JP2951975B2 - Antitumor agent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to the following formula (I) An antitumor agent comprising a compound represented by the formula or a salt thereof as an active ingredient.

In the above formula, R is -Z-OPO (OH) ₂ or Wherein Z, Z ¹ and Z ² are each an alkylene group having 1 to 6 carbon atoms, and R ¹ and R ³ are each independently -COR ⁵ or R ² and R ⁴ are each independently -CO (CH ₂ ) _n NH-COR ⁵ ,
R ⁵ is a linear or branched alkyl group having 5 to 20 carbon atoms,
Q represents an alkyl group having 1 to 6 carbon atoms, and n represents an integer of 1 to 6.

The compounds of the formula (I) and salts thereof exhibit excellent antitumor activity and low toxicity, and are useful as antitumor agents.

<Prior art> Natural lipid A has mitogen activity, i.e., it has the effect of stimulating lymphocytes, transforming them into lymphocytes, promoting the increase of lymphoid cells and enhancing immune function, and the action of inducing tumor necrosis factor. However, it is promising as a prophylactic / therapeutic agent for various diseases caused by a decrease in immune function, for example, various infectious diseases, and an antitumor agent.

As a derivative of the natural lipid A, JP-A-59-4849
No. 7, 61-53295, 61-227586 and 63-183594 are known. These known compounds are still unsatisfactory in toxicity or antitumor activity, and are not sufficiently satisfactory from the viewpoint of practicality. Therefore, compounds having useful antitumor activity and reduced toxicity are desired.

<Problems to be solved by the invention> As a result of intensive studies to find an antitumor agent having excellent antitumor activity and low toxicity, the present inventors have completed the present invention.

<Constitution of the Invention> The present invention relates to an antitumor agent comprising a compound of the formula (I) or a salt thereof as an active ingredient.

The substituents in the compound of the formula (I) will be described in detail below.

The alkylene group means methylene, polymethylene, or methylene or polymethylene substituted with an alkyl group having 1 to 6 carbon atoms, and examples thereof include methylene, ethylene, propylene, trimethylene, ethylethylene, tetramethylene, and 2-methyltetramethylene. Examples include methylene and 2,3-dimethyltetramethylene. As the alkylene group, Z, Z ¹ and Z ² in the formula (I), those having 1 to 4 carbon atoms can be mentioned as preferred. Examples of the linear or branched alkyl group include methyl, ethyl, propyl, tertiary butyl, hexyl, nonyl, decyl, 3-ethylundecyl, 3-ethyl-4-methyl-tridecyl, tetradecyl and nonadecyl. I can give it. The alkyl group R ⁵ in the formula (I) has 8 carbon atoms.
To 14 are preferred.

The compound of the formula (I) has α and β isomers derived from the substituent OR, and an antitumor agent containing these isomers or a mixture thereof as an active ingredient is also included in the present invention.

Examples of the salt of the compound of the formula (I) include a phosphoric acid group or a carboxyl group and an organic amine such as triethylamine, pyridine, N-methylamine, N-methylglucamine, trisaminomethane, arginine, ethanolamine, ethylenediamine, diethanolamine or the like. Salts with inorganic bases such as ammonia, sodium, potassium, calcium, magnesium and the like can be mentioned.

The compound of the formula (I) can be produced by various methods, examples of which are shown below.

Wherein R ⁶ is -ZOPO (OR ⁸ ) or R ⁷ and R ⁸ represent protecting groups for a phosphono group that can be removed by catalytic reduction, R ⁹ represents a protecting group for a carboxyl group that can be removed by catalytic reduction, R ¹ , R ² , R ³ and R ⁴ are the same as above. The protecting group in the above reaction formula will be described. Examples of the carboxyl-protecting group which can be removed by catalytic reduction include a benzyl group which may have a substituent such as a halogen atom, a nitro group and a lower alkoxy group. or,
Examples of the phosphono group which can be eliminated by catalytic reduction include a phenyl group and a benzyl group which may have a substituent such as a halogen atom, a nitro group and a lower alkoxy group.

The compound of formula (II) is catalytically reduced in an inert solvent such as tetrahydrofuran, methanol, ethanol, acetic acid, water or a mixture thereof under a hydrogen gas atmosphere using a catalyst such as palladium black, palladium carbon, or platinum dioxide. The compound of the formula (I) can be produced by removing the protecting group and, if necessary, purifying using silica gel chromatography or the like. The reduction reaction is usually at room temperature (0 to 30 ° C) to 60 ° C
For 1 to 12 hours. The amounts of the solvent and the catalyst used are not particularly limited.

Adding the required amount of base to the resulting compound of formula (I),
Then, a salt of the compound of the formula (I) can be produced by using a precipitation method, a lyophilization method or the like.

The starting compound of the formula (II) shown above can be produced as follows.

[Wherein, R ¹⁰ represents a hydroxyl-protecting group, and R ¹ , R ² , R ³ , R
^4, R ⁶ and R are as defined above. In the above reaction formula, the protecting group for a hydroxyl group includes a protecting group removable by catalytic reduction such as a benzyl group which may have a substituent such as a halogen atom, a nitro group or a lower alkoxy group; a trichloroethoxycarbonyl group; Tertiary butoxycarbonyl group and the like can be mentioned.

Dissolving the compound of formula (IV) in a solvent that does not participate in the reaction containing hydrogen bromide gas, for example, a single or mixed solvent such as methylene chloride and acetic acid, and reacting at 0 ° C. to room temperature for several tens minutes to 24 hours. Can replace the acetyl group at the 1-position of the sugar of the present compound with bromine. The obtained bromine-substituted product in a dry solvent,
Preferably, it is dissolved in methylene chloride, chloroform or the like, and is used alone or as a mixture of mercuric cyanide, mercury bromide, silver carbonate, silver oxide, silver perchlorate or mercuric nitrate, etc., and dehydrated such as anhydrous calcium sulfate. The compound of formula (V) can be obtained by condensation with a compound of formula (III) in the presence of an agent for several hours to 2 days under reaction conditions such as room temperature to reflux.

The zinc dust addition dissolved or suspended in acetic acid compound of formula (V), the protecting group and R ¹⁰ at the 2 'position amino group by reaction can be desorbed. R ³
The compound of formula (II) can be prepared by using a condensation method commonly used in the field of peptide chemistry with -OH.

The elimination reaction of the protecting group is usually performed at room temperature for several tens minutes to 24 hours. Examples of the condensation reaction include a carbodiimide method, an eintop method, an active ester method and the like.

In the above elimination reaction, as for R ¹⁰ which is a protecting group for a hydroxyl group, trichloroethoxycarbonyl and trichlorotert-butoxycarbonyl are preferable.

Next, a method for producing the starting compounds of the above formulas (IV) and (III) will be described.

The compound of the formula (IV) can be prepared by a known method or JP-A-61-53295.
Can be manufactured according to the method disclosed in the above-mentioned publication.

The compound of the formula (III) can be produced by the following method.

[Wherein, X is a halogen atom, Y is a lower acyl group,
The trichloroethoxycarbonyl group or trichloromethyl tert-butoxycarbonyl, W is -ZOW ² acid or A, W ¹ is ZOW ⁴ or R ⁶ is -ZOPO (OR ⁸ ) ₂ or The, Y ¹ denotes a trichloroethoxycarbonyl or trichloro-tert-butoxycarbonyl, W ² is deacetylated, benzoyl, benzyl or p- chlorobenzyl, W ³ is an alkyl group or a catalytic reduction of 1 to 6 carbon atoms And W ⁴ represents hydrogen, benzyl or p-chlorobenzyl, and R ¹ , R ² , R ⁸ , R ⁹ , Z, Z ¹ and Z ² are the same as described above.

Compounds of formula (VII) can be obtained by reacting a compound of formula (VIa) with HOW in the presence of a Lewis acid or by reacting a compound of formula (VIb) with HOW, mercuric cyanide, silver carbonate, mercury bromide, It can be produced by condensation in the presence of silver perchlorate, mercuric nitrate or a mixture thereof. Also, the formula (VII)
Wherein W is -ZO-acetyl, the compound of formula (VIc) is converted to HO in the presence of hydrogen chloride, p-toluenesulfonic acid or the like.
It can be produced by reacting with ZOH followed by acetylation.

In the formula (VII), a compound in which Y is a lower acyl group is treated with a Mayerbain reagent, or a compound in which Y is trichloroethoxycarbonyl or trichlorotert-butoxycarbonyl in the presence of hydrochloric acid, acetic acid or the like. The compound of formula (VIII) is prepared by treating with zinc powder to release the amino group at the 2-position and then condensing it with R ¹ OH using an acid chloride method, a carbodiimide method, an eintop method or an active ester method. be able to.

In the formula (VIII), W is In a method and above in formula (VIII) the desorbed then the product acetyl group and an alkyl group is reacted with the presence X-R ⁹ of an organic amine such as triethylamine to compound by hydrolysis with sodium hydroxide The compound of formula (IX) can be produced by a method of hydrolyzing a compound other than the above using aqueous ammonia.

The compound of the formula (X) can be produced by protecting the 4- and 6-positions of the compound of the formula (IX) with isopropylidene according to a conventional method.

In the formula (X), W ¹ is -ZO-benzyl or -ZO-p
Compounds which are -chlorobenzyl is R ² OH and after condensation, the catalytic reduction was then triethylamine, 4-dimethylaminopyridine, method and formula is reacted with the presence X-PO (OR ⁸⁾ ₂ in an organic amine such as pyridine (X) In W ¹ The compound of formula (XI) can be produced by a method of condensing the compound with R ² OH.

The compound of formula (XIa) is hydrolyzed in aqueous acetic acid such as a 50-90% acetic acid aqueous solution or treated with p-toluenesulfonic acid in methanol, ethanol, water or a mixture thereof to give the compound of formula (IIIa) Can be produced.

Further, the compound (IIIa ′) in which R ⁶ is —ZOPO (OR ⁸ ) ₂ in the formula (IIIa) can be produced as follows.

That is, in the formula (VII), W is -ZO-acetyl or -ZO
The compound of formula (XII) can be produced by treating a compound in which Y is trichloroethoxycarbonyl or trichlorotert-butoxycarbonyl with -benzoyl, with aqueous ammonia. By protecting this with isopropylidene, a compound of the formula (XIII) can be obtained.
This can be condensed with XPO- (OR ⁸ ) _{2 in} the presence of an organic amine and then condensed with R ² OH to produce a compound of formula (XIV). The compound of formula (XVa) can be produced by removing isopropylidene in the same manner as described above. This is desorbed from Y ^{1 in the same} manner as described above, and then condensed with the formula R ¹ OH to give a compound of the formula (II
The compounds of Ia ') can be prepared. Also, the formula (XI
The compound of V) is eliminated from Y ^{1 in the same} manner as described above,
The compound of formula (XVb) can be produced by condensation with ¹ OH. By removing isopropylidene in the same manner as described above, the compound of the formula (IIa ') can be produced.

The compound of the formula (I) or a salt thereof is usually administered intravenously, subcutaneously or locally, and the dose is usually 30 mg per adult.
The range is from 0.1 to 0.1 mg / day, which may be appropriately increased or decreased according to the patient's symptoms.

The compound of the formula (I) or a salt thereof can be formulated into an injection together with additives such as a stabilizer and an isotonic agent by a known formulation technique.

<Effect of the Invention> The compound of the formula (I) and a salt thereof have excellent antitumor activity and very low toxicity.

 Therefore, the present invention is excellent as an antitumor agent.

Hereinafter, the present invention will be further described with reference examples and examples, but the present invention is not limited thereto.

Reference Example I-1 Step 1 2-acetoxyethyl 3,4,6-tri-O-acetyl-2-deoxy-2- (2,2,2-trichloroethoxycarbonylamino) -α-D-glucopyranoside 5.00 g To 2-deoxy-2- (2,2,2-trichloroethoxycarbonylamino) -D-glucose, add 5.0 ml of ethylene glycol and dioxane containing 0.5 ml of hydrogen chloride gas, and heat to 90 ° C. for 4 hours. Stir. The reaction solution is ice-cooled, 75 ml of pyridine is added, 30.6 g of acetic anhydride is added, and the mixture is stirred. After 20 minutes, return to room temperature and stir for another 16 hours. The reaction solution is poured into 350 ml of ice water and stirred, and the precipitated solid is collected by filtration and washed with water.

The obtained solid was dissolved in chloroform, 1N hydrochloric acid,
Wash sequentially with saturated saline and dry over anhydrous sodium sulfate. The residue obtained by evaporation of the solvent under reduced pressure is recrystallized from ethanol to give 4.96 g of the title compound as colorless prism-like crystals.

Melting point 138-140 ° C ▲ [α] ²⁵ _D ▼ + 74.0 ° (C 1.2, chloroform) Step 2 2-acetoxyethyl 3,4,6-tri-O-acetyl-2-deoxy-2-tetradecanoylamino −
α-D-Glucopyranoside 4.96 g of the compound obtained in Step 1 is dissolved in 60 ml of acetic acid, and 7 g of zinc powder is added little by little while stirring at room temperature, followed by stirring for 1 hour. The insolubles are removed by filtration, the solvent is distilled off under reduced pressure, toluene is added to the obtained residue, and the solvent is again distilled off under reduced pressure. The obtained residue is dissolved in dioxane, dioxane containing hydrogen chloride gas is added, and the solvent is distilled off under reduced pressure, followed by drying.

Dissolve the resulting oil in 70 ml of anhydrous methylene chloride,
Under ice cooling, 2.88 ml of N-methylmorpholine and 3.24 g of tetradecanoic acid chloride are added, and the mixture is stirred for 1 hour. After adding 10 ml of methanol and stirring at room temperature for 10 minutes, the mixture is diluted with chloroform and washed successively with 1 N hydrochloric acid and saturated saline. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was separated on a silica gel column (elution solvent; a mixed solution of benzene and ethyl acetate (initial ratio was 9/1 (v / v), then 1/1). (V / v)) to give 4.77 g of the title compound as a colorless oil.

Step 3 2-Hydroxyethyl 2-deoxy-2-tetradecanoylamino-α-D-glucopyranoside 4.77 g of the compound obtained in Step 2 is dissolved in 80 ml of anhydrous methanol, and 9 mmol of methanol of sodium methylate is added under ice-cooling. Add the solution and stir at room temperature for 30 minutes. After dissolving the precipitated insoluble matter by adding tetrahydrofuran, the solution is neutralized with a strongly acidic ion exchange resin Dowex-50 (H ⁺ type), and the resin is removed by filtration. The solvent of the filtrate was evaporated under reduced pressure, and the residue obtained was washed with ether and collected by filtration to give the title compound 3.
02 g are obtained as a white solid. Recrystallized with ethanol-water.

Melting point 158-160 ° C ▲ [α] ²⁵ _D ▼ + 82.1 ° (C 0.8, tetrahydrofuran: water = 4/1 v / v) Step 4 2-Hydroxyethyl 2-deoxy-4,6-
O-isopropylidene-2-tetradecanoylamino-
α-D-Glucopyranoside 0.87 g of the compound obtained in Step 3 was dissolved in 20 ml of dimethylformamide, and 0.62 g of 2,2-dimethoxypropane and 38 mg of p-toluenesulfonic acid-hydrate were added at room temperature to obtain 1.
Stir for 5 hours. Neutralize with 5% aqueous sodium bicarbonate and distill off the solvent under reduced pressure. Dissolve the residue in ethyl acetate,
The extract is washed successively with water and saturated saline, and dried over anhydrous sodium sulfate. The residue obtained by distilling off the solvent under reduced pressure is subjected to a silica gel column (elution solvent; initially, a mixed solution of chloroform and acetone).
19/1 v / v) and mixed chloroform and methanol
The mixture was purified by purification (19/1 v / v) to give 0.78 g of the title compound as a colorless viscous oil.

Step 5 2- (Diphenylphosphonoxy) ethyl 2-deoxy-4,6-O-isopropylidene-2-tetradecanoylamino-α-D-glucopyranoside 0.77 g of the compound obtained in Step 4 was added to 15 ml of anhydrous methylene chloride. And 0.48 g of diphenyl phosphorochloridate, 0.19 ml of pyridine and 0.30 g of dimethylaminopyridine are added under ice cooling, and the mixture is stirred for 1 hour. After returning to room temperature and stirring for 1 hour, add 0.17 g of diphenyl phosphorochloridate and stir for 30 minutes. After adding 3 ml of methanol to the reaction solution and stirring for a while, the solvent is distilled off under reduced pressure. The residue is subjected to a silica gel column (elution solvent: chloroform: acetone = 19/1, v
/ v) to give 0.81 g of the title compound as a colorless viscous oil.

Step 6 2- (diphenylphosphonoxy) ethyl-2-
Deoxy-3-O- (N-dodecanoylglycyl) -2
-Tetradecanoylamino-α-D-glucopyranoside 0.51 g of the compound obtained in Step 5 is dissolved in 5 ml of anhydrous methylene chloride, and 0.22 g of N-dodecanoylglycine, 44 m
g of dimethylaminopyridine and 0.18 g of dicyclohexylcarbodiimide are added, and the mixture is stirred for 30 minutes under ice-cooling, then returned to room temperature and stirred for 2 hours. The insolubles are removed by filtration, and the filtrate is washed with 1N hydrochloric acid, water and saturated saline in this order, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue obtained
A 90% acetic acid aqueous solution is added, and the mixture is stirred for 30 minutes while heating to 90 ° C. The solvent was distilled off under reduced pressure, the operation of adding toluene to the residue and distilling the solvent under reduced pressure was repeated twice, and the obtained residue was subjected to a silica gel column (elution solvent; initially, a mixed solution of chloroform and acetone (19/1, v / v ) And then purified with a mixture of chloroform and methanol (19/1, v / v)) to give the title compound 0.1.
51 g are obtained as a colorless oil.

▲ [α] ²⁵ _D ▼ + 46.2 ° (C 1.1, chloroform) NMR (CDCl ₃ ), δ (ppm): 0.88 (6H, t), 1.26 (S), 2.07 (2H, t), 2.27 (2H ,
t), 4.84 (1H, d), 5.18 (1 H, m), 7.2 to 7.4 (10 H, m) Reference Example I-2 Step 1 1,3- (diethoxycarbonyl) isopropyl 2-deoxy-3,4, 6-O-acetyl-2- (2,2,2-
Trichloroethoxycarbonylamino) -α-D-glucopyranoside 8.00 g of 1,3,4,6-tetra-O-acetyl-2-deoxy-2- (2,2,2-trichloroethoxycarbonylamino) -D-gluco A 25% hydrogen bromide-acetic acid solution cooled at room temperature is added to the pyranose, and the mixture is stirred for 1 hour. It is diluted with chloroform, washed successively with a 5% aqueous sodium hydrogen carbonate solution and saturated saline, and then dried over anhydrous magnesium sulfate.

The residue obtained by distilling off the solvent under reduced pressure was dissolved in 72 ml of anhydrous methylene chloride, and under ice-cooling, 8 g of anhydrous calcium sulfate, 4.12 g of silver perchlorate and 6.24 g of diethyl-suspended in 40 ml of anhydrous benzene were dissolved. Add 3-hydroxyglutarate.

After reacting at room temperature for 3 hours, 5% aqueous sodium hydrogen carbonate solution is added to neutralize. The insolubles are removed by filtration, and the filtrate is washed with water and dried over anhydrous magnesium sulfate.

The solvent is distilled off under reduced pressure, and the obtained residue is purified by a silica gel column (eluent; chloroform: acetone = 30: 1) to obtain 7.36 g of the title compound as an oil.

▲ [α] ²⁵ _D ▼ + 42.8 ° (C 0.7, chloroform) Step 2 1,3- (diethoxycarbonyl) isopropyl 2-deoxy-2-tetradecanoylamino-3,4,6
-Tri-O-acetyl-α-D-glucopyranoside 4.00 g of the compound obtained in step 1 is dissolved in 20 ml of acetic acid,
Add 3 g of zinc powder and stir at room temperature for 2 hours. The insoluble material is removed by filtration, and the solvent is distilled off. Dissolve the residue in benzene and
% Sodium hydrogencarbonate solution and saturated saline, and dried over magnesium sulfate. The solvent is distilled off and the residue is dissolved in 30 ml of tetrahydrofuran.

On the other hand, 1.64 g of tetradecanoic acid, 1.28 g of 1-hydroxybenzotriazole and 1.48 g of dicyclohexylcarbodiimide were added to 20 ml of tetrahydrofuran, reacted, and the insoluble material was removed by filtration. Stir. After filtering off the insoluble matter,
The filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent: a mixture of chloroform and acetone (10/1)) to give 3.78 g of the title compound as an oil from the desired fraction.

▲ [α] ²⁵ _D ▼ + 46.9 ° (C 0.16, chloroform) Step 3 1,3- (dibenzyloxycarbonyl) isopropyl 2-deoxy-2-tetradecanoylamino-
α-D-Glucopyranoside 1.80 g of the compound obtained in step 2 is dissolved in 30 ml of dioxane, and 10 ml of water is added. After cooling to 5 ° C., 15 ml of a 1N aqueous potassium hydroxide solution is added, and the mixture is stirred for 6 hours.

The residue obtained by concentrating to dryness under reduced pressure is suspended in 100 ml of dimethylformamide, and 1 ml of benzyl bromide is added. After stirring at 40 ° C. for 3 hours, most of the dimethylformamide is distilled off under reduced pressure, and the resulting residue is extracted with benzene.
The benzene layer is sequentially washed with a 5% aqueous citric acid solution, a saturated saline solution, a 5% aqueous sodium hydrogen carbonate solution and a saturated saline solution, and then dried over anhydrous magnesium sulfate.

The solvent was distilled off under reduced pressure and silica gel chromatography (elution solvent: chloroform: methanol: acetone = 50: 1: 5 then 50: 5)
1:15) to give 0.65 g of the title compound as a white waxy solid.

▲ [α] ²⁵ _D ▼ + 13.2 ° (C 0.51, chloroform) Step 4 1,3- (Dibenzyloxycarbonyl) isopropyl 2-deoxy-2-deoxy-4,6-O-isopropylidene-2-tetra Decanoylamino-α-D-
Glucopyranoside 0.64 g of the compound obtained in Step 3 is dissolved in 10 ml of acetone, and treated in the same manner as in Step 4 of Reference Example I-1 to obtain 0.54 g of the title compound as an oil.

▲ [α] ²⁵ _D ▼ + 3.3 ° (C 0.7, chloroform) Step 5 1,3- (Dibenzyloxycarbonyl) isopropyl 2-deoxy-3-O- (N-dodecanoylglycyl) -2-terola Decanoylamino-α-D-glucopyranoside 0.60 g of the compound obtained in Step 4 was reacted with N-dodecanoylglycine in the same manner as in Step 6 of Reference Example I-1.
% Aqueous acetic acid to give 0.63 g of the title compound as a waxy solid.

▲ [α] ²⁵ _D ▼ + 36.9 ° (C 1.3, chloroform) NMR (CDCl ₃ ), δ (ppm): 0.89 (6H, t), 1.26 (s), 2.1 to 2.3 (4H, m), 2.5
~ 2.9 (4H, m), 4.50 (1H, m), 4.97 (1H, d), 5.70 (1
H, m), 5.18 (4H), 7.40 (10H, s) The following formula (II) was obtained in the same manner as in the methods of Reference Examples I-1 to I-2.
A compound of Reference Example represented by IA) was obtained.

Reference Example II-1 Step 1 2- (diphenylphosphonoxy) ethyl 2-deoxy-6-O- [2-deoxy-4-O-diphenylphosphono-3-O- (N-dodecanoylglycyl)- 6-O- (2,2,2-trichloroethoxycarbonyl) -2- (2,2,2-trichloroethoxycarbonylamino) -β-D-glucopyranosyl] -3-O- (N-
Dodecanoylglycyl) -2-[(N-dodecanoyl-
N-methylglycyl) amino] -α-D-glucopyranoside 370 mg of 1-O-acetyl-2-deoxy-4-O-
Diphenylphosphono-3-O- (N-dodecanoylglycyl) -6-O- (2,2,2-trichloroethoxycarbonyl) -2- (2,2,2-trichloroethoxycarbonylamino) -D- Glucopyranose is dissolved in 2 ml of anhydrous methylene chloride and 6 ml of cooled 25% hydrogen bromide at room temperature.
Add acetic acid solution and stir for 1 hour. The mixture is diluted with chloroform, washed successively with ice-water, 5% aqueous sodium hydrogen carbonate and saturated saline, and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure and 344 mg of 2- (diphenylphosphonoxy) ethyl 2-deoxy-3-O- (N-dodecanoylglycyl) -2-[(N-dodecanoyl-N
-Methylglycyl) amino] -α-D-glucopyranoside was dissolved in 5 ml of anhydrous methylene chloride, and 0.5 g of activated calcium sulfate and 182 mg of mercuric cyanide were added.
C. and stir for 3 hours. The insolubles were removed by celite filtration, and the filtrate was washed successively with a 5% aqueous potassium iodide solution and saturated saline, and dried over anhydrous sodium sulfate. The residue obtained by distilling off the solvent under reduced pressure was subjected to a silica gel column (elution solvent, first chloroform: acetone = 10: 1, then chloroform: methanol = 50: 1, finally chloroform: methanol = 20: 1). Purify to give 599 mg of the title compound as an oil.

▲ [α] ²⁵ _D ▼ + 20.0 ° (C 1.0, chloroform) Step 2 2- (diphenylphosphonoxy) ethyl 2-deoxy-6-O- ｛2-deoxy-4-O-
(Diphenylphosphono) -3-O- (N-dodecanoylglycyl) -2-[(N-dodecanoyl-N-methylglycyl) amino] -β-D-glucopyranosyl} -3-
O- (N-dodecanoylglycyl) -2-[(N-dodecanoyl-N-methylglycyl) amino] -α-D-glucopyranoside 587 mg of the compound obtained in Step 1 is dissolved in 8 ml of acetic acid, and 0.6 g is dissolved.
And suspend it for 2 hours at room temperature. The insoluble matter is removed by filtration, washed with chloroform, and the solvent is distilled off under reduced pressure. The operation of dissolving the residue in toluene and distilling off the solvent is repeated three times, and the obtained residue is dissolved in chloroform. The chloroform layer was washed with 1N hydrochloric acid, 5% aqueous sodium hydrogen carbonate,
Wash sequentially with saturated saline. After drying over anhydrous sodium sulfate, the solvent is distilled off under reduced pressure to obtain an oil.

On the other hand, 122 mg of N-dodecanoyl-N-methylglycine was dissolved in 3 ml of anhydrous tetrahydrofuran, and 77 mg of 1-
Hydroxybenzotriazole and 103 mg of dicyclohexylcarbodiimide are added and stirred. After 30 minutes, the mixture is returned to room temperature and stirred for 3 hours, and the precipitated crystals are removed by filtration.
The oily substance is dissolved in 5 ml of anhydrous methylene chloride, and the filtrate is added under ice cooling. Return to room temperature and stir for 1 hour and 30 minutes. The reaction solution is diluted with chloroform, washed with 1N hydrochloric acid, dried over anhydrous sodium sulfate, and the solvent is distilled off. The obtained residue was purified by a silica gel column (elution solvent; first, chloroform: acetone = 10: 1, then, chloroform: methanol = 50: 1, finally, chloroform: methanol = 20: 1), and 445 mg of the title compound was obtained. Obtained as an oil.

▲ [α] ²⁵ _D ▼ + 19.2 ° (C 1.0, chloroform) Step 3 2-phosphonoxyethyl 2-deoxy-6
O- {2-deoxy-3-O- (N-dodecanoylglycyl) -2-[(N-dodecanoyl-N-methylglycyl) amino] -4-O-phosphono-β-D-glycopyranosyl} -3- O- (N-dodecanoylglycyl) -2
-[(N-dodecanoyl-N-methylglycyl) amino] -α-D-glucopyranoside 424 mg of the compound obtained in Step 2 is dissolved in a mixture of 50 ml of tetrahydrofuran and 2.5 ml of water, 0.2 g of platinum dioxide is added, and a hydrogen stream is added. Stir for 2 hours below. The catalyst is filtered and the residue is washed with a mixture of chloroform: methanol: water (lower layer of 8: 3: 1). The filtrate and the washing solution are combined, and the solvent is distilled off under reduced pressure.
The obtained residue is thin-layer chromatography [elution solvent; developed with chloroform: methanol: water = 6: 4: 0.7 (v / v)]
And purified by strong acid ion exchange resin Dowex 50 (H ⁺
(Manufactured by Dow Chemical Co., Ltd.).

The residue obtained by distilling off the solvent under reduced pressure is suspended in dioxane and freeze-dried to obtain 204 mg of the title compound as a white powder.

Melting point 165-170 ° C (slowly becomes colored candy) ▲ [α] ²⁵ _D ▼ + 4.6 ° (C 0.7, chloroform: methanol = 3: 1 (v / v) ▲ IRν ^KBr _max ▼ cm ^-1 ; 3400,2930,2850,1750,1675,1650NM
R (CDCl ₃ −CD ₃ OD), δ (ppm): 0.90 (12H, t), 1.30 (s), 2.29 (4H, m), 2.44 (4H,
t), 2.94 and 3.11 (total 6H, each s), 4.84 (1H, d), 5.18 (1
H, m), 5.34 (1H, m) A part of the title compound was converted to chloroform-methanol (3:
Dissolve the mixture in 1), adjust the pH to about 9 with triethylamine, and concentrate under reduced pressure. The obtained residue is dissolved in a 0.1% aqueous solution of triethylamine, filtered through a Millipore filter, and freeze-dried to obtain a triethylamine salt of the title compound as a white powder.

The compound of Reference Example represented by the following formula (IA) was produced in the same manner as in Reference Example II-1.

Example 1 Methylcholanthrene-induced 2 × 10 ⁵ fibrosarcoma cells (Meth A) of BALB / C mice were transplanted into the flank skin of syngeneic BALB / C mice (7 mice / group). The triethylamine salt of the compound of Reference Example II was dissolved or suspended in an aqueous 0.1% triethylamine solution (v / v%), and a solution prepared at 500 μg / ml was prepared on the 7th day after transplantation at 100 μg / mouse.
On the 21st and 21st days, the compound was administered into the tail vein, and the antitumor activity was examined 21 days later.

 The results are shown in Table 1.

As is clear from the above table, it was confirmed that the compound of the formula (I) and a salt thereof have excellent antitumor activity.

Example 2 A solution prepared by dissolving or suspending the triethylamine salt of the compound of Reference Example II in a 5% dextrose solution containing 0.1% triethylamine in a group of three NZW male rabbits at a concentration of 1000 μg / ml was prepared.
0.5 ml / kg (body weight) was administered to the ear vein for 3 days every day. The determination was made based on the number of dead rabbits / the number of rabbits used 24 hours after the final administration. The results are shown in Table 2.

As can be seen from the above table, the compound of formula (I) and its salt were very low toxic. Further, blood was released under pentobarbital anesthesia 24 hours after the administration of the compound of Reference Example II,
Histological examination of the liver, spleen, kidney, heart, and lung was performed, and the liver, spleen, kidney, heart, and lung were examined. No changes were considered to be caused by the compound of Reference Example II.

Therefore, it was confirmed that the compound of the formula (I) and a salt thereof were excellent in safety.

(56) References JP-A-63-183594 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 31/70 C07H 15/04 CA (STN) REGISTRY ( STN)

Claims (1)

(57) [Claims] (1) Expression An antitumor agent comprising a compound represented by the formula or a salt thereof as an active ingredient. In the above formula, R is -Z-OPO (OH) ₂ or Wherein Z, Z ¹ and Z ² are each an alkylene group having 1 to 6 carbon atoms, and R ¹ and R ³ are each independently -COR ⁵ or R ² and R ⁴ are each independently -CO (CH ₂ ) _n NH-COR ⁵ ,
R ⁵ is a linear or branched alkyl group having 5 to 20 carbon atoms,
Q represents an alkyl group having 1 to 6 carbon atoms, and n represents an integer of 1 to 6.