 nm (ε, 7200)

Xmax (CH OH) 270 nm (ε, 7100)

Xmax (0.1 N NaOH in CH 3 OH) 270 nm (ε, 8300) Example 19 O4- (2,3-4,6-tetra-O-acetyl-g-D-glucopyranosyl) -5-fluorouracil.

34

DK 155331 B

Ϊ X i

° Y'K

AcOCH2 I fI

J-0 0 / (OÅC)

Vj / I / OAcr- ^ Y 1 OAc 7

Ac = acetyl 5.795 g (10.5 mmol) of 04- (2,3,4,6-tetra-O-acetyl-β-D-glu-copyranosyl) -O2-benzyl-5-fluorouracil was dissolved in 130 ml of distilled tetrahydrofuran. The atmosphere in the reaction vessel was then replaced with nitrogen gas, after which 526 mg of 10 wt% palladium on coal was added to the solution. Hydrogen gas was introduced into the vessel and the mixture was reduced by shaking at room temperature and atmospheric pressure until 254 ml of hydrogen had been absorbed. The atmosphere in the vessel was then again replaced with hydrogen gas which was added further.

DK 155331 B

35 mg of palladium on coal and an additional 14 ml of hydrogen gas were absorbed. When the reaction was complete, the atmosphere in the vessel was replaced with hydrogen gas and the catalyst was filtered off. The filtrate was concentrated by evaporation under reduced pressure and then dried to give a crystalline residue to which methanol was added. The resulting crystals were filtered off to give 4,318 g of the desired O- (2,3,4,6-tetra-O-acetyl-8-D-glucopyranosyl) -5-o fluorouracil (Compound 15), m.p. .: 154-155 C.

Element Analysis:

Calcd for C 18 H 21 O 11 N 2 F: C 46.96 - H 4.60 - N 6.09 - F 4.13 Found: C 46.87 - H 4.67 - N 5.93 - F 3.94

Ultraviolet absorption spectrum:

Λmax (CH 3 OH) 288 nm (ε, 4200).

O4- (β-D-glucopyranosyl) -5-fluorouracil

DK 155331B

EXAMPLE 20 36 | "I \ γκ, HQCH2 S; ] A '' * VS- ^

Hh 2.3 g (5 mmol) of O4- (2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl) -5-fluorouracil was dissolved in 125 ml of anhydrous methanol and the solution was saturated with dry ammonia under ice-cooling, stirred for 6 hours and then allowed to stand overnight in the refrigerator. When the reaction was complete, the solvent was distilled off under reduced pressure, anhydrous ethanol was added and then this was distilled off. The gelatinous substance thus obtained was filtered off and dried to give 1.40 g of the desired O 4 - (β-D-glucopyranosyl) -5-fluorouracil (compound 10) in the form of a white powder which melted at 115 ° C.

Element Analysis:

Calculated for C CiøHuOy ^FOO.SEtøO: C 39.87 - H 4.68 - N 9.30 - P 6.31 Found: C 39.36 - H 5.02 - N 9.31 - P 6.13.

DK 155331B

37

Ultraviolet absorption spectrum: µmax (0.02M phosphate buffer, pH 6.86) 286 nm (ε, 4600).

Example 21 O 2 -benzyl-04- (8-D-glucopyranosyl) -5-fluorouracil

D0CH2 F

/ K '.

fr ^ '/

OH

550 mg of 04- (2,3,4,6-tetra-O-acetyl-8-D-glucopyranosyl) - O2-benzyl-5-fluorouracil (prepared as described in Example 18) was dissolved in 100 ml of anhydrous methanol, and the mixture was saturated with dry ammonia at room temperature and after tightly closing the reaction vessel stirred for 2 hours. When the reaction was complete, the solvent was distilled off under reduced pressure leaving 410 mg of a caramel-like substance, which was subjected to thin layer chromatography using a fractionation silica gel plate, eluted and purified with a chloroform containing 10% by volume methanol. 310 mg of the desired O 2 -benzyl-04- (β-D-glucopyranosyl) -5-fluorouracil was obtained in the form of a caramel-like substance which gave a single spot by thin layer chromatography and had a melting point of EXAMPLE 22 38

DK 155331 B

82-83 ° C.

O4- (β-D-glucopyranosyl) -5-fluorouracil 310 mg of 2-benzyl-04- (BD-glucopyranosyl) -5-fluorouracil was dissolved in 20 ml of anhydrous methanol and 50 mg of 10 wt% palladium on charcoal was added . The mixture was stirred at room temperature and atmospheric pressure under a stream of hydrogen gas. When 16 ml of hydrogen had been absorbed, the catalyst was filtered off after 5 minutes and then the solvent was distilled off under reduced pressure to give a caramel-like substance which was dissolved in ethanol. The ethanol solution was allowed to stand, thereby precipitating crystals. These crystals were filtered off to give the desired 04- (8-D-glucopyranosyl) -5-fluorouracil as a white powder. The properties of this product were identical to those of the compound prepared in Example 20.

EXAMPLE 23

DK 15533 1 B

Methy1-1-0- (2-benzyloxy-5-fluoro-pyrimidin-4-yl) -2,3,4-tri-O-acetyl-g-D-qalactopyranuronat.

LX in CH30.CO γ F 'v OÅC / OÅc 7.9 g of the silver salt of O 2 -benzyl-5-fluorouracil was suspended in 320 ml of dry toluene and then about 90 ml of the solvent was removed by distillation using a water trap. to remove the water »The remaining suspension was allowed to cool. Then 9.1 g of methyl 2,3,4-tri-O-acetyl-1-bromo-1-deoxy-α-D-gamma-lactopyranuronate was added and the mixture was heated under reflux for 30 minutes. After completion of the reaction, insolubles were removed by filtration and the solvent was distilled off under reduced pressure.

The residue was dissolved in 300 ml of ethyl acetate and the resulting solution was washed successively with 200 ml of water, 300 ml of 5% by weight aqueous solution of sodium hydroxide.

DK 155331 B

gene carbonate and 200 ml 20% by weight aqueous solution of sodium chloride. Each of these aqueous washing solutions was extracted with 200 ml of ethyl acetate and the ethyl acetate solutions combined. The combined solutions were dried over anhydrous magnesium sulfate, then the solvent was distilled off, leaving a caramel-like residue. This residue was purified by column chromatography on silica gel using chloroform as the eluent to give 10.1 g of the desired methyl-1- (2-benzyloxy-5-fluoropyrimidin-4-yl) - 2,3,4-tri-O-acetyl - $ - D-galactopyranuronate as a caramel-like substance which exhibited a single spot by thin layer chromatography.

Element Analysis:

Calcd for C24H25O11N2F: C 53.73 - H 4.70 - N 5.22 - F 3.54 'Found: C 53.96 - H 4.55 - N 5.20 - F 3.52.

Ultraviolet absorption spectrum:

XmaxiO / OiN HCl, CH3OH) 271 nm W (CH3OH)

XmaxiO / 1N NaOH, CH 3 OH) 270 nm.

EXAMPLE 24

DK 155331 B

41

Methyl-1-0- (2-benzyloxy-5-fluoropyrimidine-4-yl) -6 ~ D-QA-lactopyranuronat.

CiHSCftlk ^ IL

CK30.C0 or— ° \ j ° \ 0H y OH.

A solution of sodium methoxide in methanol was prepared by dissolving 0.55 g of metallic sodium in 160 ml of anhydrous methanol and the solution was cooled to -30 ° C. To this solution was added 6.44 g of methyl 10- (2-benzyloxy-5-fluoropyrimidin-4-yl) -2,3,4-tri-O-acetyl-β-D-galactopyranuronate and the mixture allowed to react. under anhydrous conditions at -30 for 6 hours. Then 2.01 g of acetic acid dissolved in 10 ml of anhydrous methanol was added and the neutrality of the mixture was confirmed. The mixture was then evaporated to dryness under reduced pressure to give a caramel-like residue which was then dissolved in 90 ml of ethyl acetate and washed once with 100 ml of an aqueous phosphate buffer solution (pH 7.0) and twice with 60 ml of a wt% aqueous solution 42

DK 155331 B

sodium chloride. Each aqueous wash solution was extracted with 50 ml of ethyl acetate and the ethyl acetate solutions were combined and then dried over anhydrous magnesium sulfate. Then, the solvent was removed by distillation under reduced pressure, and after most of the ethyl acetate was removed, 50 ml of isopropyl alcohol was added to the remaining mixture and this solvent was gradually removed. After the odor of ethyl acetate had completely disappeared, the precipitated white powdered substance was collected by filtration and dried to give 1.8 g of the desired methyl-10- (2-benzyloxy-5-fluoropyrimidin-4-yl) - β-D-ga-lactopyranuronat.

Element Analysis:

Calcd for CigHigO 3 F: C 52.68 - H 4.67 - N 6.83 - F 4.63 Found: C 52.81 - H 4.50 - N 6.91 - F 4.42.

Ultraviolet Absorption Spectrum: ^ maxi®2®) 270 nm.

EXAMPLE 25 43

DK 155331 B

Methyl-L-0- (5-fluoro-lH-2-oxopyrimidine-4-yl) -g-D-galacto-pyranuronat.

H

CII30.C0 'Y ^ F

\, \ ®, / /

OH

2.46 g of methyl 1-O- (2-benzyloxy-5-fluoropyrimidin-4-yl) -β-D-galactopyranuronate was dissolved in 90 ml of anhydrous methanol and the mixture was stirred for 15 minutes in a stream of nitrogen. The atmosphere in the reaction vessel was then replaced with nitrogen gas. 300 mg of 10 wt% palladium on charcoal was added to the solution and 134 ml of hydrogen gas were absorbed at room temperature and atmospheric pressure.

Then, the catalyst was removed by filtration and the solvent was evaporated under reduced pressure at a bath temperature of less than 30 θ After most of the methanol was evaporated, 30 ml of ethanol was added to the residue and evaporation continued. This procedure was repeated twice and then a white powdered product was collected by filtration to give 1.1 g of the desired methyl-10- (5-fluoro-1H-2-oxopyrimidin-4-yl) β-D-galactopyranuronat.

44

DK 155331 B

Element Analysis:

Calcd for C 11 H 13 O 8 N 2 F: C 41.26 - H 4.09 - N 8.75 - F 5.93 Found: C 41.55 - H 4.30 - N 8.77 - F 5.99.

Ultraviolet absorption spectrum:

Xmax (° 02M phosphate buffer, pH 6.86) 288 nm.

EXAMPLE 26 1-O- (2-Benzyloxy-5-fluoropyrimidin-4-yl) -β-Ρ-qalactopyranuronic acid.

5 CH2Q Nv.

(x / xc coo;) γ f, OH-0 0 y 'OH / 1.0 g of methyl-1- (2-benzyloxy-5-fluoropyrimidin-4-yl) -8-D-galactopyranuronate (prepared as described in Example 24) was dissolved in 23 ml of a mixture of methanol, water and concentrated ammonia water in a volume ratio of 6: 3.9: 0.1, and then the solution was allowed to stand for 24 hours with stirring. After the reaction was complete, the solvent was distilled off from the reaction mixture under reduced pressure, leaving 1.1

DK 155331 B

g of a caramel-like residue. The residue was subjected to thin-layer preparative chromatography and a fraction containing the desired compound was separated using chloroform containing 35% by volume of methanol as the developing agent. This fraction was extracted with methanol and then the methanol was evaporated from the extract to give a caramel-like residue. Ethanol was added to the residue and allowed to stand in the refrigerator to precipitate a powdered substance which was collected by filtration to give the desired 1-O- (2-benzyloxy-5-fluoropyrimidin-4-yl) ) -β-D-galactopyranuronsyre.

Element Analysis:

Calcd for C 17 H 17 O 8 N 2 F 2 H 2 O: C 47.23 - H 4.90 - N 6.48 - F 4.39 Found: C 47.56 - H 4.51 - N 6.32 - F 4.21.

Ultraviolet Absorption Spectrum: ^ maxi ^ O) 269 nm.

EXAMPLE 27 46

DK 155331 B

1- O- (5-Fluoro-1H-2-oxopyrimidin-4-yl) -6-D-galactopyranuronic acid.

! • Y «\ ohJ - \! I

Get '

1 OH

396 mg of 1-O- (2-benzyloxy-5-fluoropyrimidin-4-yl) -B-D-gamma-lactopyranuronic acid was dissolved in 20 ml of anhydrous methanol and the atmosphere in the reaction vessel was replaced with nitrogen gas. 50 mg of 10 wt% palladium on charcoal was added to the solution and the mixture was stirred at room temperature under atmospheric pressure while passing hydrogen through it until 22 ml of hydrogen gas was absorbed.

After the reaction was complete, the catalyst was removed by filtration and then the methanol was distilled off under reduced pressure. After most of the methanol was removed, anhydrous ethanol was added to the remaining mixture and then distillation was continued. The white powdery substance obtained was collected by filtration, washed with anhydrous ethanol and dried

47 DK 155302 B

in the form of yellow crystals, from 2,4-dichloro-3-methylaniline [described by Cohen and Dakin in J. Chem. Soc., 81, 1332 (1902)], 2,4-dichloro-3-methoxyphenylhydrazine, m.p. 88-80 ° C, in the form of brown crystals, from 2,4-dichloro-3-methoxyaniline, 5 3-cyano ~ 2,4-Dichlorophenylhydrazine, m.p. 199-200 ° C, in the form of a yellow powder, from 3-cyano-2,4-dichloroaniline [described by NVPhilips Gloelampen factory in Chem. Abs., 75, P 5527K ( 1971)], 4-bromo-2,3-dichlorophenylhydrazine, mp 135-137 ° C, in the form of a cream-colored powder from 4-bromo-2,3-dichloroaniline [described by Hurtley in J. Chem. Soc., 79, 1302 (1901)], 2,3-dichloro-4-methylphenylhydrazine, mp 111-114 ° C, in the form of colorless crystals, from 2,3-dichloro-4-methylaniline [described by Sylvester and Wynne in J. Chem. Soc., 1936, 691)], 4-bromo-2-chloro-3-methylphenylhydrazine, m.p. 100-103 ° C, in the form of colorless solid, from 4-bromo-2-chloro-3-methylaniline, 2-chloro-3,4-dimethylphenylhydrazine, m.p. 71-73 ° C, in the form of a brownish yellow powder, from 2 ° chloro-3,4-dimethylaniline (described by Hinkel et al in J. Chem. Soc. 1934, 286), 2-chloro-3-cyano-4-methylphenylhydrazine, mp 168-170 ° C, in the form of a colorless powder, from 2-chloro-3 = cyano-4-methyl-aniline, 3-chloro 2,4-dibromophenylhydrazine, mp 148-150 ° C, in the form of brown crystals, after crystallization from ethanol, 25 from 3-chloro-2,4-dibromaniline [described by Hurtley in J. Chem. Soc., 79, 1304 (1901)], 3-chloro-2,4-dimethylphenylhydrazone, mp 104 ° C, in the form of a yellow powder, from 3-chloro-2,4-dimethylaniline [described by Staskun in Tetrahedron bd. 28, 5069 (1972)], 3Q 2-bromo-4-chloro-3-methylphenylhydrazine, m.p. 72-73 ° C, in the form of an off-white powder, from 2-bromo = 4 = chloro-3-methyl-aniline 4-Chloro-2,3-dimethylphenylhydrazine, m.p. 69-71 ° C, in the form of a pale yellow powder, from 4-chloro-2,3-dimethylaniline (described by Hinkel et al in J. Chem Soc., 123, 2968 (1923), 48

DK 155331 B

distillation. Then, the suspension was cooled to room temperature, to which was added a solution of 33.6 g (71 mmol) of 3,4,6-tri-O-acetyl-2-deoxy-2-trichloroethoxycarbonylamino-2-aD-glucopyranosyl bromide in 100 ml. dry toluene and the mixture was heated under reflux for 15 minutes with stirring. After the reaction was complete, insolubles were filtered off and the filtrate was concentrated by evaporation under reduced pressure and dried to give a caramel-like residue which was dissolved in 500 ml of ethyl acetate and then washed successively with 300 ml of a 5% by weight aqueous solution. solution of sodium bicarbonate and 500 ml of a saturated aqueous solution of sodium chloride. After filtering off insolubles, the organic phase was dried over anhydrous magnesium sulfate and the solvent was evaporated under reduced pressure to give 43.2 g of the desired O 2 -benzyl-04- (3,4,6-tri -O-acetyl-2-deoxy-2-trichloroethoxycarbonylamino-[(D-gluco-pyranosyl) -5-fluorouracil as a caramel-like substance.

This substance was used without further purification in the following Example 29, but for analysis purposes a sample was purified by silica gel column chromatography using chloroform containing 1% by volume of methanol as eluent.

Ultraviolet Absorption Spectrum: * max (° 1N HCl in CH 3 OH) 270 nm (ε, 6950) xmax (CH 3 OH) 271 nm (ε, 7260)

XmaxiO / 1N NaOH in CH 3 OH 272 nm (ε, 7310).

NMR Spectrum (CDCl 3) δ ppm: 6.30 (doubled, H at 1 'position, = 8.5 Hz).

49

DK 155331 B

Element Analysis:

Calcd for C26H27O11Cl3F.I / 3 CHCl3: C 43.77 - H 3.81 - N 5.81 - F 2.56 Found: C 43.88 - H 3.66 - N 6.04 - F 2.56.

EXAMPLE 29; OA ^ f!

nk2 I

Ac = acetyl 43.2 g (63.0 mmol) of the O 2 -benzyl-O 2 - (3,4,6-tri-O-acetyl-2-deoxy-2-trichloroethoxy-carbonylamino) 8-D-glucopyranosyl) -5-fluorouracil was dissolved in 200 ml of 80% acetic acid. At the same time, 20.4 g (31.5 mmol) of zinc dust was suspended in 330 ml of 80% acetic acid, stirred for about 5 minutes, and cooled to about 20 ° C by ice cooling. Then, with stirring, the solution was added to the suspension and the mixture was stirred for an additional 1 hour and 10 minutes after addition of 80% acetic acid. After completion of the reaction, insoluble matter was filtered off and then the solvent was removed.

DK 155331 B

The mixture was lightly stripped under reduced pressure to give a residue which was dissolved in 300 ml of ethyl acetate and then washed successively with 300 ml of ice water, 500 ml of 5% by weight aqueous solution of sodium bicarbonate, 200 ml of water and 200 ml of saturated aqueous solution of sodium chloride.

Each of the aqueous washing solutions was extracted with 100 ml of ethyl acetate and all of the ethyl acetate solutions were combined and dried over anhydrous magnesium sulfate, after which the solvent was distilled off. The residue was dissolved in 200 ml of ethanol and the solution was distilled to a residual volume of about 100 ml, after which it was allowed to stand overnight at about 5 ° C. The precipitated crystals were filtered off and dried to give 14.38 g of the desired O 2 -benzyl-04- (3,4,6-tri-O-acetyl-2-amino-2-deoxy- | 3-D-glucopyranosyl) -5-fluorouracil in the form of colorless crystals, melting at 141-142.

Ultraviolet Absorption Spectrum: * max (° 1N HCl 1 CH 3 OH) 270 nm (ε, 6900)

Xmax (CH 3 OH) 271 nm (ε, 6990)

Xmax (O, 1N NaOH in CH 3 OH) 270 nm (ε, 6880) NMR Spectrum (CDCl 3) δ ppm: 8.24 (doubled, H at 6 position, J6jj - F = 2.0 Hz) 5.90 ( doubled, H at the 1 'position, = 8.5 Hz) 5.40 (singlet, -CH2 "CgH5).

Element Analysis:

Calculated for C 23 H 26 O 9 N 3 F: C 54.44 - H 5.16 - N 8.28 - F 3.74 Found: C 54.18 - H 5.44 - N 8.28 - F 3.45.

EXAMPLE 30 51

DK 155331 B

O4- (3,4 / 6-tri-O-acetyl "2-amino-2-deoxy-" B-D-glucopyranosyl) -5-fluorouracil ♦

AcO.CHz Y P

ΥΝΙ sweep. Ki.'2

Ac = acetyl 517 mg of 2-benzyl-04- (3,4,6-tri-O-acetyl-2-amino-2-deoxy-β-D-glucopyranosyl) -5-fluorouracil was dissolved in 50 ml of anhydrous ethanol. Then, the atmosphere in the reaction vessel was replaced by dry nitrogen gas, 50 mg 10% by weight palladium on charcoal was added and the solution was stirred under introduction of hydrogen gas at atmospheric pressure until 20 ml of hydrogen gas was absorbed. After the reaction was complete, the catalyst was filtered off and the filtrate was concentrated by evaporation under reduced pressure. The residue was dissolved in 5 ml of anhydrous methanol, 20 ml of anhydrous ethanol, 52

DK 155331 B

and the solution was concentrated by evaporation under reduced pressure to obtain crystals which were filtered off and dried. 250 mg of the desired O- (3,4,6-tri-O-acetul-2-amino-2-deoxy-8-D-glu-copyranosyl) -5-fluorouracil (Compound 12) was obtained, m.p. 129-130 ° C (during decomposition).

Ultraviolet absorption spectrum:

Λmax (CH 3 ° H) 291 nm (ε, 5010).

NMR spectrum (dimethyl sulfoxide-d) δ ppm: 8.13 (doublet, H at 6-position, J6H-P = 5.5 Hz) 6.06 (doublet, H at 1'-position, J1'-2 ' = 8.0 Hz).

Element Analysis:

Calcd for C 16 H 20 O 9 N 3 F * H 2 O: C 44.14 - H 5.09 - N 9.65 - F 4.36 Found: C 44.05 - H 4.77 - N 9.39 - F 3.87.

Example 31 O 2 -benzyl-O - (3,4,6-tri-O-acetyl-2-amino-2-deoxy-6-D-glucopyranosyl) -5-fluorouracil hydrochloride.

3.04 g (6 mmol) of O-benzyl-O- (3,4,6-tri-O-acetyl-2-amino-2-deoxy-8-D-glucopyranosyl) -5-flururacil, prepared as described in Example 29, was dissolved in 160 ml of acetone and a solution of 1.96 ml of diethyl ether containing 0.95 equivalents of hydrogen chloride in 40 ml of acetone was added dropwise. After the reaction was complete, the resulting white precipitate was filtered off, washed with acetone and dried to give 2.67 g of the desired white crystals product. 0 a melting point of 171-172 C (during decomposition). Fil- 53

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the funnel was concentrated to give a crystalline residue to which 200 ml of acetone was added. By filtration of the residue, washing with acetone and drying an additional 378 g of the desired product was obtained, which also had a melting point of 171-172 ° C (during decomposition).

Ultraviolet absorption spectrum:

Λmax (CH 3 ° H) 270 nm (ε, 7030).

NMR Spectrum (Dimethyl Sulfoxide-dg) δ ppm: 8.63 (doubled, H at 6-position, Jg 2 -p = 2 * 7 Hz) 6.78 (doubled, H at 1'-position, J'-2 = 8.5 Hz).

Element Analysis:

Calcd for C 23 H 26 O 9 N 3 F.HCl: C 50.79 - H 5.00 - N 7.73 - Cl 6.52 - F 3.49 Found: C 50.84 - H 4.98 - N 7.80 - Cl 6, 55 - F 3.30.

Example 32 04- (3,4,6-tri-O-acetyl-2-amino-2-deoxy-8-D-glucopyranosyl) -5-fluorouracil hydrochloride.

54.4 mg of O 2 -benzyl-04- (3,4,6-tri-O-acetyl-2-amino-2-deoxy) (SD-glucopyranosyl) -5-fluorouracil hydrochloride was dissolved in 50 ml of anhydrous methanol. Then the atmosphere in the reaction vessel was replaced with dry nitrogen gas.

50 mg of 10 wt% palladium on charcoal was added and hydrogen gas was passed through the solution with stirring at atmospheric pressure until 20 ml of hydrogen gas was absorbed. After the reaction was complete, the catalyst was filtered off and the filtrate was concentrated to give crystals which were filtered off, washed 54

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with anhydrous ethanol and dried to give white crystals of the desired 04- (3,4,6-tri-O-acetyl-2-amino-2-deoxy-8-D-glucopyranosyl) -5-fluorouracil hydro o chloride, m.p. 112-116 ° C (during decomposition). Element Analysis:

Calcd for C 16 H 20 O 9 N 3 F.HCl.H 2 O: C 40.73 - H 4.70 - N 8.91 - Cl 7.51 - F 4.03 Found: C 40.66 - H 4.67 - N 9.55 - Cl 7.23 - F 3.57.

Example 33 O 2 -benzyl-04- (2-amino-2-deoxy-8-D-glucopyranosyl) -5-fluorouracil.

C5H5CH2O 'Ssy ^^' sN

2.06 g (4 mmol) O 2 -benzyl-04- (3,4,6-tri-O-acetyl-2-amino-2-deoxy-8-D-glucopyranosyl) -5-fluorouracil was dissolved in a mixture of 30 ml of anhydrous methanol and 20 ml of methylene chloride, and the solution was cooled to -35 ° C. 8 ml of a 1N methanolic solution of sodium was added.

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and the mixture was kept at a temperature of between -30 ° C and -25 ° C for 2 hours. After this time o, the solution was cooled to -40 ° C, 10 ml of anhydrous methanol in 528 mg of acetic acid was added and then a solution of anhydrous methanol in 0.1 ml of acetic acid was added dropwise to adjust the pH to 6-7, after which the solvent was evaporated under reduced pressure to give a gel-like residue. To this residue was added 50 ml of a 0.1 N phosphoric acid buffer solution and 50 ml of ethyl acetate, and the mixture was stirred thoroughly to give a precipitate which was crushed, filtered, washed with 20 ml of ethyl acetate and 20 ml of water and dried to give 597 mg of the desired O during decomposition).

Ultraviolet absorption spectrum:

Xmax (cH3 ° H) 270 nm (e, 6930).

NMR spectrum (dimethyl sulfoxide-dg) δ ppm: 8.50 (doubled, H at 6 position, J6H-P = 2.5 Hz) 5.83 (doubled, H at 1 'position, i 2' = 8, 0 Hz) 5.40 (singlet, -CH 2 -C 6 H 5).

Element Analysis:

Calcd for C 17 H 20 Cl 6 N 3 F: C 53.54 - H 5.29 - N 11.02 - F 4.93 Found: C 53.50 - H 5.45 - N 11.03 - F 4.51.

EXAMPLE 34 56

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O4- (2-amino-2-deoxy-6-D-glucopyranosyl) -5-fluorouracil.

¥ \ VY \

CH2OH J F

r N ° fe /.

1 12 762 mg (2 mmol) of O 2 -benzyl-04- (2-amino-2-deoxy- $ - D-gluco-pyranosyl) -5-fluorouracil was dissolved in 60 ml of anhydrous methanol. The atmosphere in the reaction vessel containing the solution was replaced with dry nitrogen gas and then 100 mg of 10 wt% palladium on charcoal was added. Hydrogen was passed through the solution with stirring at room temperature until 46 ml of hydrogen was absorbed. After the reaction was complete, the catalyst was filtered off and then the filtrate was concentrated by evaporation under reduced pressure to obtain crystals. A small volume of anhydrous ethanol was added to the crystals and then filtered off and dried to give the desired 04- (2-amino-2-deoxy-fS-D-glucopyrosyl) -5-fluorouracil (Compound 11) white crystals having a melting point of 120-130 ° C (below -57 ° C)

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composing).

Element Analysis:

Calc'd for C 10 H 14 O 6 N 3 F.H 2 O.CH 3 OH: C 38.71 - H 5.91 - N 12.31 - F 5.57 Found: C 38.59 - H 5.62 - N 12.06 - F 5.26.

Claims (1)

DK-155331B 1. 5-fluorouracil derivatives for use as intermediates in the preparation of antitumorally active 5-fluoro-uracil compounds of the general formula 2 pyr [1] H (1) H 2 wherein R 2 R4qC0 / = \ | or <RTI ID = 0.0> OR </RTI> OR · HR · 5 HR · 5 wherein R is hydrogen, an aliphatic acyl group of 2-4 carbon atoms or an aromatic acyl group of 7-11 carbon atoms, R3 is -OR where R has the aforementioned meaning, or amino, benzyloxycarbonylamino, optionally having a nitro or halogen substituent in the phenyl ring, or β-haloethoxycarbonylamino, and R aralkyl having 7-12 carbon atoms or aryl having 6-10 carbon atoms, wherein the aromatic ring in the aralkyl or aryl group optionally has a substituent selected from alkyl of 1-4 carbon atoms, alkoxy of 1-4 carbon atoms and halogen, characterized in that they have the general formula 3R2 r705S RQ wherein R2 has the above meaning, and? 7 means benzyl optionally having a nitro or halogen substituent in the phenyl ring, or β-halogenethyl, or acid addition salts of such compounds where in R 3 is amino.