JP2706666B2 - 2'-deoxy-5-fluorouridine derivatives and salts thereof - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a novel 2′-deoxy-deoxy-5-fluorouridine derivative and a salt thereof. These compounds exhibit excellent anticancer and antiviral effects, and are useful as antitumor and antiviral agents.

2. Prior Art and its Problems 2'-deoxy-5-fluorouridine (FudR) exhibits excellent antitumor activity, but has the problems of high toxicity and a narrow safety margin.

Various derivatives have been developed to solve the above problems. For example, JP-A-60-61591 and JP-A-60-106593 disclose a large number of 2'-deoxy-5-fluorouridine derivatives. Although these compounds have alleviated the above problems to some extent, their antitumor activity is not satisfactorily potent, and their therapeutic index, absorption and the like are insufficient.

Means for Solving the Problems The present inventor has conducted intensive studies in view of the above-mentioned problems of the prior art, and as a result, has a stronger antitumor activity than conventional 2'-deoxy-5-fluorouridine derivatives. And succeeded in obtaining a novel 2'-5-fluorouridine derivative which is highly safe for the human body, and completed the present invention.

That is, the present invention provides a 2′-formula represented by the following general formula (I).
It provides a deoxy-5-fluorouridine derivative.

(In the formula, _one of R ₁ and R ₂ may have a group selected from the group consisting of an alkyl group, an alkoxy group, a halogenated alkyl group, a halogen atom, a hydroxyl group and a nitro group as a substituent on a phenyl ring. The compound of the present invention represented by the general formula (I) is useful as an antitumor agent and an antiviral agent, has low toxicity, and shows a benzyl group and the other forms an amino acid residue.
It is easily soluble in water and has a wide safety margin.

The groups represented by R ₁ and R ₂ in the general formula (I) are more specifically as follows.

Examples of the alkyl group that is a substituent of a benzyl group that may have a substituent on the phenyl ring include, for example, methyl,
1 to 1 carbon atoms such as ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl
And 6 linear or branched alkyl groups.

Examples of the alkoxy group include a linear or branched alkoxy group having 1 to 6 carbon atoms such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, sec-butoxy, tert-butoxy, pentyloxy, and hexyloxy. it can.

Examples of the halogen atom include fluorine, chlorine, bromine and iodine.

Examples of the halogenated alkyl group include alkyl having 1 to 3 carbon atoms such as chloromethyl, bromomethyl, trifluoromethyl, 1,2-dichloroethyl, 1,2-difluoroethyl, and 2,2,2-trifluoroethyl. Examples thereof include a halogenated alkyl group in which 1 to 3 halogen atoms have been substituted.

When the benzyl group represented by R ₁ or R ₂ has a substituent, the number is preferably one, two or three.

The benzyl group which may have 1 to 3 substituents includes benzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2-bromobenzyl, 3-bromobedyl, 4-bromobenzyl, 2-fluoro Benzyl, 3-fluorobenzyl, 4fluorobenzyl, 2,4
-Dichlorobenzyl, 3,4-dichlorobenzyl, 2-chloro-4-bromobenzyl, 2-trifluoromethylbenzyl, 4-trifluoromethylbenzyl, 4-nitrobenzyl, 2,4-dinitrobenzyl, 2-methyl Benzyl, 4-methylbenzyl, 4-ethylbenzyl, 4-te
An alkyl group, an alkoxy group, an alkyl halide on a phenyl group such as rt-butylbenzyl, 4-methoxybenzyl, 3-ethoxybenzyl, 4-hydroxybenzyl, 2,4-dihydroxybenzyl, and 2-methyl-3-nitrobenzyl A benzyl group which may have one or two groups selected from the group consisting of a group, a halogen atom, a hydroxyl group and a nitro group is preferred.

Generally, a benzyl group having one or two halogen atoms on the phenyl ring is more preferred.

An amino acid residue refers to a monovalent group obtained by removing an OH group from a carboxy group of an amino acid. The amino acid referred to in the present invention is of natural or synthetic origin, and is a compound having at least one amino group and one carboxyl group in one molecule, each having 2 to 20 carbon atoms. I have.

Examples of naturally occurring amino acids include, for example, alanine, isoleucine, glycine, serine, threonine, valine, leucine, arginine, lysine, asparagine, aspartic acid, glutamine, glutamic acid, cystine, cysteine, methionine, which are component amino acids of biological proteins. Examples include tyrosine, phenylalanine, tryptophan, histidine, hydroxyproline, proline and the like. It is not a protein component of the living body, but also plays an important role in the living body, and includes other naturally occurring amino acids and amino acids of synthetic origin, such as sarcosine, creatine, homocysteine, cysteine sulfonic acid, Norleucine, isoserine, homoserine, oxylysine, norvaline, dehydrovaline, ornithine,
Arginosuccinic acid, doper, 3-monoiodotyrosine, 3,5-diiodothyronine, thyroxine, α, γ-
Diaminobutyric acid, 2,3-diaminosuccinic acid, α-aminoadipic acid, α, β-diaminopropionic acid, saccharopine, β-alanine, γ-aminobutyric acid, β-aminobutyric acid,
Examples of each amino acid such as ε-aminocaproic acid, acediasulfone, agaristin, alanosine, hadacidin, melphalan, ibotenic acid and the like can be given. Furthermore, oxyamino acids such as serine and threonine, basic amino acids having a plurality of amino groups such as asparagine, ornithine and lysine, and hydroxyl, amino or carboxyl groups of acidic amino acids having a plurality of carboxyl groups such as glutamic acid and aspartic acid are acetyl. Also included are amino acids substituted with a group, an ethoxycarbonyl group, a benzyloxycarbonyl group or a benzyl group, for example, acetylserine, acetylthreonine, benzylaspartic acid and the like.

The amino acid residue represented by R ₁ or R ₂ is preferably an α-amino acid residue, and more preferably a component amino acid residue of a protein. Specific examples of such amino acid residues include alanine, isoleucine, glycine, serine, threonine, valine, leucine, arginine, hydroxylysine, lysine, asparagine, aspartic acid, glutamine, glutamic acid, cystine, cysteine, methionine, tyrosine, phenylalanine, Residues such as tryptophan, histidine, hydroxyproline and proline can be exemplified.

Most preferred amino acid residues are alanine, isoleucine, glycine, O-acetylserine, vario, leucine, lysine, aspartic acid-β-benzyl ester,
It is a residue of phenylalanine, histidine, proline and the like.

The amino acid residue according to the present invention is an amino acid residue represented by the following formula: -CO-B-NR ¹⁰ R ¹¹ (f) wherein B is a lower alkylene which may have a phenyl group or a lower alkylthio group. Represents a group, R ¹⁰ and R
¹¹ represents a hydrogen atom or a lower alkyl group, respectively, or may form a piperidine ring together with the nitrogen atom to which they are bonded. )], And the amino acid residue may further include a pyrrolidinylcarbonyl group or a piperidylcarbonyl group (however, the nitrogen atom forming the heterocycle and the carbonyl group are not bonded).

Here, the lower alkylene group which may have a phenyl group or a lower alkylthio group represented by B and which is ring with the amino group residue of the general formula (f) includes, for example, methylene, methylmethylene, ethylmethylene, isopropylmethylene, Isobutylmethylene, tert-butylmethylene,
Ethylene, trimethylene, tetramethylene, 2-methyltrimethylene, pentamethylene, hexamethylene, phenylmethylene, benzylmethylene, 5-phenylpentylmethylene, 3-phenylhexamethylene, methylthiomethylene, 2-methylthioethylmethylene, hexylthiomethylene , Isopropylthiomethylmethylene, 3-te
A phenyl group such as rt-butylthiohexamethylene group or a linear or branched alkylthio group having 1 to 6 carbon atoms, and an alkylene moiety having a linear or branched chain having 1 to 6 carbon atoms. Can be exemplified. R ¹⁰
And lower alkyl groups represented by R ¹¹ include, for example, methyl, ethyl, propyl, isopropyl, butyl, tert.
A straight-chain or branched-chain alkyl group having 1 to 6 carbon atoms, such as -butyl, pentyl and hexyl groups.

As the salt of the 2'-deoxy-5-fluoro-uridine derivative of the present invention, an acid addition salt of an amino group of an amino acid with an acid capable of forming a salt, and a carboxyl group of an acidic amino acid and a base of a salt capable of forming a salt Salts are included. Examples of the acid include inorganic acids such as hydrogen chloride, hydrogen bromide, phosphoric acid, nitric acid, sulfuric acid, and sulfurous acid, p-toluenesulfonic acid, methanesulfonic acid, tartaric acid, phthalic acid, fumaric acid, citric acid, maleic acid, and malonic acid. Organic acids such as acid, lactic acid, succinic acid, ascorbic acid, linoleic acid, oleic acid, nicotinic acid, and picryl sulfonic acid, and examples of bases include sodium hydroxide, potassium hydroxide, and sodium carbonate.

The compound of the present invention represented by the general formula (I) is produced by the method shown in the following reaction schemes 1 and 2.

[Reaction process formula 1] (Wherein R ₁ and R ₂ are the same as above, one of R ₃ and R ₄ represents a hydrogen atom, and the other is an alkyl group, an alkoxy group, a halogenated alkyl group, a halogen atom, a hydroxyl group And a benzyl group which may have a group selected from the group consisting of nitro and nitro, wherein one of R ₅ and R ₆ is N
Represents an amino acid residue having a protecting group at the position, and the other has, as a substituent, a group selected from the group consisting of an alkyl group, an alkoxy group, a halogenated alkyl group, a halogen atom, a hydroxyl group and a nitro group on the phenyl ring. A represents an amino acid residue having a protecting group at the N-position. That is, compound (IV) is produced by ester-condensing amino acid (III) with compound (II). The amino acids are used as such or in the form of their reactive derivatives such as halides and acid anhydrides. As the protecting group at the N-position of an amino acid, those generally used in amino acid chemistry may be used, and specifically, for example, t-butoxycarbonyl (Boc), trichloroethoxycarbonyl, benzyloxycarbonyl, methoxy, methyl, nitro, etc. And a benzyloxycarbonyl group having 1 to 3 substituents. The ester condensation of the compound (II) and the compound (III) is usually performed in an anhydrous solvent in the presence of a basic compound and a condensing agent. As the solvent, for example, benzene, toluene, aromatic hydrocarbons such as xylene, dichloromethane, chloroform, trichloromethane, halogenated hydrocarbons such as dichloroethane, ethyl ether, dioxane, ethers such as tetrahydrofuran, pyridine, nitromethane, Dimethylformamide and the like can be used. Examples of the basic compound include, for example, trialkylamine, pyridine, dialkylaminopyridine, picoline,
Examples thereof include tertiary amines such as lutidine, and metal carbonates such as sodium carbonate and barium carbonate. Usually, dialkylaminopyridine, particularly di (C ₁ -C ₃ alkyl) aminopyridine such as dimethylaminopyridine and diethylaminopyridine Is preferably used. Examples of the condensing agent include p-toluenesulfonyl chloride, 2,4,6-triisopropylbenzenesulfonyl chloride (TPS chloride)
Arylsulfonyl chloride, alkylsulfonyl chloride (particularly, C ₁ -C ₃ alkylsulfonyl chloride such as methanesulfonyl chloride, ethanesulfonyl chloride), N, N-dicyclohexylcarbodiimide (DCC),
Examples thereof include thionyl chloride and phosphorus oxychloride. Usually, DCC and TPCS chloride are preferably used. The use ratio of the above compound (III) is 1 to 1 mol of the compound (II).
The molar ratio of the basic compound and the condensing agent is about 1 to 5 moles per 1 mole of the compound (II). The reaction temperature is usually about −50 ° C. to room temperature.

The compound (IV) thus obtained is optionally subjected to a deprotection reaction. The reaction may be performed according to a conventional method, for example, by adding a 0.5 to 5N hydrochloric acid solution in the presence of a solvent. As a result, the protecting group of the amino acid residue is removed, and the hydrochloride (I ′) of the compound (I) is obtained. As the solvent, for example, ethylene chloride, chloroform, tetrahydrofuran, dioxane and the like are used.

[Reaction process formula 2] (Wherein R ₁ and R ₂ are the same as above. A ′ represents an amino acid residue having a protecting group at the N-position, X represents a halogen atom,
One of R ₇ and R ₈ is a hydrogen atom, the other is an amino acid residue having a protecting group at the N-position, and R ₉ is an alkyl group, an alkoxy group, a halogenated alkyl group as a substituent on the phenyl ring,
It indicates a benzyl group which may have a group selected from the group consisting of a halogen atom, a hydroxyl group and a nitro group. That is, the compound (VI) is obtained by reacting FudR with an amino acid halide (V) in a solvent in the presence of a basic substance to obtain a substituted or unsubstituted benzylimidate (VI).
By reacting the compound with I) in the presence of trifluoromethanesulfonic acid, the protecting group of the amino acid residue is removed, and the compound (I ″) which is a trifluoromethanesulfonic acid salt of the above compound is obtained.

In the general formula (V), examples of the halogen atom represented by X include chlorine and bromine.

As the solvent and the basic substance, those exemplified in the reaction scheme 1 can be used. The use ratio is about 1 to 1.5 times mol of amino acid halide and 0.1 to 0.1 mol of basic substance per mol of FudR.
It may be about 0.5 times the mole. The reaction is completed at a temperature of about −50 ° C. to about room temperature for about 2 to 12 hours.

In the reaction of the compound (VI) with the substituted or unsubstituted benzylimidate (VII), the use ratio of the compound (VII) and trifluoromethanesulfonic acid is 1 to 5 moles per 1 mole of the compound (VI). Degree and 0.01-0.5
It is about twice the mole. As the solvent, those similar to the aforementioned reaction can be used, and the reaction temperature is from room temperature to around the boiling point of the solvent,
The reaction time is about 2 to 12 hours.

In order to derive the hydrochloride of the compound of the present invention and the trifluoromethanesulfonate obtained from the reaction schemes 1 and 2 into other desired salts, a known and commonly used salt exchange reaction is carried out in the field of chemistry. It should be attached to. For example, Reaction Scheme 1
And dissolving the compound obtained in step 2 in an aqueous solution of a base such as sodium hydroxide, potassium hydroxide, sodium carbonate, sodium hydrogen carbonate, neutralizing the acid addition salt, adding the desired acid component, and freeze-drying. Good.

The compound of the present invention produced by the above method can be isolated and purified by generally known separation and purification means, specifically, recrystallization, column chromatography and the like.

When the derivative of the present invention is used as a medicament, it is usually used in combination with a suitable pharmacologically acceptable carrier.
It is prepared in a formulation suitable for the administration route. As the carrier to be used, known and commonly used excipients, binders, lubricants,
Coloring agents, disintegrants and the like may be used, and the dosage forms thereof are suitable for oral administration, such as tablets, capsules, granules, powders, liquids, etc., suitable for parenteral administration such as intravenous injection. Examples include injections, and suppositories suitable for rectal administration. The content of the active ingredient (compound of the present invention) per unit form of each preparation may be appropriately determined according to the form, and is not significantly different from those of ordinary pharmaceuticals. The preferred active ingredient content is generally about 25-500 mg per unit. The preparation method for each of the above-mentioned preparation forms may be in accordance with a conventional method.

The dosage of each of the preparations thus obtained naturally depends on the condition, weight, age, etc. of the patient to which the preparation is administered, and cannot be unconditionally limited.In general, about 50 to 2000 mg of the active ingredient is usually administered per adult day. It can be administered once a day or divided into two to four times a day.

Examples The following examples, pharmacological test results and formulation examples are given to further clarify the present invention.

In addition, regarding the NMR measurement results in the following description, the numbers attached to the right shoulder of H and the numbers attached to the lower right of C and N indicate the positions in the compound. Accordingly, for example, “H ⁶ ” or “C ₆ -H” indicates a hydrogen atom bonded to the carbon atom at the 6-position. Similarly, "H ³ ', H ⁴ ',
“H ⁵ ′” or “C ₃ ′, ₄ ′, ₅ ′ -H” refers to both 3 ′
It represents a hydrogen atom bonded to carbon atoms at the-, 4'- and 5'-positions.

Example 1 O ³ '- trityl-2'-deoxy-5-fluorouridine 2.44g was dissolved in tetrahydrofuran (THF) 15 ml. To this solution was added 0.5 g of sodium hydride, and at 60 ° C,
After stirring for 30 minutes, 0.63 g of benzyl chloride and 0.375 g of sodium iodide were added and reacted at 60 ° C. for 6 hours. The reaction mixture was neutralized with aqueous 1N HCl and then concentrated.
20 ml of 0.5N-HCl methanol solution was added to the obtained residue, followed by stirring at room temperature overnight. The reaction mixture was neutralized with a saturated aqueous solution of NaHCO ₃ and then concentrated.
Was added to collect a precipitate. This was dissolved in ethyl acetate, washed with water, dried over茫硝, then concentrated, O ⁵ '- to give the benzyl-2'-deoxy-5-fluorouridine.
This was suspended in 15 ml of methylene chloride, 0.84 g of Boc glycine,
1.24 g of DCC and 0.05 g of dimethylaminopyridine were added, and the mixture was stirred at room temperature overnight. The reaction solution was passed, and the solution was concentrated.
To the obtained residue, 4N-HCl dioxane (5 ml) was added. After stirring for 30 minutes, diethyl ether (20 ml) was added, and the precipitated crystals were collected and purified with methanol-ethyl ether to obtain Compound 1 shown in Table 1 below. (Yield 1.28 g) was obtained.

Example 2 Compounds 2 to 1 shown in Table 1 below were prepared in the same manner as in Example 1 except that the raw materials used were changed to those corresponding to the target compound.
1. Compounds 28-30 were prepared.

Example 3 O4-trityl-2'-deoxy-5-fluorouridine (2.44 g) was dissolved in THF ( ¹⁵ ml). To this solution was added 0.5 g of powdered sodium hydroxide and powdered molecular sieves 3A0.
After adding 225 g and heating under reflux for 2 hours, 0.845 g of p-chlorobenzyl chloride and 0.15 g of sodium iodide were added, and the mixture was heated under reflux for 2 hours. After allowing to cool, the reaction mixture was filtered, and the solution was neutralized by adding 15 ml of a saturated aqueous solution of ammonium chloride, washed with a saturated aqueous solution of sodium chloride, and the organic layer was dried over sodium sulfate and concentrated. To the obtained residue 0.5N-HCl.methanol solution 20 ml
Was added and stirred overnight. The reaction solution was neutralized by adding a saturated aqueous solution of NaHCO ₃ and then concentrated. 20 ml of THF was added to the obtained residue, and the residue was washed with a saturated aqueous solution of NaCl, and then dried over sodium sulfate. After concentration, recrystallization from ethanol gave O ³ '-p-chlorobenzyl-2'.
1.32 g of -deoxy-5-fluorouridine was obtained. THF12
After dissolving 1.07 g of DCC and 0.73 g of Boc-glycine in ml, O ³ ′ -p-
Chlorobenzyl-2'-deoxy-5-fluorouridine (1.32 g) and dimethylaminopyridine (0.05 g) were added, the mixture was stirred at room temperature for 5 hours, filtered, and concentrated. To the obtained residue, 4N-HCl dioxane (5 ml) was added, and after stirring for 30 minutes, diethyl ether (20 ml) was added to collect a precipitated crystal. Recrystallize with ethanol-isopropyl alcohol to give the compound
12 (yield 0.83 g) was obtained.

Example 4 Compounds 13 to 22 shown in Table 1 below were prepared in the same manner as in Example 3 except that the raw materials used were changed to those corresponding to the target compound.
Was manufactured.

Example 5 O ³ '-p- chlorobenzyl -O ^5' - alanyl-2'
0.10 g of deoxy-5-fluorouridine hydrochloride was added to water /
It was dissolved in 4 ml of a 1: 1 mixture of chloroform. To this solution was added 0.018 g of NaHCO ₃ with stirring, and after stirring for 10 minutes, the chloroform layer was separated. Add 0.10 g of tosylic acid to 2 ml of methanol
The solution was added dropwise, and a white precipitate was collected and recrystallized from methanol-ether to obtain Compound 23 shown in Table 1 below (yield: 0.2%).
105 g).

Example 6 Compounds 24 to 27 shown in Table 1 below were prepared in the same manner as in Example 5 except that the raw materials used were changed to those corresponding to the target compound.
Was manufactured.

Example 7 2.44 g of 2'-deoxy-5-fluorouridine was dissolved in 25 ml of pyridine, 0.97 g of Boc-glycyl chloride was added, and the mixture was reacted under ice cooling for 3 hours. After completion of the reaction, the solvent was distilled off, water was added to the residue, extracted with methylene chloride, dried over sodium sulfate and concentrated. The obtained residue was dissolved in 25 ml of methylene chloride, 2.14 g of p-chlorobenzylimidate and 0.75 g of trifluoromethanesulfonic acid were added, and the mixture was reacted at room temperature for 3 hours. A saturated aqueous solution of NaHCO ₃ was added to the obtained residue to make it alkaline, and the mixture was extracted with ethyl ether, dried over sodium sulfate, passed through hydrogen chloride, and the precipitated crystals were collected and recrystallized from methanol-ethyl ether to give the compound.
12 (0.75 g yield) was obtained.

The chemical structures and physicochemical constants of Compounds 1 to 30 obtained in Examples 1 to 7 are shown in Table 1 below.

Example 8 (A) Preparation of 5'-O- [t-butoxycarbonyl) glycyl] -3'-O- (4-chlorobenzyl) -2'-deoxy-5-fluorouridine N- (t (butoxycarbonyl) ) 6.13 g of dicyclohexylcarbodiimide was added to a solution of 9.45 g of glycine in 60 ml of acetonitrile under ice-cooling and stirring, and after 3 hours, the formed dicyclohexylurea was removed and the solution was concentrated.The oily residue was dissolved in 15 ml of anhydrous pyridine, 3'-O- (4-
After adding 5.00 g of (chlorobenzyl) -2'-deoxy-5-fluorouridine, the mixture was left at room temperature for 3 hours and then at 40 ° C for 30 minutes. Next, 5 ml of water was added at room temperature, the mixture was left for 30 minutes, and then concentrated. The residue was dissolved in 50 ml of ethyl acetate, and three times with 30 ml of a saturated aqueous solution of sodium hydrogencarbonate, and 30 ml of a saturated saline solution.
Wash twice with ml, dry over anhydrous magnesium sulfate,
Concentrated. The residue was purified using a silica gel column with 1% methanol-chloroform, and 5'-O- [N- (t
-Butoxycarbonyl) glycyl] -3'-O- (4-
5.33 g (75%) of (chlorobenzyl) -2'-deoxy-5-fluorouridine were obtained.

_{NMR (DMSCO-d 6) δ} : 11.85 (1H, bs, NH) 7.94 (1H, d, J = 7Hz, C 6 -H) 7.40 (4H, s, C 6 H 4 Cl) 7.20 (1H, t, J = 6Hz, OCONH) 6.14 ( 1H, t, J = 6Hz, C 1 '-H) 4.54 (2H, s, C H 2 C 6 H 4 Cl) 4.30-4,16 (4H, m, C 3' _{, 4 ', 5' -H)} 3.74 (2H, d, J = 6Hz, CH 2 CO) 2.38-2D32 (2H, m, C 2 '-H) 1.37 (9H, s, CH 3 × 3) (B ) And (C) In the same manner as in Example 8 (A), the compounds shown in Table 2 were obtained.

However, in Table 2, the compounds are represented by the general formula (I)
And R ₂ and R ₁ groups. The same applies to the following tables.

(D) Preparation of 5'-O- (N-benzyloxycarbonyl) glycyl-3'-O- (4-chlorobenzyl) -2'-deoxy-5-fluorouridine 3'-O- (4-chlorobenzyl ) To a solution of 1.00 g of -2'-deoxy-5-fluorouridine in 20 ml of pyridine,
1.23 g of N-benzyloxycarbonylglycine chloride was added and reacted at room temperature for 2 days.

The reaction solution was concentrated under reduced pressure, and 30 ml of ethyl acetate and water were added to the residue.
20 ml was added. The organic layer was separated, dried over anhydrous magnesium sulfate, and the solvent was distilled off. The resulting oily residue was purified using a silica gel column, eluting with 1% methanol-chloroform, to obtain the desired 5'-O- (N-benzyloxycarbonyl) glycyl-3'-O- (4-chlorobenzyl). ) -2'-Deoxy-5-fluorouridine 0.26
g (17%) was obtained.

This was identified using the compound obtained in Example 8 (B) and thin layer chromatography (developing solvent; chloroform: methanol = 39: 2, detection; irradiation with UV light).

(E) Production of 3'-O- (4-chlorobenzyl) -2'-deoxy-5-fluoro-5'-O-glycyluridine hydrochloride 5 '-[N obtained in Example 8 (A) -(T-butoxycarbonyl) glycyl] -3'-O- (4-chlorobenzyl) -2'-deoxy-5-fluorouridine 0.88 g
Was dissolved in 3 ml of 4N-HCl-dioxane and left at room temperature for 30 minutes. After evaporating the solvent, a silica gel column was used to give 4-
Elution with 6% methanol-chloroform yielded 3'-O
-(4-chlorobenzyl) -2'-deoxy-5-fluoro-5'-O-glycyuridine hydrochloride 0.36 g (45
%).

Properties: hygroscopic powder NMR (DMSO-d ₆ ) δ: 11.83 (1H, bs, NH) 8.68 (3H, bs, NH ₂ , HCl) 8.04 (1H, d, J = 7Hz, C ₆ -H) 7.41 _{(4H, s, C 6,} H 4 Cl) 6.15 (1H, t, J = 6Hz, C 1 '-H) 4.56 (2H, s, C H 2 C 6 H 4 Cl) 4.41-4.15 (4H, m _{, C 3 ', 4',} 5 '-H) 3.83 (2H, s, CH 2 CO) 2.44-1.99 (2H, m, C 2' -H) elemental analysis _{(%): C 18 H 19} ClFN 3 As O ₆ .HCl.H ₂ O, CH N Calculated: 44.83, 4.60, 8.71 Found: 44.64, 4.91, 8.43 Examples 9-17 Same as Example 8 (A), then Example 8 (E) Thus, each compound shown in the following Table 3 was obtained. However, Example 12
Was obtained in the same manner as in Example 8 (A).

Example 18 Preparation of 5'-O-benzyl-2'-deoxy-5-fluoro-3'-O-glycyluridine hydrochloride 1.00 g of 5'-O-benzyl-2'-deoxy-5-fluorouridine To a solution of 30 ml of anhydrous pyridine was added N- (t-
0.78 g of (butoxycarbonyl) glycine and 1.80 g of 2,4,6-triisopropylbenzenesulfonyl chloride were added, and reacted at room temperature overnight.

The reaction solution was concentrated under reduced pressure, and the residue was partitioned between ethyl acetate and water. The organic layer was washed with a saturated aqueous solution of sodium hydrogen carbonate, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified using a silica gel column and eluted with 1% methanol-chloroform to give the desired 5'-O-benzyl-3'-O- (N- (t-butoxycarbonyl) glycyl] -2'-deoxy. 1.06 g of -5-fluorouridine
I got

Next, the above compound (reaction intermediate) was dissolved in 10% hydrochloric acid-dioxane (10 ml) and stirred at room temperature for 2 hours.

The reaction solution was concentrated under reduced pressure, and the residue was purified using a silica gel column, eluting with 10% methanol-chloroform.
The desired 5'-O-benzyl-2'-deoxy-5-
0.77 g of fluoro-3'-O-glycyluridine hydrochloride (6
0%).

Properties: hygroscopic powder NMR (DMSO-d ₆ ) δ: 9.10 (3H, bs, NH ₂ , HCl) 7.97 (1H, d, J = 7 Hz, C ₆ -H) 7.35 (5H, s, C ₆ H) _{5) 6.22 (1H, t,} J = 6Hz, C 1 '-H) 5.36 (1H, bs, C 3' -H) 4.58 (2H, s, C H 2 C 6 H 5) 4.27 (1H, bs, _{C 4 '-H) 3.81 (2H} , s, CH 2 CO) 3.77-3.71 (2H, m, C 5' -H) 2.55-2.31 (C 2 '-H, overlap with DMSO) example 19-26 embodiment In the same manner as in Example 18, the compounds shown in Table 4 below were obtained.

Example 27 3'-O- (4-chlorobenzyl) -2'-deoxy-5'-O- (3-diethylaminopropanoyl) -5
-Production of fluorouridine hydrochloride 3-diethylaminopropanoic acid hydrochloride
A solution of 0.74 g in 30 ml of tetrahydrofuran was cooled to −15 ° C., and 0.56 ml of triethylamine and 0.50 ml of isobutyl chloroformate were added to prepare a mixture anhydride.

1.00 g of 3'-O- (4-chlorobenzyl) -2'-deoxy-5-fluorouridine and 0.56 ml of triethylamine were added thereto, and the mixture was stirred at room temperature overnight. The insolubles were removed, the solution was concentrated, and the residue was purified using a silica gel column.
And purified by eluting with
3'-O- (4-chlorobenzyl) -2'-deoxy-
5'-O- (3-diethylaminopropanoyl) -5
0.10 g (7%) of fluorouridine hydrochloride was obtained.

Sexual shape: hygroscopic powder _{NMR (DMSO-d 6) δ} : 11.96 (1H, bs, NH) 7.98 (1H, d, J = 7Hz, C 6 -H) 7.41 (4H, s, C 5 H 4 Cl) 6.16 (1H, t, J = 6Hz, C 1 '-H) 4.55 (2H, s, C H 2 C 6 H 4 Cl) 4.27-4.23 (4H, m, C 3', 4 ', 5' -H ) 3.19-2.94 (8H, m, CH 2 CH 2 CO, C H 2 CH 3 × 2) 2.41-2.33 (2H, m, C 2 '-H) 1.21 (6H, t, J = 7Hz, CH 3 × 2) Example 28 Preparation of 3'-O- (4-chlorobenzyl) -2'-deoxy-5-fluoro-5'-O-glycyluridine L-tartrate 5 obtained in Example 8 (A) '-O- [N- (t-butoxycarbonyl) glycyl] -3'-O- (4-chlorobenzyl) -2'-deoxy-5-fluorouridine 0.50
2 g of 4N hydrochloric acid-dioxane was added to g, and the mixture was left at room temperature for 15 minutes. Then the solvent was distilled off by evaporation, and the residue was dissolved in a small amount of water. A saturated aqueous solution of sodium hydrogen carbonate was added to make the mixture weakly basic, and the mixture was extracted three times with 40 ml of ethyl acetate. After drying the ethyl acetate layer over anhydrous magnesium sulfate,
A solution of 9.14 g of L-tartaric acid in ethyl acetate was added, and the mixture was concentrated to about 40 ml. The resulting precipitate was removed and 3'-O- (4-
(Chlorobenzyl) -2'-deoxy-5-fluoro-
0.38 g (65%) of 5'-O-grilluridine L-tartrate
I got

Sexual shape: hygroscopic powder _{NMR (DMSO-d 6) δ} : 7.97 (1H, d, J = 7Hz, C 6 -H) 7.40 (4H, s, C 6 H 4 Cl) 7.26 (7H, bs, NH, _{NH 2, OH × 2, COOH} × 2) 6.14 (1H, t, J = 6Hz, C 1 '-H) 4.54 (2H, s, C H 2 C 6 H 4 Cl) 4.43-4.20 (4H, m, _{C 3 ', 4', 5} '-H) 4.06 (2H, s, HOOCC H OHC H OHCOOH) 3.75 (2H, s, CH 2 CO) 2.41-2.28 (2H, m, C 2' -H) example 29 and 30 The compounds shown in Table 5 were obtained in the same manner as in Example 28.

Pharmacological Test Example I The test results of the antitumor effect of the compound of the present invention are shown below, and the usefulness of the compound of the present invention will be described.

5 × 10 ⁶ mouse transplantable tumor sarcoma 180 cells were implanted subcutaneously on the back of male ICR / JCL mice (27-30 g). The sample was a weakly acidic aqueous solution, and the solution was applied 1.0 m to a group of 7 mice.
1/100 g body weight, 1 volume from the day after tumor implantation
Oral administration once daily for 7 consecutive days. To the control group, 1.0 ml / 100 g body weight of the above solution containing no specimen was similarly orally administered once a day for 7 consecutive days.

The average tumor weight at each dose for each sample was measured 10 days after the transplantation, and these were compared with the average tumor weight in the control group. From the dose-response curve, a dose (ED ₅₀ ) that suppresses the increase in tumor weight by 50%, and a dose (IB ₅₀ ) that suppresses the weight gain by 50% from the dose-weight change curve,
To determine the therapeutic index _{TI (IB 50 / ED 50)} . The results are shown in Table 6.

From the results shown in Table 6, Compound 12 is more potent than O3'-p-chlorobenzyl-2'-deoxy-5-fluorouridine and has a large therapeutic index, indicating an excellent anticancer effect.

Pharmacological Test Example II Passaged Sarcoma (Sar
coma) -180 was diluted with saline to 2 × 10 ⁷ per animal
An individual amount was subcutaneously transplanted into the back of a syngeneic mouse and used for the experiment. Twenty-four hours after tumor implantation, the following test compound dissolved in sterile physiological saline was administered by intravenous injection from the tail vein every day for seven days once a day.

On the 10th day after tumor implantation, solid tumor under the back was excised and the tumor weight was measured, and the ratio (T / T) of the tumor weight (T) of the drug-administered group to the tumor weight (C) of the drug-untreated control group was measured seeking C), the dose of drug dose and the ratio (T / C) - T / C from the reaction curve was determined 50% tumor inhibition dose becomes 0.5 (ED ₅₀ value).

The results obtained using each of the following test compounds are shown in the following VII.
It is shown in the table.

Next, preparation examples of the compound of the present invention will be shown.

Formulation Example 1 Capsule Compound 3, lactose, crystalline cellulose, and corn starch were mixed in the following proportions, and magnesium stearate was further added and mixed in the following proportions. This mixture was filled using an appropriate capsule filling machine so as to be about 293 mg per capsule to obtain a product.

Capsule formulation mg / capsule compound 3 200.0 Lactose 30.0 Crystalline cellulose 50.0 Maize starch 10.0 Magnesium stearate 3.0 293.0 Formulation example 2 Granules Compound 7, lactose, crystalline cellulose and corn starch were mixed in the following proportions. A 10% ethanol solution of hydroxypropylcellulose was added to the mixture and kneaded, and then granulated by using an appropriate granulator. After drying, this was sized to 12 to 42 mesh. The sized product was coated with hydroxypropylmethylcellulose at the following ratio using an appropriate coating device. 12-42
The product was sized after meshing.

Formulation of granules mg / compound in one package 7 200.0 Lactose 200.0 Microcrystalline cellulose 311.0 Maize starch 200.0 Hydroxypropylloose 10.0 Hydroxypropyl methylcellulose 70.0 Fatty acid monoglyceride 3.5 Titanium dioxide 5.5 1000.0 Formulation example 3 Tablets Compound 1, corn starch and calcium cellulose glycolate Are mixed in the following proportions. A 10% ethanol solution of hydroxypropylcellulose is added to the mixture, and the mixture is kneaded. The mixture is granulated with an appropriate granulator, dried, and the following mixture of magnesium stearate and silicic anhydride is added to a suitable tableting machine. The tablet is coated with hydroxypropyl methylcellulose using
The product.

Tablets Formulation mg / Tablet compound 1 200.0 Maize starch 5.0 Calcium cellulose glycolate 20.0 Hydroxypropylcellulose 2.0 Magnesium stearate 2.5 Silicic anhydride 2.5 Hydroxypropylmethylcellulose 19.999 Macrogol 6000 0.001 Titanium oxide 2.0 254 Formulation example 4 Suppository Witepsol W -35 (trade name, manufactured by Dynamite Nobel) was melted at about 60 ° C, and then kept at about 45 ° C. After mixing Compound 5 in the following ratio, the mixture was molded into a 1 g suppository using an appropriate suppository manufacturing apparatus.

Preparing suppository prescription mg / suppository Compound 5 400.0 Witepsol W-35 600.0 1000.0 Formulation Example 5 Compound of Example 13 5 mg Lactose 97mg capsules of crystalline cellulose 50mg Magnesium stearate 3mg The above mixing ratio at 1 per capsule 200mg .

Formulation Example 6 Compound of Example 14 10 mg Lactose 184 mg Crystalline cellulose 100 mg Magnesium stearate 6 mg A 300 mg / capsule is prepared at the above mixing ratio.

Formulation Example 7 Compound of Example 19 10 mg Lactose 240 mg Corn starch 340 mg Hydroxypropylcellulose 10 mg A granule of 600 mg per packet is prepared in the above mixing ratio.

Formulation Example 8 Compound of Example 8 (E) 10 mg Macrogol 300 500 mg Distilled water for injection Appropriate amount 5 mg per ampoule for injection is prepared in the above mixing ratio.

Formulation Example 9 1000 tablets for oral use each containing 10 mg of the compound of Example 22 are prepared according to the following formula.

Compound of Example 22 10g Lactose (Japanese Pharmacopoeia) 45g Corn Starch (Japanese Pharmacopoeia) 25g Microcrystalline Cellulose (Japanese Pharmacopoeia) 25g Methylcellulose (Japanese Pharmacopoeia) 1.5g Magnesium Stearate (Japanese Pharmacopoeia) 1 g The compound of Example 22, lactose, corn starch and microcrystalline cellulose are thoroughly mixed, granulated with a 5% aqueous solution of methylcellulose and carefully dried through a 200 mesh sieve. The dried granules are passed through a 200 mesh screen, mixed with magnesium stearate and pressed into tablets.

Formulation Example 10 Compound of Example 9 10 mg Macrogol 300 500 mg Distilled water for injection Appropriate amount 5 mg per ampoule for injection is prepared in the above mixing ratio.

Formulation Example 11 Compound of Example 8 (E) 0.4 g Polyethylene glycol 1000 30 g Polyethylene glycol 6000 50 g Refined water 20 g 2.5 g of suppositories are prepared per unit at the above mixing ratio.

Claims (1)

(57) [Claims] (1) General formula (In the formula, _one of R ₁ and R ₂ may have a group selected from the group consisting of an alkyl group, an alkoxy group, a halogenated alkyl group, a halogen atom, a hydroxyl group and a nitro group as a substituent on a phenyl ring. A benzyl group; the other is a monovalent group having at least one amino group and one carboxyl group in one molecule, and excluding the OH group from the carboxyl group of an amino acid having 2 to 20 carbon atoms. 2) -Deoxy-5-fluorouridine derivatives represented by the formula: