JP3051213B2 - Maltooligosaccharide derivatives, their synthetic intermediates and their production - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel maltooligosaccharide derivative and a method for producing the same, and more particularly to a maltooligosaccharide derivative having two amino groups at both ends and a method for synthesizing the same.

[0002]

2. Description of the Related Art Maltooligosaccharides and derivatives thereof are α-
Amylase, β-amylase, glucoamylase and α-
Many carbohydrases such as glucosidases are important as substrates for diagnostics in research and clinical use, and their need is increasing as their mechanism of action is elucidated.

The present inventors have previously developed a method for synthesizing maltooligosaccharides having a degree of polymerization of 6 to 8 or a derivative thereof with high selectivity (see Japanese Patent Application Laid-Open No. 2-235898).

[0004]

By the way, unlike rigid cyclic oligosaccharides such as cyclodextrins, if malto-oligosaccharides having a flexible and closable ring structure via a metal coordination bond can be constructed, heavy metal It is expected to be applicable to anticancer agents and the like by forming a complex with a scavenger and platinum ions.

Accordingly, an object of the present invention is to provide a maltooligosaccharide derivative having a metal coordination ability at both ends, which is essential for the construction of a flexible and closable ring structure via a metal coordination bond, and a method for synthesizing the same. is there.

[0006]

Means for Solving the Problems The present inventors disclosed in Japanese Patent Laid-Open No.
Using the maltooligosaccharide derivative disclosed in Japanese Patent No. 235898 as a starting material, the sugar residues at both reduced and non-reduced terminals were chemically modified to successfully synthesize a maltooligosaccharide derivative having two amino groups at both terminals. Thus, the present invention has been completed.

That is, the present invention provides: (1) a maltooligosaccharide derivative represented by the formula (I):

[0008]

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Wherein R represents a hydroxyl group which may be independently protected, and n represents an integer of 4 to 6. (2) a synthetic intermediate of a maltooligosaccharide derivative represented by the formula (II):

[0010]

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(Wherein, R ¹ and R ² each independently represent an alkyl group or an aryl group which may be substituted, R represents each independently a hydroxyl group which may be protected, and n represents 4
(3) an intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (III):

[0012]

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(Wherein, R ¹ and R ³ each independently represent an optionally substituted alkyl group or aryl group, R represents each independently an optionally protected hydroxyl group, and n represents 4
(4) an intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (IV):

[0014]

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(Wherein, R ¹ and R ⁴ each independently represent an optionally substituted alkyl group or an aryl group, R represents each independently an optionally protected hydroxyl group, and n represents 4
(5) An intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (V).

[0016]

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(Wherein R ⁴ and R ⁵ each independently represent an optionally substituted alkyl or aryl group, R represents each independently an optionally protected hydroxyl group, and n represents 4
(6) an intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (VI), and

[0018]

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Wherein R represents a hydroxyl group which may be independently protected, and n represents an integer of 4 to 6. (7) Starting from a maltooligosaccharide derivative,
It is intended to provide a method for synthesizing a maltooligosaccharide having two amino groups at both ends by chemically modifying the sugar residues at both ends of the non-reducing terminal.

R in the above formula represents a hydroxyl group which may be protected, and examples of the protecting group include an alkyl group. R is preferably a hydroxyl group or a linear or branched alkyl group having 1 to 10 carbon atoms. R ¹ and R ²
Examples of the group include an alkyl group and an aryl group. Among them, a linear or branched alkyl group or a phenyl group having 1 to 6 carbon atoms, and a substituted phenyl group are preferable.

Examples of R ³ include an alkyl group and an aryl group.
A linear or branched alkyl group or phenyl group of 0, or a substituted phenyl group is preferred. R ⁴ and R ⁵
Examples thereof include an alkyl group and an aryl group, and among them, a linear or branched alkyl group or phenyl group having 1 to 6 carbon atoms, and a substituted phenyl group are preferable.

Hereinafter, typical reaction examples for synthesizing the maltooligosaccharide derivative of the present invention are shown in the scheme, but the present invention is not limited to the following reaction scheme. In the scheme, compound ( 1 ) can be synthesized, for example, according to the method described in JP-A-2-235898.

