JPH0631303B2 - Novel 6-Substituted Aldohexopyranos Derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention has the general formula (1) [Wherein R is a 1-substituted indole, isatin, indoline, 5-fluorouracil, pyrrolidine, or imidazole residue, R ₁ is hydrogen, an acetyl group, a benzyl group, or a benzoyl group, and R ₂ is a hydroxyl group. , Acetoxy group, benzyloxy group, benzoyloxy group, amino group, N-acetylamino group or N-benzoylamino group, and R ₃ represents hydrogen or a methyl group. ] The novel 6-position substituted aldohexopyranose derivative represented by the following and an acid addition salt of the above derivative in which R ₂ is an amino group.

Examples of the acid used when forming the acid addition salt include inorganic acids and organic acids.

These series of novel 6-substituted aldhexopyranose derivatives are a group of compounds useful for medicines, agricultural chemicals and the like. For example, it can be used as an antitumor agent or an immunostimulant as a medicine.

Conventionally, the sugar moiety of a physiologically active nucleic acid-related compound is DNA or RN.
As represented by A, it is limited to ribose and deoxyribose, which are natural pentoses. Most of these nucleobases are linked to the active 1-position of the sugar moiety. A group of compounds in which an adenine derivative is slightly added to the 5-position of ribose or deoxyribose is known as "reversed" nucleic acid. (J. Heterocycl.Chem :, 3 (4), 485〜
9, 1966 ) I have no memory of its activity. on the other hand,
The hexoses (glucose, mannose, galactose, etc.), which are more widely present in nature, have been shown to exist for the first time in recent years from mouse cancer cells.
Its presence and role are drawing attention as shown in glucose contained in A).

Many studies on this hexose sugar derivative have also been carried out by adding a nucleobase at the 1-position of the sugar, and there are few remarkable examples of physiological activity. However, we utilize the specificity of the conformation by using the 6-position of this hexose and the glycoside as a glycosyl donor, and its affinity as a carrier for permeation through biological membranes. I thought about that.

As compounds to be added to the 6-position of hexoses, compounds that have a simple structure without using the so-called nucleic acid bases found in nature, for example, various investigations on a group of compounds that can pass through biological membranes and react with cancer cells etc. did. As a result, a series of novel aldohexopyranose derivatives in which indole, imidazole, and their derivatives, which show almost no antitumor activity by themselves, are added to the 6-position of hexose, They have found that they exhibit an immune promoting action and the like.

That is, the present invention relates to a 6-substituted aldohexopyranose (glucose, galactose), an aldohexopyranosylamine (glucoseamine) derivative, which is a novel compound group having useful applications, and a method for producing them.

About the production of these compounds, the following methods were used. First, various 6-deoxy-s that are synthetic intermediates
The 6-halogeno aldohexopyranose derivative was synthesized according to a generally known method (as a representative reference example, J. Amer. Chem. Soc., 70, 2785 (1948); Ann. Chem., 8
25 (1980); Bull. Chem. Soc. Japan, 46, 3165 (1973);
55-122796 and the like), which can be represented by, for example, the following reaction formulas I to IV.

Reaction formula I Reaction formula II Reaction formula III Reaction formula IV However, the symbols in the formulas have the following meanings.

Ac: acetyl, Tr: trityl, Ms: methanesulfonyl,
Bn: benzyl, Bz: benzoyl, Ts: p-toluenesulfonyl, Py: pyridine, DMF: dimethylformamide, which can be obtained by such a method as the general formula (2). [Wherein X is chlorine, bromine, or iodine, R ₁ ′ is an acetyl group, a benzyl group, or a benzoyl group, and R ₂ ′ is
Is an acetoxy group, a benzyloxy group, a benzoyloxy group, an amino group, an N-acetylamino group, an N-benzoylamino group or an N-p-methoxybenzylideneamino group, and P ₂ ′ is a methyl group, an acetyl group or a benzyl group. Or represents a benzoyl group. A 6-deoxy-6-halogeno-aldhexopyranose derivative or an acid addition salt of the above derivative in which R ₂ is an amino group and an indole, isatin, indoline, 5-fluorouracil, pyrrolidine, or imidazole One of them is reacted in the presence of a phase transfer catalyst under the condition of an aqueous alkali solution, or is reacted in the presence of an alkali metal hydride in an aprotic solvent and then subjected to usual post-treatment operations and purification. The 6-position substituted aldohexopyranose derivative represented by the general formula (1) can be produced by deprotecting