[0023]

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[0024]

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[0025]

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[0026]

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In the above reaction scheme, reagents, reaction conditions and the like which can be suitably used are described below. 1. 1 → 2 (first step) Solvent: water, alcohol, aqueous acetic acid, aqueous tetrahydrofuran, etc. Temperature: -50 to 100 ° C Time: 1 to 24 hours Reagents: acetic acid, trifluoroacetic acid, sulfuric acid, etc., acid catalyst (second step) 1.) Solvent: pyridine or an aprotic solvent such as dichloromethane containing amines such as pyridine and triethylamine. Temperature: -50 to 100 ° C. Time: 1 to 48 hours. Reagents: sulfonic acid chlorides such as methanesulfonyl chloride and p-toluenesulfonyl chloride. 2 → 3 solvent: aprotic polar solvent such as N, N-dimethylformamide, dimethylsulfoxide, hexamethylphosphoramide temperature: 50 to 150 ° C. time: 5 to 100 hours reagent: carboxylate such as sodium acetate, sodium benzoate 3. 3 → 4 (First step) Solvent: Protic solvent such as water, alcohol, hydrated tetrahydrofuran Temperature: 0 to 100 ° C. Time: 1 to 24 hours Reagent: Base such as sodium methoxide, sodium hydroxide, ammonia (Second step) 3.) Solvent: aprotic solvent such as dichloromethane containing amines such as pyridine or pyridine and triethylamine Temperature: -50 to 100 ° C. Time: 1 to 48 hours Reagent: Sulfon chloride such as methanesulfonyl chloride, p-toluenesulfonyl chloride 4 → 5 Solvent: aprotic solvent such as anhydrous diethyl ether, dichloromethane, acetonitrile, etc. Temperature: −80 to 50 ° C. Time: 5 to 100 hours Reagent: alkylating agent such as methyltrifluoromethanesulfonate or oxidizing agent such as N-bromosuccinimide 5 . 5 → 6 Solvent: aprotic polar solvent such as N, N-dimethylformamide, dimethylsulfoxide, hexamethylphosphoramide, etc. Temperature: 50 to 150 ° C. Time: 5 to 100 hours Reagent: sodium azide, lithium azide, tetraethylammonium 5. Hydrogen azide salt such as azide 6 → 7 Solvent: hydrated methanol, ethanol, alcohols such as 2-methoxyethanol or ethers such as diethyl ether and tetrahydrofuran Temperature: -50 to 50 ° C. Time: 1 to 100 hours Reagent: Palladium carbon and acid catalyst such as hydrochloric acid, acetic acid Lower hydrogen gas or sodium borohydride, lithium aluminum hydride, hydrogen sulfide, or other reducing agent To further extend the oligosaccharide of maltooligosaccharide, use a maltooligosaccharide derivative as a starting material in which the oligosaccharide is further extended. Alternatively, the maltooligosaccharide derivative produced as described above is extended with a sugar chain by the method described in Japanese Patent Application No. 2-204723, and then the above reaction is repeated.

[0028]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a preferred embodiment of the present invention will be described in detail with reference to examples. The scope of the present invention is not limited to this embodiment. The compound of 1a of this example is
It was produced by the method described in JP-A-2-235898. (Example 1)

[0029]

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[0030]1a→2a Phenyl 2¹, 2^Two, 2^Three, 2^Four, 2^Five, 2⁶,
3¹, 3^Two, 3^Three, 3^Four, 3 ^Five, 3⁶, 6¹, 6^Two, 6
^Three, 6^Four, 6^Five-Heptadeca-O-benzyl-1-thio
-Β-maltohexaside (2aPreparation of phenyl-2¹, 2^Two, 2^Three, 2^Four, 2^Five, 2⁶,
3¹, 3^Two, 3^Three, 3^Four, 3^Five, 3⁶, 6¹, 6^Two, 6
^Three, 6^Four, 6^Five-Heptadeca-O-benzyl-4⁶, 6
⁶-O-benzylidene-1-thio-β-maltohexao
Sid (1a) (10 g, 3.7 mmol) in acetic acid (80 ml)
Dissolve and stir at 80-90 ° C while adding water (20 ml).
added. After heating and stirring for 4 hours, the solvent was distilled off under reduced pressure.
And the residue is purified by silica gel column chromatography.
Purification with ruene-ethyl acetate 19: 1)2a(7.5
g, 78% yield).

[Α] _D ²⁴ + 58 ° (c 0.14, CHCl ₃ ) Elemental analysis: measured values; C, 73.77; H, 6.47; S, 1.26.
Calcd _{C 161 H 168 O 30 S;} C, 73.94; H, 6.
48; S, 1.23. (Example 2)

[0032]

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[0033]2a→3a Phenyl 2¹, 2^Two, 2^Three, 2^Four, 2^Five, 2⁶,
3¹, 3^Two, 3^Three, 3^Four, 3 ^Five, 3⁶, 6¹, 6^Two, 6
^Three, 6^Four, 6^Five-Heptadeca-O-benzyl-4⁶, 6
⁶-Di-O-mesyl-1-thio-β-maltohexaoshi
Do (3a) Production of compounds2a(6.0 g, 2.3 mmol) in pyridine (40 ml)
Methanesulfonyl chloride while stirring at 0 ° C.
(2 ml) was added dropwise. After stirring at 0 ° C for 14 hours,
The reaction solution was poured into ice water and extracted with chloroform. Extract
1M hydrochloric acid, saturated aqueous sodium hydrogen carbonate solution, and saturated
Wash with brine, dry over anhydrous sodium sulfate, and
The solvent was distilled off. The obtained syrup is applied to a silica gel column.
Chromatography (benzene-ethyl acetate 97: 3)
To purify3a(5.3 g, yield 83%).

[Α] _D ²⁴ + 67 ° (c 0.67, chloroform) ¹ H-NMR: δ (CDCl ₃ ) 2.76 (s, 3H, Ms), 2.82 (s, 3H, Ms),
5.49 (d, 1H, J3.1 Hz, H-1), 5.55 (d, 1H, J 3.7 H
z, H-1), 5.63 (d, 1H, J 3.7 Hz, H-1), 5.67 (d, 1H,
J 3.4 Hz, H-1), 5.69 (d, 1H, J 2.9 Hz, H-1) Elemental analysis: found; C, 70.45; H, 6.26; S, 3.59.
Calcd _{C 163 H 172 O 34 S 3} ; C, 70.64; H,
6.26; S, 3.47. (Example 3)

[0035]

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[0036]3a→4a Phenyl 2¹, 2^Two, 2^Three, 2^Four, 2^Five, 3¹,
3^Two, 3^Three, 3^Four, 3^Five, 6 ¹, 6^Two, 6^Three, 6^Four, 6
^Five-Pentadeca-O-benzyl-4^Five-O- (4,6-
Di-O-benzoyl-2,3-di-O-benzyl-α-
D-galactopyranosyl) -1-thio-β-maltopene
Taoside (4a) Production of compounds3a(5.5 g, 2.1 mmol) and sodium benzoate
(2 g) to hexamethyl phosphoroamide (40 ml)
Heated at 100 ° C. for 2 days. Dilute with diethyl ether
And washed three times with saturated saline, and then with anhydrous magnesium sulfate.
After drying, the mixture was concentrated under reduced pressure. Put the resulting syrup on silica gel
Ram chromatography (toluene-ethyl acetate 97:
Attached to 3)4a(4.25 g, yield 72%) was obtained.

[Α] _D ²⁴ + 66 ° (c1.47, chloroform) ¹ H-NMR: δ (CDCl ₃ ); 5.48 (d, 1H, J 3.7 Hz, H-1), 5.
54 (d, 1H, J 3.6Hz, H-1), 5.58 (d, 1H, J 3.0 Hz, H-
1), 5.67 (m, 2H, H-1, 4), 5.72 (d, 1H, J 3.2 Hz, H
-1). Elemental analysis: found: C, 73.90; H, 6.30; S, 1.04.
Calcd _{C 175 H 176 O 32 S ·} H 2 O; C, 73.88;
H, 6.31; S, 1.13. (Example 4)

[0038]

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[0039]4a→5a Phenyl 2¹, 2^Two, 2^Three, 2^Four, 2^Five, 3¹,
3^Two, 3^Three, 3^Four, 3^Five, 6 ¹, 6^Two, 6^Three, 6^Four, 6
^Five-Pentadeca-O-benzyl-4^Five-O- (2,3-
Di-O-benzyl-4,6-di-O-mesyl-α-D-
Galactopyranosyl) -1-thio-β-maltopentao
Sid (5a) Production of compounds4a(4.1 g, 1.45 mmol) in methanol-tet
Dissolve in lahydrofuran (1: 1, 100 ml) and add 5M
Add thorium methoxide (0.2 ml) and stir at room temperature overnight
did. Strong acidic ion exchange resin Dowex 50w x 8 (H
⁺The solvent was distilled off under reduced pressure. Obtained whitebait
Was dissolved in pyridine (50 ml) and stirred at 0 ° C.
Add methanesulfonyl chloride (0.5 ml) at room temperature overnight
Stirred. Compound3aPost-processing similar to the synthesis of5a
(2.8 g, 70% yield).

[Α] _D ²⁷ + 57 ° (c 0.17, chloroform) ¹ H-NMR: δ (CDCl ₃ ); 3.81 (s, 3H, Ms), 3.95 (s, 3H,
Ms), 5.50 (d, 1H, J 3.2 Hz, H-1), 5.54 (d, 1H, J
3.6 Hz, H-1), 5.59 (d, 1H, J 3.3 Hz, H-1), 5.68
(d, 1H, J 3.7 Hz, H-1), 5.72 (d, 1H, J 3.6 Hz, H-1). Elemental analysis: found; C, 70.35; H, 6.27; S, 3.50.
Calcd _{C 163 H 172 O 34 S 3} ; C, 70.64; H,
6.26; S, 3.47. (Example 5)

[0041]

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[0042] 5a → 6a 1,3-di tosyloxy-2-yl 2 ^1, 2
^2, 2 ^3, 2 ^4, 2 ^5, 3 ^1, 3 ^{2, 3} 3, 3 ^4,
3 ^5, 6 ^1, 6 ^2, 6 ^3, 6 ^4, 6 ⁵ - pentadeca -O
- benzyl -4 ⁵ -O- (2,3-di -O- benzyl -
Preparation of 4,6-di-O-mesyl-α-D-galactopyranosyl) -α-maltopentaoside ( 6a ) Compound 5a (0.58 g, 0.21 mmol), 1,3-ditosyloxy-2 Methyl triflate (0.13 ml) was added to a suspension of propanol (250 mg) and molecular sieves 4A (2 g) in diethyl ether (10 ml) at 0 ° C. under an argon atmosphere, followed by stirring at room temperature for 3 days. Triethylamine (0.1 ml) and methanol (0.2 ml)
Was added to stop the reaction, and the mixture was filtered. The filtrate was diluted with diethyl ether, washed with 1M hydrochloric acid, saturated sodium bicarbonate, and then with saturated saline, and dried over anhydrous sodium sulfate. The syrup obtained by distilling off the solvent was separated by silica gel column chromatography (toluene-ethyl acetate 94: 6) to give 6a (0.29 g, yield 5).
2%).

[Α] _D ²⁴ + 83 ° (c 0.21, chloroform) ¹ H-NMR: δ (CDCl ₃ ); 2.34 (s, 3H, Me Ph); 2.42 (s, 3
H, Me Ph), 2.80 (s, 3H, Ms), 2.94 (s, 3H, Ms), 5.58
(d, 1H, J 3.6 Hz, H-1), 5.60 (d, 1H, J 3.4 Hz, H-
1), 5.64 (d, 1H, J 3.4 Hz, H-1), 5.65 (d, 1H, J 3.
4 Hz, H-1), 5.70 (d, 1H, J 3.4 Hz, H-1).

Elemental analysis: measured values; C, 68.27; H, 6.1
5; S, 4.17. C ₁₇₄ H ₁₈₆ O ₄₁ S ₄ Calculated; C,
68.26; H, 6.12; S, 4.19. (Example 6)

[0045]

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[0046] 6a → 7a 1,3-diazide-2-yl 2 ^{1, 2} 2, 2
^3, 2 ^4, 2 ^5, 3 ^1, 3 ^{2, 3} 3, 3 ^4, 3 ^5,
6 ^1, 6 ^2, 6 ^3, 6 ^4, 6 ⁵ - pentadeca -O- benzyl -4 ⁵ -O- (4,6-Jiajito 2,3--O
Preparation of -benzyl-4,6-dideoxy-α-D-glucopyranosyl) -α-maltopentaoside ( 7a ) Compound 6a (140 mg, 0.046 mmol) and lithium azide (200 mg) were converted to N, N-dimethylformamide. (5 ml), and the mixture was heated and stirred at 100 ° C. for 24 hours.
After cooling, the mixture was diluted with diethyl ether, washed five times with a saturated saline solution, and dried over anhydrous sodium sulfate. The residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (toluene-ethyl acetate 49: 1) to give 7a (1
22 mg, yield 99%).

[Α] _D ²⁴ + 77 ° (c 0.38, chloroform) ¹ H-NMR: δ (CDCl ₃ ); 3.04 (dd, 2H, J 3.0, 12.8 Hz, C
H ₂ ), 3.18 (d, 2H, J 12.3Hz, CH ₂ ), 4.90 (d, 1H, J
3.4 Hz, H-1 '), 5.56 (d, 1H, J 3.3 Hz, H-1), 5.62
(d, 1H, J 3.3 Hz, H-1), 5.63 (d, 1H, J 3.4 Hz, H-
1), 5.66 (d, 1H, J 3.3 Hz, H-1), 5.67 (d, 1H, J 3.
8 Hz, H-1).

IR: νmax (film) 2100 cm ^-1
(N ₃ ) Elemental analysis: found; C, 70.10; H, 6.23; N, 6.06.
Calculated for C ₁₅₈ H ₁₆₆ O ₂₉ N ₁₂ ; C, 70.36; H,
6.20; N, 6.23. (Example 7)

[0049]

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[0050] 7a → 8a 1,3-diaminopropane-2-yl 4 ⁵ - (4,6
Preparation of -diamino-4,6-didequioxy-α-D-glucopyranosyl) -α-maltopentaoside ( 8a ) Compound 7a (230 mg) was treated with 2-methoxyethanol (1
0M), 1M hydrochloric acid (0.4 ml) and 10% palladium on carbon (140 mg) were added, and the mixture was added under hydrogen atmosphere at normal pressure for 24 hours.
Shake time. The catalyst was filtered and washed with water (5ml). The filtrate and the washing solution are combined, and 10% palladium carbon (15%) is added.
0 mg) and shaken under a hydrogen atmosphere for a further 18 hours. The catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was subjected to Amberlite CG-50 (H ⁺ ) column chromatography (elution solvent: water → 0.5 M aqueous ammonia) to give 8a (7
8 mg, 87%).

[Α] _D ²⁴ + 149 ° (c 0.22, water) ¹ H-NMR: δ (D ₂ o) 3.30 (t, 1H, J9.0 Hz), 5.28 (m, 6
H). Elemental analysis: found; C, 42.88; H, 6.02; N, 5.31.
C ₃₉ H ₆₂ O ₂₉ N ₄ · H ₂ CO ₃ Calculated; C, 43.17;
H, 5.80; N, 5.03.

[0052]

The maltooligosaccharide derivative of the present invention forms a complex with a metal ion and is therefore useful as a heavy metal scavenger. It is also useful as a protein stabilizer.
In addition, since the shape changes from a chain to a ring depending on the presence or absence of metal ions and can be opened and closed, it is useful for construction of a drug delivery system and the like.

Further, by forming a complex with platinum ions,
Useful as an anticancer agent. The method of the present invention for synthesizing a maltooligosaccharide having two amino groups at both ends of the present invention using a maltohexaose derivative as a starting material can be achieved by adding a metal ligand or the like to both reduced and non-reduced residues of the maltooligosaccharide. Useful for introducing functional groups.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI C08B 37/00 C08B 37/00 G J K

Claims (7)

(57) [Claims] 1. A maltooligosaccharide derivative represented by the formula (I). Embedded image (Wherein, R represents a hydroxyl group which may be independently protected, and n represents an integer of 4 to 6) 2. An intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (II). Embedded image (Wherein, R ¹ and R ² each independently represent an alkyl group or an aryl group which may be substituted, R each independently represents a hydroxyl group which may be protected, and n is an integer of 4 to 6. Show) 3. An intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (III). Embedded image (Wherein, R ¹ and R ³ each independently represent an alkyl group or an aryl group which may be substituted, R each independently represents a hydroxyl group which may be protected, and n is an integer of 4 to 6. Show) 4. An intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (IV). Embedded image (In the formula, R ¹ and R ⁴ each independently represent an alkyl group or an aryl group which may be substituted, R each independently represents a hydroxyl group which may be protected, and n is an integer of 4 to 6. Show) 5. An intermediate for synthesizing a maltooligosaccharide derivative represented by the formula (V). Embedded image (Wherein, R ⁴ and R ⁵ each independently represent an alkyl group or an aryl group which may be substituted, R each independently represents a hydroxyl group which may be protected, and n is an integer of 4 to 6. Show) 6. A synthetic intermediate of a maltooligosaccharide derivative represented by the formula (VI). Embedded image (Wherein, R represents a hydroxyl group which may be independently protected, and n represents an integer of 4 to 6) 7. A maltooligosaccharide derivative as a starting material,
A method of synthesizing maltooligosaccharides having two amino groups at both ends by chemically modifying sugar residues at both ends of reduction and non-reduction.