Here, as the phase transfer catalyst, a quaternary salt such as a quaternary ammonium salt, a quaternary phosphonium salt, or a quaternary pyridinium salt is used, but a quaternary ammonium salt is preferably used. Examples include triethylbenzylammonium halide, cetyltrimethylammonium halide, cetyldimethylbenzylammonium halide and the like. The amount used is about 0.1 to 10 mol% with respect to the substrate (2). A concentrated aqueous solution of sodium hydroxide, potassium hydroxide or the like is used as the alkaline aqueous solution, but a preferable concentration is 30 to 70% aqueous solution. 1-1 for substrate (2)
Use an excess of 5 equivalents. In the reaction using a phase transfer catalyst, the organic phase is not particularly limited as long as it is an organic solvent inert to the reaction and capable of extracting the 6-substituted aldohexopyranose derivative of the product, but preferably benzene, toluene, etc. The aromatic hydrocarbons, halogenated hydrocarbons such as methylene chloride, ethers such as dimethoxyethane and diethoxymethane, etc. are used alone or in combination.

The reaction temperature is not particularly limited, but usually from room temperature to the reflux temperature in the reaction system is used. The reaction using the phase transfer catalyst is usually completed in 3 to 10 hours.

When a phase transfer catalyst is used here, the reaction rate and the yield are improved, and in addition, N-substituted products of a base group represented by indole are preferentially obtained.

Further, as the aprotic solvent, N, N-dimethylformamide, dimethylsulfoxide, hexamethylphosphoric triamide and the like are preferably used. It is desirable to use these aprotic solvents after dehydration treatment by a conventional method.

Sodium hydride as an alkali metal hydride,
Lithium hydride, potassium hydride and the like are used. The amount used is 1.0 to 1.2 equivalents relative to one kind of indole, isatin and the like.

The reaction temperature is room temperature to about 200 ° C. The reaction is usually completed in 5 to 20 hours. The progress of these reactions can be detected by thin layer chromatography plate, gas chromatography or the like. The product is purified by silica gel column or silica gel thin layer chromatography or the like. On the other hand, as a starting material, an indole derivative of the 6-position substituted aldohexopyranose derivative represented by the general formula (1) can be produced from the uronic acid derivative represented by the following formula (3) or (4).

[In the formula, Z represents a benzyl group or a benzoyl group. That is, these uronic acid derivatives are reacted with indoline in the presence of a condensing agent such as dicyclohexylcarbodiimide (DCC) to obtain a compound of formula [In formula, Z is as above-mentioned. ] Indoline amide represented by the following formula and further oxidized to indole amide. Thereafter, reduction with diborane or the like is carried out to convert the carbonyl group into a methylene group, whereby a 6-position substituted aldohexopyranose-indole derivative can be produced. In this case, depending on the target compound, the protecting group can be deprotected or further converted into another protecting group to obtain the target compound. However, the target compound is a compound represented by the general formula (1) in which R ₁ is a benzyl group and R ₂ is a benzyloxy group,
Alternatively, when R ₁ is a benzoyl group and R ₂ is a benzoyloxy group, a compound represented by the general formula (4) is used as a starting material, and the target compound is obtained as it is without deprotecting the protecting group. You can These can be represented by, for example, the following reaction formulas V to VI.

Reaction formula V (However, DDQ represents dicyclodicyanoquinone.) Reaction formula VI The various novel compounds synthesized as described above have a good tumor cell growth inhibitory action on tumor cells such as P388. For example, the effect on P388 mouse leukemia cells was tested by the following method. That is, P38
8 Leukemia cells were taken from the abdominal cavity of DBA / 2NCrj mouse, and were collected from the abdominal cavity of the mouse.
Rosewell with 2-hydroxyethyl disulfide
Park Memorial Institute Medium 1640 (hereinafter RPMT
1640 medium) and the cell concentration is 1.0 x 1
It was adjusted to 0 ⁵ cells / ml. On the other hand, RPMI 16
It was adjusted to 20,2,0.2 μg / ml in 40 medium. 0.5 ml each of the cell suspension and the drug solution was placed in a culture test tube, and the final cell concentration was 5.0 × 10 ⁴ cells / ml and the drug concentration was 10,1,0.1 μg / ml. The culture tube was covered with an aluminum cap and cultured at 37 ° C. for 48 hours in a humid atmosphere containing 5% carbon dioxide.

Count the number of cells before and after culturing with a Coulter counter.
Calculate the number of cells grown over time, take the ratio to the control,
It was defined as the growth rate. The growth rate was subtracted from 1 to obtain the growth inhibition rate. I.e. These results indicate that indole, isatin derivative, etc., which by themselves show no antitumor activity or only weak activity, have a remarkable anti-tumor effect on mouse leukemia as a 6-position substitution product of aldohexopyranose derivative. It is a new finding showing that it has usefulness that could not have been predicted until now.

The antibody-producing action of these novel compounds (general formula (1)) was determined by the following test method, for example.

In vivo antibody production [Method] Each compound was intraperitoneally administered to 5 ICR mice (♂, 8 weeks old) at 100 mg / kg each day for 3 days, and 10% sheep red blood cells (SRBC) were administered on the 3rd day. Immunization was performed by 0.2 ml intravenous injection.

On the 5th day of SRBC sensitization, the number of antibody-producing cells per 10 ₆ nucleated cells of mouse spleen was calculated by the method of Cunningham et al.

[Results] The antibody production of each of the compounds shown in the table below was statistically significantly enhanced as compared with the control.

Specific examples of the compound of the present invention include the following compounds.

Next, the present invention will be described in more detail with reference to Examples, but these are only a part and the present invention is not limited to these.

Example 1 1,2: 3,4-di-O-isopropylidenegalacturonic acid-
Water salt 4.40 g (15 mmol), indoline 1.87 g (15.8
Mmol) was dissolved in 80 ml of methylene chloride and cooled to 0 ° C. A solution of 3.09 g (15 mmol) of dicyclohexylcarbodiimide in methylene chloride (5 ml) was added dropwise, and the mixture was left at room temperature overnight. 100 ml of ethyl acetate was added and the precipitated crystals were separated. The solution was concentrated and then crystallized from 10 ml of isopropanol to obtain 3.481 g of indoline amide of 1,2: 3,4-di-O-isopropylidenegalacturonic acid. Yield 6
It was 1.8%. NMR peak is δ (CDCl ₃ , ppm) = 1.33
(6H, singlet), 1.47 (3H, singlet), 1.55 (3H, singlet), 3.07 (2H, triplet, J = 12Hz), 4.0 to 5.0 (6H, multiplet), 5.63 (1H, doublet, J = 7.5) Hz), 6.7
It was ~ 7.4 (3H, Maruplet) and 8.1-8.4 (1H, multiplet).

Example 2 1.489 g (3.97 mmol) of indoline amide of 1,2: 3,4-di-O-isopropylidenegalacturonic acid obtained in Example 1 and 1.36 g (6 mmol) of dichlorodicyanoquinone were dissolved in 20 ml of xylene. And reacted under reflux for 3.5 hours. After separating the solid, the liquid was concentrated and purified by a silica gel column to obtain 1.039 g of indole amide of 1,2: 3,4-di-O-isopropylidenegalacturonic acid. Yield 70.
It was 1%.

The NMR peak δ is (CDCl ₃ , ppm) = 1.33 (6H, singlet), 1.47 (3H, singlet), 1.55 (3
H, singlet), 4.40 (1H, doublet, J = 6)
Hz), 4.70 (2H, singlet), 4.93 (1H, singlet), 5.70 (1H, doublet, J = 6Hz),
6.53 (1H, doublet, J = 5Hz), 7.2 to 7.7 (3
H, multiplet), 7.95 (1H, doublet, J =
5 Hz) and 8.4 to 8.6 (1H, multiplet).

Example 3 0.551 g (1.48 mmol) of indole amide of 1,2: 3,4-di-O-isopropylidenegalacturonic acid obtained in Example 2 is dissolved in 7 ml of tetrahydrofuran and cooled to 0 ° C. 6 ml of a THF solution of diborane (0.5 molar concentration) was added dropwise, and the mixture was reacted at 0 ° C. for 30 minutes and at room temperature for 2 hours. Post-treatment was carried out by a conventional method and purified by silica gel thin layer chromatography to obtain 6-deoxy-6-indolyl-1,2: 3,4-di-O.
0.368 g of isopropylidene galactopyranose was obtained. The yield was 69.4%.

The NMR peak δ is (CDCl ₃ , ppm) = 1.10 (3H, singlet), 1.20 (3H, singlet), 1.43 (6)
H, singlet), 3.47 (1H, doublet, J = 1
2Hz), 3.9-4.5 (4H, multiplet), 5.47
(1H, doublet, J = 7.5Hz), 5.87 (1H, doublet, J = 12Hz), 6.38 (1H, doublet, J
= 5Hz), 7.0-7.2 (2H, multiplet), 7.15
(1H, doublet, J = 5Hz) and 7.4 to 7.8 (2H, multiplet). IR is ν (neat, cm ^-1 ) = 300
It was 0,1460,1380,1260,1220,1080,1010,910,740.

Example 4 1,2: 3,4-di-O-isopropylidenegalacturonic acid-
6-Deoxy-6-indolyl-1, was carried out in the same manner as in Example 1, Example 2 and Example 3, except that 1,2,3,4-tetrabenzoylgalacturonic acid was used in place of the water salt.
2,3,4-Tetrabenzoylgalactopyranose was obtained.

The NMR peaks are δ (CDCl ₃ , ppm) = 4.2 to 5.0 (4H, multiplet), 5.8 to 6.1 (3H, multiplet),
6.4 to 6.5 (1H, multiplet), 6.5 to 8.1 (25
H, multiplet).

Example 5 6-deoxy-6-indolyl-obtained in Example 4
To 161 mg (0.231 mmol) of 1,2,3,4-tetrabenzoylgalactopyranose was added 5 ml of a 1% sodium hydroxide solution in methanol, and the mixture was stirred at room temperature for 3.5 hours. After concentration, the product was purified by silica gel thin layer chromatography to give 6-deoxy-6-
47 mg of indolyl-galactose was obtained. Yield 72.8%
Met.

The NMR peak is δ (d ₄ -methanol, ppm) = 4.0-5.1
(10H, multiplet), 6.35 (1H, broad), and 6.8 to 8.1 (6H, multiplet).

Optical rotation is ▲ [α] ²⁰ _D ▼ + 8.6 ° (C8.88, methanol)
Met.

Example 6 1,2: 3,4-di-O-isopropylidenegalacturonic acid-
Methyl 6-deoxy-6-indolyl-2 was prepared by the same procedure as in Example 1, Example 2 and Example 3 except that 2,3,4-tribenzoylglucopyranosideuronic acid was used in place of the water salt. 3,4-Tribenzoylglucopyranoside was obtained.

The NMR peaks are δ (CDCl ₃ , ppm) = 2.87 (3H, singlet), 4.0 to 4.6 (3H, multiplet), 5.0 to
The values were 6.3 (4H, multiplet), 6.35 (1H, doublet, J = 8Hz), and 6.8 to 8.0 (20H, multiplet).

Example 7 Methyl 6-deoxy-6-indolyl-2,3,4-tribenzoylglucopyranoside 344 mg obtained in Example 6
To (0.568 mmol), 5 ml of a 1% sodium hydroxide solution in methanol was added, and the mixture was stirred at room temperature for 1 hour. After concentration, the residue was purified by silica gel thin layer chromatography to obtain 182 mg of methyl 6-deoxy-6-indolylglucopyranoside. 100% yield
Met.

The NMR peak is δ (d ₄ -methanol, ppm) = 2.90 (3
H, singlet), 3.0-4.9 (10H, multiplet), 6.33 (1H, doublet, J = 5Hz), 6.8-
It was 8.1 (5H, multiplet).

IR was ν (KBr disc, cm ¹ ) = 3370,1060,745.

Example 8 To 662 mg (4.5 mmol) of isatin, 9 ml of hexamethylphosphoric triamide was added, and 216 mg (4.5 mmol) of 50% sodium hydride was further added. 1 at room temperature
After stirring for 6 hours, 6-deoxy-6-iodo-1,2: 3,4-
Di-O-isopropylidene galactopyranose 1.11 g
(3 mmol) was added, and the mixture was reacted at 130 ° C. for 14.5 hours. After cooling, water was added and the mixture was extracted with ethyl acetate. After drying with mirabilite, ethyl acetate was distilled off under reduced pressure, and the residue was purified by silica gel thin layer chromatography to obtain 666 mg of a substituted isatin at the 6-position of 1,2: 3,4-di-O-isopropylidenegalactopyranose. The yield was 57%.

The NMR peaks are δ (CDCl ₃ , ppm) = 1.23 (3H, singlet), 1.33 (3H, singlet), 1.37 (3H, 3H, singlet).
Singlet), 1.50 (3H, singlet), 3.8-
The values were 4.8 (6H, multiplet), 5.40 (1H, doublet, = 6Hz), and 6.9 to 7.8 (4H, multiplet).

The IR was ν (KBr disk, cm ⁻¹ ) = 1740, 1615, 1170.

Example 9 80 mg was added to 559 mg of the 6-position isatin substitution product obtained in Example 8.
% Trifluoroacetic acid (1 ml) was added, and the mixture was left at room temperature for 1.5 hours. After concentration under reduced pressure, ethanol was added and the mixture was concentrated again. As a result, red crystals of the 6-position isatin-substituted galactose were red.
89 mg were obtained. The yield was 100%.

The NMR peak is δ (d ₆ -DMSO, ppm) = 3.3 to 4.3 (7H,
Multiplet), 4.20 (4H, Singlet), 7.0
˜7.8 (4H, multiplet).

IR is ν (KBr disk, cm ^-1 ) = 3300,1740,1730,161
It was 0,1460.

Examples 10 to 11 Under the same conditions as in Example 8, the reaction was carried out using N, N-dimethylformamide as a solvent. After the same post-treatment, the residue was purified by silica gel column chromatography and then the deprotecting group operation was performed, or the target product was obtained as it was. The reaction starting materials, yields and physical properties are shown in Table I.

Example 12 4 ml of benzene and 2 ml of 2,2-dimethoxyethane were added to 109 mg (0.93 mmol) of indole and further cetyl dimethylbenzyl ammonium chloride 1 was used as a phase transfer catalyst.
8.4 mg (0.047 mmol) was added. A 30% aqueous sodium hydroxide solution (0.37 g NaOH dissolved in 0.86 g of water) was added thereto, and the mixture was stirred at room temperature for 2 hours, then heated to 50 ° C. and stirred for 1 hour. Here, a solution prepared by dissolving 450 mg (0.84 mmol) of N- (p-methoxybenzylidene) -triacetyl-6-deoxy-6-iodoglucosamine in 3 ml of benzene and 1.5 ml of 2,2-dimethoxyethane was vigorously stirred. After dropping into the above reaction solution at 50 ° C. for about 1 hour, the reaction was carried out at the same temperature for 3 hours.

After cooling, excess water was added and the mixture was extracted twice with ethyl acetate. The organic layer was washed with saturated brine and dried over magnesium sulfate, and the ethyl acetate layer was evaporated under reduced pressure. The residue was purified twice on a silica gel column to give 81 mg of N- (p-methoxybenzylidene) -triacetyl-6-deoxy-6-indolylglucosamine. The yield was 18.5%.
To this product was added 1 ml of a 2% sodium methylate methanol solution, and the mixture was stirred at room temperature for 3 hours and then treated with an acidic ion exchange resin to obtain 6 mg of 6-deoxy-6-indolyl-glycoseamine. When a 5N hydrochloric acid solution was added to an acetone solution of this product, 6-indolyl-glucosamine hydrochloride was quantitatively obtained.

Examples 13 and 14 The same procedure as in Example 8 or Example 12 was carried out. The structural formulas of the products together with their physical properties are shown in Table II.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Shinya Morisada 4-2-1 Takashi, Takarazuka-shi, Hyogo Sumitomo Kagaku Kogyo Co., Ltd. (72) Inventor Yu Fukui 4-2-1 Takashi, Takarazuka-shi, Hyogo No. Sumitomo Kagaku Kogyo Co., Ltd. (72) Inventor Keiichi Kadota 4-2-1 Takashi, Takarazuka-shi, Hyogo Sumitomo Kagaku Kogyo Co., Ltd. (72) Inventor Takao Okuda 4-2-1 Takashi, Takarazuka-shi, Hyogo No. Sumitomo Chemical Industry Co., Ltd.

Claims (3)

[Claims] 1. A general formula [In the formula, R represents a 1-substituted indole, isatin, indoline, 5-fluorouracil, pyrrolidine, or imidazole residue, R ₁ represents hydrogen, an acetyl group, a benzyl group, or a benzoyl group, and R ₂ represents a hydroxyl group. , Acetoxy group, benzyloxy group, benzoyloxy group, amino group, N-acetylamino group or N-benzoylamino group, and R ₃ represents hydrogen or a methyl group. ] The 6-position substituted aldohexopyranose derivative shown by these, or the acid addition salt of the said derivative whose R < ₂ > is an amino group. 2. General formula Wherein, X is chlorine, bromine or iodine,, R _'1 is an acetyl group, a benzyl group or a benzoyl group, R' ₂ is an acetoxy group, a benzyl group, a benzoyloxy group, an amino group, N- acetylamino group, N- benzoylamino group or N-p-methoxy benzylidene amino group,, R _'3 represents a methyl group, an acetyl group, a benzyl group or a benzoyl group. ] 6-deoxyhalogeno aldohexopyranose compound represented by or an acid addition salt of the above compound in which R ₂ is an amino group, and one of indole, isatin, 5-fluorouracil, indoline, pyrrolidine and imidazole
After reacting the seed with a phase transfer catalyst in the presence of an aqueous alkaline solution or in the presence of an alkali metal hydride in an aprotic solvent, the protecting group may or may not be deprotected. Characteristic general formula [In the formula, R represents a 1-substituted indole, isatin, indoline, 5-fluorouracil, pyrrolidine, or imidazole residue, R ₁ represents hydrogen, an acetyl group, a benzyl group, or a benzoyl group, and R ₂ represents a hydroxyl group. , Acetoxy group, benzyloxy group, benzoyloxy group, amino group, N-acetylamino group or N-benzoylamino group, and R ₃ represents hydrogen or a methyl group. ] The 6-position substituted aldohexopyranose derivative shown by these, or the manufacturing method of the acid addition salt of the said derivative whose R < ₂ > is an amino group. 3. General formula Or [In the formula, Z represents a benzyl group or a benzoyl group. ] By reacting the uronic acid derivative of 2 with indoline Or [In formula, Z is as above-mentioned. ] Indoline amide represented by the above, and further reduced to indolamide by oxidation, and then reduced, and then the protecting group is not deprotected, or deprotected, or further converted to another protecting group, formula [In the formula, R ₁ represents hydrogen, an acetyl group, a benzyl group, or a benzoyl group, R ″ ₂ represents a hydroxyl group, an acetoxy group, a benzyloxy group, or a benzoyloxy group, and R ₃ represents a hydrogen or a methyl group.] A method for producing a 6-substituted aldohexopyranose derivative represented by: